摘要
BACKGROUND Inhibition of liver fibrosis plays a crucial role in curbing the advancement of chronic disease to cirrhosis and even liver cancer.However,modern medicine currently lacks direct anti-fibrotic drugs.He-He-Shu-Yang formula(HHSY)is a renowned Chinese medicine for the treatment of liver fibrosis.However,its mechanism of action has not been fully unraveled.AIM To explore the efficacy and mechanism of action of HHSY through in vitro and in vivo experiments.METHODS A liver fibrosis rat model(carbon tetrachloride-induced)was treated with low-or high-dose HHSY(10.42 g/kg or 20.84 g/kg)or with colchicine(1 mg/kg)for 9 weeks.In vitro,LX-2 human hepatic stellate cells(HSCs)were activated using transforming growth factor-β1 and subsequently treated with HHSY-containing serum or a nicotinamide adenine dinucleotide phosphate oxidase 4(NOX4)inhibitor.Through high-performance liquid chromatography,histopathology(hematoxylin and eosin,Masson),immunohistochemistry,western blot,and quantitative reverse transcription polymerase chain reaction analyses,we demonstrated that HHSY inhibited HSC activation and suppressed the NOX4/reactive oxygen species(ROS)/nucleotide binding oligomerization domain-like receptor family pyrin domain-containing 3(NLRP3)pathway.RESULTS In vivo,HHSY improved liver function and alleviated liver pathology,including reducing inflammatory cell infiltration,and liver fibrosis in carbon tetrachloride rats.with more significant effects at higher doses.Immunohistochemistry revealed that HHSY could decrease alpha-smooth muscle actin,NOX4,and NLRP3 expression,as well as serum ROS levels(O_(2)-and H_(2)O_(2),P<0.05).Western blot analysis confirmed HHSY also reduced NLRP3 protein levels(P<0.05).In vitro,HHSY at 1.25%or 2.5%reduced the levels of ACTA2 mRNA,NOX4 protein and NOX4 mRNA,ROS production,and NLRP3 and IL-1βmRNA in activated LX-2 cells(P<0.05).CONCLUSION HHSY effectively treats liver fibrosis,likely by inhibiting HSC activation through the NOX4/ROS/NLRP3 pathway.This underscores HHSY’s clinical relevance as a potential therapeutic option for liver fibrosis.
基金
Supported by the Guangdong Basic and Applied Basic Research Fund Project,No.2022A1515110825 and No.2023A1515011092
the Incubation Program for the Science and Technology Development of Chinese Medicine Guangdong Laboratory,No.HQL2024PZ033
the Chi Xiaoling National Famous Traditional Chinese Medicine Expert Inheritance Studio,Teaching Letter from the State Traditional Chinese Medicine Office[2022],No.75
the Advantage Disease Project of Guangdong Provincial Hospital of Traditional Chinese Medicine[2020],No.37
the Science and Technology Planning Project of Guangdong Province,No.2023B1212060063
the Project of Guangdong Provincial Key Laboratory of Chinese Medicine for Prevention and Treatment of Refractory Chronic Diseases,No.YN2023MB04
the Clinical Research Projects of Guangdong Provincial Hospital of Chinese Medicine,No.YN2022QN05
the Guangzhou Basic Research Program for Young Doctors in Basic and Applied Basic Research,No.2024A04J3004.
