目的 基于鞘氨醇激酶1(Sphk1)/1-磷酸鞘氨醇受体2(S1PR2)信号通路研究雷公藤多苷片(GTW)改善免疫球蛋白A肾病(IgAN)大鼠肾损伤的作用机制.方法 通过牛血清白蛋白灌胃+蓖麻油和四氯化碳皮下注射+脂多糖尾静脉注射等手段建立IgAN大鼠模型.将造模成功大鼠随机分为模型组、对照组和实验组,每组9只;另取10只正常大鼠作为空白组.对照组灌胃给予6.25 mg·kg-1·d-1泼尼松;实验组灌胃给予9.375 mg·kg-1·d-1 GTW;空白组和模型组均灌胃给予0.5 mL·100 g-1·d-1 0.9％NaCl.各组大鼠每天给药1次.于15周末留尿取材,测定各组大鼠血白蛋白(ALB)、尿素氮(BUN)、24 h尿蛋白定量(24 h-UTP)、尿红细胞数,用蛋白质印迹法检测各组大鼠Sphk1/S1PR2蛋白的表达水平.结果 苏木精-伊红染色及免疫荧光见对照组与实验组肾病理损伤均较模型组明显减轻.空白组、模型组、对照组和实验组的ALB分别为(32.49±2.23)、(22.98±0.51)、(26.01±1.33)和(26.53±1.92)g·L1,BUN 分别为(6.11±1.71)、(13.75±2.96)、(6.71±1.35)和(4.77±0.99)mmol·L-1,24 h-UTP 分别为(5.72±1.96)、(9.12±2.15)、(5.78±2.05)和(4.75±1.50)mg·24 h-1,尿红细胞数分别为(9.73±2.40)、(14.62±2.60)、(9.90±1.59)和(9.46±2.94)cell·μL-1,Sphk1 蛋白相对表达水平分别为 0.85±0.02、1.47±0.02、1.06±0.02 和 1.09±0.02,S1PR2 蛋白相对表达水平分别为 0.27±0.02、0.88±0.01、0.43±0.02 和 0.42±0.02.模型组的上述指标与对照组和实验组比较,在统计学上差异均有统计学意义(均P＜0.01).结论 GTW可能通过抑制Sphk1/S1PR2信号通路,减少肾小球系膜细胞增殖,从而减轻IgAN大鼠的肾损伤.
Objective To study the mechanism of the amelioration of renal injury in immunoglobulin A nephropathy(IgAN)rats by multi-glycosides of Tripterygium wilfordii(GTW)based on the sphingosine kinase 1(Sphk1)/sphingosine 1-phosphate receptor 2(S1PR2)signalling pathway.Methods An IgAN rat model was established by means of bovine serum albumin gavage+castor oil and carbon tetrachloride subcutaneous injection+lipopolysaccharide tail vein injection.The rats were randomly divided into the model,control and experimental groups,with 9 rats in each group,and 10 normal rats were taken as the blank group.In the control group,6.25 mg·kg-1·d-1 prednisone was given by gavage;in the experimental group,9.375 mg·kg-1·d-1GTW was given by gavage;and in the blank and model groups,0.5 mL·100 g-1·d-1 0.9％NaCl was given by gavage,and the drugs were administered to the rats once a day in each group.At the end of the 15th week,urine samples were collected and blood albumin(ALB),blood urea nitrogen(BUN),24 hour-urine protein quantification(24 h-UTP),and urine erythrocyte counts were determined in each group,and the expression levels of Sphk1/S1PR2 proteins in each group were detected by Western blotting.Results The renal pathological changes in the control and experimental groups were significantly reduced compared with those in the model group by hematoxylin-eosin staining and immunofluorescence.The levels of ALB in the blank,model,control and experimental groups were(32.49±2.23),(22.98±0.51),(26.01±1.33)and(26.53±1.92)g·L-1;the levels of BUN were(6.11±1.71),(13.75±2.96),(6.71±1.35)and(4.77±0.99)mmol·L-1;the levels of 24 h-UTP were(5.72±1.96),(9.12±2.15),(5.78±2.05)and(4.75±1.50)mg·24 h-1;the urine erythrocyte counts were(9.73±2.40),(14.62±2.60),(9.90±1.59)and(9.46±2.94)cell·μL-1;the relative expression levels of Sphk1 protein were 0.85±0.02,1.47±0.02,1.06±0.02 and 1.09±0.02;the relative expression levels of S1PR2 protein were 0.27±0.02,0.88±0.01,0.43±0.02,and 0.42±0.02,respectively.The above indexes in the model group were statistically significant when compared with those of the control group and the experimental group(all P＜0.01).Conclusion GTW may reduce the proliferation of mesangial cells by inhibiting the Sphk1/S1PR2 signalling pathway,thus attenuating kidney injury in IgAN rats.
孟子璐;宋纯东;王耀献;张霞;丁樱;任献青;翟文生
河南中医药大学第一附属医院儿科,河南郑州 450000||河南中医药大学儿科医学院,河南郑州 450000河南中医药大学儿科医学院,河南郑州 450000
中医学
雷公藤多苷片;免疫球蛋白A肾病;肾损伤;鞘氨醇激酶1;1-磷酸鞘氨醇受体2
multi-glycosides of Tripterygium wilfordii;immunoglobulin A nephropathy;renal injury;sphingosine kinase 1;sphingosine 1-phosphate receptor 2
《中国临床药理学杂志》 2024 (006)
879-883 / 5
国家自然科学基金资助项目(82074493);河南省卫健委国家中医临床研究基地科研专项基金资助项目(2022JDZX003);河南省中医药科学研究重大专项基金资助项目(2023ZYZD02);河南省中医药学科领军人才基金资助项目(豫卫中医函[2021]8号)
10.13699/j.cnki.1001-6821.2024.06.020
评论