[1]牛丽娜,窦 婧,马 燕,等.和血柔肝方通过PI3K/Akt/NF-κB信号通路调控自噬相关蛋白改善大鼠肝纤维化机制研究[J].陕西中医,2024,(2):165-170.[doi:DOI:10.3969/j.issn.1000-7369.2024.02.004]
　NIU Lina,DOU Jing,MA Yan,et al.Mechanism of Hexue Rougan decoction in relieve liver fibrosis in rats by regulating autophagy-related proteins through PI3K/Akt/NF-κB signaling pathway[J].,2024,(2):165-170.[doi:DOI:10.3969/j.issn.1000-7369.2024.02.004]
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和血柔肝方通过PI3K/Akt/NF-κB信号通路调控自噬相关蛋白改善大鼠肝纤维化机制研究
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《陕西中医》[ISSN:1000-7369/CN:61-1281/TN]

卷:
期数:
2024年2期
页码:
165-170
栏目:
基础研究
出版日期:
2024-02-05

文章信息/Info

Title:
Mechanism of Hexue Rougan decoction in relieve liver fibrosis in rats by regulating autophagy-related proteins through PI3K/Akt/NF-κB signaling pathway
作者:
牛丽娜1; 窦 婧2; 马 燕2; 张 冰1; 王 尹1; 王晓忠2
(1.新疆医科大学第四临床医学院,新疆 乌鲁木齐 830054; 2.新疆维吾尔自治区中医医院,新疆 乌鲁木齐 830099)
Author(s):
NIU Lina; DOU Jing; MA Yan; ZHANG Bing; WANG Yin; WANG Xiaozhong
(The Fourth Clinical College of Xinjiang Medical University,Urumqi 830054,China)
关键词:
肝纤维化;  和血柔肝方;  磷脂酰肌醇3激酶/蛋白激酶B/核因子-κB信号通路;  自噬;  大鼠
Keywords:
Liver fibrosis;  Hexue Rougan decoction;  PI3K/Akt/NF-κB signaling pathway;  Autophagy;  Rats
分类号:
R 256.4
DOI:
DOI:10.3969/j.issn.1000-7369.2024.02.004
文献标志码:
A
摘要:
目的:探讨和血柔肝方通过调控磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(Akt)/核因子-κB(NF-κB)信号通路调控自噬相关蛋白改善大鼠肝纤维化的作用机制。方法:将40只SPF级SD雄性大鼠随机分为空白对照组、模型组、秋水仙碱组与和血柔肝方最佳剂量组,每组各10只。除空白对照组外,其余各组均以四氯化碳(CCl4)构建肝纤维化大鼠模型。造模8周后,空白对照组、模型组采用0.9%氯化钠溶液灌胃; 秋水仙碱组采用秋水仙碱灌胃; 和血柔肝方最佳剂量组以和血柔肝方灌胃。灌胃结束后,取各组大鼠血清及肝脏组织。生化检测各组大鼠血清天门冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)水平; HE及Masson染色评估大鼠肝纤维化情况。RT-qPCR法检测大鼠肝组织PI3K、Akt、NF-κB以及NF-κB下游自噬相关蛋白微管相关蛋白轻链3Ⅱ(LC3-Ⅱ)、Beclin-1的mRNA的表达水平; 蛋白免疫印迹(Western blot)检测大鼠肝组织中PI3K、p-PI3K、Akt、p-Akt、NF-κB P65、p-NF-κB P65、LC3-Ⅱ、Beclin-1蛋白表达水平。结果:与模型组比较,和血柔肝方最佳剂量组血清ALT、AST水平明显降低(P<0.05)。与模型组比较,和血柔肝方最佳剂量组大鼠肝脏病理受损可见明显缓解。与模型组比较,和血柔肝方最佳剂量组大鼠肝组织PI3K、Akt、NF-κB、LC3-Ⅱ、Beclin-1基因表达水平明显降低(P<0.05)。与模型组比较,和血柔肝方最佳剂量组的PI3K、Akt、p-Akt、NF-κB P65、p-NF-κB P65、LC3-Ⅱ、Beclin-1蛋白表达Western blot结果均显著降低(P<0.05)。结论:和血柔肝方可通过抑制肝纤维化大鼠肝组织PI3K/Akt/NF-κB信号通路的表达以降低自噬相关蛋白活化,从而改善大鼠肝脏炎症,减缓肝脏病理损害,改善肝纤维化。
Abstract:
Objective:To investigate the mechanism of Hexue Rougan decoction in improving liver fibrosis in rats by regulating phosphatidylinositol 3 kinase/protein kinase B/ nuclear factor-κB signaling pathway and regulating autophagy-related proteins.Methods:Forty SPF male SD rats were randomly divided into blank control group,model group,colchicine group and optimal dose of Hexue Rougan decoction group,with 10 rats in each group.Except for the blank control group,the rats models of liver fibrosis were established with carbon tetrachloride(CCl4)in the other groups.After 8 weeks of modeling,the blank control group and the model group were given normal saline by gavage.The mice in the colchicine group were treated with colchicine by gavage.The optimal dose of Hexue Rougan decoction group was gavaged with Hexue Rougan decoction.At the end of gavage,serum and liver tissue were collected from each group.The serum levels of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)were detected by biochemical methods.HE and Masson staining were used to evaluate liver fibrosis in rats.RT-qPCR was used to detect the mRNA expression levels of PI3K,Akt,NF-κB and NF-κB downstream autophagy-related proteins microtubule-associated protein light chain 3Ⅱ(LC3-Ⅱ)and Beclin-1 in rat liver tissue.Western blot was used to detect the protein expression levels of PI3K,p-PI3K,Akt,p-Akt,NF-κB P65,p-NF-κB P65,LC3-Ⅱ and Beclin-1 in rat liver tissue.Results:Compared with the model group,the levels of ALT and AST in the optimal dose of Hexue Rougan decoction group were significantly decreased(P<0.05).Compared with the blank control group,the liver pathological damage of the optimal dose of Hexue Rougan decoction group was significantly alleviated.Compared with the model group,the expression levels of PI3K,Akt,NF-κB,LC3-Ⅱ and Beclin-1 genes in liver tissue of rats in the optimal dose of Hexue Rougan decoction group were significantly decreased(P<0.05).Compared with the model group,PI3K,Akt,p-Akt,NF-κB P65,p-NF-κB P65,LC3-Ⅱ and Beclin-1 in the optimal dose of Hexue Rougan decoction group were significantly decreased(P<0.05).Conclusion:Hexue Rougan decoction can inhibit the expression of PI3K/Akt/NF-κB signaling pathway in liver tissue of liver fibrosis rats and reduce the expression of autophagy-related proteins,thereby improving liver inflammation,reducing liver pathological damage and liver fibrosis in rats.

参考文献/References:

[1] 田艳茹.miR-133b与慢性乙型肝炎肝纤维化相关性研究[J].陕西医学杂志,2023,52(4):448-451.
[2] 蔡碧莲,文亦磊,罗伟生.大鼠肝纤维化实验动物模型的构建[J].陕西医学杂志,2020,49(6):659-662.
[3] JI D,ZHAO Q,QIN Y,et al.Germacrone improves liver fibrosis by regulating the PI3K/AKT/mTOR signalling pathway[J].Cell Biol Int,2021,45(9):1866-1875.
[4] 王燕,肖雄,郭峰,等.和血柔肝方对肝纤维化大鼠SphK1/S1P/S1PR信号通路的影响[J].中国中医药信息杂志,2022,29(3):85-91.
[5] 延国威,王晓忠,范灵芝,等.和血柔肝方治疗慢性乙型肝炎肝纤维化临床研究[J].中国中医药信息杂志,2019,26(8):24-29.
[6] WANG Y,XIAO X,WANG X B,et al.Identification of differentially expressed long noncoding RNAs and pathways in liver tissues from rats with hepatic fibrosis[J].PLoS One,2021,16(10):258194.
[7] 侯宝峰,寇小妮.肝纤维化的中医病机浅析[J].陕西中医,2008,29(6):761.
[8] FENG J,CHEN K,XIA Y,et al.Salidroside ameliorates autophagy and activation of hepatic stellate cells in mice via NF-κB and TGF-β1/Smad3 pathways[J].Drug Des Devel Ther,2018,12:1837-1853.
[9] 周志朋,薛娟娣,武琰琼,等.基于PI3K/AKT和MAPK信号通路探讨防风桃红四物汤抗肝纤维化的作用机制[J].中医临床研究,2022,14(11):19-22.
[10] LEE C,BAK J,YOON S,et al.Protective effect of oligonol on dimethylnitrosamine-induced liver fibrosis in rats via the JNK/NF-κB and PI3K/Akt/Nrf2 signaling pathways[J].Antioxidants(Basel),2021,10(3):366.
[11] YANG J,PI C,WANG G.Inhibition of PI3K/Akt/mTOR pathway by apigenin induces apoptosis and autophagy in hepatocellular carcinoma cells[J].Biomed Pharmacother,2018,10(3):699-707.
[12] FANG B,WEN S,LI Y,et al.Prediction and verification of target of helenalin against hepatic stellate cell activation based on miR-200a-mediated PI3K/Akt and NF-κB pathways[J].Int Immunopharmacol,2021,92:107208.
[13] WU L,ZHANG Q,MO W,et al.Quercetin prevents hepatic fibrosis by inhibiting hepatic stellate cell activation and reducing autophagy via the TGF-β1/Smads and PI3K/Akt pathways[J].Sci Rep,2017,7(1):9289.
[14] YANG L,FU W L,ZHU Y,et al.T beta 4 suppresses lincRNA-p21-mediated hepatic apoptosis and fibrosis by inhibiting PI3K-Akt-NF-κB pathway[J].Gene,2020,758:144946.
[15] 李硕熙,王炳予,杨磊,等.芪参汤通过PI3K/Akt/mTOR信号通路抗肝纤维化的机制研究[J].海南医学报,2020,26(12):905-909,914.
[16] 周翔,顾达,童聪,等.鳖甲煎丸对CCl4诱导小鼠肝纤维化的治疗作用机制研究[J].陕西中医,2022,43(2):151-156.
[17] 孙廷杰,周晓玲.中药复方治疗乙肝肝纤维化临床研究进展[J].实用中医药杂志,2023,39(3):632-634.
[18] MOHAMMED S,NICKLAS E H,THADATHIL N,et al.Role of necroptosis in chronic hepatic inflammation and fibrosis in a mouse model of increased oxidative stress[J].Free Radic Biol Med,2021,164:315-328.
[19] NIXON R A.The role of autophagy in neurodegenerative disease[J].Nat Med,2013,19(8):983-997.
[20] GAO J,WEI B,ASSUNCAO T M,et al.Hepatic stellate cell autophagy inhibits extracellular vesicle release to attenuate liver fibrosis[J].J Hepatol,2023,73(5):1144-1154.
[21] TSUCHIDA T,FRIEDMAN S L.Mechanisms of hepatic stellate cell activation[J].Nat Rev Gastroenterol Hepatol,2017,14(7):397-411.
[22] 吕艳杭,吴姗姗,王振常,等.中医药调控活性氧(ROS)抗肝纤维化的研究进展[J].中华中医药学刊,2021,39(6):117-121.
[23] MIHAYLOVA M M,SHAW R J.The AMPK signalling pathway coordinates cell growth,autophagy and metabolism[J].Nat Cell Biol,2011,13(9):1016-1023.
[24] LIU C,WANG X,WANG X,et al.A new LKB1 activator,piericidin analogue S14,retards renal fibrosis through promoting autophagy and mitochondrial homeostasis in renal tubular epithelial cells[J].Theranostics,2022,12(16):7158-7179.
[25] 王宝玉.益气活血方调节肝脏细胞自噬及逆转肝纤维化机制的研究[D].石家庄:河北医科大学,2017.

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备注/Memo

备注/Memo:
基金项目:国家自然科学基金资助项目(81760832); 新疆维吾尔自治区自然科学基金资助项目(2022D01C173); 新疆医科大学附属中医医院院级重点专项课题; 乐德行全国名中医传承工作室创新项目
更新日期/Last Update: 2024-02-07