Yinchenhao Tang alleviates high fat diet induced NAFLD by increasing NR1H4 and APOA1 expression

10.1016/j.jtcme.2023.02.010

Li Xu；Hongliang Cui

NAFLD ； Yinchenhao tang ； Lipid metabolism ； NR1H4 ； APOA1

Journal of Traditional and Complementary Medicine

13卷4期（2023 / 07 / 01）

325 - 336

英文

Background and aim: Traditional Chinese medicine Yinchenhao Tang (YCHT) demonstrated benefits when treating nonalcoholic fatty liver disease (NAFLD), but the dose effects and potential targets are still ambiguous. In this study, different concentrations of YCHT were employed to treat NAFLD and the underlying therapeutic targets were investigated. Experimental procedure: Kunming mice were fed with high fat diet (HFD) for 8 weeks to induce NAFLD, then treated with 3 different concentrations of YCHT. Hepatic pathological changes and serum lipid levels were examined. Network pharmacology was applied to screen the potential targets of YCHT for NAFLD modulation. NR1H4 and APOA1 expression was evaluated by QPCR and western blotting. Immunohistochemistry (IHC) staining was conducted to visualize the localization pattern of NR1H4 and APOA1 in the liver. Results: YCHT significantly reduced liver lipid storage and improved the liver pathological status of NAFLD mice. The serum lipid levels, as well as alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, were remarkably reduced by the middle and high dose YCHT. There are 35 potential targets for YCHT to regulate NAFLD. HFD suppressed both RNA and protein expression of NR1H4 and APOA1, while YCHT elevated NR1H4 and APOA1 expression. IHC staining indicated that NR1H4 was mainly located in the cell nucleus and the APOA1 signal was observed at the liver sinusoid or cytoplasm. Conclusion: YCHT can effectively ameliorate HFD induced NAFLD by modulating the promising targets of NR1H4 and APOA1.

1. Apweiler, R,Bairoch, A,Wu, CH(2004).UniProt: the universal protein knowledgebase.Nucleic Acids Res,32,115-119.
2. Asrani, SK,Devarbhavi, H,Eaton, J(2019).Burden of liver diseases in the world.J Hepatol,70(1),151-171.
3. Cai, FF,Bian, YQ,Wu, R(2019).Yinchenhao decoction suppresses rat liver fibrosis involved in an apoptosis regulation mechanism based on network pharmacology and transcriptomic analysis.Biomed Pharmacother,114,108863.
4. Cai, FF,Wu, R,Song, YN(2018).Yinchenhao decoction alleviates liver fibrosis by regulating bile acid metabolism and TGF-β/smad/ERK signalling pathway.Sci Rep,8(1),15367.
5. Cariou, B,van Harmelen, K,Duran-Sandoval, D(2006).The farnesoid X receptor modulates adiposity and peripheral insulin sensitivity in mice.J Biol Chem,281(16),11039-11049.
6. Chen, SD,Fan, Y,Xu, WJ(2011).Effects of yinchenhao decoction for non-alcoholic steatohepatitis in rats and study of the mechanism.Journal of traditional Chinese medicine ,31(3),220-223.
7. Collaboratio, NRF(2016).Trends in adult body-mass index in 200 countries from 1975 to 2014: a pooled analysis of 1698 population-based measurement studies with 19$2 million participants.Lancet,387(10026),1377-1396.
8. Cooke, AL,Morris, J,Melchior, JT(2018).A thumbwheel mechanism for APOA1 activation of LCAT activity in HDL.J Lipid Res,59(7),1244-1255.
9. Dong, H,Lu, FE,Zhao, L(2012).Chinese herbal medicine in the treatment of nonalco-holic fatty liver disease.Chin J Integr Med,18(2),152-160.
10. Elshourbagy, NA,Meyers, HV,Abdel-Meguid, SS(2014).Cholesterol: the good, the bad, and the ugly-therapeutic targets for the treatment of dyslipidemia.Med Princ Pract : international journal of the Kuwait University, Health Science Centre,23(2),99-111.
11. Flores, R,Jin, X,Chang, J(2019).LCAT, ApoD, and ApoA1 expression and review of cholesterol deposition in the cornea.Biomolecules,9(12),785.
12. Guo, Y,Li, JX,Wang, YL(2017).Yinchen linggui zhugan decoction ameliorates nonalcoholic fatty liver disease in rats by regulating the Nrf2/ARE signaling pathway.Evid Based Complement Alternat Med,2017,6178358.
13. Huang, J,Cheung, F,Tan, HY(2017).Identification of the active compounds and significant pathways of yinchenhao decoction based on network pharmacology.Mol Med Rep,16(4),4583-4592.
14. Kong, B,Wang, L,Chiang, JY(2012).Mechanism of tissue-specific farnesoid X receptor in suppressing the expression of genes in bile-acid synthesis in mice.Hepatology,56(3),1034-1043.
15. Lee, TY,Chang, HH,Kuo, JJ(2009).Changes of hepatic proteome in bile duct ligated rats with hepatic fibrosis following treatment with Yin-Chen-Hao-Tang.Int J Mol Med.,23(4),477-484.
16. Lee, TY,Chang, HH,Lo, WC(2010).Alleviation of hepatic oxidative stress by Chinese herbal medicine Yin-Chen-Hao-Tang in obese mice with steatosis.Int J Mol Med.,25(6),837-844.
17. Levene, AP,Kudo, H,Armstrong, MJ(2012).Quantifying hepatic steatosis - more than meets the eye.Histopathology,60(6),971-981.
18. Levene, AP,Kudo, H,Thursz, MR(2010).Is oil red-O staining and digital image analysis the gold standard for quantifying steatosis in the liver?.Hepatology,51(5),1859-1860.
19. Li, J,Wu, H,Liu, Y(2020).High fat diet induced obesity model using four strains of mice: Kunming, C57BL/6, BALB/c and ICR.Exp Anim,69(3),326-335.
20. Liu, Z,Guo, F,Wang, Y(2016).BATMAN-TCM: a bioinformatics analysis tool for molecular mechANism of traditional Chinese medicine.Sci Rep,6,21146.
21. Ma, K,Saha, PK,Chan, L(2006).Farnesoid X receptor is essential for normal glucose homeostasis.J Clin Invest,116(4),1102-1109.
22. Ru, J,Li, P,Wang, J(2014).TCMSP: a database of systems pharmacology for drug discovery from herbal medicines.J Cheminf,6,13.
23. Santhekadur, PK,Kumar, DP,Sanyal, AJ(2018).Preclinical models of non-alcoholic fatty liver disease.J Hepatol,68(2),230-237.
24. Sarwar, R,Pierce, N,Koppe, S.(2018).Obesity and nonalcoholic fatty liver disease: current perspectives.Metab Syndrome Obes Targets Ther,11,533-542.
25. Schmittgen, TD,Livak, KJ(2008).Analyzing real-time PCR data by the comparative C(T) method.Nat Protoc,3(6),1101-1108.
26. Stelzer, G,Rosen, N,Plaschkes, I(2016).The GeneCards suite: from gene data mining to disease genome sequence analyses.Current protocols in bioinformatics,54,1.30.31-31.30.33.
27. Stofan, M,Guo, GL(2020).Bile acids and FXR: novel targets for liver diseases.Front Med,7,544.
28. Szklarczyk, D,Gable, AL,Lyon, D(2019).STRING v11: protein-protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets.Nucleic Acids Res,47(D1),d607-d613.
29. Takahashi, Y,Soejima, Y,Kumagai, A(2014).Japanese herbal medicines shosai-koto, inchinkoto, and juzentaihoto inhibit high-fat diet-induced nonalcoholic steatohepatitis in db/db mice.Pathol Int,64(10),490-498.
30. Wu, YL,Li, ZL,Zhang, XB(2019).Yinchenhao decoction attenuates obstructive jaundice-induced liver injury and hepatocyte apoptosis by suppressing protein kinase RNA-like endoplasmic reticulum kinase-induced pathway.World J Gastroenterol,25(41),6205-6221.
31. Xue, R,Fang, Z,Zhang, M(2013).TCMID: traditional Chinese Medicine integrative database for herb molecular mechanism analysis.Nucleic Acids Res,41,1089-1095.
32. Yan, J,Xie, G,Liang, C(2017).Herbal medicine Yinchenhaotang protects against α-naphthylisothiocyanate-induced cholestasis in rats.Sci Rep,7(1),4211.
33. Yan, T,Yan, N,Wang, P(2020).Herbal drug discovery for the treatment of nonalcoholic fatty liver disease.Acta Pharm Sin B.,10(1),3-18.
34. Yang, JM,Sun, Y,Wang, M(2019).Regulatory effect of a Chinese herbal medicine formula on non-alcoholic fatty liver disease.World J Gastroenterol,25(34),5105-5119.
35. Younossi, Z,Tacke, F,Arrese, M(2019).Global perspectives on nonalcoholic fatty liver disease and nonalcoholic steatohepatitis.Hepatology,69(6),2672-2682.
36. Zhang, A,Sun, H,Qiu, S,Wang, X(2013).Advancing drug discovery and development from active constituents of yinchenhao tang, a famous traditional Chinese medicine formula.Evid Based Complement Alternat Med,2013,257909.
37. Zhang, H,Wang, X,Hu, P(2016).Serum metabolomic characterization of liver fibrosis in rats and anti-fibrotic effects of yin-chen-hao-tang.Molecules,21,126.
38. Zhou, Y,Zhou, B,Pache, L(2019).Metascape provides a biologist-oriented resource for the analysis of systems-level datasets.Nat Commun,10(1),1523.