尿毒康合剂对缺血再灌注损伤大鼠的肾保护作用及其机制研究  

Protective effect and mechanism of Niaodukang mixture for renal ischemia reperfusion injury in rats

作　　者：安海文[1] 李李 李燕林[1] 刘琳娜[1] 黄琳[1] 李锦山[1] 杨文钦[1] AN Hai-wen;LI Li;LI Yan-lin;LIU Lin-na;HUANG Lin;LI Jin-shan;YANG Wen-qin(Department of Nephrology,Zhongshan TCM Hospital,Zhongshan 528400,China)

机构地区：[1]中山市中医院肾内科,广东中山528400

出　　处：《实用药物与临床》2020年第12期1063-1068,共6页Practical Pharmacy and Clinical Remedies

基　　金：中山市第一批科学事业费项目(2015B1045)。

摘　　要：目的探究尿毒康合剂对大鼠肾脏缺血再灌注损伤(RIRI)的保护作用及其机制。方法将73只SD大鼠随机分为模型组、假手术组、尿毒康合剂(低剂量4.95 g/kg、中剂量9.9 g/kg、高剂量19.8 g/kg)组,银杏叶提取物(100 mg/kg)组(阳性药组),每组12只(模型组13只)。除假手术组以外,另外5组SD大鼠建立RIRI模型。建立RIRI模型前,连续7 d给予尿毒康合剂各剂量组及阳性药大鼠相应剂量药物灌胃。24 h后,检测各组大鼠血清中肌酐值(Scr)和尿素氮值(BUN)以及白介素-6(IL-6)、肿瘤坏死因子(TNF-α)和干扰素-γ(IFN-γ)指标;通过TUNEL试剂盒上的方法检测大鼠肾小管上皮细胞的凋亡率;通过免疫组化法及Western blot法检测FAS、FASL、Bax、Bad、Caspase-3、Caspase-8、Caspase-9、Bcl-2和VEGF这9种细胞凋亡相关蛋白的表达情况。结果与假手术组比较,模型组大鼠血清中的Scr、BUN、IL-6、TNF-a、IFN-γ水平及肾小管上皮细胞凋亡率均明显升高(P<0.01);模型组大鼠FAS、FASL、Bax、Bad、Caspase-3、Caspase-8、Caspase-9蛋白表达量显著升高,Bcl-2、VEGF蛋白的表达降低,差异均有统计学意义(P<0.05)。尿毒康合剂各剂量组及阳性药组大鼠血清中的Scr、BUN、IL-6、TNF-a、IFN-γ及肾小管上皮细胞凋亡率低于模型组(P<0.05);尿毒康合剂各剂量组大鼠FAS、FASL、Bax、Bad、Caspase-3、Caspase-8及Caspase-9蛋白表达量有降低趋势,Bcl-2、VEGF蛋白表达有升高的趋势。结论尿毒康合剂对RIRI大鼠肾脏具有保护作用,其机制可能是调节炎症因子,抑制炎症反应,或是通过对细胞凋亡相关蛋白表达进行干预,从而抑制其凋亡。Objective To investigate the protective effect and mechanism of Niaodukang Mixture for rats with renal ischemia-reperfusion injury(RIRI).Methods Totally 73 SD rats were randomly divided into model group,sham operation group,Niaodukang Mixture groups(4.95 g/kg for low-dose group,9.9 g/kg for medium-dose group,19.8 g/kg for high-dose group)and ginkgo biloba extract group(100 mg/kg,positive drug group),12 rats in each group(13 in model group).Except for the sham operation group,in the other 5 groups of SD rats RIRI model was established.Before the establishment of the RIRI model,each dose group of Niaodukang Mixture and positive drug group were given the corresponding dose of the drug for 7 days.Serum creatinine(Scr),urea nitrogen(BUN),interleukin-6(IL-6),tumor necrosis factor(TNF-α)and interferon-γ(IFN-γ)were measured at 24 h after RIRI modeling.The apoptosis rate of renal tubular epithelial cells was detected by the method on TUNEL kit;the expression of apoptosis-related proteins of FAS,FASL,Bax,Bad,Caspase-3,Caspase-8,Caspase-9,Bcl-2 and VEGF of apoptosis-related proteins was detected by immunohistochemistry and Western blot.Results Compared with the sham-operation group,the levels of Scr,BUN,IL-6,TNF-α,IFN-γ and the apoptosis rate of renal tubular epithelial cells were higher(P<0.01)in model group.Furthermore,the expression levels of FAS,FASL,Bax,Bad,Caspase-3,Caspase-8 and Caspase-9 in the model group were also significantly increased,while the expression of Bcl-2 and VEGF protein was significantly decreased,and the differences were statistically significant(P<0.05).Compared with model group,the levels of Scr,BUN,IL-6,TNF-α and IFN-γ and the apoptosis rate of renal tubular epithelial cells in Niaodukang Mixture group and positive drug group were decreased(P<0.05).Furthermore,the expression of FAS,FASL,Bax,Bad,Caspase-3,Caspase-8 and Caspase-9 in Niaodukang Mixture group tended to decrease,while the expression of Bcl-2 and VEGF protein tended to increase.Conclusion Niaodukang Mixture has protective effect

关 键 词：尿毒康合剂 大鼠 肾缺血再灌注损伤 保护作用 

分 类 号：R285.5[医药卫生—中药学]