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  本文選題：青蒿琥酯 + 腎小球系膜細(xì)胞�。� 參考：《桂林醫(yī)學(xué)院》2017年碩士論文

【摘要】：目的:觀察高糖環(huán)境下大鼠腎小球系膜細(xì)胞增殖的情況,探討不同濃度青蒿琥酯是否可抑制高糖環(huán)境培養(yǎng)的大鼠腎小球系膜HBZY-1細(xì)胞增殖和炎癥因子TNF-α、IL-6的分泌、并誘導(dǎo)其凋亡。方法:將大鼠腎小球系膜細(xì)胞分成7組:正常培養(yǎng)對照組、高糖培養(yǎng)模型組、青蒿琥酯微劑量組、青蒿琥酯小劑量組、青蒿琥酯中劑量組、青蒿琥酯大劑量組、依那普利陽性藥物對照組。倒置顯微鏡下觀察各組腎小球系膜細(xì)胞形態(tài)變化。采用MTT法觀察青蒿琥酯對各組腎小球細(xì)胞細(xì)胞24h、48h、72h增殖的抑制情況,并通過流式細(xì)胞儀檢測各組細(xì)胞凋亡率,ELISA法觀察各組細(xì)胞上清液中腫瘤壞死因子α(TNF-α)、白細(xì)胞介素6(IL-6)的分泌量變化。結(jié)果:(1)青蒿琥酯細(xì)胞12h左右貼壁,24h伸展成紡錘狀或不規(guī)則形狀,48h進(jìn)入對數(shù)期生長,3-4天可長至融合。細(xì)胞核成卵圓形或圓形。加入藥物后,各組細(xì)胞融合度有差異。(2)MTT結(jié)果顯示:正常培養(yǎng)對照組細(xì)胞增殖抑制率明顯高于高糖培養(yǎng)模型組(P0.01),各濃度青蒿琥酯藥物組的抑制率均高于高糖組(P0.05),并成一定時間、劑量依賴性。與依那普利組對比,青蒿琥酯微劑量組抑制率無統(tǒng)計學(xué)意義(P0.05)。(3)青蒿琥酯對HBZY-1細(xì)胞凋亡有誘導(dǎo)作用(P0.05),并呈一定劑量依賴性;(4)ELISA結(jié)果顯示:與正常對照組相比,高糖培養(yǎng)模型組細(xì)胞培養(yǎng)的上清液中TNF-α、IL-6的含量明顯增高(P0.01);與高糖培養(yǎng)模型組比較,各青蒿琥酯藥物組的上清液中TNF-α、IL-6的含量均降低(P0.05),大劑量青蒿琥酯組比依那普利組作用明顯(P0.01)。結(jié)論:高糖環(huán)境可促進(jìn)HBZY-1細(xì)胞增殖和炎癥因子TNF-α、IL-6的分泌,可誘導(dǎo)機(jī)體炎癥反應(yīng);青蒿琥酯能明顯逆轉(zhuǎn)高糖刺激下GMCs的增殖和凋亡并且對高糖環(huán)境下HBZY-1細(xì)胞分泌TNF-α、IL-6有明顯抑制作用。由此推論青蒿琥酯可以作為治療、防治糖尿病腎病的有效藥物,并減輕高糖環(huán)境對組織器官的損害。
[Abstract]:Aim: to observe the proliferation of rat glomerular Mesangial cells in high glucose environment, and to investigate whether artesunate at different concentrations can inhibit the proliferation of glomerular Mesangial HBZY-1 cells and the secretion of inflammatory factor TNF- 偽 and IL-6, and induce apoptosis in rat glomerular Mesangial HBZY-1 cells cultured in high glucose environment. Methods: rat glomerular Mesangial cells were divided into 7 groups: normal culture control group, high glucose culture model group, artesunate microdose group, artesunate small dose group, middle dose group of artesunate, high dose group of artesunate. Enalapril positive drug control group. The morphological changes of glomerular Mesangial cells were observed under inverted microscope. The inhibitory effects of artesunate on proliferation of glomerular cells at 24 h and 48 h and 72 h were observed by MTT assay. The expression of TNF- 偽 TNF- 偽 and IL-6 in the supernatant of each group was detected by flow cytometry and Elisa. Results (1) artesunate cells extended into spindle-like or irregularly shaped cells for 12 h or so for 24 h. The cells could grow to fusion in 3 to 4 days after entering logarithmic phase. The nucleus becomes oval or round. The results of MTT showed that the inhibition rate of cell proliferation in normal culture group was significantly higher than that in high glucose culture model group (P 0.01), and the inhibition rate of artesunate drug group in each concentration was higher than that in high glucose group, and the inhibition rate of artesunate drug group was higher than that of high glucose group, and the inhibition rate of artesunate group was higher than that of high glucose group. Dose dependence. In comparison with enalapril group, the inhibitory rate of artesunate microdose group was not statistically significant (P 0.05. 0. 3) artesunate induced apoptosis of HBZY-1 cells. The results of Elisa showed that artesunate could induce apoptosis of HBZY-1 cells in a dose-dependent manner, and the results showed that: compared with normal control group, artesunate could induce apoptosis of HBZY-1 cells in a dose-dependent manner. The content of TNF- 偽 and IL-6 in the supernatant of high glucose culture model group was significantly higher than that in the high glucose culture model group, and the content of TNF- 偽 IL-6 in the supernatant of each artesunate drug group was lower than that of enalapril group, and the effect of high dose artesunate group was more obvious than that of enalapril group. Conclusion: high glucose environment can promote the proliferation of HBZY-1 cells and the secretion of TNF- 偽 IL-6, and induce inflammatory reaction. Artesunate could significantly reverse the proliferation and apoptosis of GMCs stimulated by high glucose, and inhibit the secretion of TNF- 偽-偽 IL-6 by HBZY-1 cells under high glucose. Artesunate can be used as an effective drug to prevent and treat diabetic nephropathy and reduce the damage to tissues and organs caused by high glucose environment.
【學(xué)位授予單位】：桂林醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R587.2;R692.9

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