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

doi:10.11843/j.issn.0366-6964.2022.07.030

摘要/Abstract

摘要： 为探讨茵山莲水提物对四氯化碳(CCl₄)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CCl₄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。

关键词: 茵山莲水提物, 急性肝损伤, 四氯化碳, 机制

Abstract: 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 (CCl₄) 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% CCl₄ 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 CCl₄, 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.

Key words: aqueous extract of Yinshanlian, acute liver injury, CCl₄, mechanism

中图分类号:

S853.74

魏媛媛, 樊艺萌, 袁燕燕, 尕玉, 张艳楠, 韩俊成, 郝智慧. 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[J]. 畜牧兽医学报, 2022, 53(7): 2333-2342.

WEI Yuanyuan, FAN Yimeng, YUAN Yanyan, GA Yu, ZHANG Yannan, HAN Juncheng, HAO Zhihui. Protective Effect of Aqueous Extract of Yinshanlian on Acute Liver Injury Induced by Carbon Tetrachloride in Mice[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(7): 2333-2342.

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