Intermedin通过抑制氧化应激减轻单侧输尿管梗阻大鼠肾脏间质纤维化

发布时间：2018-11-04 15:37

【摘要】：目的:探讨intermedin(IMD)是否通过抑制氧化应激效应来缓解单侧输尿管梗阻(UUO)后大鼠的肾脏纤维化。方法:雄性Wistar大鼠,随机分为5组:假手术组(sham组)、UUO模型组、UUO+IMD组、UUO+IMDshRNA组、UUO+空质粒组。转染组的UUO模型在质粒转染后7d来制做,各组分别于术后1d、3d、7d、14d取6只,Masson染色来观察肾脏纤维化的水平;化学荧光法检测大鼠肾组织局部活性氧类(ROS)物质的表达,比色法去检测超氧化物歧化酶(SOD)的活性及脂质过氧化产物丙二醛(MDA)含量,real time RT-PCR检测肾组织血红素加氧酶-1(HO-1)的mRNA表达,免疫组化法检测其蛋白质表达。结果:Masson染色结果显示,Sham组检测各时间点肾组织无明显纤维化表现;UUO各组伴随着梗阻时间延长而肾间质纤维化进行性加重;UUO+IMD组各时间点肾间质纤维化程度较相应UUO组明显减轻(P0.05);而UUO+IMDshRNA组肾间质纤维化程度较相应UUO组更重(P0.05);UUO+空质粒组与UUO相比无明显区别。与sham组相比,UUO组大鼠各检测时间点肾组织ROS和MDA含量明显增高(P0.05)。UUO+IMD组较相应时间点UUO组ROS和MDA含量明显降低;而UUO+IMDshRNA组较相应UUO组ROS和MDA含量更高(P0.05);UUO+空质粒组则与相应UUO相比无明显区别(P0.05)。与sham组相比,UUO组SOD活性、HO-1的mRNA和蛋白表达在第1d明显升高,以后逐渐下降,SOD活性第3d开始低于sham组,而HO-1的mRNA和蛋白表达各检测时间点均高于sham组(P0.05),UUO+IMD组较相应时间点UUO组SOD活性和HO-1的表达更高(P0.05),而UUO+IMDshRNA组SOD活性和HO-1的表达较相应UUO组显著下降(P0.05),UUO+空质粒组与UUO相比无明显区别(P0.05)。结论:IMD是通过抑制了氧化应激来减轻UUO引起的大鼠肾脏纤维化。
[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.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R692

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