發(fā)布時間：2018-08-16 10:19

【摘要】：【目的】研究阿托伐他�。ˋTO）對在無血清和缺氧條件下對比劑（CM）誘導(dǎo)人腎小管上皮細(xì)胞凋亡的影響，以及氧化應(yīng)激在該過程中的可能作用機(jī)制。 【方法】以人腎小管上皮細(xì)胞（HKC）為研究對象，進(jìn)行培養(yǎng)，在無血清、缺氧條件下,分別與不同濃度（100、120、150、200mgI/mL）碘海醇孵育12h；碘海醇（120mgI/mL）分別處理細(xì)胞2、6、9、12h；不同濃度ATO（10-11、10-10、10-9、10-8、10-7、10-6、10-5mol/L）先與細(xì)胞孵育24h，同一濃度ATO（10-7mol/L）先處理細(xì)胞6、12、24、36、48h，二者分別再與碘海醇（120mgI/mL）在無血清、缺氧條件下共刺激12h；不同濃度ATO（10-10、10-9、10-8、10-7mol/L）與細(xì)胞共孵育24h后，再與碘海醇（120mgI/mL）在無血清、缺氧條件下共刺激12h。采用CCK-8法檢測細(xì)胞增殖能力； TUNEL法觀察細(xì)胞凋亡情況；采用化學(xué)法檢測丙二醛（MDA）含量和超氧化物歧化酶（SOD）活性； Western blot檢測gp91phox、P22phox、bax及bcl-2等蛋白表達(dá)。 【結(jié)果】 CCK-8法示與正常組(6.06±0.18)%相比，各組細(xì)胞增殖能力均受不同程度抑制，其中CM+缺氧組受影響最明顯[（57.86±0.28）%, P0.05]，ATO處理后有所改善，且與濃度有關(guān)。 TUNEL檢測顯示與正常組（0.06±0.01）相比，CM+缺氧組細(xì)胞凋亡情況最明顯（0.88±0.03, P0.05），，而ATO可改善凋亡情況。檢測MDA含量與SOD活性顯示， CM使MDA含量增加及SOD活性降低，而ATO可使MDA含量減少及SOD活性升高（P0.05）。 Western blot法檢測顯示， CM使gp91phox、 p22phox、 bax表達(dá)上調(diào)，bcl-2表達(dá)下調(diào)，而ATO使gp91phox、p22phox、bax表達(dá)下調(diào)，而bcl-2表達(dá)上調(diào)（P0.05）。 【結(jié)論】在無血清缺氧條件下，碘海醇呈濃度和時間依賴性降低細(xì)胞增殖能力，并誘導(dǎo)細(xì)胞凋亡，而他汀可減輕細(xì)胞凋亡情況，且與濃度、時間有關(guān)。同時，碘海醇可能通過增強(qiáng)氧化應(yīng)激水平，且可能與NADPH氧化酶途徑相關(guān)，以誘導(dǎo)細(xì)胞凋亡，而ATO能改善這一氧化應(yīng)激損傷作用，起到細(xì)胞保護(hù)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 鹵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 鹵0.28), P0.05] after ATO treatment, the expression of Gp91phoxP22phoxnbax and bcl-2 were detected. [results] compared with the normal group (6.06 鹵0.18)%, the proliferation ability of the cells was inhibited to some extent, especially in the CM hypoxia group [(57.86 鹵0.28), P0.05]. And it is related to the concentration. Compared with the normal group (0.06 鹵0. 01), TUNEL showed that apoptosis was the most obvious (0. 88 鹵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.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R692.6

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