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姜宣羽,王思为,楼丽君,钟松阳,胡泽富.土茯苓活性分子落新妇苷抗顺铂诱导的人肾小管上皮细胞凋亡作用研究[J].浙江中西医结合杂志,2018,28(9):
土茯苓活性分子落新妇苷抗顺铂诱导的人肾小管上皮细胞凋亡作用研究
Anti-apoptotic effect of Astilbin from Smilax glabra Roxb. on cisplatin-induced HK-2 cells
投稿时间:2018-03-26  修订日期:2018-04-20
DOI:
中文关键词:  土茯苓  落新妇苷  顺铂肾毒性  凋亡  OCT2
英文关键词:Smilax glabra Roxb.  astilbin  cisplatin nephrotoxicity  apoptosis  OCT2
基金项目:浙江省自然科学(LYY18H280001);浙江省中西医结合学会科研(No 2016LYK011)
作者单位E-mail
姜宣羽 衢州市人民医院 药学部 262530238@qq.com 
王思为 衢州市人民医院 药学部  
楼丽君 衢州市人民医院 药学部  
钟松阳 衢州市人民医院 药学部  
胡泽富 衢州市人民医院 药学部  
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中文摘要:
      目的:研究土茯苓活性分子落新妇苷抗顺铂诱导的人肾小管上皮细胞(HK-2)凋亡的作用。方法:采用MTT法分别观察不同浓度落新妇苷联合顺铂(50 μM)对HK-2细胞活性的影响。将HK-2细胞分为对照组、顺铂组、落新妇苷(100、200 μM)组。药物干预24 h后, AnnexinV/PI 流式细胞术检测各组细胞凋亡情况;Western blot法检测凋亡相关蛋白bax、bcl-2和cleaved-caspase-3表达情况;RT-PCR法和Western blot法检测药物转运蛋白OCT2 mRNA和蛋白质的表达情况。结果:MTT结果显示,落新妇苷能够增强顺铂诱导的HK-2细胞活性,且呈一定剂量依赖关系。流式细胞凋亡结果显示,落新妇苷能够显著抑制顺铂诱导的HK-2细胞凋亡,尤其200 μM剂量组显著(P<0.01)。Western blot结果显示,落新妇苷能够显著上调bax、cleaved-caspase-3,下调bcl-2蛋白表达。RT-PCR和Western blot结果显示,落新妇苷能够显著抑制HK-2细胞药物转运蛋白OCT2 mRNA和蛋白质表达。结论:落新妇苷具有抑制顺铂诱导的HK-2细胞凋亡的作用,其机制可能与抑制OCT2蛋白表达有关。
英文摘要:
      Objective: To study the anti-apoptotic effect of astilbin from Smilax Glabra Roxb. on cisplatin-induced HK-2 cells. Methods: MTT assay was used to observe the effects of different concentrations of astilbin and cisplatin (50 μM) on the growth of HK-2 cells. HK-2 cells were divided into control group, cisplatin group and astilbin (100, 200 μM) group. After 24 h drug intervention, Annexin V/PI flow cytometry was used to detect the apoptosis of HK-2 cells; Western blot was used to detect the apoptotic protein of bax, bcl-2 and cleaved-caspase-3 expression; RT-PCR and Western blot was used to detect the mRNA and protein expressions of drug transporter OCT2. Results: MTT results showed that astilbin could enhance the activity of cisplatin-induced HK-2 cells in a dose-dependent manner. Annexin V/PI flow cytometry results showed astilbin significantly inhibited cisplatin-induced apoptosis in HK-2 cells, especially in the 200 μM dose group (P <0.01). Western blot results showed that astilbin could up-regulate bax, cleaved-caspase-3 and down-regulate bcl-2 protein expression. RT-PCR and Western blot analysis showed that astilbin could inhibit the mRNA and protein expression of OCT2 in HK-2 cells. Conclusion: Astilbin has protective effect on the apoptosis of HK-2 cells induced by cisplatin, which mechanism may be related to the inhibition of OCT2 expression .
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