【摘要】：目的:探討intermedin(IMD)是否通過抑制氧化應(yīng)激效應(yīng)來緩解單側(cè)輸尿管梗阻(UUO)后大鼠的腎臟纖維化。方法:雄性Wistar大鼠,隨機分為5組:假手術(shù)組(sham組)、UUO模型組、UUO+IMD組、UUO+IMDshRNA組、UUO+空質(zhì)粒組。轉(zhuǎn)染組的UUO模型在質(zhì)粒轉(zhuǎn)染后7d來制做,各組分別于術(shù)后1d、3d、7d、14d取6只,Masson染色來觀察腎臟纖維化的水平;化學(xué)熒光法檢測大鼠腎組織局部活性氧類(ROS)物質(zhì)的表達,比色法去檢測超氧化物歧化酶(SOD)的活性及脂質(zhì)過氧化產(chǎn)物丙二醛(MDA)含量,real time RT-PCR檢測腎組織血紅素加氧酶-1(HO-1)的mRNA表達,免疫組化法檢測其蛋白質(zhì)表達。結(jié)果:Masson染色結(jié)果顯示,Sham組檢測各時間點腎組織無明顯纖維化表現(xiàn);UUO各組伴隨著梗阻時間延長而腎間質(zhì)纖維化進行性加重;UUO+IMD組各時間點腎間質(zhì)纖維化程度較相應(yīng)UUO組明顯減輕(P0.05);而UUO+IMDshRNA組腎間質(zhì)纖維化程度較相應(yīng)UUO組更重(P0.05);UUO+空質(zhì)粒組與UUO相比無明顯區(qū)別。與sham組相比,UUO組大鼠各檢測時間點腎組織ROS和MDA含量明顯增高(P0.05)。UUO+IMD組較相應(yīng)時間點UUO組ROS和MDA含量明顯降低;而UUO+IMDshRNA組較相應(yīng)UUO組ROS和MDA含量更高(P0.05);UUO+空質(zhì)粒組則與相應(yīng)UUO相比無明顯區(qū)別(P0.05)。與sham組相比,UUO組SOD活性、HO-1的mRNA和蛋白表達在第1d明顯升高,以后逐漸下降,SOD活性第3d開始低于sham組,而HO-1的mRNA和蛋白表達各檢測時間點均高于sham組(P0.05),UUO+IMD組較相應(yīng)時間點UUO組SOD活性和HO-1的表達更高(P0.05),而UUO+IMDshRNA組SOD活性和HO-1的表達較相應(yīng)UUO組顯著下降(P0.05),UUO+空質(zhì)粒組與UUO相比無明顯區(qū)別(P0.05)。結(jié)論:IMD是通過抑制了氧化應(yīng)激來減輕UUO引起的大鼠腎臟纖維化。
[Abstract]:Aim: to investigate whether intermedin (IMD) can alleviate renal fibrosis after unilateral ureteral obstruction (UUO) by inhibiting oxidative stress. Methods: male Wistar rats were randomly divided into 5 groups: sham operation group (sham group,), UUO model group, UUO IMD group, UUO IMDshRNA group, UUO empty plasmid group). The UUO model of the transfection group was made 7 days after transfection, and 6 rats were taken from each group on the 1st day, 3d, 7d, 14d, respectively. The level of renal fibrosis was observed by Masson staining. The activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA), the products of lipid peroxidation were detected by chemical fluorescence method and colorimetric method respectively. The mRNA expression of heme oxygenase-1 (HO-1) and the protein expression of heme oxygenase-1 (HO-1) in renal tissue were detected by real time RT-PCR and immunohistochemistry respectively. Results: the results of Masson staining showed that there was no obvious fibrosis in renal tissue at different time points in Sham group, and the renal interstitial fibrosis in UUO group was aggravated with the prolongation of obstruction time. The degree of renal interstitial fibrosis in UUO IMD group was significantly less than that in corresponding UUO group at each time point (P0.05), while the degree of renal interstitial fibrosis in UUO IMDshRNA group was more serious than that in corresponding UUO group (P0.05). There was no significant difference between); UUO empty plasmid group and UUO group (P0.05). Compared with sham group, the content of ROS and MDA in renal tissue in UUO group was significantly higher than that in UUO group at different time points (P0.05), but the content of ROS and MDA in UUO IMDshRNA group was higher than that in UUO group (P0.05). There was no significant difference between UUO empty plasmid group and corresponding UUO group (P0.05). Compared with sham group, SOD activity, HO-1 mRNA and protein expression in UUO group increased significantly on day 1, then decreased gradually, SOD activity began to be lower than that in sham group on day 3, while mRNA and protein expression in HO-1 group was higher than that in sham group at all time points (P0.05). SOD activity and HO-1 expression in UUO IMD group were higher than those in UUO group (P0.05), while SOD activity and HO-1 expression in UUO IMDshRNA group were significantly lower than those in UUO group (P0.05). There was no significant difference between UUO empty plasmid group and UUO group (P0.05). Conclusion: IMD attenuates renal fibrosis induced by UUO by inhibiting oxidative stress.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R692

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