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目的:研究血竭素高氯酸盐对高糖环境下肾小管上皮细胞(HKC)产生结缔组织生长因子(connective tissue growthfactor,CTGF)和纤维连接蛋白(fibronectin,FN)的影响作用,探讨血竭素高氯酸盐在糖尿病肾病(diabetlc nephropathy,DN)肾小管间质纤维化中的作用机制。方法:将体外培养的HKC细胞分为正常组(NG组,5.5 mmol·L~(-1) D-葡萄糖)、高糖培养组(HG组,25 mmol·L~(-1) D-葡萄糖),血竭素高氯酸盐+正常组(DNG组,7.5μmol·L~(-1) 血竭素高氯酸盐+5.5 mmol·L~(-1) D-葡萄糖)、血竭素高氯酸盐+高糖组(DHG组,7.5μmol·L~(-1)血竭素高氯酸盐+25 mmol·L~(-1) D-葡萄糖),48 h和72 h后,采用半定量RT-PcR法检测4组培养细胞中CTGF mRNA与FN mRNA的表达差异,并采用免疫组织化学染色法检测FN蛋白表达的变化。结果:常规培养(低糖环境)的HKc细胞有基础量的CTGF mRNA和FN mRNA及FN蛋白的表达;与NG组比较,高糖环境下HKC细胞CTGF mRNA和FN mRNA及FN蛋白表达均增加(P<0.01,P<0.05);与HG组比较,经血竭素高氯酸盐干预48 h和72 h后,高糖培养组的CTGF mRNA和FN的mRNA及FN蛋白表达量明显下调,其差异有显著性(P<0.01,P<0.05);与NG组比较,DNG组的CTGF mRNA和FN的mRNA及FN蛋白质的表达量亦下调,但差异无显著性(P>0.05)。结论:血竭素高氯酸盐能抑制高糖环境下的HKC中CTGF及FN的表达从而产生抗肾小管间质纤维化的作用,有望在DN的肾纤维化的治疗中发挥作用。
Objective: To study the effect of DRG on the expression of connective tissue growth factor (CTGF) and fibronectin (FN) in renal tubular epithelial cells (HKC) under high glucose condition Mechanism of perchlorate in tubulointerstitial fibrosis of diabetic nephropathy (DN). Methods: HKC cells cultured in vitro were divided into normal group (NG group, 5.5 mmol·L -1 D-glucose), high glucose group (HG group, 25 mmol·L -1 D-glucose ), Dracorhodin perchlorate + normal group (DNG group, 7.5μmol·L -1 dracorhodin perchlorate +5.5 mmol·L -1 D-glucose), dracorhodin Perchlorate + 25 mmol·L -1 D-glucose perfused with 7.5 μmol·L -1 DHG group), after 48 h and 72 h, Semi-quantitative RT-PcR method was used to detect the expression of CTGF mRNA and FN mRNA in four groups of cultured cells. Immunohistochemical staining was used to detect the expression of FN protein. Results: Compared with NG group, the amount of CTGF mRNA, FN mRNA and FN protein in HKc cells were significantly increased in HKc cells cultured in the low glucose condition (P <0.01, P <0.05). Compared with HG group, mRNA and FN protein expression of CTGF mRNA and FN in high glucose group were significantly decreased after 48 h and 72 h treatment (P <0.01, P <0.05). Compared with NG group, the expression of CTGF mRNA, FN mRNA and FN protein were also decreased in DNG group, but the difference was not significant (P> 0.05). CONCLUSION: Dracorhodin perchlorate inhibits the expression of CTGF and FN in HKC under high glucose conditions and thus produces anti-tubulointerstitial fibrosis, which is expected to play a role in the treatment of renal fibrosis in DN.