茵山莲水提物对四氯化碳诱导小鼠急性肝损伤的保护作用

Protective Effect of Aqueous Extract of Yinshanlian on Acute Liver Injury Induced by Carbon Tetrachloride in Mice

为探讨茵山莲水提物对四氯化碳(CCl_(4))诱导的小鼠急性肝损伤的保护作用及可能机制,以对后续茵山莲的深入研究及临床转化提供理论指导。将60只ICR小鼠随机分为6组,每组10只,分别为空白对照组、模型对照组、联苯双酯对照组、茵山莲低(1.17 mg·g^(-1))、中(2.34 mg·g^(-1))、高(4.68 mg·g^(-1))剂量组,按不同剂量药物每天灌胃1次,给药7 d。在末次给药1 h后,除空白对照组以外,其余各组小鼠腹腔注射0.2%四氯化碳构建急性肝损伤模型。12 h后小鼠眼球采血并分离血清,收集肝组织。采用生化分析检测血清总蛋白(TP)和谷胱甘肽S转移酶(GST)活性,肝组织中GST、琥珀酸脱氢酶(SDH)、过氧化氢酶(CAT)和总超氧化物歧化酶(T-SOD)活性,利用HE染色和Masson染色观察肝组织病理及肝纤维化情况,采用蛋白免疫印迹法和免疫组织化学法检测肝组织白介素1beta(IL-1β)、肿瘤坏死因子alpha(TNF-α)、血管生成因子A(VEGFA),金属基质蛋白酶9(MMP9)及转录因子NF-κBp65、蛋白激酶IKK、p38MAPK等蛋白表达及定位情况。结果表明,与模型对照组相比,2.34 mg·g^(-1)茵山莲水提物能显著降低小鼠血清GST水平,显著升高肝内CAT水平,各组间T-SOD无显著差异,但2.34 mg·g^(-1)茵山莲水提物组小鼠与空白对照组、模型对照组在肝GST、SDH和血清TP水平上无显著差异。2.34 mg·g^(-1)茵山莲水提物能缓解CCl_(4)诱导的小鼠肝病理损伤和纤维化病变,抑制肝IL-1β、TNF-α、VEGFA、NF-κBp65和IKK的表达。提示茵山莲水提物可能通过NF-κB和p38MAPK通路抑制肝氧化应激和炎症水平,从而缓解小鼠急性肝损伤。

The aim of this study was to explore the protective effect and possible mechanism of aqueous extract of Yinshanlian(YSL)on acute liver injury induced by carbon tetrachloride(CCl_(4))in mice,so as to provide theoretical guidance for further research and clinical transformation of YSL.Sixty ICR mice were randomly divided into 6 groups(n=10):control group,model group,low(1.17 mg·g^(-1)),medium(2.34 mg·g^(-1))and high(4.68 mg·g^(-1))YSL dosage groups.The drugs were administered by gavage once a day for 7 days.One hour after the last administration,except the control group,the mice in other groups were injected intraperitoneally with 0.2%CCl_(4) to establish the model of acute liver injury.After 12 hours,blood was collected from mouse eyeballs,serum was isolated,and liver tissue was collected.The serum total protein(TP)and activity of glutathione S transferase(GST),as well as the activity of GST,succinate dehydrogenase(SDH),catalase(CAT)and total superoxide dismutase(T-SOD)in liver tissue were detected by biochemical analysis.The liver pathology and liver fibrosis were observed by HE staining and Masson staining.Expression and localization of IL-1β,tumor necrosis factor alpha(TNF-α),angiogenesis factor A(VEGFA),matrix metalloproteinase 9(MMP9)and transcription factor NF-κBp65,protein kinase IKK,and p38MAPK in liver tissue were detected by Western blot and immunohistochemistry.The results showed that 2.34 mg·g^(-1) YSL aqueous extract could significantly reduce the level of serum GST and significantly increase the level of intrahepatic CAT compared with the model group.There was no significant difference in T-SOD among these groups.There was no significant difference in the levels of liver GST,SDH and serum TP between 2.34 mg·g^(-1) YSL aqueous extract group and control group or model group.Besides,2.34 mg·g^(-1) YSL aqueous extract could alleviate liver pathological injury and fibrosis induced by CCl_(4),and decrease the expression of liver IL-1β,TNF-α,VEGFA,NF-κBp65 and IKK.It is suggested that the aqueous extract of YSL probably inhibits liver oxidative stress and inflammation through NF-κB and p38MAPK pathways to alleviate acute liver injury in mice.