脾多肽对阿奇霉素与对乙酰氨基酚联用所致小鼠急性肝损伤的保护作用

Protective effect of spleen polypeptide on acute liver injury induced by azithromycin combined with acetaminophen in mice

目的探究脾多肽对阿奇霉素与对乙酰氨基酚联用所致小鼠急性肝损伤的影响。方法小鼠随机分成6组,正常对照组、模型组、脾多肽25、50和100 mg/(kg·d)剂量组、水飞蓟素组[阳性药组,70 mg/(kg·d)]。正常对照组小鼠实验中均注射生理盐水;模型组、脾多肽剂量组和阳性药组小鼠连续4 d分别给予尾静脉注射生理盐水、脾多肽和灌胃水飞蓟素,第4天模型组、脾多肽剂量组和阳性药组给药2 h后,均给予对乙酰氨基酚(350 mg/kg)及阿奇霉素(280 mg/kg)灌胃造模,连续7 d。观察小鼠的一般状况,计算脏器指数,检测血浆中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、尿素氮(BUN)、肌酐(CREA)、尿酸(UA)、总蛋白(TP)、白蛋白(ALB)、球蛋白(GLO)和白球比(A/G);肝组织匀浆中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)活性、肝组织病理结构;蛋白免疫印迹法检测肝细胞中Bax及Bcl-2蛋白的表达。结果与正常对照组相比,模型组的肝指数显著升高(P<0.01),脾指数和肾指数显著降低(P<0.05,P<0.01);与模型组相比,脾多肽100 mg/(kg·d)组肝指数显著下降(P<0.01),脾多肽25、50和100 mg/(kg·d)组脾指数和肾指数升高(P<0.01),阳性药组的肝指数显著下降(P<0.01)。与正常对照组相比,模型组小鼠血浆中ALT、AST、TP、GLO、UA、BUN和CREA显著上升(P<0.05,P<0.01),ALB、A/G和SOD显著下降(P<0.01,P<0.05);与模型组相比,脾多肽25、50、100 mg/(kg·d)组和阳性对照组血浆的AST均显著降低(P<0.01),脾多肽100 mg/(kg·d)组和阳性对照组的ALT、AST、TP、GLO、UA、BUN和CREA均显著降低(P<0.05,P<0.01),ALB和A/G显著升高(P<0.05)。与正常对照组相比,模型组小鼠肝组织中GSH-Px和CAT水平及Bal-2表达均显著下降(P<0.01),Bax表达显著升高(P<0.01);与模型组相比,脾多肽25、50、100 mg/(kg·d)组和阳性对照组肝组织中GSH-PX、CAT和SOD水平及Bal-2表达均显著升高(P<0.01),Bax表达显著下调(P<0.01)。结论脾多肽具有改善阿奇霉素与对乙酰氨基酚联用所致小鼠急性肝损伤的作用。

Objective To investigate the effect of spleen polypeptide on liver injury induced by azithromycin combined with acetaminophen in mice. Methods The mice were randomly divided into 6 groups：the normal control group,model group,spleen polypeptide dose groups[25,50,and 100 mg/（kg·d）],and silybin positive drug group[70 mg/（kg·d）]. The mice in the normal control group were injected with normal saline in the whole animal experiment. The mice in the model group,spleen polypeptide dose group and positive group were treated respectively with normal saline,spleen polypeptide by caudal vein injection and silybin by gavage for 4 d. After 2 hours of administration on day 4,all mice except those in the normal control group were given acetaminophen（350 mg/kg）and azithromycin（280 mg/kg）by gavage for 7 d. The organ index of mice was calculated,and the plasma alanine aminotransferase（ALT）,aspartate aminotransferase（AST）,ALT/AST,urea nitrogen（BUN）,creatinine（CREA）,uric acid（UA）,total protein（TP）,albumin（ALB）,globulin（GLO）and albumin/globulin（A/G）in mice were measured. The superoxide dismutase（SOD）,catalase（CAT）and glutathione peroxidase（GSH-PX）activity of liver tissue homogenate were measured. The expression of Bax and Bcl-2 protein in hepatocytes was measured by Western blot. Results Compared with the normal control group,the liver index of the model group was increased significantly（P〈0.01）,and the spleen and kidney indices were decreased（P〈0.05,P〈0.01）. Compared with the model group,the liver index of the spleen polypeptide 100 mg/（kg·d）group was decreased significantly（P〈0.01）,the spleen and the kidney index of the spleen polypeptide 25,50 and 100 mg/（kg·d）groups were increased significantly（P〈0.01）,and the liver index of positive drug group was decreased significantly（P〈0.01）. Compared with the normal control group,the plasma levels of ALT,AST,TP,GLO,UA,BUN and CREA were increased significantly in the model mice（P〈0.05,P〈0.01）,and the plasma levels of ALB and A/G were decreased（P〈0.05,P〈0.01）. Compared with the model group,the plasma AST levels of the spleen polypeptide 25,50,100 mg/（kg·d）and positive drug groups were decreased significantly（P〈0.01）,and the plasma ALT,AST,TP,GLO,UA,BUN and CREA levels of the spleen polypeptide 100 mg/（kg·d）and positive drug groups were decreased（P〈0.05,P〈0.01）,and the plasma levels of ALB and A/G were increased（P〈0.05）. Compared with the normal control group,hepatic levels of SOD,CAT,GSH-Px activity and the expression of Bcl-2 were decreased significantly（P〈0.01）,and the expression of Bax was increased significantly（P〈0.01）. Compared with the model group,the activity of GSH-Px,CAT,SOD and the expression of Bal-2 in liver of the spleen polypeptide[25,50 and 100 mg/（kg·d）]groups and the positive group were increased significantly（P〈0.01）,and the expression of Bax decreased significantly（P〈0.01）. Conclusion Spleen polypeptide could improve acute liver injury induced by azithromycin combined with acetaminophen in mice.