CXCR4在草酸钙晶体致肾损伤及纤维化中的作用和机制  

The role and mechanism of CXCR4 in renal injury and fibrosis caused by calcium oxalate crystals

作　　者：叶泽华 夏煜琦 李柏均 颜信州 余伟民[1] 饶婷[1] 阮远[1] 周向军[1] 程帆[1] Ye Zehua;Xia Yuqi;Li Bojun;Yan Xinzhou;Yu Weimin;Rao Ting;Ruan Yuan;Zhou Xiangjun;Cheng Fan(Department of Urology,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院泌尿外科,武汉430060

出　　处：《中华泌尿外科杂志》2022年第4期285-290,共6页Chinese Journal of Urology

基　　金：国家自然科学基金资助项目(82170775)。

摘　　要：目的探讨C-X-C趋化因子受体4(CXCR4)在小鼠草酸钙晶体致肾损害和纤维化中的作用和机制。方法2021年6月将15只雄性C57/BL6小鼠按随机数字表法分为对照组5只、模型组5只、AMD3100干预组5只。模型组和AMD3100干预组连续7 d腹腔注射乙醛酸水溶液(100 mg/kg)制备草酸钙结石动物模型。AMD3100干预组同时连续7 d腹腔注射AMD3100(1 mg/kg)。对照组连续7 d腹腔注射与模型组等量生理盐水。处理7 d后取各组小鼠外周血测定血清尿素氮(BUN)、肌酐(Scr)水平评估肾功能;同时取左侧肾脏行HE、Von-Kossa、Picrosirius Red染色,观察肾组织的病理改变;采用免疫组化法染色和蛋白质印迹法检测CXCR4、转化生长因子β1(TGF-β1)的表达水平;采用蛋白质印迹法检测PI3K/AKT通路相关蛋白的表达水平。结果生化指标检测结果显示,模型组较对照组血清Scr[(108.03±13.56)μmol/L与(39.50±4.48)μmol/L,P<0.01]和BUN[(5.66±0.48)mmol/L与(0.77±0.10)mmol/L,P<0.01]水平显著上升;AMD3100干预组较模型组Scr[(65.77±3.27)μmol/L与(108.03±13.56)μmol/L,P<0.05]和BUN[(2.97±0.44)mmol/L与(5.66±0.48)mmol/L,P<0.05]水平显著下降。病理检查结果显示,模型组较对照组肾组织的肾小管明显扩张,伴炎性细胞浸润,同时出现大量草酸钙晶体及胶原纤维沉积;AMD3100干预组较模型组肾脏的损伤程度、草酸钙晶体及胶原纤维沉积均显著缓解。蛋白质印迹法检测结果显示,模型组较对照组CXCR4(0.639±0.019与0.158±0.012,P<0.01)和TGF-β1(0.698±0.018与0.314±0.015,P<0.05)的相对表达量明显增多;AMD3100干预组较模型组CXCR4(0.322±0.231与0.639±0.019,P<0.05)和TGF-β1(0.445±0.017与0.698±0.018,P<0.05)的相对表达量明显下降。免疫组化染色检查结果显示各组的CXCR4和TGF-β1表达趋势与蛋白质印迹法检测结果一致。蛋白质印迹检测结果显示,模型组较对照组p-PI3K(0.613±0.016与0.213±0.011,P<0.01)和p-AKT(0.149±0.013与0.047±0.014,P<0.0Objective To investigate the role and mechanism of chemokine receptor type 4(CXCR4)in renal injury and fibrosis caused by calcium oxalate crystals in mice.Methods In June 2021,Fifteen male C57/BL6 mice were divided into control group(5 mice),model group(5 mice),and AMD3100 intervention group(5 mice)by random number table method.In model group and AMD3100 intervention group,glyoxylate(100 mg/kg)was injected intraperitoneal for 7 consecutive days for modeling.Meanwhile,the AMD3100 intervention group was also given intraperitoneal injection of AMD3100(1 mg/kg)for 7 days.The control group was continuously injected with equal volume saline intraperitoneally.After 7 days,peripheral blood was collected from each group to determine the levels of serum urea nitrogen(BUN)and creatinine(Scr)to assess the renal function;HE,Von-Kossa,Picrosirius Red staining was also taken from the left kidney to observe the pathological changes of renal tissue.CXCR4,transforming growth factorβ1(TGF-β1)were detected by immunohistochemistry and western blot.The expression levels of PI3K/AKT pathway-related proteins were detected by western blot.Results The results of biochemical indexes showed that the serum Scr[(108.03±13.56)μmol/L vs.(39.50±4.48)μmol/L,P<0.01]and BUN[(5.66±0.48)mmol/L vs.(0.77±0.10)mmol/L,P<0.01]levels were significantly increased in model group compared with the control group.The AMD3100 intervention group was significantly lower than the model group in terms of Scr[(65.77±3.27)μmol/L vs.(108.03±13.56)μmol/L,P<0.05]and BUN[(2.97±0.44)mmol/L vs.(5.66±0.48)mmol/L,P<0.05]levels.The results of kidney pathology in mice showed that renal tubules were significantly dilated with inflammatory cell infiltration in the model group compared with the control group,and a large number of calcium oxalate crystals and collagen fibers were deposited.The extent of kidney damage,calcium oxalate crystals and collagen fibers deposition were significantly reduced in the AMD3100 intervention group compared with the model group.The re

关 键 词：草酸钙 C-X-C趋化因子受体4 肾结石 纤维化 损伤 

分 类 号：R692[医药卫生—泌尿科学]