胆南星对四氯化碳致小鼠急性肝损伤的保护作用及机制研究

Study on protective effect and mechanism of Arisaema Cum Bile on acute liver injury induced by CCl4 in mice

目的研究胆南星对四氯化碳(CCl_(4))致小鼠急性肝损伤的保护作用及可能的作用机制。方法将50只小鼠按体质量随机分为正常组、模型组、阳性对照组(联苯双酯滴丸,150 mg/kg)和胆南星低、高剂量组(0.78、2.34 g/kg),每组10只。每天灌胃给药1次,连续7 d。末次灌胃2 h后,除正常组外的其余各组小鼠均腹腔注射0.2%CCl_(4)-橄榄油溶液复制急性肝损伤模型。腹腔注射17 h后,按照试剂盒方法检测小鼠血清中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)和肝组织中丙二醛(MDA)、超氧化物歧化酶(SOD)水平,测定小鼠的肝脏指数,采用苏木精-伊红染色法观察小鼠肝组织的病理形态学变化并对肝损伤程度进行量化评分,采用实时荧光定量PCR法检测小鼠肝组织中IL-6、TNF-αmRNA表达水平,采用Western blot法检测小鼠肝组织中酪氨酸激酶2(JAK2)、信号传导及转录激活因子3(STAT3)、核因子κB p65(NF-κB p65)蛋白表达水平。结果与正常组比较,模型组小鼠ALT、AST、IL-6、TNF-α、MDA水平和肝脏指数显著升高(P<0.05),SOD水平显著降低(P<0.05);IL-6、TNF-αmRNA及JAK2、STAT3、NF-κB p65蛋白表达显著上调(P<0.05);肝索结构严重紊乱,肝细胞有较多炎症细胞浸润,肝损伤程度评分显著升高(P<0.05)。与模型组比较,胆南星低、高剂量组小鼠肝组织病理变化及上述指标水平均显著改善(P<0.05)。结论胆南星对CCl_(4)致小鼠急性肝损伤具有保护作用,其机制可能与抑制JAK2/STAT3/NF-κB信号通路介导的炎症反应及抗氧化应激作用有关。

OBJECTIVE To investigate the protective effect and potential mechanism of Arisaema Cum Bile on acute liver injury induced by carbon tetrachloride(CCl_(4))in mice.METHODS Fifty mice were randomly divided into normal group,model group,positive control group(Biphenyl diester dropping pills,150 mg/kg),Arisaema Cum Bile low-dose and high-dose groups(0.78,2.34 g/kg),with 10 mice in each group.The mice in each group were given relevant medicine intragastrically,once a day,for 7 consecutive days.Two hours after the last administration,those groups were given intraperitoneal injection of 0.2%CCl_(4)-olive oil solution to induce acute liver injury model except for normal group.Seventeen hours after intraperitoneal injection,the serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and malondialdehyde(MDA),superoxide dismutase(SOD)in liver tissue were measured with kit method.The hepatic index was detected.The pathological changes of liver tissue were observed by HE staining,and the degree of liver injury was scored quantitatively.The mRNA expressions of TNF-αand IL-6 in liver tissue were detected by real-time fluorescence quantitative PCR;the protein expressions of Janus kinase 2(JAK2),signal transducer and activator of transcription protein 3(STAT3)and nuclear factor-κB p65(NF-κB p65)in liver tissue were detected by Western blot assay.RESULTS Compared with normal group,the levels of ALT,AST,IL-6,TNF-αand MDA,the hepatic index were increased significantly(P<0.05),while the level of SOD was decreased significantly(P<0.05);the mRNA expressions of IL-6 and TNF-α,and the protein expressions of JAK2,STAT3 and NF-κB p65 were up-regulated significantly(P<0.05);the pathological observation of liver tissue showed that the structure of hepatic cord was seriously disordered,there were many inflammatory cells infiltration of liver cells,and the liver injury score was significantly increased(P<0.05).Compared with model group,pathological changes and above indexes in mice were improved significantly in Arisaema Cum Bile low-dose and high-dose groups(P<0.05).CONCLUSIONS Arisaema Cum Bile has a protective effect on CCl_(4)-induced acute liver injury in mice,which may be related to the inhibition of inflammatory response mediated by JAK2/STAT3/NF-κB signal pathway and antioxidant stress.