Effect of the extract of cordyceps militaris on senescence in cultured human umbilical vein endothelial cells treated with high glucose

Abstract

Abstract: AIM: To determine the effect of the ethyl acetate extract of cordyceps militaris (CME) on senescence in cultured human umbilical vein endothelial cells (HUVECs) treated with high glucose. METHODS: The HUVECs senescence model was established by incubation with high glucose(40 mmol/L) , the extract of Cordyceps Sinensis(CSE) (50 μg/mL) was treated as a positive control, and then CME (12.5-100 μg/mL) was treated to observe its effect on cell senescence. Cell proliferation was determined by the methylthiazolyl tetrazolium (MTT) reduction assay , SA-β-galactosidase (SA-β-gal) positive cells were detected by X-gal, and phase distribution of cell cycles and cellular reactive oxygen species (ROS) were detected by flow cytometry. RESULTS: Compared with unintervented cells, cell OD value significantly decreased in cells treated with high glucose for 24, 48, and 72 h (P<0.01). The percentage of cells positive for SA-β-gal, the percentage in G₀/G₁ phase and ROS levels notably increased after cells incubated by high glucose for 96 or 48 h (P<0.01). Compared with high glucose intervention cells, cell OD value significantly increased in cells intervented by CME (25-50 μg/mL) for 48 h and 72 h ( P<0.05); CME(12.5-100 μg/mL)stimulating for 96 h significantly lowered the percentage of cells positive for SA-β-gal (P<0.01); CME(12.5-100 μg/mL)incubation for 48 h significantly lowered the percentage of G₀/G₁ phase of HUVECs (P<0.01) and ROS production (P<0.05); All these effects of CME 25, 50 μg/mL groups were better than those12.5, 100 μg/mL groups (P<0.05), and were similar to CSE 50 μg/mL group(P>0.05). CONCLUSION: Ethyl acetate extract of cordyceps militaris may promote the HUVECs growth and proliferation, protecting from high glucose-stimulated vascular endothelial senescence. The effects may be associated with ethyl acetate extract of cordyceps militaris inhibiting reactive oxygen species and alleviating oxidative stress injury.

Key words: Human umbilical vein endothelial cells, High glucose, Cell senescence, Antioxidative activity, Ethyl acetate extract of cordyceps militaris

CLC Number:

R965

ZHU Peng-li, YE Zhang-zheng, YU Hui-zhen, HUANG Hua-wei, RUAN Jing-ming, LIN Fan. Effect of the extract of cordyceps militaris on senescence in cultured human umbilical vein endothelial cells treated with high glucose[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2012, 17(11): 1216-1222.

References

[1] Minamino T, Komuro I.Vascular cell senescence:contribution to atherosclerosis[J].Circ Res,2007,100(1):15-26.
[2] Wanger M, Hampel B, Bernhard D, et al.Replicative senescence of human endothelial cells in vitro involves G₁ arrest, Polyploization and senescence-associated apoptosis[J].Exp Gerontol,2001,36(8):1327-1347.
[3] Unterluggauer H, Hampel B, Zwerschke W, et al.Senescence-associated cell death of human endothelial cells: the role of oxidative stress[J].Exp Gerontol,2003,38(10):1149-1160.
[4] Ceriello A.New insights on oxidative stress and diabetic complications may lead to a “Causal” antioxidant therapy[J].Diabetes Care,2003,26(5):1589-1596.
[5] Hayashi T, Matsui Hirai H, Miya zaki Akita A, et al. Endothelial cellular senescence is inhibited by nitric oxide: Implications in atherosclerosis associated with menopause and diabetes[J].Proc Natl Acad Sci USA,2006,103(45):17018-17023.
[6] Das SK, Masuda M, Sakurai A, et al.Medicinal uses of the mushroom Cordyceps militaris: Current state and prospects[J].Fitoterapia,2010,81(8):961-968.
[7] Gan CP, Hamid S, Hor SY, et al.Decreased proliferation of aging tenocytes is associated with down-regulation of cellular senescence-inhibited gene and up-regulation of p27[J].J Orthop Res,2011,9(10):1598-1603.
[8] Dimri GP, Lee X, Basile G, et al.A biomarker that identifies senescent human cells in culture and in aging skin in vivo[J].Proc Natl Acad Sci USA,1995,92(20):9363-9367.
[9] Lee MH, Han DW, Hyon SH, et al.Apoptosisof human fibrosarcoma HT-1080 cells by epigallocatechin-3-O-gallate via induction of p53 and caspases as well as suppression of Bcl-2 and phosphorylated nuclear factor-kappaB[J].Apoptosis, 2011,16(1):75-85.
[10] Kao CL, Chen LK, Chang YL, et al.Resveratrol protects human endothelium from H₂O₂-induced oxidative stress and senescence via SirT1 activation[J]. J Atheroscler Thromb, 2010, 17(9): 9970-9979.
[11] Fox CS, Coady S, Sorlie PD, et al.Trends in cardiovascular complications of diabetes[J]. J Am Med Assoc,2004,292(20):2495-2499.
[12] Resnick HE, Shorr RI, Kuller L, et al.Prevalence and clinical implications of American Diabetes Association-defined diabetes and other categories of glucose dysregulation in older adults[J]. J Clin Epidemiol,2001,54(9):869-876.
[13] Ross R.The pathogenesis of atherosclerosis: a perspective for the 1990s[J].Nature, 1993,362(6423):801.
[14] Avogaro A, Fadini GP, Gallo A, et al.Endothelial dysfunction in type 2 diabetes mellitus[J].Nutr Metab Cardiovasc Dis,2006,16(Suppl 1):S39-S45.
[15] Ungvari Z, Kaley G, de Cabo R, et al. Mechanisms of vascular aging: new perspectives[J].J Gerontol A Biol Sci Med Sci,2010,65(10):1028-1041.
[16] Yokoi T, Fukuo K, Yasuda O, et al.Apoptosis signal-regulating kinase 1 mediates cellular senescence induced by high glucose in endothelial cells[J].Diabetes,2006,55(6):1660-1665.
[17] 顾宇翔,宋聿文,范立强,等. 虫草减轻自由基引起的细胞氧化损伤[J]. 食品科学, 2008,29(2):387-390.
[18] Harman D.The free radical theory of aging[J].Antioxid Redox Signal,2003,5(5):557-561.
[19] Yu HM, Wang BS, Huang SC, et al.Comparison of protective effects between cultured cordyceps militaris and natural cordyceps sinensis against oxidative damage[J].J Agric Food Chem,2006,54(8):3132-3138.