Kangtaizhi Granule Alleviated Nonalcoholic Fatty Liver Disease in High-Fat Diet-Fed Rats and HepG2 Cells via AMPK/mTOR Signaling Pathway

Abstract

Kangtaizhi granule (KTZG) is a Chinese medicine compound prescription and has been proven to be effective in nonalcoholic fatty liver disease (NAFLD) treatment clinically. However, the underlying mechanisms under this efficacy are rather elusive. In the present study, network pharmacology and HPLC analysis were performed to identify the chemicals of KTZG and related target pathways for NAFLD treatment. Network pharmacology screened 42 compounds and 79 related targets related to NAFLD; HPLC analysis also confirmed six compounds in KTZG. Further experiments were also performed. In an in vivo study, SD rats were randomly divided into five groups: control (rats fed with normal diet), NAFLD (rats fed with high-fat diet), and KTZG 0.75, 1.5, and 3 groups (NAFLD rats treated with KTZG 0.75, 1.5, and 3 g/kg, respectively). Serum lipids were biochemically determined; hepatic steatosis and lipid accumulation were evaluated with HE and oil red O staining. In an in vitro study, HepG2 cells were incubated with 1 mM FFA to induce lipid accumulation with or without KTZG treatment. MTT assay, intracellular TG level, oil red O staining, and glucose uptake in cells were detected. Western blotting and immunohistochemical and immunofluorescence staining were also performed to determine the expression of lipid-related genes PPAR-γ, SREBP-1, p-AKT, FAS, and SIRT1 and genes in the AMPK/mTOR signaling pathway. In high-fat diet-fed rats, KTZG treatment significantly improved liver organ index and serum lipid contents of TG, TC, LDL-C, HDL-C, ALT, and AST significantly; HE and oil red O staining also showed that KTZG alleviated hepatic steatosis and liver lipid accumulation. In FFA-treated HepG2 cells, KTZG treatment decreased the intracellular TG levels, lipid accumulation, and attenuated glucose uptake significantly. More importantly, lipid-related genes PPAR-γ, SREBP-1, p-AKT, FAS, and SIRT1 expressions were ameliorated with KTZG treatment in high-fat diet-fed rats and FFA-induced HepG2 cells. The p-AMPK and p-mTOR expressions in the AMPK/mTOR signaling pathway were also modified with KTZG treatment in high-fat diet-fed rats and HepG2 cells. These results indicated that KTZG effectively ameliorated lipid accumulation and hepatic steatosis to prevent NAFLD in high-fat diet-fed rats and FFA-induced HepG2 cells, and this effect was associated with the AMPK/mTOR signaling pathway. Our results suggested that KTZG might be a potential therapeutic agent for the prevention of NAFLD.

Comprehensive RNA-Seq Analysis of Potential Therapeutic Targets of Gan–Dou–Fu–Mu Decoction for Treatment of Wilson Disease Using a Toxic Milk Mouse Model

“…Specifically, we found that GDFMD could effectively reverse the abnormal expression of 20 genes in WD; among these the expression of 18 genes (Apoa2, Acaa1b, Acsl1, Acox1, Ppara, Fads2, Scd2, Slc27a2, Pck1, Hmgcs2, Cyp7a1, Cyp4a31, Cyp4a14, Slc27a5, Slc27a1, Scd1, Cyp8b1, and Fabp1) was low and that of 2 genes (Cyp4a32 and Cyp4a10) was high in the Model group. A large body of evidence suggests that PPAR signaling pathway is involved in different liver diseases (Panebianco et al, 2017;Zhang et al, 2020), especially liver fibrosis (Chen et al, 2015;Pawlak et al, 2015;Chhimwal et al, 2020), as was also confirmed in the present study. Our results show that GDFMD can alleviate liver damage in WD by reversing the changes in the PPAR signaling pathway (Figure 8).…”

11β-HSD1 Inhibitor Alleviates Non-Alcoholic Fatty Liver Disease by Activating the AMPK/SIRT1 Signaling Pathway

“…Recently published studies have demonstrated that acute glucocorticoid exposure increases cardiac-fatty-acid oxidation, by activating AMPK [ 34 ]. In contrast, in mice chronically exposed to glucocorticoids, AMPK phosphorylation levels and lipid metabolism gene levels have been shown to decrease with activation of 11β-HSD1 [ 19 , 35 , 36 ]. These phenomena may be due to AMPK inhibition associated with the 11β-HSD1-induced time-dependence of cortisol.…”

“…HepG2 cells (hepatocellular carcinoma) were purchased from the Procell Life Science Technology Co., Ltd., China, (CL-0103); these cells are useful for constructing the conditions of lipid deposition [ 19 , 20 ]. The cells were cultured in DMEM (Gibco, Carlsbad, CA, USA), comprising 10% fetal bovine serum (FBS) (Gibco) and 1% antibiotic (Invitrogen, Carlsbad, CA, USA), at 37 °C in a humidified 5% CO 2 atmosphere.…”

Transcriptome reveals key microRNAs involved in fat deposition between different tail sheep breeds

“…Studies have shown that some drugs ameliorate lipid accumulation and inflammation in nonalcoholic fatty liver disease through the AMPK pathway, such as LB100 [ 48 ], Ursolic acid [ 49 ], Allyl isothiocyanate [ 50 ], Ginsenoside Rk3 [ 51 ] and Kangtaizhi Granule [ 52 ]. As reported, miR-122 promotes lipogenesis via inhibiting the AMPK pathway by targeting SIRT1 in HepG2 and Huh-7 cells [ 53 ]. In our study, SIRT1 was also enriched in this pathway.…”