特异性敲除巨噬细胞CXCR4基因对单侧输尿管梗阻小鼠肾间质纤维化的缓解作用及机制  

Mitigative effect of specific CXCR4 gene knockout in macrophages on renal interstitial fibrosis in mice with unilateral ureteral obstruction

作　　者：张翠薇[1,2] 熊小明 刘勇 阮思蓓 马跃荣[1] ZHANG Cuiwei XIONG Xiaoming LIU Yong RUAN Sibei MA Yuerong(Chengdu University of Traditional Chinese Medicine, Chengdu 610000, China)

机构地区：[1]成都中医药大学,成都610000 [2]西南医科大学附属医院

出　　处：《山东医药》2016年第37期11-14,共4页Shandong Medical Journal

基　　金：国家自然科学基金面上项目(81473522)

摘　　要：目的观察特异性敲除巨噬细胞的CXCR4基因对单侧输尿管梗阻(UUO)小鼠肾间质纤维化的缓解作用,并探讨其作用机制。方法构建巨噬细胞CXCR4基因敲除小鼠模型(KO小鼠)16只并随机分为KO UUO组、KO Sham组各8只;将16只野生型C57小鼠(WT小鼠)随机分为WT UUO组、WT Sham组各8只。构建UUO模型,造模后第12天处死小鼠取左肾组织。用HE染色法评估肾小管病变程度;用Masson染色法评估肾间质胶原沉积程度;用免疫组化染色法评估肾组织巨噬细胞浸润情况并行M1、M2型巨噬细胞计数,计算两型细胞比值(M1/M2);用Western blotting法检测肾组织中基质细胞衍生因子1(SDF-1)、诱导型一氧化氮合酶(i NOS)及CD206蛋白表达。结果与KO Sham组、WT Sham组比较,KO UUO组、WT UUO组肾小管损伤程度减轻、肾间质胶原沉积程度及浸润的巨噬细胞数量减少,M1/M2升高,肾组织中i NOS、SDF-1蛋白表达升高(P均<0.05),KO UUO组以上指标比WT UUO组变化更明显(P均<0.05)。结论特异性敲除巨噬细胞的CXCR4基因可缓解UUO小鼠的肾间质纤维化程度,其机制可能为阻止巨噬细胞通过SDF-1/CXCR4轴趋化到肾间质中,减少M1型巨噬细胞向M2型转化。Objective To investigate the mitigative effect and mechanism of specific CXCR4 gene knockout of macrophages on renal interstitial fibrosis in mice with unilateral ureteral obstruction（ UUO）. Methods We set up the mice models of CXCR4 gene knockout（ KO mice） and divided them into KO UUO group and KO sham group with 8 rats in each group. Meanwhile,we divided 16 wild type C57 mice（ WT mice） into WT UUO group and WT sham group with 8 rats in each group. On the twelfth day after UUO model was made,we sacrificed the mice to obtain the left kidney tissues. Then HE staining was used to evaluate the degree of renal tubular lesions,Masson staining to evaluate the degree of renal interstitial collagen deposition,immunohistochemical staining to evaluate the infiltration of macrophages in renal tissues and calculate the number and cell proportion of M1 / M2 macrophages,and Western blotting to detect the expression of stromal cell derived factor 1（ SDF-1）,inducible nitric oxide synthase（ i NOS） and CD206 protein in renal tissues. Results Compared with the KO sham group and WT sham group,the degree of renal tubular lesions and interstitial collagen deposition was reduced,the number of macrophages was decreased in KO UUO group and WT UUO group. Meanwhile,M1 / M2 and the i NOS,SDF-1 protein expression in kidney tissues was increased（ all P〈0. 05）. In addition,the above indicators in the KO UUO group changed more significantly than those of the WT UUO group（ all P〈0. 05）. Conclusion Specific CXCR4 gene knockout of macrophages could prevent the macrophages from chemotaxis to the renal interstitial via the SDF-1 /CXCR4 pathway,reducing M1 macrophage changing into M2 macrophage,thereby alleviating the degree of renal interstitial fibrosis of UUO mouse.

关 键 词：输尿管梗阻 肾间质纤维化 基质细胞衍生因子1/趋化因子受体轴 巨噬细胞 基因敲除 小鼠 

分 类 号：R693[医药卫生—泌尿科学]