CXCR6介导的NKT细胞激活在急性肾损伤小鼠肾纤维化中的作用  

Role of CXCR6-mediated activation of natural killer T cells in renal fibrosis following acute kidney injury in mice

作　　者：徐枫[1] 张涛[1] 钟超群 刘本铨 梁桦[1] 王汉兵[1] Xu Feng;Zhang Tao;Zhong Chaoqun;Liu Benquan;Liang Hua;Wang Hanbing(Department of Anesthesiology,The First People′s Hospital of Foshan,Foshan 528000,China)

机构地区：[1]佛山市第一人民医院麻醉科,528000

出　　处：《中华麻醉学杂志》2021年第8期1005-1009,共5页Chinese Journal of Anesthesiology

摘　　要：目的CXC趋化因子受体6(CXCR6)介导的自然杀伤T(NKT)细胞激活在急性肾损伤小鼠肾纤维化中的作用。方法雄性野生型C57BL/6小鼠和CXCR6基因敲除型C57BL/6小鼠各18只,8~10周龄,体重20~30 g,采用随机数字表法分为6组(n=6):野生型小鼠对照组(WT-CON组)、CXCR6基因敲除型小鼠对照组(CXCR6^(-/-)-CON组)、野生型小鼠急性肾损伤组(WT-AKI组)、CXCR6基因敲除型小鼠急性肾损伤组(CXCR6^(-/-)-AKI组)、野生型小鼠急性肾损伤+NKT细胞过继转移组(WT-AKI-NKT组)和CXCR6基因敲除型小鼠急性肾损伤+NKT细胞过继转移组(CXCR6^(-/-)-AKI-NKT组)。腹腔注射叶酸250 mg/kg制备急性肾损伤小鼠肾纤维化模型。WT-AKI-NKT组和CXCR6^(-/-)-AKI-NKT组分别于注射叶酸后第4和9天时尾静脉注射NKT细胞悬液250μl(1×10^(6)个)。注射叶酸后第14天时,采取眼眶血标本,检测血清BUN和Cr浓度。处死小鼠取肾组织,采用天狼星红染色观察肾纤维化面积,采用HE染色观察肾损伤情况,并行肾损伤评分,采用流式细胞术测定CD1d Tetramer阳性(CD1d Tetramer^(+))细胞比例,采用免疫荧光双染法行CD206和α-平滑肌肌动蛋白(α-SMA)双阳性(CD206^(+)-α-SMA+)细胞计数,采用RT-PCR法检测IL-4和IL-13的mRNA表达水平。结果与WT-CON组比较,WT-AKI组和WT-AKI-NKT组血清BUN、Cr浓度和肾损伤评分升高,肾纤维化面积增加,肾组织CD1d Tetramer^(+)百分比和CD206^(+)-α-SMA+细胞计数升高,IL-4和IL-13的mRNA表达上调(P<0.05);与WT-AKI组比较,WT-AKI-NKT组血清BUN、Cr浓度和肾损伤评分升高,肾纤维化面积增加,肾组织CD1d Tetramer+细胞百分比和CD206^(+)-α-SMA+细胞计数升高,IL-4和IL-13的mRNA表达上调,CXCR6^(-/-)-AKI组血清BUN、Cr浓度和肾损伤评分降低,肾纤维化面积减少,肾组织CD1d Tetramer^(+)细胞百分比和CD206^(+)-α-SMA+细胞计数降低,IL-4和IL-13的mRNA表达下调(P<0.05);与CXCR6^(-/-)-CON组比较,CXCR6^(-/-)-AKI组和CXCR6^(-/-)-AKI-NKT组血清BUN、Cr浓度�Objective To evaluate the role of CXC chemokine receptor 6(CXCR6)-mediated activation of natural killer T(NKT)cells in renal fibrosis following acute kidney injury(AKI)in mice.Methods Eighteen male wild-type C57BL/6 mice and 18 CXCR6 knockout C57BL/6 mice,aged 8-10 weeks,weighing 20-30 g,were divided into 6 groups(n=6 each)using a random number table method:wild-type mouse control group(group WT-CON),CXCR6 knockout mouse control group(group CXCR6^(-/-)-CON),wild-type mouse with AKI group(group WT-AKI),CXCR6 knockout mouse with AKI group(group CXCR6^(-/-)-AKI),wild-type mouse with AKI+NKT cell adoptive transfer group(group WT-AKI-NKT)and CXCR6 knockout mouse with AKI+NKT cell adoptive transfer group(group CXCR6^(-/-)-AKI-NKT).Folic acid 250 mg/kg was intraperitoneally injected to establish the model of renal fibrosis in mice with AKI.NKT cellsuspension 250μl(1×10^(6)cells)was injected through the tail vein on the 4th and 9th days after folic acid injection in group WT-AKI-NKT and group CXCR6^(-/-)-AKI-NKT,respectively.Blood samples were taken from orbital at day 14 after folic acid injection for determination of the concentrations of serum blood urea nitrogen(BUN)and creatinine(Cr).The animals were sacrificed,and renal tissues were obtained for observation of the area of renal fibrosis(by Sirius red staining)and renal injury(using H&E staining)which was scored and for determination of the proportion of CD1d Tetramer+cells(by flow cytometry),the number of CD206 andα-smooth muscle actin(α-SMA)double positive(CD206^(+)-α-SMA+)cells(by immunofluorescence)and expression of interleukin(IL)-4 and IL-13 mRNA(by real-time polymerase chain reaction).Results Compared with group WT-CON,the BUN and Cr levels,renal injury scores,area of renal fibrosis,proportion of CD1d Tetramer+cells and CD206^(+)-α-SMA+cell count were significantly increased,and the expression of IL-4 and IL-13 mRNA was up-regulated in group WT-AKI and WT-AKI-NKT(P<0.05).Compared with group WT-AKI,the BUN and Cr levels,renal injury scores,area of renal

关 键 词：受体 CXCR 自然杀伤T细胞 急性肾损伤 纤维化 

分 类 号：R692[医药卫生—泌尿科学]