摘要
目的探讨非诺贝特对游离脂肪酸诱导的肾小管上皮细胞(HK-2)凋亡的影响。方法通过MTT法检测细胞的增殖,分光光度法检测细胞MDA表达和SOD活性;ELISA法检测细胞上清中的MCP-1和IL-8;Real-time PCR法及Western blot分别检测Bax和Bcl-2的m RNA和蛋白表达水平。结果游离脂肪酸的刺激呈剂量和时间依赖性抑制HK-2细胞的增殖,并使细胞Bax的m RNA与蛋白表达显著上升,Bcl-2的m RNA和蛋白的表达显著下降。非诺贝特的加入显著减轻游离脂肪酸对HK-2细胞增殖的抑制作用;并使细胞bax的m RNA与蛋白表达下降,而Bcl-2的m RNA与蛋白的表达上升。同时细胞SOD表达水平上升,MDA的表达水平下降;炎性介质MCP-1和IL-8表达水平均下降。结论游离脂肪酸诱导的肾小管上皮细胞凋亡具有时间和剂量依赖性,非诺贝特可通过减少氧化应激,抑制炎性反应,上调Bcl-2以及下调Bax的表达来抑制游离脂肪酸诱导的细胞凋亡,从而减少其肾毒性。
Objeetive To explore the influence of fenofibrate on apoptasis of human renal proximal tubular ceUs （HK-2） induced by free fatty acid （FFAs）. Methods Methyl aso thiazole blue（determined by MTr）was applied for deteetian Of IlK- 2 eell proliferation capacity; Spectmphotometry was used to determine the expression of MDA and SOD; MCP- 1 and IL-8 in culture supematant of cells were detected by ELISA; Real-time PCR and Western blot were used to evaluate w.RNA and protein expression of Bax and Bcl-2 respectively. Reslflts FFAs inhibited HK-2 cell prolifera- tion in a dose and lime dependence, mRNA and protein expression of Bax signdleanfly increased compared with control, but mRNA and protein ex- pression of Bel- 2 decreased. After preincubafian of fenofibrate, the inhibition of HK- 2 eel] proliferation by FFAs was alleviated; expression of SOD increased and expression of MDA, MCP- 1 and IL-8 were decreased; mRNA and protein expression of Bax was signiiieanfly decreased, but mRNA and protein expression of Bcl-2 was increased. Conclusion FFAs can induce apoptosis of renal proximal tubular cell in a dose and time dependent manner. Fenofibrate can inhibite HK-2 cell apoptesis by decreasing oxidative stress, inhibiting inflammation, up-regulating expression of Bcl-2 and down-regulating expression of Bax,which alleviated nephrotexicity of FFAs.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2015年第2期127-131,142,共6页
Journal of China Medical University
基金
辽宁省教育厅科学研究一般项目(L2011132)
关键词
非诺贝特
游离脂肪酸
肾小管上皮细胞
凋亡
fenofibrate
free fatty acids
renal proximal tubular cell
apoptosis
