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目的探讨醌型二氢生物喋呤还原酶(Quinoid dihydropteridine reductase,QDPR)基因改变对高糖环境下肾小管上皮细胞系NRK-52Eα-辅肌动蛋白(α-actinin)的影响及其在糖尿病肾病(Diabetic nephropathy,DN)中的可能作用机制。方法 Western-blot检测DN动物(OLETF大鼠)以及对照LETO大鼠肾皮质的α-actinin蛋白含量。构建过表达及敲低QDPR基因的NRK-52E及其对照模型,分别给予正常糖(5.4mmol/L)和高糖(30mmol/L)培养基培养72h,Western-blot检测α-actinin在细胞模型各组的表达水平。结果 OLETF大鼠肾皮质内α-actinin含量降低[(0.86±0.32)vs(0.31±0.22),P<0.01];NRK-52E高糖组的α-actinin含量降低[(0.80±0.13)vs(0.44±0.10),P<0.05];与空载过表达病毒对照组(0.66±0.04)比,空载过表达病毒对照高糖组(0.42±0.06)及QDPR基因过表达组(0.49±0.06)的α-actinin蛋白表达含量降低,差异有统计学意义(P<0.05);与QDPR基因过表达组相比,QDPR基因过表达高糖组的α-actinin含量降低[(0.49±0.06)vs(0.21±0.02),P<0.05];与空载过表达病毒对照高糖组比,QDPR基因过表达高糖组的α-actinin含量降低[(0.42±0.06)vs(0.21±0.02),P<0.05];与敲低随机序列对照组比,敲低随机序列对照高糖组的α-actinin降低[(0.93±0.09)vs(0.69±0.08),P<0.05];敲低QDPR基因不影响α-actinin蛋白表达量。结论α-actinin在DN模型中含量减少,QDPR基因可能通过α-actinin影响DN的发生、发展。
Objective To investigate the effect of Quinoid dihydropteridine reductase (QDPR) gene alteration on renal tubular epithelial cell line NRK-52Eα-actinin and its effect on diabetic nephropathy (Diabetic nephropathy, DN) in the possible mechanism of action. Methods Western-blot was used to detect the protein content of α-actinin in renal cortex of DN animals (OLETF rats) and control LETO rats. NRK-52E and its control model were overexpressed and knocked down QDPR gene and cultured in normal glucose (5.4mmol / L) and high glucose (30mmol / L) medium for 72h respectively. The expression of α-actinin in cell model The expression level of each group. Results The content of α-actinin in the renal cortex of OLETF rats was significantly lower than that of the control rats [(0.86 ± 0.32) vs (0.31 ± 0.22), P <0.01] 0.44 ± 0.10), P <0.05]. Compared with the control group without empty vector (0.66 ± 0.04), the control group without empty vector overexpression (0.42 ± 0.06) and the overexpression of QDPR gene (0.49 ± 0.06) (P <0.05). Compared with QDPR overexpression group, the content of α-actinin in QDPR overexpression high glucose group decreased ([0.49 ± 0.06] vs ( 0.21 ± 0.02), P <0.05]. The level of α-actinin in QDPR overexpressing high glucose group was significantly lower than that in control overexpression high glucose group [(0.42 ± 0.06) vs (0.21 ± 0.02), P < 0.05]. Compared with knocking down random sequence control group, the knockdown of α-actinin in high glucose group with random sequence was significantly lower than that in control group [(0.93 ± 0.09) vs (0.69 ± 0.08), P <0.05] -actinin protein expression. Conclusion α-actinin decreased in DN model, QDPR gene may affect the occurrence and development of DN through α-actinin.