解毒凉血方对急性肝衰竭小鼠的治疗作用及机制  

Study on treatment effect of Jiedu liangxue Formula for acute liver failure in mice and its mechanism

作　　者：刘慧敏[1] 高方媛[1] 李玉鑫 王宪波[1] 江宇泳[1] LIU Hui-min;GAO Fang-yuan;LI Yu-xin;WANG Xian-bo;JIANG Yu-yvng(Center for Integration of Chinese and Western Medicine,Beijing Ditan Hospital Affiliated to the Capital Medical University,Beijing 100015,China)

机构地区：[1]首都医科大学附属北京地坛医院中西医结合中心,北京100015

出　　处：《北京中医药》2022年第3期248-254,共7页Beijing Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(81503428,81804009);北京市医院管理局“青苗”计划(QML20161804);北京市医院管理局“培育”计划(PZ2021030)。

摘　　要：目的探讨解毒凉血方对D氨基半乳糖(D-GalN)联合脂多糖(LPS)致急性肝衰竭小鼠的保护作用及机制。方法将30只小鼠随机分为正常组、模型组、解毒凉血方组,每组10只。模型组、解毒凉血方组腹腔注射D-GalN(400 mg/kg)和LPS(0.25 mg/kg)诱导急性肝衰竭模型。解毒凉血方组于造模剂注射后立即给予解毒凉血方500μL灌胃,模型组、正常组给予同体积双蒸水灌胃,造模5 h后心脏取血并留取肝组织样本。采用全自动生化分析仪检测肝功能[包括血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、总胆红素(TBIL)、直接胆红素(DBIL)、白蛋白(ALB)、胆碱酯酶(CHE)],苏木素-伊红(HE)染色观察肝组织病理学变化,原位末端转移酶标记法(TUNEL)检测肝细胞的凋亡率,蛋白免疫印迹法检测肝脏细胞线粒体凋亡通路中Cleaved Caspase-9和Caspase-3、硫酸糖蛋白2(SGP-2)、Bcl-2、Bax蛋白表达,反转录聚合酶链反应检测SGP-2、Bcl-2、Bax mRNA表达。结果与正常组比较,模型组血清ALT、AST、TBIL及DBIL水平高,CHE水平低(P<0.05);肝组织病理损伤明显,凋亡率高(P<0.05);SGP-2、Bax、Cleaved Caspase-9和Caspase-3高表达,Bcl-2低表达(P<0.05)。与模型组比较,解毒凉血方组血清ALT、AST、TBIL及DBIL水平低(P<0.05),肝组织病理损伤轻,肝细胞凋亡率低(P<0.05);SGP-2、Bax、Cleaved Caspase-9、Cleaved Caspase-3呈低表达,Bcl-2高表达(P<0.05)。结论解毒凉血方对D-GalN/LPS诱导的急性肝衰竭小鼠有显著的治疗作用,其作用机制可能与抑制线粒体凋亡信号通路介导的肝细胞凋亡有关。Objective To study the protective effect of Jiedu Liangxue Formula on D-galactose(D-GalN)combined with lipopolysaccharide(LPS)induced acute liver failure in mice and its mechanism.Methods 30 mice were randomly divided into normal group,model group and Jiedu Liangxue Formula group with 10 mice in each group.Except for normal group,acute liver failure model was induced by intraperitoneal injection of D-GalN(400 mg/kg)and LPS(0.25 mg/kg)in the other two groups.The Jiedu Liangxue Formula group was gavaged 500μL Jiedu Liangxue Formula immediately after the injection of the model agent,and the model group and the normal group were gavaged with the same volume of double steamed water.After 5 hours of modeling,the heart blood was collected and liver tissue samples were kept.Liver function including serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL),direct bilirubin(DBIL),albumin(propagated),cholinesterase(CHE)were tested by automatic biochemical analyzer,the liver tissue pathology change was observed by wood grain-eosin(HE)staining,liver cell apoptosis rate was detected by TUNEL,the protein expressions of caspase-9,caspase-3,glycoprotein 2(SGP-2),Bcl-2 and Bax in mitochondrial apoptosis pathway of liver cells were detected by Western blot,and mRNA expressions of SGP-2,Bcl-2 and Bax were detected by reverse transcriptase polymerase chain reaction(PCR).Results Compared with normal group,serum levels of ALT,AST,TBIL and DBIL in model group were significantly increased,CHE was significantly decreased(P<0.05),the liver pathological damage was obvious,apoptosis cells were significantly increased(P<0.05),the expression of SGP-2,Bax and cleaved caspase-9 and caspase-3 were high,and the expression of Bcl-2 was low(P<0.05);compared with model group,the levels of ALT,AST,TBIL and DBIL in serum of Jiedu Liangxue Formula group were significantly decreased(P<0.05),the pathological damage of liver tissue was mild,and the apoptosis rate of liver cells was low(P<0.05),SGP-2,Bax and cleaved caspase-

关 键 词：急性肝衰竭 线粒体凋亡信号通路 解毒凉血方 小鼠 

分 类 号：R285.5[医药卫生—中药学]