Involvement of Fas-dependent pathway in rhein-induced apoptosis of HK-2 cells

SUN Hao; YANG Jiapei; MAO Yong; WANG Dandan; YU Feng

doi:10.11665/j.issn.1000-5048.20150414

Journal of China Pharmaceutical University > 2015 > 46(4): 469-475. > DOI: 10.11665/j.issn.1000-5048.20150414

SUN Hao, YANG Jiapei, MAO Yong, WANG Dandan, YU Feng. Involvement of Fas-dependent pathway in rhein-induced apoptosis of HK-2 cells[J]. Journal of China Pharmaceutical University, 2015, 46(4): 469-475. DOI: 10.11665/j.issn.1000-5048.20150414

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Involvement of Fas-dependent pathway in rhein-induced apoptosis of HK-2 cells

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Abstract

To investigate the effect and mechanism of cytotoxicity by rhein in human renal tubular epithelial HK-2 cells, HgCl₂ was choosen as positive control. Cell viability was determined by MTT assay. LDH release assay was used to evaluate cell membrane damage. The activity of caspase-3, and -8 was measured by assay kit. Real-Time qPCR was employed to determine the mRNA expressions of Fas, FasL, FADD, caspase-3 and caspase-8. The protein expressions of Fas, FasL, cytoplasmic cytochrome C, caspase-3, caspase-8, caspase-9 were detected by Western blot. The results demonstrated that rhein inhibited cell viability and increased LDH release in a dose-dependent manner. The activity of caspase-3 and caspase-8 was significantly enhanced by rhein. The mRNA expression of Fas, FasL, FADD, caspase-3, caspase-8 was remarkably up-regulated by rhein. Rhein also elevated protein expressions of Fas, FasL, cytoplasmic cytochrome C, cleaved caspase-3, caspase-8 and reduced expressios of Pro caspase-8. There was no significant difference in caspase-9 expression. These results indicate that rhein has a cytotoxic effect and apoptosis-inducing effect in HK-2 cells. The Fas-dependent pathway is involved in rhein-induced apoptosis.
- rhein,
- HK-2 cell,
- cytotoxicity,
- Fas,
- apoptosis

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[1]	Xiong XF.On the main active ingredient of TCM-rhubard in phamacology[J].Clin J Chin Med(中医临床研究 ),2014,6(10):143-146.
[2]	Su H,Leng J,Wang SD,et al.Development of pharmacological effects of rhein and its conjugates[J].China Pharm(中国药业),2011,20(15):92-94.
[3]	Hsia TC,Yang JS,Chen GW,et al.The roles of endoplasmic reticulum stress and Ca²⁺ on rhein-induced apoptosis in A-549 human lung cancer cells[J].Anticancer Res,2009,29(1):309-318.
[4]	Lin ML,Chen SS,Lu YC,et al.Rhein induces apoptosis through induction of endoplasmic reticulum stress and Ca²⁺-dependent mitochondrial death pathway in human nasopharyngeal carcinoma cells[J].Anticancer Res,2007,27(5A):3313-3322.
[5]	Du Q,Bian XL,Xu XL,et al. Role of mitochondrial permeability transition in human hepatocellular carcinoma Hep-G2 cell death induced by rhein[J].Fitoterapia,2013,91:68-73.
[6]	National Toxicology Program.NTP toxicology and carcinogenesis studies of EMODIN(CAS NO.518-82-1)feed studies in F344/N rats and B6C3F1 mice[J].Natl Toxicol Program Tech Rep Ser,2001,493:1-278.
[7]	Zhang LY,Jiang ZZ,Pu CH,et al.Six-month oral toxicity study of total anthraquinone in radix et rhizoma rhei in SD rats[J].Chin J Biochem Pharm(中国生化药物杂志),2004,25(4):206-209.
[8]	Ren HB,Wang YY,Wang TJ.Rhubarb total anthraquinone to rat acute renal toxicity research[J].J Liaoning Univ Tradit Chin Med(辽宁中医药大学学报),2012,14(1):69-71.
[9]	Zhu S,Wang Y,Jin J,et al.Endoplasmic reticulum stress mediates aristolochic acid I-induced apoptosis in human renal proximal tubular epithelial cells[J].Toxicol In Vitro,2012,26(5):663-671.
[10]	Da HY,Jiang ZZ,Wang CF,et al.The toxic effects of rhein and emodin on human renal tubular epithelial cells in vitro[J].Chin Tradit Herb Drug(中草药),2009,40(1):102-105.
[11]	Du AP,Zou WZ,Yang JP.Apoptosis in injury and regeneration of tubular epithelial cells[J].J Beijing Med Univ(北京医科大学学报),1997,29(6):481-484.
[12]	Tang W,Wang W,Zhang Y,et al.Tumour necrosis factor-related apoptosis-inducing ligand(TRAIL)-induced chemokine release in both TRAIL-resistant and TRAIL-sensitive cells via nuclear factor kappa B[J].FEBS J,2009,276(2):581-593.
[13]	Zhao YC,Gu G. Advance research of apoptosis pathway[J].Modern Med J(现代医学),2013,41(4):285-288.
[14]	Yu XL,Wang H,Zhang B,et al.Relationship between apoptosis of renal tubular epithelial cells and the change of expression of Bc-l 2 and Fas/ FasL proteins in rats after renal ischemia/reperfusion injury[J].J Fourth Mil Med Univ(第四军医大学学报),2003,24(8):723-726.
[15]	Reimer T,Herrnring C,Koczan D,et al.FasL:Fas ratio — a prognostic factor in breast carcinomas[J].Cancer Res,2000,60(4):822-828.
[16]	Kober A MM,Legewie S,Pforr C,et al.Caspase-8 activity has an essential role in CD95/Fas-mediated MAPK activation[J].Cell Death Dis,2011,2(10):e212.
[17]	Dasmahapatra G,Almenara JA,Grant S.Flavopiridol and histone deacetylase inhibitors promote mitochondrial injury and cell death in human leukemia cells that overexpress Bcl-2[J].Mol Pharmacol,2006,69(1):288-298.
[18]	Chen J,Mehta J L,Haider N,et al.Role of caspases in Ox-LDL-induced apoptotic cascade in human coronary artery endothelial cells[J].Circ Res,2004,94(3):370-376.

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