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Ginsenoside Rg-1 prevents elevated cytosolic Ca2+ via store-operated Ca2+ entry in high-glucose–stimulated vascular endothelial and smooth muscle cells

Abstract

Background

Ginsenoside Rg-1 (Rg-1), a triterpenoid saponin abundantly present in Panax ginseng, is a type of naturally occurring steroid with known anti-diabetic and anti-inflammatory effects. In this study, we sought to confirm the effects and mechanisms of action of Rg-1 on store-operated Ca2+ entry (SOCE) in human vascular endothelial cell line (EA) and murine aortic vascular smooth muscle cell line (MOVAS) cells exposed to high glucose.

Methods

Cytosolic Ca2+ concentrations in EA and MOVAS cells were measured by monitoring fluorescence of the ratiometric Ca2+-indicator, Fura-2 AM.

Results

High glucose significantly increased Ca2+ influx by abnormally activating SOCE in EA and MOVAS cells. Notably, this high glucose-induced increase in SOCE was restored to normal levels in EA and MOVAS cells by Rg-1. Moreover, Rg-1 induced reductions in SOCE in cells exposed to high glucose were significantly inhibited by the plasma membrane Ca2+ ATPase (PMCA) blocker lanthanum, the Na+/K+-ATPase blocker ouabain, or the Na+/Ca2+ exchanger (NCX) blockers Ni2+ and KB-R7943. These observations suggest that the mechanism of action of Rg-1 inhibition of SOCE involves PMCA and Na+/K+-ATPase, and an increase in Ca2+ efflux via NCXs in both EA and MOVAS cells exposed to high glucose.

Conclusions

These findings indicate that Rg-1 may protect vascular endothelial and smooth muscle cells from Ca2+ increases following exposure to hyperglycemic conditions.

Peer Review reports

Background

Ca2+, a second messenger involved in vast array of cellular processes such as metabolic signals, energy production, cell viability, and apoptosis [1]. Ca2+ signals are generated by Ca2+ influx from the extracellular space via plasma membrane Ca2+ channels and through intracellular release from the endoplasmic reticulum (ER)/sarcoplasmic reticulum (SR) via Ca2+-release channels [2]. Store-operated Ca2+ entry (SOCE) is an important extracellular Ca2+-influx mechanism in vascular endothelial and smooth muscle cells [3]. In this mechanism, depletion of ER/SR Ca2+ causes stromal interaction molecule 1 (STIM1) to bind to the ER-plasma membrane and signal the Orai channel to trigger Ca2+ influx into the cytosol [4]. SOCE is critical to the primary Ca2+ signaling pathway of cells and plays an essential role in a wide range of physiological functions, including extracellular excretion, enzyme activity, gene transcription, cell proliferation, and apoptosis [5].

High glucose causes an imbalance in cytosolic Ca2+ homeostasis. High glucose upregulates STIM1 and increases Orai1 protein levels, thereby activating SOCE and increasing Ca2+ influx [6, 7]. Previous studies have reported that STIM1 and Orai1 proteins are overexpressed in human aortic endothelial cells exposed to high glucose, resulting in increased Ca2+ influx through SOCE [8], and have further shown that chronic exposure to high glucose significantly increases apoptosis in human umbilical vein endothelial cells owing to increased hydrogen peroxide production and SOCE-mediated activation of calcineurin [9]. Similar to what is observed in vascular endothelial cells, SOCE and Orai1 protein levels were significantly increased in human aortic smooth muscle cells exposed to high glucose [10].

Ca2+ is intimately involved in the regulation of most cellular functions, such that even relatively small disturbances in Ca2+ homeostasis can have fatal consequences for cellular function [11]. Disruption of Ca2+ homeostasis causes vascular endothelial dysfunction by pathologically activating endothelial nitric oxide synthase [12] and increases the frequency of Ca2+ spikes in vascular smooth muscle cells, leading to excessive vasoconstriction [13]. Therefore, an imbalance in cytosolic Ca2+ homeostasis induced by high glucose is thought to have crucial effects on vascular health.

Panax ginseng is widely used in traditional herbal medicine owing to the pharmacological action of its ginsenoside saponins [14]. Among the 150 types of ginsenoside saponins, ginsenosides Rg-1 (Fig. 1), Rb-1 and Rg3 have demonstrated various pharmacological actions on cardiovascular, neuronal, and immune systems [15]. Moreover, ginsenosides are metformin mimetics with anti-diabetes properties [16]; among them, ginsenoside Rg-1 (hereafter, Rg-1) in particular, is known to have an excellent anti-diabetes profile. Rg-1 has also been demonstrated to directly protect pancreatic β-cell function and viability through the PI3K/Akt pathway [17]. Many studies have reported that Rg-1 not only inhibits the deterioration in the viability of cells exposed to high glucose, but also protect against cell damage. However, there are no reports of the effects of Rg-1 on SOCE or the underlying mechanism in vascular cells exposed to high glucose. Accordingly, we herein sought to investigate the preventive effects and related mechanisms of Rg-1 on SOCE in a human endothelial cell line (hereafter, EA) and a murine aortic vascular smooth muscle cell line (MOVAS) under high glucose.

Fig. 1
figure 1

Molecular structure of ginsenoside Rg-1

Methods

Materials

Rg-1, Rb-1, glucose, lanthanum (La3+), ouabain, nickel (Ni2+) and KB-R7943 were purchased from Sigma-Aldrich (St. Louis, MO, USA). MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) solution was from Amresco (Solon, OH, USA), and Fura-2 AM was supplied by Molecular Probes (Eugene, OR, USA).

Cell culture and hyperglycemia induction

EA cells, originally isolated from human umbilical veins, were purchased from the American Type Culture Collection (Manassas, VA, USA). MOVAS cells, isolated from aortic smooth muscle cells of C57BL6 mice, were purchased from Charles River Laboratories (Wilmington, MA, USA). EA and MOVAS cells were maintained in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum, 1% MEM non-essential amino acids, penicillin, and streptomycin. For culture under high-glucose conditions, cells were exposed to medium containing 30 mM glucose for 48 hours [18].

Cell viability assay

The optimal concentration of Rg-1 was determined by first confirming cell viability using the MTT technique, according to the procedure described in a previous study [19]. Briefly, cells were seeded in 96-well plates at 1 × 104 cells per well in 200-μl aliquots. Cells were incubated with 0 to 50 μM Rg-1 at 37 °C for 48 hours. Each well was then washed with phosphate-buffered saline, followed by addition of a pure-grade MTT (5 mg/ml) solution to each well. After incubation for 3 hours, 100 ml dimethyl sulfoxide (DMSO) was added to each well to dissolve formazan crystals generated by reduction of MTT by metabolically active cells. Plates were incubated for an additional 3 hours in the dark, and the optical density of each well was measured at 540 nm using a plate reader (Spectrostar Nano; BMG LABTECH, Ortenberg, Germany).

Cytosolic Ca2+ measurement

EA and MOVAS cells were exposed to high glucose with or without Rg-1 for 48 hours. Cytosolic Ca2+ was then measured using Fura-2 AM, a membrane-permeable, fluorescent ratiometric Ca2+-binding dye [20]. Approximately 1 × 106/ml cells were incubated with 2.5 μM Fura-2 AM in the dark for 30 minutes at room temperature. Dye remaining in the extracellular fluid was removed by exchanging the supernatant twice. Cells were placed in a quartz cuvette, and fluorescence at 510 nm was measured using a fluorescence spectrometer (Photon Technology Instruments) with alternating wavelengths of 340 nm and 380 nm with a chopper wheel (50 Hz). Cytosolic Ca2+ was expressed as the ratio of fluorescence at 340 nm to fluorescence at 380 nm (F340/F380).

Data analysis and statistics

All statistical analyses were performed using SPSS Statistics software version 26 (IBM, IL, USA). Data are expressed as means ± standard error of the mean (SEM). Differences among groups were analyzed by analysis of variance and Fisher’s Least Significant Difference post-hoc test. p-values < 0.05 were considered statistically significant.

Results

Effects of ginsenosides Rg-1 and Rb-1 on high-glucose–exposed EA and MOVAS cells

First, we compared the effects of Rg-1 and Rb-1 on SOCE in EA and MOVAS cells exposed to high glucose. In the normal glucose group, the effects of Rg-1 and Rb-1 on SOCE did not significantly differ between EA and MOVAS cell. However, the SOCE-inhibitory effects of Rg-1 and Rb-1 were different in the high-glucose group, with Rg-1 significantly reducing SOCE in both EA and MOVAS cells and Rb-1 significantly decreasing SOCE only in MOVAS cells (Fig. 2A-C). These findings confirmed that, of the ginsenosides evaluated, Rg-1 had a greater effect than Rb-1 on Ca2+ homeostasis in EA and MOVAS cells exposed to high glucose.

Fig. 2
figure 2

Effects of Rg-1 on SOCE in EA and MOVAS cells exposed to high glucose compared with those of Rb-1. A Representative traces showing the effects of normal glucose, high glucose, and high glucose with 10 μM Rg-1 or 10 μM Rb-1, on SOCE in EA cells. B Representative traces showing the effects of normal glucose, high glucose, and high glucose with 10 μM Rg-1 or 10 μM Rb-1, on SOCE in MOVAS cells. C Summary data showing area under the curve (AUC) values for 500 seconds for EA and MOVAS cells. Data are means ± SEM (n = 6–14; #p < 0.05, ###p < 0.001 compared with normal glucose; *p < 0.05, **p < 0.01 compared with high glucose). The symbol () indicates the time point at which 30 μM BHQ was applied

Optimal Rg-1 concentration inhibiting SOCE in high-glucose–exposed EA and MOVAS cells

The optimal Rg-1 concentration inhibiting SOCE was determined by the MTT method. Incubation of EA and MOVAS cells with 0, 0.01, 0.1, 1, 10, 20 or 50 μM Rg-1 showed that Rg-1 at concentrations above 20 μM significantly reduced cell viability compared with controls (data not shown). In addition, the effect of Rg-1 on the degree of SOCE inhibition was assessed by incubating EA and MOVAS cells exposed to high glucose with 0.1, 1, or 10 μM Rg-1. In both cell types, 10 μM Rg-1 concentration significantly reduced the high glucose-induced increase in SOCE, restoring normal glucose levels, whereas neither 0.1 nor 1 μM Rg-1 had any effect (Fig. 3A-C). Thus, Rg-1 was used at a concentration of 10 μM in subsequent experiments.

Fig. 3
figure 3

Effects of various concentrations of Rg-1 on SOCE in EA and MOVAS cells exposed to high glucose. A & B Representative traces showing effects of normal glucose, high glucose, and high glucose plus 0.1, 1, or 10 μM Rg-1 on SOCE in (A) EA cells and (B) MOVAS cells. C Summary data showing area under the curve (AUC) values for 500 seconds for EA and MOVAS cells. The symbol () indicates the time point at which 30 μM BHQ was applied. Data are means ± SEM (n = 6–11; #p < 0.05, ##p < 0.01, ###p < 0.001 compared with normal glucose; *p < 0.05, ***p < 0.001 compared with high glucose)

Mechanisms of SOCE inhibition by ginsenoside Rg-1 on high-glucose–exposed EA and MOVAS cells

The mechanism of SOCE inhibition by Rg-1 was analyzed by comparing its effect with the effects of the plasma membrane Ca2+ ATPase (PMCA) blocker La3+; the Na+/K+ ATPase blocker ouabain; and the Na+/Ca2+ exchanger (NCX) blockers, Ni2+ and KB-R7943. Specifically, KB-R7943 is an inhibitor of the reverse mode of Na+/Ca2+ exchanger.

SOCE was more substantially inhibited in cells pretreated with La3+ plus Rg-1 compared with cells pretreated with La3+ alone. La3+ alone significantly decreased SOCE in high-glucose–exposed EA cells, but not MOVAS cells. Notably, La3+ plus Rg-1 resulted in a significantly greater increase in SOCE than Rg-1 alone in both EA and MOVAS cells exposed to high glucose (Fig. 4A-C). These results suggest that the mechanism by which Rg-1 decreases SOCE in EA and MOVAS cells exposed to high glucose may be associated with the PMCA pathway.

Fig. 4
figure 4

Effects of La3+ on Rg-1–induced increases in SOCE in EA and MOVAS cells exposed to high glucose. A & B Representative traces showing the effects of high glucose, high glucose plus 125 μM La3+, high glucose plus 10 μM Rg-1, and high glucose plus 10 μM Rg-1 and 125 μM La3+ on SOCE in (A) EA and (B) MOVAS cells. C Summary data showing area under the curve (AUC) values for 500 seconds for EA and MOVAS cells. The symbol () indicates the time point at which 30 μM BHQ was applied. Data are means ± SEM (n = 7–11; #p < 0.05, ##p < 0.01 compared with high glucose; **p < 0.01 compared with high glucose plus Rg-1)

To determine whether the SOCE inhibitory effect of Rg-1 was related to Na+/K+ channels, EA and MOVAS cells exposed to high glucose were pretreated with Rg-1, the Na+/K+ ATPase blocker ouabain, or both. Treatment of cells exposed to high glucose with ouabain or Rg-1 alone reduced SOCE compared with controls. Compared with Rg-1 alone, however, Rg-1 plus ouabain significantly increased SOCE (Fig. 5A-C). This suggests that Rg-1 increases Ca2+ efflux in EA and MOVAS cells exposed to high glucose via the Na+/K+ ATPase pathway.

Fig. 5
figure 5

Effects of ouabain on Rg-1–induced increases in SOCE in EA and MOVAS cells exposed to high glucose. A & B Representative traces showing the effects of high glucose, high glucose plus 10 nM ouabain, high glucose plus 10 μM Rg-1, and high glucose plus 10 μM Rg-1 and 10 nM ouabain on SOCE in (A) EA and (B) MOVAS cells. C Summary data showing area under the curve (AUC) values for 500 seconds for EA and MOVAS cells. The symbol () indicates the time point at which 30 μM BHQ was applied. Data are means ± SEM (n = 6–11; #p < 0.05 compared with high glucose; *p < 0.05, **p < 0.01 compared with high glucose plus Rg-1)

The NCX pathway has been reported to be effective in coping with increased intracellular Ca2+ in pancreatic cells [21]. Therefore, we assessed whether the Rg-1-induced reduction in SOCE was related to the NCX pathway. Treatment of EA and MOVAS cells with Ni2+ or KB-R7943 inhibited the high glucose-induced increase in SOCE to a level similar to that of Rg-1. However, SOCE was more substantially increased in cells pretreated with Rg-1 plus Ni2+ or Rg-1 plus KB-R7943 than in cells pretreated with Ni2+ or KB-R7943 alone, respectively (Fig. 6A-C).

Fig. 6
figure 6

Effects of Ni2+ and KB-R7943 on Rg-1–induced increases in SOCE in EA and MOVAS cells exposed to high glucose. A & B Representative traces showing the effects of high glucose, high glucose plus 100 nM Ni2+, high glucose plus 10 μM KB-R7943, high glucose plus 10 μM Rg-1, high glucose plus 10 μM Rg-1 and 100 nM Ni2+, and high glucose plus 10 μM Rg-1 and 10 μM KB-R7943 on SOCE in (A) EA and (B) MOVAS cells. C Summary data showing area under the curve (AUC) values for 500 seconds for EA and MOVAS cells. The symbol () indicates the time point at which 30 μM BHQ was applied. Data are means ± SEM (n = 6–12; #p < 0.05, ##p < 0.01 compared with high glucose; *p < 0.05, **p < 0.01 compared with high glucose plus Rg-1)

Collectively, these findings indicate that high glucose significantly increases Ca2+ influx by abnormally activating SOCE in EA and MOVAS cells, and that Rg-1 reverses SOCE by increasing Ca2+ efflux through PMCA, Na+/K+ ATPase, and NCX.

Discussion

In vascular endothelial and smooth muscle cells, the SOCE pathway, activated by depletion of intracellular Ca2+ stores, plays an important role in regulating intracellular functions [3]. High glucose exposure promotes enhanced permeability and proliferation of human coronary artery endothelial cells through increases in SOCE via Orai-mediated, Ca2+ release-activated Ca2+ channels [22]. Notably in this context, it has been reported that diabetic hyperglycemia increases the expression of Orai1 and SOCE in vascular smooth muscle cells [10]. Our findings demonstrated that exposure to high glucose resulted in a significant increase in SOCE in EA and MOVAS cells; this effect was inhibited by Rg-1 treatment, which restored Ca2+ to levels found under normal glucose conditions; this Rg-1-associated inhibition of SOCE in EA and MOVAS cells exposed to high glucose might be related to the PMCA, Na+/K+ ATPase, and NCX pathways (Fig. 7). Notably, Rg-1 did not have a significant SOCE-inhibitory effect under normoglycemic conditions, but only exhibited this action under hyperglycemic conditions. In a similar vein, a study investigating the blood pressure effects of Codonopsis lanceolata, a natural plant rich in triterpenoid saponins, showed no effect on blood pressure in the normotensive group, but exerted a significant blood pressure-lowering effect in the hypertensive group [23].

Fig. 7
figure 7

Action mechanisms of Rg-1 on SOCE in EA and MOVAS cells exposed to high glucose. The red and blue colors indicate changes in the regulation of intracellular Ca2+ levels by exposure to high glucose or Rg-1 treatment, respectively. The thicknesses of the arrows reflect the amount of Ca2+ influx or efflux

In this study, treatment with Ca2+ antagonists such as La3+, ouabain, Ni2+, and KB-R7943 reduced SOCE in cells exposed to high glucose concentrations. However, incubation of cells with both Rg-1 and Ca2+ antagonists resulted in a rapid increase in SOCE. Increased intracellular Ca2+ is returned to the extracellular compartment by the action of the PMCA. This was shown, for example, by treatment of cells with 125 μM La3+, which blocked PMCA [24]. Treatment of pancreatic beta cells with 22.2 mM glucose resulted in significantly greater reductions in PMCA1 and PMCA2 mRNAs than treatment with 2.8 mM glucose [25]. These findings, showing that PMCA activity was inhibited under high glucose conditions, suggest that the inhibition of PMCA may have affected the increase in SOCE under high glucose than normal glucose conditions. In addition, the inhibition of SOCE by a relatively high concentration of La3+ under high glucose conditions may be due to its blocking of non-selective cation channels. Conversely, the increase in SOCE induced by treatment with both La3+ and Rg-1 under high glucose conditions may be related to the Rg-1-induced increase of Ca2+ efflux through PMCA, which inhibits SOCE under high glucose conditions.

Exposure of pancreatic β-cells to glucose leads to conversion of a PMCA-based, low-efficiency Ca2+ efflux mechanism to an NCX-based, high-efficiency system that is better able to cope with glucose-induced Ca2+ influx [21]. As the glycemic load in pancreatic β-cells increases, it induces a concentration-dependent decrease in PMCA activity, while significantly increasing NCX activity [25]. NCX is a reversible transporter of Ca2+ across plasma membranes. Exposure to abnormally high glucose concentrations has been reported to increase NCX activity and contribute to diabetic microvascular complications [26, 27]. This increase in NCX activity was inhibited by KB-R7943, suggesting that hyperglycemia-induced NCX activity is strongly associated with the NCX reverse mode [28]. In the present study, Rg-1 treatment inhibited the abnormally increased SOCE induced by high glucose concentrations, suggesting that Rg-1 may inhibit Ca2+ influx through the NCX reverse mode. In contrast, treatment with both Rg-1 and KB-R7943 under high glucose conditions resulted in a greater increase in SOCE than high glucose alone. Because pretreatment with KB-R7943 was found to increase the intracellular basal sodium concentration in cardiomyocytes of diabetic rats [27], the increased potential difference may depolarize cells and stimulate voltage-dependent calcium channels. When Ca2+ influx is excessively disturbed, various ion channels, such as NCXs or voltage-operated Ca2+ channels, play an important role in generating the Ca2+ influx underlying oscillations [29,30,31]. Future studies needed to assess the effects of Rg-1 on the expression of Ca2+ channel proteins, especially the expression of proteins that regulate SOCE, such as Orai1, Orai2, Orai3, STIM1, and STIM2.

Hyperglycemia-induced metabolic dysfunctions, such as enhanced formation of advanced glycation end products (AGEs) and increased production of reactive oxygen species, have been suggested as factors for developing diabetic vascular complications [32]. In mouse mesangial cells, high glucose enhanced the formation of methylglyoxal (the major precursor of AGEs) and the level of 8-OHdG, suggesting increased oxidative stress [33]. In line with these reports, catalase treatment of platelets from patients with type 2 diabetes mellitus increased the PMCA tyrosine phosphorylation induced by thapsigargin plus ionomycin, suggesting that oxidative stress is involved in the reduced platelet PMCA activity seen in diabetic patients [34]. High glucose also significantly increased NCX activity and malondialdehyde production in human umbilical vascular endothelial cells, and this effect was abolished by KB-R7943-mediated inhibition of the reverse mode of NCX, indicating that increased reverse-mode NCX activity plays an important role in high glucose-induced endothelial dysfunction [28]. Rg-1 has demonstrated several pharmacological actions including antioxidant effects [35]. Rg-1 reduces reactive oxygen species (ROS) by modulating Nrf2 (nuclear factor-erythrocyte 2-related factor 2) and NLRP3 (nucleotide-binding oligomerization domain-like receptor protein 3) signaling pathways [36]. Several studies have shown that hyperglycemia induces lipid peroxidation, nitrite production, and intracellular ROS formation [37, 38]. In the current study, the reduction in SOCE is probably related to the antioxidant activity of Rg-1.

In vascular endothelial cells, Rg-1 produced a more prominent SOCE-reducing effect than Rb-1. Although few studies have investigated the regulation of intracellular Ca2+ in vascular cells by Rb-1, one study in rat cortical synaptosomes showed that it decreased intracellular Ca2+ by increasing Na+/K+-ATPase and Ca2+/Mg2+-ATPase activity [39]. In addition, Rb-1 protects cardiomyocytes by reducing the high-Ca2+–induced delayed afterdepolarizations of cardiomyocytes. On the other hand, Rg-1 has been reported to have Ca2+-regulating effects in various cell types. In an ischemia-reperfused hippocampal cell model, it was shown to have neuroprotective effects by blocking excess Ca2+ influx and decreasing neuronal nitric oxide synthase activity [40]. In myocardial cells, it has been shown that, Rg-1 restores cellular homeostasis by reducing ROS and inhibiting Ca2+ influx [41]. In neurons, Rg-1 shows a cell-protective effect through a reduction in intracellular free Ca2+ [42] and exerts a Ca2+ homeostasis-maintenance effect by reducing ROS and oxidative stress and protecting against neuroinflammation [43]. In lymphocytes, Rg-1 has been shown to inhibit Ca2+ influx attributable to H2O2-induced damage and significantly reduce apoptosis and lymphocyte damage caused by oxidative stress, indicating protective effects on immune cells [44]. Therefore, Rg-1 exerts cytoprotective effects under pathological conditions by maintaining Ca2+ homeostasis. However, further studies are needed to show the different mechanism of Rg-1 in different cell types such as endothelial cell and smooth muscle cells.

Conclusions

Present study demonstrated that treatment with Rg-1 may be a new approach to protecting vascular endothelial and smooth muscle cells in patients with hyperglycemia through maintenance of Ca2+ homeostasis.

Availability of data and materials

All data generated or analyzed during this study are included in this published article.

Abbreviations

AGE:

Advanced glycation end product

AUC:

Area under the curve

DMSO:

Dimethyl sulfoxide

EA:

Human endothelial cell line

ER:

Endoplasmic reticulum

MOVAS:

Murine aortic vascular smooth muscle cell line

NCX:

Na+/Ca2+ exchanger

PMCA:

Plasma membrane Ca2+ ATPase

SEM:

Standard error of the mean

SOCE:

Store-operated Ca2+ entry

SR:

Sarcoplasmic reticulum

STIM:

Stromal interaction molecule

ROS:

Reactive oxygen species

References

  1. Marchi S, Patergnani S, Missiroli S, Morciano G, Rimessi A, Wieckowski MR, et al. Mitochondrial and endoplasmic reticulum calcium homeostasis and cell death. Cell Calcium. 2018;69:62–72. https://0-doi-org.brum.beds.ac.uk/10.1016/j.ceca.2017.05.003.

    CAS  Article  PubMed  Google Scholar 

  2. Hogan PG, Rao A. Store-operated calcium entry: mechanisms and modulation. Biochem Biophys Res Commun. 2015;460:40–9. https://0-doi-org.brum.beds.ac.uk/10.1016/j.bbrc.2015.02.110.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  3. Avila-Medina J, Mayoral-González I, Galeano-Otero I, Redondo PC, Rosado JA, Smani T. Pathophysiological significance of store-operated calcium entry in cardiovascular and skeletal muscle disorders and angiogenesis. Adv Exp Med Biol. 2020;1131:489–504. https://0-doi-org.brum.beds.ac.uk/10.1007/978-3-030-12457-1_19.

    CAS  Article  PubMed  Google Scholar 

  4. Shen Y, Thillaiappan NB, Taylor CW. The store-operated Ca2+ entry complex comprises a small cluster of STIM1 associated with one Orai1 channel. Proc Natl Acad Sci U S A. 2021;118:e2010789118. https://0-doi-org.brum.beds.ac.uk/10.1073/pnas.2010789118.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  5. Parekh AB, Putney JW Jr. Store-operated calcium channels. Physiol Rev. 2005;85:757–810. https://0-doi-org.brum.beds.ac.uk/10.1152/physrev.00057.2003.

    CAS  Article  PubMed  Google Scholar 

  6. Chaudhari S, Wu P, Wang Y, Ding Y, Yuan J, Begg M, et al. High glucose and diabetes enhanced store-operated Ca2+ entry and increased expression of its signaling proteins in mesangial cells. Am J Physiol Renal Physiol. 2014;306:F1069–F80. https://0-doi-org.brum.beds.ac.uk/10.1152/ajprenal.00463.2013.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  7. Xu Z, Xu W, Song Y, Zhang B, Li F, Liu Y. Blockade of store-operated calcium entry alleviates high glucose-induced neurotoxicity via inhibiting apoptosis in rat neurons. Chem Biol Interact. 2016;254:63–72. https://0-doi-org.brum.beds.ac.uk/10.1016/j.cbi.2016.05.025.

    CAS  Article  PubMed  Google Scholar 

  8. Daskoulidou N, Zeng B, Berglund LM, Jiang H, Chen G-L, Kotova O, et al. High glucose enhances store-operated calcium entry by upregulating ORAI/STIM via calcineurin-NFAT signalling. J Mol Med. 2015;93:511–21. https://0-doi-org.brum.beds.ac.uk/10.1007/s00109-014-1234-2.

    CAS  Article  PubMed  Google Scholar 

  9. Tamareille S, Mignen O, Capiod T, Rücker-Martin C, Feuvray D. High glucose-induced apoptosis through store-operated calcium entry and calcineurin in human umbilical vein endothelial cells. Cell Calcium. 2006;39:47–55. https://0-doi-org.brum.beds.ac.uk/10.1016/j.ceca.2005.09.008.

    CAS  Article  PubMed  Google Scholar 

  10. Ma K, Sukkar B, Zhu X, Zhou K, Cao H, Voelkl J, et al. Stimulation of ORAI1 expression, store-operated Ca2+ entry, and osteogenic signaling by high glucose exposure of human aortic smooth muscle cells. Pflug Arch Eur J Physiol. 2020;472:1093–102. https://0-doi-org.brum.beds.ac.uk/10.1007/s00424-020-02405-1.

    CAS  Article  Google Scholar 

  11. Petersen OH, Michalak M, Verkhratsky A. Calcium signalling: past, present and future. Cell Calcium. 2005;38:161–9. https://0-doi-org.brum.beds.ac.uk/10.1016/j.ceca.2005.06.023.

    CAS  Article  PubMed  Google Scholar 

  12. Goligorsky MS. Vascular endothelium in diabetes. Am J Physiol Renal Physiol. 2017;312:F266–F75. https://0-doi-org.brum.beds.ac.uk/10.1152/ajprenal.00473.2016.

    CAS  Article  PubMed  Google Scholar 

  13. Navedo MF, Takeda Y, Nieves-Cintrón M, Molkentin JD, Santana LF. Elevated Ca2+ sparklet activity during acute hyperglycemia and diabetes in cerebral arterial smooth muscle cells. Am J Physiol Cell Physiol. 2010;298:C211–C20. https://0-doi-org.brum.beds.ac.uk/10.1152/ajpcell.00267.2009.

    CAS  Article  PubMed  Google Scholar 

  14. Yu T, Yang Y, Kwak Y-S, Song GG, Kim M-Y, Rhee MH, et al. Ginsenoside Rc from Panax ginseng exerts anti-inflammatory activity by targeting TANK-binding kinase 1/interferon regulatory factor-3 and p38/ATF-2. J Ginseng Res. 2017;41:127–33. https://0-doi-org.brum.beds.ac.uk/10.1016/j.jgr.2016.02.001.

    Article  PubMed  Google Scholar 

  15. Mohanan P, Subramaniyam S, Mathiyalagan R, Yang D-C. Molecular signaling of ginsenosides Rb1, Rg1, and Rg3 and their mode of actions. J Ginseng Res. 2018;42:123–32. https://0-doi-org.brum.beds.ac.uk/10.1016/j.jgr.2017.01.008.

    Article  PubMed  Google Scholar 

  16. Aliper A, Jellen L, Cortese F, Artemov A, Karpinsky-Semper D, Moskalev A, et al. Towards natural mimetics of metformin and rapamycin. Aging (Albany NY). 2017;9:2245–68. https://0-doi-org.brum.beds.ac.uk/10.18632/aging.101319.

    CAS  Article  Google Scholar 

  17. Li N, Zhang H, Li X. Ginsenoside Rg1 protects pancreatic INS-1 cell survival in β-cell through the PI3K/Akt signaling pathway. Metab Clin Exp. 2020;104:154078. https://0-doi-org.brum.beds.ac.uk/10.1016/j.metabol.2019.12.024.

  18. Zhang R-J, Hao H-Y, Liu Q-J, Zuo H-Y, Chang Y-N, Zhi Z-J, et al. Protective effects of Schisandrin on high glucose-induced changes of RhoA and eNOS activity in human umbilical vein endothelial cells. Pathol Res Pract. 2018;214:1324–9. https://0-doi-org.brum.beds.ac.uk/10.1016/j.prp.2018.02.010.

    CAS  Article  PubMed  Google Scholar 

  19. Han AY, Lee HS, Seol GH. Foeniculum vulgare mill. Increases cytosolic Ca2+ concentration and inhibits store-operated Ca2+ entry in vascular endothelial cells. Biomed Pharmacother. 2016;84:800–5. https://0-doi-org.brum.beds.ac.uk/10.1016/j.biopha.2016.10.013.

    CAS  Article  PubMed  Google Scholar 

  20. Kim MK, Han AY, Shin YK, Lee K-W, Seol GH. Codonopsis lanceolata contributes to Ca2+ homeostasis by mediating SOCE and PLC/IP3 pathways in vascular endothelial and smooth muscle cells. Planta Med. 2020;86:1345–52. https://0-doi-org.brum.beds.ac.uk/10.1055/a-1214-6718.

    CAS  Article  PubMed  Google Scholar 

  21. Herchuelz A, Kamagate A, Ximenes H, Van Eylen F. Role of Na/ca exchange and the plasma membrane Ca2+−ATPase in β cell function and death. Ann N Y Acad Sci. 2007;1099:456–67. https://0-doi-org.brum.beds.ac.uk/10.1196/annals.1387.048.

    CAS  Article  PubMed  Google Scholar 

  22. Bai S, Wei Y, Hou W, Yao Y, Zhu J, Hu X, et al. Orai-IGFBP3 signaling complex regulates high-glucose exposure-induced increased proliferation, permeability, and migration of human coronary artery endothelial cells. BMJ Open Diabetes Res Care. 2020;8:e001400. https://0-doi-org.brum.beds.ac.uk/10.1136/bmjdrc-2020-001400.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Han AY, Lee YS, Kwon S, Lee HS, Lee K-W, Seol GH. Codonopsis lanceolata extract prevents hypertension in rats. Phytomedicine. 2018;39:119–24. https://0-doi-org.brum.beds.ac.uk/10.1016/j.phymed.2017.12.028.

  24. Chen J, McLean PA, Neel BG, Okunade G, Shull GE, Wortis HH. CD22 attenuates calcium signaling by potentiating plasma membrane calcium-ATPase activity. Nat Immunol. 2004;5:651–7. https://0-doi-org.brum.beds.ac.uk/10.1038/ni1072.

    CAS  Article  PubMed  Google Scholar 

  25. Ximenes HM, Kamagate A, Van Eylen F, Carpinelli A, Herchuelz A. Opposite effects of glucose on plasma membrane Ca2+−ATPase and Na/ca exchanger transcription, expression, and activity in rat pancreatic β-cells. J Biol Chem. 2003;278:22956–63. https://0-doi-org.brum.beds.ac.uk/10.1074/jbc.M212339200.

    CAS  Article  PubMed  Google Scholar 

  26. Li J, Jin H-B, Sun Y-M, Su Y, Wang L-F. KB-R7943 inhibits high glucose-induced endothelial ICAM-1 expression and monocyte-endothelial adhesion. Biochem Biophys Res Commun. 2010;392:516–9. https://0-doi-org.brum.beds.ac.uk/10.1016/j.bbrc.2009.12.183.

    CAS  Article  PubMed  Google Scholar 

  27. Nishio S, Teshima Y, Takahashi N, Thuc LC, Saito S, Fukui A, et al. Activation of CaMKII as a key regulator of reactive oxygen species production in diabetic rat heart. J Mol Cell Cardiol. 2012;52:1103–11. https://0-doi-org.brum.beds.ac.uk/10.1016/j.yjmcc.2012.02.006.

    CAS  Article  PubMed  Google Scholar 

  28. Liu D, Cui K-Z, Sun Y-M, Liu J-W, Li Y-B, Su Y. Protective effects of the sodium/calcium exchanger inhibitor on endothelial dysfunction induced by high glucose. Exp Clin Endocrinol Diabete. 2015;123:7–10. https://0-doi-org.brum.beds.ac.uk/10.1055/s-0034-1385924.

    CAS  Article  Google Scholar 

  29. Country MW, Campbell BF, Jonz MG. Spontaneous action potentials in retinal horizontal cells of goldfish (Carassius auratus) are dependent upon L-type Ca2+ channels and ryanodine receptors. J Neurophysiol. 2019;122:2284–93. https://0-doi-org.brum.beds.ac.uk/10.1152/jn.00240.2019.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  30. Fodor J, Matta C, Oláh T, Juhász T, Takács R, Tóth A, et al. Store-operated calcium entry and calcium influx via voltage-operated calcium channels regulate intracellular calcium oscillations in chondrogenic cells. Cell Calcium. 2013;54:1–16. https://0-doi-org.brum.beds.ac.uk/10.1016/j.ceca.2013.03.003.

    CAS  Article  PubMed  Google Scholar 

  31. Luo L, Song S, Ezenwukwa CC, Jalali S, Sun B, Sun D. Ion channels and transporters in microglial function in physiology and brain diseases. Neurochem Int. 2021;142:104925. https://0-doi-org.brum.beds.ac.uk/10.1016/j.neuint.2020.104925.

    CAS  Article  PubMed  Google Scholar 

  32. Yamagishi S-i, Maeda S, Matsui T, Ueda S, Fukami K, Okuda S. Role of advanced glycation end products (AGEs) and oxidative stress in vascular complications in diabetes. Biochim Biophys Acta. 2012;1820:663–71. https://0-doi-org.brum.beds.ac.uk/10.1016/j.bbagen.2011.03.014.

    CAS  Article  PubMed  Google Scholar 

  33. Kim KM, Kim YS, Jung DH, Lee J, Kim JS. Increased glyoxalase I levels inhibit accumulation of oxidative stress and an advanced glycation end product in mouse mesangial cells cultured in high glucose. Exp Cell Res. 2012;318:152–9. https://0-doi-org.brum.beds.ac.uk/10.1016/j.yexcr.2011.10.013.

    CAS  Article  PubMed  Google Scholar 

  34. Jardin I, Redondo PC, Salido GM, Pariente JA, Rosado JA. Endogenously generated reactive oxygen species reduce PMCA activity in platelets from patients with non-insulin-dependent diabetes mellitus. Platelets. 2006;17:283–8. https://0-doi-org.brum.beds.ac.uk/10.1080/09537100600745187.

    CAS  Article  PubMed  Google Scholar 

  35. Gao Y, Li J, Wang J, Li X, Li J, Chu S, et al. Ginsenoside Rg1 prevent and treat inflammatory diseases: a review. Int Immunopharmacol. 2020;87:106805. https://0-doi-org.brum.beds.ac.uk/10.1016/j.intimp.2020.106805.

    CAS  Article  PubMed  Google Scholar 

  36. Gao Y, Li J, Chu S, Zhang Z, Chen N, Li L, et al. Ginsenoside Rg1 protects mice against streptozotocin-induced type 1 diabetic by modulating the NLRP3 and Keap1/Nrf2/HO-1 pathways. Eur J Pharmacol. 2020;866:172801. https://0-doi-org.brum.beds.ac.uk/10.1016/j.ejphar.2019.172801.

    CAS  Article  PubMed  Google Scholar 

  37. Jayakumar T, Chang C-C, Lin S-L, Huang Y-K, Hu C-M, Elizebeth AR, et al. Brazilin ameliorates high glucose-induced vascular inflammation via inhibiting ROS and CAMs production in human umbilical vein endothelial cells. Biomed Res Int. 2014;2014:403703. https://0-doi-org.brum.beds.ac.uk/10.1155/2014/403703.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  38. Zhuang X, Maimaitijiang A, Li Y, Shi H, Jiang X. Salidroside inhibits high-glucose induced proliferation of vascular smooth muscle cells via inhibiting mitochondrial fission and oxidative stress. Exp Ther Med. 2017;14:515–24. https://0-doi-org.brum.beds.ac.uk/10.3892/etm.2017.4541.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  39. Jiang X, Zhang J, Shi C. Mechanism of action of ginsenoside Rb1 in decreasing intracellular Ca2+. Acta Pharm Sin. 1996;31:321–6.

    CAS  Google Scholar 

  40. He Q, Sun J, Wang Q, Wang W, He B. Neuroprotective effects of ginsenoside Rg1 against oxygen–glucose deprivation in cultured hippocampal neurons. J Chin Med Assoc. 2014;77:142–9. https://0-doi-org.brum.beds.ac.uk/10.1016/j.jcma.2014.01.001.

    CAS  Article  PubMed  Google Scholar 

  41. Zhu D, Wu L, Li CR, Wang XW, Ma YJ, Zy Z, et al. Ginsenoside Rg1 protects rat cardiomyocyte from hypoxia/reoxygenation oxidative injury via antioxidant and intracellular calcium homeostasis. J Cell Biochem. 2009;108:117–24. https://0-doi-org.brum.beds.ac.uk/10.1002/jcb.22233.

    CAS  Article  PubMed  Google Scholar 

  42. Zhang Y-F, Fan X-J, Li X, Peng L-L, Wang G-H, Ke K-F, et al. Ginsenoside Rg1 protects neurons from hypoxic–ischemic injury possibly by inhibiting Ca2+ influx through NMDA receptors and L-type voltage-dependent Ca2+ channels. Eur J Pharmacol. 2008;586:90–9. https://0-doi-org.brum.beds.ac.uk/10.1016/j.ejphar.2007.12.037.

    CAS  Article  PubMed  Google Scholar 

  43. Han Y, Li X, Yang L, Zhang D, Li L, Dong X, et al. Ginsenoside Rg1 attenuates cerebral ischemia-reperfusion injury due to inhibition of NOX2-mediated calcium homeostasis dysregulation in mice. J Ginseng Res. 2021;46:515–25. https://0-doi-org.brum.beds.ac.uk/10.1016/j.jgr.2021.08.001.

  44. Bi S, Ma X, Wang Y, Chi X, Zhang Y, Xu W, et al. Protective effect of ginsenoside Rg1 on oxidative damage induced by hydrogen peroxide in chicken splenic lymphocytes. Oxidative Med Cell Longev. 2019;2019:8465030. https://0-doi-org.brum.beds.ac.uk/10.1155/2019/8465030.

    CAS  Article  Google Scholar 

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Funding

This work was supported by a grant from the Basic Science Research Program through the National Research Foundation of Korea (NRF-2021R1A2C2004118, 2020R1I1A1A0107450513) and the Institute of Nursing Research, Korea University Grant. This manuscript is a revision of SMH’s master’s thesis from Korea University.

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Conceptualization, G.H.S.; methodology, G.H.S.; formal analysis, A.Y.H., S.M.H., Y.K.S. and G.H.S.; funding acquisition, G.H.S.; investigation, A.Y.H., S.M.H. and Y.K.S.; project administration, G.H.S.; supervision, G.H.S.; writing—original draft preparation, A.Y.H., S.M.H., Y.K.S. and G.H.S.; writing—review and editing, G.H.S.; visualization, A.Y.H. and G.H.S. All authors have read and approved the final manuscript.

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Correspondence to Geun Hee Seol.

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Han, A.Y., Ha, S.M., Shin, Y.K. et al. Ginsenoside Rg-1 prevents elevated cytosolic Ca2+ via store-operated Ca2+ entry in high-glucose–stimulated vascular endothelial and smooth muscle cells. BMC Complement Med Ther 22, 166 (2022). https://0-doi-org.brum.beds.ac.uk/10.1186/s12906-022-03647-5

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Keywords

  • Ginsenoside Rg-1
  • Store-operated Ca2+ entry
  • High glucose
  • Vascular endothelial cells
  • Vascular smooth muscle cells