南方医科大学学报

南方医科大学学报

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基于PI3K/Akt/NF-κB信号通路探讨竹节参总皂苷对CCl4致急性肝损伤的保护作用

吴广阳，宋添力，唐 浪，王一民，刘 绪，黄 胜   

  1. 湖北民族大学医学部，风湿性疾病发生与干预实验室，恩施 湖北 445000；湖北恩施学院，恩施 湖北 445000
  • 出版日期:2024-01-27 发布日期:2024-01-27

Protective effects of total saponins of Panax japonicus on CCl4-induced acute liver injury based on PI3K/AktNF-κB signaling pathway

WU Guangyang, SONG Tianli, TANG Lang, WANG Yiming, LIU Xu, HUANG Sheng   

  1. Hubei Minzu University Department of medicine, Hubei Minzu University Hubei Provincial Key Laboratory of Occurrence and Intervention of Rheumatic diseases, Enshi 445000, China; Hubei Ensch College, Enshi 445000, China
  • Online:2024-01-27 Published:2024-01-27

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摘要： 目的 探讨土家族药用植物竹节参总皂苷对CCl4致急性肝损伤的保护作用及潜在的药理学机制。方法 将6周龄SPF级雄性SD大鼠鼠随机分为正常组、模型组、联苯双酯组（100 mg/kg）、竹节参总皂苷低、中、高（50、100、200 mg/kg）剂量组，各组8只，除空白组外，其余各组采用CCl4诱导大鼠急性肝损伤模型，处理组于造模中给予正肝方灌胃干预。比较各组大鼠的血清谷草转氨酶（AST）、谷丙转氨酶（ALT）、总胆红素（TBil）和碱性磷酸酶（ALP）水平指标；HE染色观察肝组织病理学改变；免疫组化检测肝脏组织PI3K/Akt/NF-κB信号通路相关分子的表达；酶联免疫法测定肝脏组织总超氧化物歧化酶（T-SOD）、谷胱甘肽过氧化物酶（GSH-Px）、丙二醛（MDA）水平；蛋白免疫印迹法检测肝脏组织PI3K-Akt和SIRT6-NF-κB通路相关蛋白表达情况。 结果 网络药理学分析显示，竹节参总皂苷对急性肝损伤的有治疗作用，其关键的通路为PI3K/Akt等信号通路。血清学和酶联免疫学实验结果显示，与正常组相比，模型组大鼠血清AST、ALT、ALP、TBil和MDA明显增高（P<0.01），T-SOD和GSH-Px水平显著降低（P<0.01）；与模型组相比，各治疗组ALT、AST、ALP、TBIL和MDA水平显著降低（P<0.01），T-SOD和GSH-Px水平显著升高（P<0.01）。免疫组化结果显示，正常组大鼠肝组织细胞内未见p-NF-κB表达阳性，与正常组相比，模型组中p-NF-κB阳性表达则明显增加，阳性细胞数增多（P<0.01）；与模型组比较，各治疗组阳性表达明显减少（P<0.01）。免疫印迹结果显示，相对于正常组，模型组PI3K、p-Akt和SIRT6蛋白的表达水平下降，p-NF-κB、TNF-α和IL-6蛋白的表达水平升高（P<0.05）；与模型组相比，各治疗组PI3K、p-Akt和SIRT6蛋白的表达水平显著升高，p-NF-κB p65、TNF-α和IL-6的蛋白的表达水平显著降低（P<0.05）。结论 竹节参总皂苷可以通过调节PI3K/Akt和SIRT6/NF-κB通路，有效减缓CCl4诱导的大鼠急性肝损伤，发挥其抗炎、抗氧化应激和保护肝损伤的作用。

关键词: 急性肝损伤, 竹节参总皂苷, PI3K/Akt/NF-κB, 抗炎, 抗氧化

Abstract: Objective The aim of this study was to investigate the protective effect of total saponins of Drukophora ginseng against CCl4- induced acute liver injury and the potential pharmacological mechanisms. Methods SD rats were randomly divided into normal group, model group, biphenyl bisabololol group (100 mg/kg), and total saponin of Druchysanthes chinensis low, medium, and high (50, 100, and 200 mg/kg) dosage group, with 8 rats in each group. Except for the blank group, the other groups used the CCl4-induced ALI model in rats, and the treatment group was given the intervention of positive liver formula by gavage in the modeling. The AST, ALT, TBil and alkaline phosphatase ALP levels of rats in each group were compared; HE was used to observe the histopathological changes in liver tissue; IHC was used to detect the expression of molecules related to PI3K/Akt/NF-κB signaling pathway in liver tissue; ELISA was used to determine the levels of T- SOD, GSH-Px, and MDA; WB was used to detect the levels of PI3K-Akt and SIRT6-NF-κB pathway in liver tissue; and WB was used to detect the levels of PI3K-Akt and SIRT6-NF-κB pathway in liver tissue. -NF-κB pathway related protein expression in liver tissue. Results The network pharmacological analysis showed that total saponin of Druksen had therapeutic effects on ALI, and the key pathways were PI3K/Akt and other signaling pathways. The experimental results showed that serum AST, ALT, ALP, TBil and MDA were significantly increased in the model group (P<0.01), and the levels of T- SOD and GSH- Px were significantly decreased (P<0.01); the levels of ALT, AST, ALP, TBil and MDA were significantly decreased in the treatment groups (P<0.01), and the levels of T-SOD and GSH-Px were significantly increased (P<0.01). elevated (P<0.01).IHC showed that no positive expression of p-NF-κB was seen in the liver tissue cells of rats in the normal group, while the positive expression of p- NF- κB was significantly increased in the model group, and the number of positive cells was increased (P<0.01); the positive expression was significantly reduced in the treatment groups (P<0.01).WB showed that the expression of PI3K, p-Akt and SIRT6 proteins was decreased in the model group, and the level of p-NF- κB was significantly decreased in the model group, and the level of p- NF- Px was significantly decreased in the model group (P<0.01). levels decreased, and the expression levels of p-NF-κB, TNF-α and IL-6 proteins increased (P<0.05); the expression levels of PI3K, p- Akt and SIRT6 proteins in each treatment group significantly increased, and the expression levels of p- NF- κB p65, TNF- α and IL- 6 proteins significantly decreased (P<0.05). Conclusion Total saponins of bamboo ginseng can effectively slow down CCl4-induced acute liver injury in rats by regulating the PI3K/Akt and SIRT6/NF- κB pathways, exerting their anti- inflammatory, anti- oxidative stress and protective effects ALI.

Key words: Acute liver injury, Total saponins of panax japonicus, PI3K/Akt/NF-κB, anti-inflammation , anti-oxidation