Objective: To analyze the mechanism of Huanglian Jiedu Decoction (HLJDD) in the treatment of metabolism-associated fatty liver disease (MAFLD). Methods: The main components, targets, and pathways for treating MAFLD of HLJDD were screened through network pharmacology and molecular docking validation was done; HLJDD was used to intervene MAFLD model of rat, the levels of ALT, AST, TC, TG, GLU, HDL, and LDL were identified, HE staining was used to observe the pathological changes, lipid deposition in liver was detected by oil red O staining. MAFLD model of HepG2 (hepatocellular carcinoma cell line) was constructed by PA (palmitate-acid) incubating, and HLJDD was administered with drug-containing serum intervention, lipid droplets in HepG2 cells was observed by oil red O staining, TG and FFA of HepG2 were detected, the expressions of AMPK, mToR, and Beclin-1 were detected through Western blot. Results: Seventy components and 229 targets were obtained, and 85 targets were used to treat MAFLD, which focus on the signal passways of AMPK/mToR/PI3K-AKt/MAPK, NAFLD, autophagy-animal, insulin, etc. Molecular docking outcomes showed quercetin, kaempferol, and baicalein that were successfully docked with AMPK and mToR, and had good binding activity, compared with MAFLD group of rats, the levels of ALT, AST, TC, TG, GLU, HDL, and LDL were significantly decreased in Silybin group and each dos group of HLJDD, liver pathology and lipid deposition were significantly improved; the results in vitro experiments showed that drug-containing serum of HLJDD and Silybin could improve intracellular lipid accumulation and reduce the increase of TG and FFA levels in HepG2 cells, the therapeutic effect of HLJDD was significantly attenuated after application of AMPK inhibitor; the results of Western blot showed that HLJDD could up-regulate the protein expression of AMPK and Beclin-1,down-regulate the protein expression of mToR. Conclusion: Within process of MAFLD intervention, HLJDD could regulate AMPK-mToR signaling pathway to treat MAFLD.

中文翻译：

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网络药理学及实验验证解析黄连解毒汤治疗MAFLD的机制

目的：分析黄连解毒汤（HLJDD）治疗代谢相关性脂肪肝（MAFLD）的作用机制。方法：通过网络药理学筛选HLJDD治疗MAFLD的主要成分、靶点和通路，并进行分子对接验证；采用HLJDD干预大鼠MAFLD模型，测定ALT、AST、TC、TG、GLU、HDL、LDL水平，HE染色观察病理变化，油红O检测肝脏脂质沉积。染色。PA（棕榈酸）孵育构建HepG2（肝癌细胞系）MAFLD模型，给予HLJDD含药血清干预，油红O染色、HepG2 TG、FFA观察HepG2细胞内脂滴采用Western blot检测AMPK、mToR、Beclin-1的表达。结果：共获得70个成分、229个靶点，其中85个靶点用于治疗MAFLD，重点关注AMPK/mToR/PI3K-AKt/MAPK、NAFLD、自噬-动物、胰岛素等信号通路。分子对接结果显示槲皮素、山奈酚、黄芩素与AMPK、mToR成功对接，并具有良好的结合活性，与MAFLD组大鼠相比，水飞蓟宾的ALT、AST、TC、TG、GLU、HDL、LDL水平显着降低HLJDD组及各剂量组肝脏病理及脂质沉积均明显改善；结果体外实验表明，HLJDD和水飞蓟宾含药血清可改善HepG2细胞内脂质积累，降低TG和FFA水平的升高，应用AMPK抑制剂后HLJDD的治疗效果明显减弱。Western blot结果显示，HLJDD能够上调AMPK和Beclin-1蛋白表达，下调mToR蛋白表达。结论:在MAFLD干预过程中,HLJDD可通过调节AMPK-mToR信号通路发挥治疗MAFLD的作用。