真武汤对脾肾阳虚型肾炎小鼠内皮细胞功能和炎症因子的影响

Effect of Zhenwu Decoction(真武汤) on Endothelial Cell Function and Inflammatory Factors in Nephritis Mice with Spleen-Kidney Yang Deficiency Syndrome

目的 观察真武汤对脾肾阳虚型肾炎小鼠内皮细胞功能和炎症因子的影响。方法 选取BALB/c小鼠40只,雌雄各半,给予缺血再灌注制作脾肾阳虚型肾炎模型。成模后随机均分为实验组和PBS阴性对照组,其中实验组小鼠给予每日2次灌胃真武汤0.2 mL/只,PBS阴性对照组小鼠给予每日2次灌胃PBS 0.2 mL/只,同时选取20只健康小鼠作为对照组,给予每日2次灌胃生理盐水0.2 mL/只,观察4周。各组分别用药连续干预2周和4周后检测血肌酐(SCr)和血尿素氮(BUN),并计算尿蛋白和尿肌酐的比值(ACR),采用酶联免疫吸附测定(ELISA)方法检测小鼠血清血管内皮生长因子(VEGF)、单核细胞趋化蛋白-1(MCP-1)、肿瘤坏死因子-α(TNF-α)以及白细胞介素-6(IL-6)水平。结果 正常对照组小鼠精神状态良好、动作自如、反应敏捷、体毛光泽。PBS阴性对照组和实验组小鼠精神状态相对较差,体毛偏黄、相对干枯、脱落较多、欠光泽,多尿,进食量减少,活动减少。实验组动物诸情况相对PBS阴性对照组为轻。与正常组比较,0、2、4周时PBS阴性对照组和实验组小鼠的BUN、SCr以及尿ACR水平显著上调(P<0.01)。与0周时比较,PBS阴性对照组小鼠2周和4周时的BUN、SCr以及尿ACR水平有上调趋势,但差异不具有统计学意义(P>0.05)。与0周时比较,实验组小鼠2周和4周时的BUN、SCr以及尿ACR水平显著下调(P<0.01)。与正常对照组比较,干预2、4周时PBS阴性对照组和实验组小鼠的血清VEGF以及MCP-1水平均显著上调(P<0.01)。与PBS阴性对照组比较,干预2周时实验组小鼠的血清VEGF以及MCP-1水平有下调趋势,但差异不具有统计学意义(P>0.05);与PBS阴性对照组比较,干预4周时实验组小鼠的血清VEGF以及MCP-1水平显著下调(P<0.01)。与正常对照组比较,干预2、4周时PBS阴性对照组和实验组小鼠的血清TNF-α以及IL-6水平均显著上调(P<0.01)。与PBS阴性对照组比较,干预2、4周时实验组小鼠的血清TNF-α以及IL-6水平显著下调(P<0.01)。结论 真武汤能显著改善脾肾阳虚型肾炎模型小鼠肾功能,其分子机制可能与下调VEGF、MCP-1、TNF-α以及IL-6的水平,保护内皮细胞功能、抑制炎症反应有关。

Objective To observe the effects of Zhenwu Decoction(真武汤)on endothelial cell function and inflammatory factors in nephritis mice with spleen-kidney Yang deficiency syndrome.Methods Forty BALB/c mice,half male and half male,were selected and given ischemia reperfusion to make spleen-kidney Yang deficiency nephritis model.After modeling,they were randomly divided into experimental group and PBS negative control group.The mice in the experimental group were given 0.2 mL of Zhenwu Decoction twice a day,and the mice in the PBS negative control group were given 0.2 mL PBS twice a day.Each mouse was selected as the control group at the same time,and 0.2 mL of normal saline was administered twice a day for observation for 4 weeks.Blood creatinine(SCr)and blood urea nitrogen(BUN)were measured after continuous intervention for 2 and 4 weeks in each group,the ratio of urine protein to urine creatinine(ACR)was calculated,and the enzyme-linked immunosorbent assay(ELISA)to detect the serum levels of vascular endothelial growth factor(VEGF),monocyte chemoattractant protein-1(MCP-1),tumor necrosis factor-α(TNF-α)and interleukins-6(IL-6).Results The mice in the normal control group had good mental state,free movement,quick response and glossy body hair.The mental state of mice in PBS negative control group and experimental group was relatively poor,with yellowish body hair,relative dryness,more shedding,lack of luster,polyuria,reduced food intake and activity.The conditions of animals in the experimental group were lighter than those in the PBS negative control group.The conditions of the animals in the experimental group were less severe than those in the PBS negative control group.Compared with those of the normal group,the levels of BUN,SCr and urinary ACR of mice in the PBS negative control group and the experimental group were significantly up-regulated at 0,2 and 4 weeks(P<0.01).Compared with those at week 0,the levels of BUN,SCr and urinary ACR in the PBS negative control group mice at 2 and 4 weeks had an upward-regulated trend,but the difference was not statistically significant(P>0.05).Compared with those at week 0,the levels of BUN,SCr and urinary ACR of mice in the experimental group were significantly down-regulated at 2 and 4 weeks(P<0.01).Compared with those of the normal control group,the serum levels of VEGF and MCP-1 of the mice in the PBS negative control group and the experimental group were significantly up-regulated at 2 and 4 weeks of intervention(P<0.01).Compared with those of the PBS negative control group,the serum levels of VEGF and MCP-1 of the experimental group had a downward-regulation trend after 2 weeks of intervention,but the difference was not statistically significant(P>0.05).Compared with those of the PBS negative control group,after the intervention was 4 weeks,the serum levels of VEGF and MCP-1 of mice in the experimental group were significantly down-regulated(P<0.01).Compared with those of the normal control group,the serum levels of TNF-αand IL-6 of the mice in the PBS negative control group and the experimental group were significantly up-regulated at 2 and 4 weeks of intervention(P<0.01).Compared with those of the PBS negative control group,the serum levels of TNF-αand IL-6 of the mice in the experimental group were significantly down-regulated at 2 and 4 weeks of intervention(P<0.01).Conclusion Zhenwu Decoction can significantly improve the renal function of nephritis mice with spleen-kidney Yang deficiency.Its molecular mechanism may be related to down-regulating the levels of VEGF,MCP-1,TNF-αand IL-6,protecting endothelial cell function and inhibiting inflammation.