基于IL-6/JAK2/STAT3信号通路研究岩白菜素对急性肝损伤大鼠的保护作用

Study on the protective effect of bergenin on acute liver injury in rats based on IL-6/JAK2/STAT3 signaling pathway

基于IL-6/JAK2/STAT3信号通路探讨岩白菜素对四氯化碳(CCl 4)致大鼠急性肝损伤的保肝作用及其作用机制。60只SD大鼠随机分为正常组、模型组、水飞蓟素(120 mg/kg)组、岩白菜素低、中、高(20、40、80 mg/kg)剂量组,每组10只。水飞蓟素组、岩白菜素各剂量组大鼠按照10 mL/kg灌胃给药,正常组和模型组灌胃相应体积溶剂,每日1次,连续1周。末次给药2 h后,除正常组以外,其余各组腹腔注射25%四氯化碳(CCl 4)橄榄油溶液2 mL/kg,建立急性肝损伤大鼠模型。禁食不禁水,16 h后眼球取血,并收集肝脏。生化法测定血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)、白蛋白(ALB)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)以及超氧化物歧化酶(SOD)的水平或活性。酶联免疫吸附法(ELISA)测定检测肝组织中白细胞介素-6(IL-6)、肿瘤坏死因子(TNF-α)和白细胞介素-1β(IL-1β)的含量。采用HE染色观察肝组织的病理变化程度,免疫组化(IHC)观察肝组织中IL-6的表达水平,蛋白免疫印迹法(Western blot)检测IL-6/JAK2/STAT3通路相关蛋白的表达水平。实验结果表明,与模型组相比,岩白菜素各剂量组和水飞蓟素组大鼠肝组织炎症坏死和IL-6表达不同程度的减轻。岩白菜素可显著降低血清中的ALT、AST、MDA和肝组织中TNF-α、IL-6和IL-1β活性或水平,上调血清中ALB、SOD和GSH-Px的活性或水平,并且抑制IL-6/JAK2/STAT3通路相关蛋白的表达。综上所述,岩白菜素对CCl 4致急性肝损伤大鼠具有保护作用,其保肝机制可能与IL-6/JAK2/STAT信号通路介导的炎症反应和抗氧化有关。

To explore the hepatoprotective effect and mechanism of bergenin on acute liver injury induced by CCl 4 in rats based on IL-6/JAK2/STAT3 signal pathway.A total of 60 SD rats were randomly divided into normal group,model group,silymarin group(120 mg/kg),and bergenin low,middle and high dose(20,40,80 mg/kg)groups with 10 rats in each group.Rats in silymarin group and bergenin groups were given intragastric administration according to 10 mL/kg,while normal group and model group were given corresponding volume of solvent once a day for a week.Two hours after the last administration,except the normal group,the other groups were intraperitoneally injected with 25%CCl 4 olive oil solution 2 mL/kg,to establish the rat model of acute liver injury.After fasting,the blood was taken from the eyeball and the liver was collected after 16 hours of fasting.The levels or activities of alanine transaminase(ALT),aspartate transaminase(AST),albumin(ALB),malondialdehyde(MDA),glutathione peroxidase(GSH-Px)and superoxide dismutase(SOD)in serum were determined by biochemical method.The contents of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)in liver tissue were measured by enzyme-linked immunosorbent assay(ELISA).The pathological changes of liver tissue were observed by HE staining,and the expression level of IL-6 in liver tissue was observed by immunohistochemical(IHC).The protein expression levels of JAK2,p-JAK2,STAT3,p-STAT3,NF-κB p65 and p-NF-κB p65 were detected by Western blot.The results showed that inflammation,necrosis and IL-6 expression of liver tissue were reduced in different doses of bergenin and silymarin groups compared with model group,bergenin significantly reduced the activities or levels of ALT,AST,MDA in serum,TNF-α,IL-6 and IL-1βin liver tissue,up-regulated the activities or levels of ALB,SOD and GSH-Px in serum,and inhibited the expression of proteins related to IL-6/JAK2/STAT3 pathway.In conclusion,bergenin has a protective effect on acute liver injury induced by CCl 4 in rats,and the liver protection mechanism may be related to the inflammatory response and antioxidant mediated by IL-6/JAK2/STAT3 signaling pathway.