肾间质纤维化过程中HA/CD44表达变化

Changes of HA/CD44 Expression During Renal Interstitial Fibrosis

目的:观察肾间质纤维化过程中透明质酸(hyaluronic acid,HA)/CD44的表达变化及其与肾间质纤维化的关系。方法:单侧输尿管结扎(unilateral ureteral obstruction,UUO)复制小鼠肾间质纤维化模型,同时设假手术组。模型小鼠分别设手术后1周、2周、3周共3个时间观察组,同时设假手术组。检测小鼠血清肌酐、尿素氮水平,HE染色观察肾脏炎症,q Fibrosis分析胶原沉积,ELISA检测肾组织HA的含量,q-PCR分析透明质酸合成酶2(hyaluronic acid synthetase,HAS2)mRNA表达,免疫组化观察肾组织HA受体CD44表达,并进行以上指标相关性分析。结果:与假手术组相比,1周模型小鼠血清肌酐出现一过性升高,随着模型时间延长,逐渐接近正常水平;模型各时间点血清尿素氮水平无明显变化。q Fibrosis结果显示,假手术组胶原面积为(0.53±0.12)%;随着模型时间延长,胶原沉积逐渐增加,1周~3周胶原面积分别为(3.35±0.43)%、(5.87±0.71)%、(9.78±1.35)%;ELISA与q-PCR结果显示,随着肾间质纤维化进展,肾组织HA与HAS2表达均逐渐增加,3 w模型组HA与HAS的表达量分别为假手术组的7.2倍与23.8倍。免疫组化染色显示,随着纤维化发展,CD44阳染细胞数量逐渐增多,1周~3周CD44阳染面积分别为(3.50±0.91)%、(5.60±0.90)%、(14.12±2.31)%。结论:CD44分子主要表达于肾小管上皮细胞,随肾间质纤维化发展,肾组织HA、HAS2及CD44表达逐渐增加,与肾间质纤维化形成呈正相关关系。

Objective:To observe the changes of hyaluronic acid(HA)/CD44 expression and its relationship with renal interstitial fibrosis.Methods:Replicate the model of unilateral ureteral obstruction(UUO)mouse renal interstitial fibrosis and set a sham operation group.The model mice were divided into three groups:1 w,2 w,and 3 w after operation,and sham-operated group as control.Serum creatinine and urea nitrogen levels were measured to evaluate renal function,renal inflammation was observed by HE staining,renal tissue collagen deposition was analyzed by q Fibrosis,HA content in renal tissue was detected by ELISA,mRNA expression of hyaluronic acid synthase 2 was detected by q-PCR,renal tissue CD44(the receptor of HA)expression was observed by immunohistochemical staining,then analyze the correlation of the above indicators.Results:The serum creatinine appeared a transient increase,with the model time prolonged,gradually approaching normal levels;serum urea nitrogen levels did not change obviously in each group.The results of q Fibrosis showed that the collagen area in the sham operation group was(0.53±0.0.12)%;With the prolongation of the model,the renal interstitial collagen deposition gradually increased,and the collagen area of 1 w,2 w,3 w was(3.35±0.43)%,(5.87±0.71)%,(9.78±1.35)%.Conclusion:CD44 is mainly expressed in renal tubular epithelial cells.With the development of renal interstitial fibrosis,the expression of HA,HAS2 and CD44 in renal tissues is gradually increased,which is positively correlated with the formation of renal interstitial fibrosis.