阿托伐他汀对无血清和缺氧条件下对比剂诱导人肾小管上皮细胞凋亡的影响

发布时间：2018-08-16 10:19

【摘要】：【目的】研究阿托伐他汀（ATO）对在无血清和缺氧条件下对比剂（CM）诱导人肾小管上皮细胞凋亡的影响，以及氧化应激在该过程中的可能作用机制。【方法】以人肾小管上皮细胞（HKC）为研究对象，进行培养，在无血清、缺氧条件下,分别与不同浓度（100、120、150、200mgI/mL）碘海醇孵育12h；碘海醇（120mgI/mL）分别处理细胞2、6、9、12h；不同浓度ATO（10-11、10-10、10-9、10-8、10-7、10-6、10-5mol/L）先与细胞孵育24h，同一浓度ATO（10-7mol/L）先处理细胞6、12、24、36、48h，二者分别再与碘海醇（120mgI/mL）在无血清、缺氧条件下共刺激12h；不同浓度ATO（10-10、10-9、10-8、10-7mol/L）与细胞共孵育24h后，再与碘海醇（120mgI/mL）在无血清、缺氧条件下共刺激12h。采用CCK-8法检测细胞增殖能力； TUNEL法观察细胞凋亡情况；采用化学法检测丙二醛（MDA）含量和超氧化物歧化酶（SOD）活性； Western blot检测gp91phox、P22phox、bax及bcl-2等蛋白表达。【结果】 CCK-8法示与正常组(6.06±0.18)%相比，各组细胞增殖能力均受不同程度抑制，其中CM+缺氧组受影响最明显[（57.86±0.28）%, P0.05]，ATO处理后有所改善，且与浓度有关。 TUNEL检测显示与正常组（0.06±0.01）相比，CM+缺氧组细胞凋亡情况最明显（0.88±0.03, P0.05），，而ATO可改善凋亡情况。检测MDA含量与SOD活性显示， CM使MDA含量增加及SOD活性降低，而ATO可使MDA含量减少及SOD活性升高（P0.05）。 Western blot法检测显示， CM使gp91phox、 p22phox、 bax表达上调，bcl-2表达下调，而ATO使gp91phox、p22phox、bax表达下调，而bcl-2表达上调（P0.05）。【结论】在无血清缺氧条件下，碘海醇呈浓度和时间依赖性降低细胞增殖能力，并诱导细胞凋亡，而他汀可减轻细胞凋亡情况，且与浓度、时间有关。同时，碘海醇可能通过增强氧化应激水平，且可能与NADPH氧化酶途径相关，以诱导细胞凋亡，而ATO能改善这一氧化应激损伤作用，起到细胞保护作用。
[Abstract]:[objective] to study the effect of Atto vastatin (ATO) on apoptosis of human renal tubular epithelial cells induced by (CM), a contrast medium without serum and hypoxia. [methods] Human renal tubular epithelial cells (HKC) were cultured and incubated with different concentrations of 100120150200mgI/mL (100120150200mgI/mL) for 12 h without serum and hypoxia. The cells were incubated with different concentrations of ATO (10-11 10-10 -10 -10 ~ (-9) 10 ~ (-8) 10 ~ (-8) 10 ~ (-6) 10 ~ (-5) mol / L for 24 h, and ATO (10-7mol/L) with the same concentration of ATO (10-7mol/L) for 24 ~ 48 h. The cells were incubated with iodohexanol (120mgI/mL) for 12 h under anoxic condition for 12 h, and ATO at different concentration (10 ~ (-10 ~ (-9) 10 ~ (-8) mol / L) was incubated with the cells for 24 h after incubating with different concentrations of ATO (10-10 ~ (-9) and 10 ~ (-8) mol / L for 24 h, respectively. Then stimulated with 120mgI/mL for 12 h under anoxic condition without serum. Cell proliferation was detected by CCK-8 assay, apoptosis was observed by TUNEL assay, malondialdehyde (MDA) content and (SOD) activity of superoxide dismutase (SOD) were detected by chemical method. [results] compared with normal group (6.06 卤0.18)%, the proliferation ability of all groups was inhibited to some extent by Western blot. The CM hypoxia group was the most affected [(57.86 卤0.28), P0.05] after ATO treatment, the expression of Gp91phoxP22phoxnbax and bcl-2 were detected. [results] compared with the normal group (6.06 卤0.18)%, the proliferation ability of the cells was inhibited to some extent, especially in the CM hypoxia group [(57.86 卤0.28), P0.05]. And it is related to the concentration. Compared with the normal group (0.06 卤0. 01), TUNEL showed that apoptosis was the most obvious (0. 88 卤0. 03, P 0. 05) in the anoxia group, while ATO could improve the apoptosis. Detection of MDA and SOD activity showed that CM increased MDA content and decreased SOD activity, while ATO decreased MDA content and SOD activity (P0.05). Western blot assay showed that CM up-regulated the expression of gp91phox, p22phoxand bax, while ATO down-regulated the expression of gp91phoxp22phoxanbax, while the expression of bcl-2 was up-regulated (P0.05). [conclusion] under the condition of no serum hypoxia, the expression of gp91phox, p22phox and bax was down-regulated. Iodohexanol decreased cell proliferation and induced apoptosis in a concentration-and time-dependent manner, while statins alleviated apoptosis in a dose-and time-dependent manner. At the same time, iodohexanol may induce apoptosis by enhancing the oxidative stress level and may be related to the NADPH oxidase pathway, while ATO can improve the oxidative stress injury and play a cell protection role.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R692.6

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