血竭素高氯酸盐对高糖培养的肾小管上皮细胞表达结缔组织生长因子和纤维连接蛋白的影响

Effects of dracorhodin perchlorate on the expression of CTGF and FN in human renal proximal tubular epithelial cell line cultured in high glucose

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。

OBJECrlVE To investigate the role of dracorhodin perchlorate on the expression of connective tissue growth factor （CTGF） and fibronectin （FN） in human renal proximal tubular epithelial cell line（ HKC cells）cultured under high glucose circumstance, and to explore its mechanism of renal tubular interstitial fibrosis in diabetic nephropathy （DN）. METHODS HKC cells cultured in vitro were randomized into four groups： a normal glucose group （NG, 5. 5 mmol· L^-1 D-glucose）, a high glucose group （HG, 25 mmol/L D-glucose）, a dracorhodin perchlorate and normal glucose group（DNG, 7. 5 μmol/L dracorhodin perchlorate ＋ 5.5 μmmol·L^-1 D-glucose ）,an dracorhodin perchlorate and high glucose group（DHG,7. 5μmmol· L^-1 dracorhodin perchlorate ＋ 25 mmol·L^-1 D-glucose ）. After the treatment of dracorhodin perchlorate for 48 h and 72 h, the expressions of CTGF mRNA and FN mRNA were assessed by semi quantitative RT-PCR, and the presence of FN protein was measured by immunohistochemistry. RESULTS A basal expression of CTGF mRNA, FN mRNA and FN protein expression were detected in HKC cells. Compared with the NG group, the level of CTGF mRNA , FN mRNA and FN protein were signif ieantly higher in HG group after stimulating 48 h and 72 h （P〈0. 05）. These parameters were diminished both in the DHG group and in the DNG group. It exists statistically significance between the HG group and the DHG group（P〈0.05）, but none in the DNG group and the NG group （P〉0. 05）. CONCLUSION Dracorhodin perchlorate can inhibit the renal tubular intersti tial fibrosis via suppressing the synthesis of CTGF and FN, and may be applied to the prevention and treatment of renal fibrosis in diabetic nephropathy.