结缔组织生长因子在阿魏酸钠干预单侧输尿管梗阻大鼠肾脏的表达(英文)

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背景:结缔组织生长因子是转化生长因子β1的下游效应因子,其异常升高在肾纤维化发生、发展中起关键作用,干预结缔组织生长因子在肾脏的异常表达可改善肾脏的纤维化。目的:从mRNA和蛋白水平观察阿魏酸钠对单侧输尿管梗阻大鼠肾组织结缔组织生长因子的影响,以及阿魏酸钠干预后大鼠肾间质纤维化的变化,并与氯沙坦进行比较。设计:随机对照动物实验。单位:四川大学华西医院肾脏科和四川大学公共卫生学院。材料:实验选用雄性成年健康SD大鼠24只,购于四川大学实验动物中心(实验用阿魏酸钠片由成都亨达药业有限公司惠赠,批号:050302)。兔抗大鼠结缔组织生长因子多克隆抗体(Santa Cruz)。Western印迹设备购自美国BioRAD,DNA Engine OpticonTM实时荧光定量PCR仪购自美国MJ Re-search公司。方法:实验于2006-05/12在四川大学公共卫生学院医学检验研究室完成(生物安全级别:BSL-1)。将24只大鼠随机分为4组:模型组、阿魏酸钠组、氯沙坦组和假手术组,每组6只。前3组按既往文献方法建立单侧输尿管梗阻模型,假手术行假手术。术后24h开始,阿魏酸钠治疗组予阿魏酸钠溶液150mg/(kg·d)灌胃;氯沙坦组予氯沙坦溶液20mg/(kg·d)灌胃,其余2组灌胃同体积生理盐水。术后14d处死大鼠,取部分肾组织用于分析。主要观察指标:采用荧光定量PCR,Western印迹方法从mRNA和蛋白质水平检测肾组织结缔组织生长因子的表达情况;行HE,Masson染色观察肾间质病理学改变。结果:①阿魏酸钠对肾组织结缔组织生长因子表达的影响:模型组术后14d结缔组织生长因子mRNA和蛋白的表达较假手术组明显升高,而阿魏酸钠治疗组与模型组比较有明显的下降(P<0.05),同时,与氯沙坦组比较,差异无显著性意义(P>0.05)。②阿魏酸钠干预后肾组织病理学改变:单侧输尿管梗阻术后第14天苏木精-伊红染色和Masson染色可见间质中多量炎性细胞浸润和小管、间质的改变,间质胶原的沉积等肾纤维化的病变;单侧输尿管梗阻大鼠经阿魏酸钠治疗治疗后,阿魏酸钠治疗改善了肾组织的纤维化(P<0.05),与氯沙坦组比较,差异无显著性意义(P>0.05)。结论:①阿魏酸钠对单侧输尿管梗阻所致的大鼠肾脏纤维化的形成有明显的抑制作用,其作用机制可能与其降低结缔组织生长因子的表达有关。②阿魏酸钠抑制肾脏纤维化作用与氯沙坦相当。 BACKGROUND: Connective tissue growth factor is a downstream effector of transforming growth factor β1. Abnormally elevated connective tissue plays a key role in the occurrence and development of renal fibrosis. Abnormal expression of connective tissue growth factor in the kidney may improve renal fibrosis. OBJECTIVE: To observe the effects of sodium ferulate on the expression of connective tissue growth factor (TGF) in rat renal tissue after unilateral ureteral obstruction (UUO) and the changes of rat renal interstitial fibrosis after sodium ferulate intervention with losartan Compare. Design: Randomized controlled animal experiments. Unit: West China Hospital of Sichuan University Department of Nephrology and Sichuan University School of Public Health. MATERIALS: Twenty-four male adult healthy Sprague-Dawley rats were selected and purchased from Experimental Animal Center of Sichuan University (experimental ferulic acid sodium tablets were kindly donated by Chengdu Hengda Pharmaceutical Co. Ltd., lot number: 050302). Rabbit anti-rat connective tissue growth factor polyclonal antibody (Santa Cruz). Western blotting equipment purchased from the United States BioRAD, DNA Engine OpticonTM real-time fluorescence quantitative PCR instrument was purchased from the United States MJ Re-search company. Methods: The experiment was performed at the Institute of Medical Laboratory Science, School of Public Health, Sichuan University from May to December 2006 (BSL-1). 24 rats were randomly divided into 4 groups: model group, sodium ferulate group, losartan group and sham operation group, 6 rats in each group. The first three groups according to the previous literature to establish unilateral ureteral obstruction model, sham surgery sham surgery. The rats in the sodium ferulate group were administrated 150mg / (kg · d) of sodium ferulate solution by intragastric administration after 24 hours, losartan group was given 20mg / (kg · d) Stomach the same volume of saline. The rats were sacrificed 14 days after operation and some of the kidney tissue was taken for analysis. MAIN OUTCOME MEASURES: The expression of connective tissue growth factor in renal tissues was detected by quantitative PCR and Western blotting at mRNA and protein levels. The pathological changes of renal interstitium were observed by HE and Masson staining. Results: ① The effect of sodium ferulate on the expression of connective tissue growth factor in kidney: The expression of connective tissue growth factor mRNA and protein in model group at 14th day after operation was significantly higher than that in sham operation group, while the sodium ferulate group and model group (P <0.05). At the same time, there was no significant difference between losartan group and losartan group (P> 0.05). Pathological changes of renal tissue after sodium ferulate intervention: Unilateral ureteral obstruction on the fourteenth day after hematoxylin-eosin staining and Masson staining showed interstitial inflammatory cell infiltration and tubulointerstitial changes Collagen deposition and other pathological changes of renal fibrosis; unilateral ureteral obstruction rats treated with sodium ferulate, sodium ferulate improved renal fibrosis (P <0.05), compared with losartan , No significant difference (P> 0.05). Conclusion: (1) Sodium ferulate has a significant inhibitory effect on the formation of renal fibrosis induced by unilateral ureteral obstruction in rats, which may be related to the decrease of the expression of connective tissue growth factor. ② sodium ferulate inhibition of renal fibrosis and losartan quite.
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