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人参归脾丸对脾不统血证大鼠肝损伤的保护作用及其机制

[Protective effect of Ginseng Guipi Pill on liver injury in rats with spleen failing to control blood syndrome and its mechanism].

作者信息

Wang Dan, Gao Feng, Chen Hui, Xiang Wei-Ling, Luo Zhi-Xian, Jin Li-Qin

机构信息

Department of Biology School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou 325035.

Department of Acupuncture Massage, Zhejiang Provincial People's Hospital, Hangzhou 310014.

出版信息

Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2020 Nov;36(6):571-575. doi: 10.12047/j.cjap.5975.2020.120.

DOI:10.12047/j.cjap.5975.2020.120
PMID:33719260
Abstract

To investigate the therapeutic effect of Ginseng Guipi pill on rats with spleen failing to control blood syndrome and its effect on liver. Forty SPF male Sprague-Dawley rats were randomly divided into normal control group (=10) and experimental group (=30). For the first 42 days, the experimental group swam for 30 minutes every day, eating for one day and fasting for two days (diet disorder). On the 43rd to 72nd day, the rats were injected with low-molecular-weight heparin calcium to induce hemorrhage on the basis of exhaustion of swimming and diet disorder to construct a rat model of spleen failing to control blood syndrome. Those thirty rat that successfully established the model were randomly divided into 3 groups (=10), model control group (MD), natural rehabilitation group (NR) and Guipi Pill group (GP). On the 73rd to 103rd day, the MD group continued to apply factors (exhausted swimming, eating disorder, and injection of low-molecular-weight heparin calcium), the NR group and the GP group stopped applying the factors , the GP group was daily administered with Ginseng Guipi Pill solution (0.2 g/ml, 10 ml/(kg·d)) , and the NR group was given an equal dose of saline. After the 103 days, blood and liver samples were collected, and the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBil) and total protein (TP) content were detected, the number of red blood cells (RBC), hemoglobin (HGB) content and hematocrit (HCT) were detected, the expressions of IL-6 and TNF-α in serum were determined by ELISA kit, the levels of apoptosis-related proteins Bcl-2, Bax and Caspase3 in liver tissue were detected by Western blot. The expression levels of apoptosis-related proteins Bcl-2, Bax and Caspase3 mRNA in liver tissue were detected by real-time PCR. Compared with the control group, the serum levels of ALT, AST and TBil in the model group were increased significantly, while the serum level of TP was reduced significantly (<0.01), the number of RBC, HGB content and HCT were decreased significantly (<0.01), the expressions of IL-6 and TNF-α in serum were increased (<0.05), the Bcl-2 protein and mRNA levels were reduced, Bax and Caspase3 protein and mRNA levels were increased significantly (<0.01); Compared with the model group, the serum levels of ALT, AST and TBil were significantly lower in the natural rehabilitation group and the Guipi Pill group, the level of TP was increased significantly (<0.01), the blood RBC, HGB and HCT were increased significantly (< 0.01), the expressions of IL-6 and TNF-α were decreased (<0.05), the levels of Bcl-2 protein and mRNA were increased significantly, the Bax, Caspase3 protein and mRNA levels were decreased (<0.05); Compared with the natural rehabilitation group, the serum levels of ALT, AST and TBil were reduced , and the TP content was increased significantly in the Guipi Pill group (< 0.01), the number of RBC was increased significantly in the Guipi Pill group (<0.05), the levels of Bcl-2 protein and mRNA were increased (<0.05),the level of Caspase3 protein was decreased (<0.05), the expression of Bax mRNA was reduced significantly (<0.05). Ginseng Guipi Pill has protective effect on liver injury in rats with spleen failing to control blood syndrome. The mechanism may be related to the inhibition the liver cell apoptosis and the release of pro-inflammatory cytokines.

摘要

探讨归脾丸对脾不统血证大鼠的治疗作用及其对肝脏的影响。将40只SPF级雄性Sprague-Dawley大鼠随机分为正常对照组(n = 10)和实验组(n = 30)。前42天,实验组大鼠每天游泳30分钟,进食1天禁食2天(饮食紊乱)。第43至72天,在游泳疲劳和饮食紊乱的基础上,给大鼠注射低分子肝素钙诱导出血,构建脾不统血证大鼠模型。将成功造模的30只大鼠随机分为3组(n = 10),即模型对照组(MD)、自然恢复组(NR)和归脾丸组(GP)。第73至103天,MD组继续施加造模因素(疲劳游泳、饮食紊乱、注射低分子肝素钙),NR组和GP组停止施加造模因素,GP组每日灌胃归脾丸溶液(0.2 g/ml,10 ml/(kg·d)),NR组给予等量生理盐水。103天后,采集血液和肝脏样本,检测血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆红素(TBil)和总蛋白(TP)含量,检测红细胞(RBC)数量、血红蛋白(HGB)含量和血细胞比容(HCT),用ELISA试剂盒测定血清中IL-6和TNF-α的表达,用Western blot检测肝组织中凋亡相关蛋白Bcl-2、Bax和Caspase3的水平。用实时荧光定量PCR检测肝组织中凋亡相关蛋白Bcl-2、Bax和Caspase3 mRNA的表达水平。与对照组比较,模型组血清ALT、AST和TBil水平显著升高,而血清TP水平显著降低(P<0.01),RBC数量、HGB含量和HCT显著降低(P<0.01),血清中IL-6和TNF-α表达升高(P<0.05),Bcl-2蛋白和mRNA水平降低,Bax和Caspase3蛋白及mRNA水平显著升高(P<0.01);与模型组比较,自然恢复组和归脾丸组血清ALT、AST和TBil水平显著降低,TP水平显著升高(P<0.01),血液RBC、HGB和HCT显著升高(P<0.01),IL-6和TNF-α表达降低(P<0.05),Bcl-2蛋白和mRNA水平显著升高,Bax、Caspase3蛋白及mRNA水平降低(P<0.05);与自然恢复组比较,归脾丸组血清ALT、AST和TBil水平降低,TP含量显著升高(P<0.01),归脾丸组RBC数量显著增加(P<0.05),Bcl-2蛋白和mRNA水平升高(P<0.05),Caspase-Ⅲ蛋白水平降低(P<0.05),Bax mRNA表达显著降低(P<0.05)。归脾丸对脾不统血证大鼠肝损伤具有保护作用。其机制可能与抑制肝细胞凋亡及促炎细胞因子释放有关。

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