Sinomenine Attenuates Acetaminophen-Induced Acute Liver Injury by Decreasing Oxidative Stress and Inflammatory Response via Regulating TGF-β/Smad Pathway in vitro and in vivo

Hui Chen

Yao Wang

Fang-Zhou Jiao

Fan Yang

Xun Li

Lu-Wen Wang

^{Institute of Infectious Diseases, Hubei Center for Disease Control and Prevention, Wuhan, 430079, Hubei Province, People’s Republic of China}

^{Department of Infectious Diseases, Renmin Hospital of Wuhan University, Wuhan, 430060, Hubei Province, People’s Republic of China}

Correspondence: Lu-Wen Wang Department of Infectious Diseases, Renmin Hospital of Wuhan University, No. 238 Jiefang Road, Wuhan, Hubei Province, 430060, People’s Republic of China, Phone: Tel +86-15366254410, Email luwenwangmyemail@126.com

Received 2020 Feb 7; Accepted 2020 Apr 24.

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Abstract

Introduction

Liver disease is common and often life-threatening. Sinomenine (SIN) is an active ingredient extracted from Sinomenium acutum. This study investigated the protective effect and mechanism of sinomenine (SIN) on acetaminophen (APAP)-induced liver injury from in vitro and in vivo.

Methods

In vivo experiments, mice were randomly divided into six groups (n=10): control group, model group, SIN (25 mg/kg) group, SIN (50 mg/kg) group, SIN (100 mg/kg) group and SIN (100 mg/kg) + SRI-011381 group. Alanine transaminases (ALT), aspartate transaminases (AST) and alkaline phosphatase (ALP) were detected. The pathological lesion was measured by HE staining. Apoptosis was measured by TUNEL staining. In vitro experiments, BRL-3A cells were treated with APAP (7.5 mM) and then subjected to various doses of SIN (10, 50 and 100 μg/mL) at 37°C for 24 h. Inflammatory factors and oxidative stress index were measured by ELISA. The expression of proteins was detected by Western blot.

Results

The results showed that compared with the control group, the levels of ALT, AST and ALP in the serum of APAP-induced mice were significantly increased, followed by liver histological damage and hepatocyte apoptosis. Besides, APAP reduced the activity of SOD and GSH-Px, while increasing the content of MDA and LDH. Notably, APAP also promoted the expression of NLRP3, ASC, caspase-1 and IL-1β. Interestingly, SIN treatment dose-dependently reduced APAP-induced liver injury and oxidative stress, inhibited the activation of NLRP3 inflammasomes, and reduced the levels of inflammatory cytokines. In vitro studies have shown that SIN treatment significantly reduced the viability of BRL-3A cells and oxidative stress and inflammation. In addition, the Western blotting analysis showed that SIN inhibited the activation of TGF-β/Smad pathway in a dose-dependent manner in vitro and in vivo. These effects were significantly reversed by TGF-β/Smad activator SRI-011381 or TGF-β overexpression.

Discussion

The study indicates that SIN attenuates APAP-induced acute liver injury by decreasing oxidative stress and inflammatory response via TGF-β/Smad pathway in vitro and in vivo.

Keywords: Sinomenine, acetaminophen, inflammatory response, oxidative stress, TGF-β/Smad pathway, acute liver injury

References

1. Ju H-Y, Hu K-X, Zhao G-W, Tang Z-S, Song X. Design, preparation, and characterization of dioscin nanosuspensions and evaluation of their protective effect against carbon tetrachloride-induced acute liver injury in mice. Evid Based Complement Alternat Med. 2019;2019:3907915. doi:10.1155/2019/3907915 ] [
2. Liu H, Zhang Z, Hu H, et al. Protective effects of Liuweiwuling tablets on carbon tetrachloride-induced hepatic fibrosis in rats. BMC Complement Altern Med. 2018;18(1):212. doi:10.1186/s12906-018-2276-8 ] [
3. Leng J, Wang Z. NF-kappaB and AMPK/PI3K/Akt signaling pathways are involved in the protective effects of Platycodon grandiflorum saponins against acetaminophen-induced acute hepatotoxicity in mice. Phytother Res. 2018;32(11):2235–2246. [[PubMed]
4. Zhao H, Han Q, Lu N, Xu D, Tian Z, Zhang J. HMBOX1 in hepatocytes attenuates LPS/D-GalN-induced liver injury by inhibiting macrophage infiltration and activation. Mol Immunol. 2018;101:303–311. doi:10.1016/j.molimm.2018.07.021 [] [[PubMed]
5. Farzaei MH, Zobeiri M, Parvizi F, El-Senduny FF. Curcumin in liver diseases: a systematic review of the cellular mechanisms of oxidative stress and clinical perspective. Nutrients. 2018;10(7):855.
6. Kullak-Ublick GA, Andrade RJ, Merz M, et al. Drug-induced liver injury: recent advances in diagnosis and risk assessment. Gut. 2017;66(6):1154–1164. doi:10.1136/gutjnl-2016-313369 ] [
7. Al-Metwaly A, Shehatou G, Shebl A, Suddek G. Protective effects of trimetazidine against acetaminophen-induced liver injury in mice. J Pharm Sci Pharmacol. 2017;3:34–43. doi:10.1166/jpsp.2017.1074 [[PubMed]
8. Bai Q, Yan H, Sheng Y, et al. Long-term acetaminophen treatment induced liver fibrosis in mice and the involvement of Egr-1. Toxicology. 2017;382:47–58. doi:10.1016/j.tox.2017.03.008 [] [[PubMed]
9. Alonso EM, James LP, Zhang S, Squires RH. Acetaminophen adducts detected in serum of pediatric patients with acute liver failure. J Pediatr Gastroenterol Nutr. 2015;61(1):102–107. doi:10.1097/MPG.0000000000000814 ] [
10. Jaeschke H, McGill MR, Ramachandran A. Oxidant stress, mitochondria, and cell death mechanisms in drug-induced liver injury: lessons learned from acetaminophen hepatotoxicity. Drug Metab Rev. 2012;44(1):88–106. doi:10.3109/03602532.2011.602688 ] [
11. Jaeschke H, Knight TR, Bajt ML. The role of oxidant stress and reactive nitrogen species in acetaminophen hepatotoxicity. Toxicol Lett. 2003;144(3):279–288. doi:10.1016/S0378-4274(03)00239-X [] [[PubMed]
12. Lee HC, Yu HP, Liao CC, Chou AH, Liu FC. Escin protects against acetaminophen-induced liver injury in mice via attenuating inflammatory response and inhibiting ERK signaling pathway. Am J Transl Res. 2019;11(8):5170–5182.
13. Che J, Yang S, Qiao Z, et al. Schisandra chinensis acidic polysaccharide partially reverses acetaminophen-induced liver injury in mice. J Pharmacol Sci. 2019;140(3):248–254. doi:10.1016/j.jphs.2019.07.008 [] [[PubMed]
14. Ou Y, Su M, Ling Y, et al. Anti-allodynic effects of N-demethylsinomenine, an active metabolite of sinomenine, in a mouse model of postoperative pain. Eur J Pharmacol. 2018;823:105–109. doi:10.1016/j.ejphar.2018.01.044 [] [[PubMed]
15. Yue MF, Zhang XY, Dou YN, et al. Gut-sourced vasoactive intestinal polypeptide induced by the activation of α7 nicotinic acetylcholine receptor substantially contributes to the anti-inflammatory effect of sinomenine in collagen-induced arthritis. Front Pharmacol. 2018;9:675. doi:10.3389/fphar.2018.00675 ] [
16. Yao RB, Zhao ZM, Zhao LJ, Cai H. Sinomenine inhibits the inflammatory responses of human fibroblast-like synoviocytes via the TLR4/MyD88/NF-κB signaling pathway in rheumatoid arthritis. Die Pharmazie. 2017;72(6):355. [[PubMed]
17. Liu W, Zhang Y, Zhu W, et al. Sinomenine inhibits the progression of rheumatoid arthritis by regulating the secretion of inflammatory cytokines and monocyte/macrophage subsets. Front Immunol. 2018;9:2228.
18. Kondo Y, Takano F, Yoshida K, Hojo H. Protection by sinomenine against endotoxin-induced fulminant hepatitis in galactosamine-sensitized mice. Biochem Pharmacol. 1994;48(5):1050–1052. doi:10.1016/0006-2952(94)90378-6 [] [[PubMed]
19. Tian XP, Yin YY, Li X. Effects and mechanisms of Acremoniumterricola milleretal mycelium on liver fibrosis induced by carbon tetrachloride in rats. Am J Chin Med (Gard City N Y). 2011;39(3):537–550. doi:10.1142/S0192415X11009019 [] [[PubMed]
20. Yoshida K, Matsuzaki K. Differential regulation of TGF-beta/Smad signaling in hepatic stellate cells between acute and chronic liver injuries. Front Physiol. 2012;3:53. doi:10.3389/fphys.2012.00053 ] [
21. Gong S, Tian L, Zeng L, et al. Gut microbiota mediates diurnal variation of acetaminophen induced acute liver injury in mice. J Hepatol. 2018;69(1):S0168827818301466. doi:10.1016/j.jhep.2018.02.024 ] [
22. Bachmann M, Pfeilschifter J, Mühl H. A prominent role of interleukin-18 in acetaminophen-induced liver injury advocates its blockage for therapy of hepatic necroinflammation. Front Immunol. 2018;9:161. doi:10.3389/fimmu.2018.00161 ] [
23. Liu CF, Zhou WN, Lu Z, Wang XT, Qiu ZH. The associations between liver enzymes and the risk of metabolic syndrome in the elderly. Exp Gerontol. 2018;106:132–136. doi:10.1016/j.exger.2018.02.026 [] [[PubMed]
24. Kumar V, Gill KD. To Estimate the Activity of Alkaline Phosphatase in Serum; In: Basic Concepts in Clinical Biochemistry: A Practical Guide. Springer, Singapore. 2018. [PubMed]
25. Stojanović M, Todorović D, Šćepanović L, et al. Subchronic methionine load induces oxidative stress and provokes biochemical and histological changes in the rat liver tissue. Mol Cell Biochem. 2018;448(7):1–8. doi:10.1007/s11010-018-3311-2 [] [[PubMed]
26. Siegert M, Kranawetvogl A, Thiermann H, John H. Glutathione as an antidote for sulfur mustard poisoning: mass spectrometric investigations of its potency as a chemical scavenger. Toxicol Lett. 2017;293:S0378427417315230. [[PubMed]
27. He Y, Meng W, Wei Z, Jia J, Li Y, Wang H. The synthesis of the catalyst imitating SOD and its functions of scavenging oxygen free radicals. J Tongji Med Univ. 1992;21:261–264. [PubMed]
28. Arif M, Kitchen P, Conner MT, et al. Downregulation of aquaporin 3 inhibits cellular proliferation, migration and invasion in the MDA-MB-231 breast cancer cell line. Oncol Lett. 2018. doi:10.3892/ol.2018.8759 ] [
29. Lin Y, Miao LH, Pan WJ, et al. Effect of nitrite exposure on the antioxidant enzymes and glutathione system in the liver of bighead carp, Aristichthys nobilis. Fish Shellfish Immunol. 2018;76:S1050464818300779. doi:10.1016/j.fsi.2018.02.015 [] [[PubMed]
30. Gan F, Yang Y, Chen Y, Che C, Pan C, Huang K. Bush sophora root polysaccharide could help prevent aflatoxin B1-induced hepatotoxicity in the primary chicken hepatocytes. Toxicon. 2018;150:180–187. doi:10.1016/j.toxicon.2018.05.019 [] [[PubMed]
31. Zhu Q, Zhang Y, Liu Y, et al. MLIF alleviates SH-SY5Y neuroblastoma injury induced by oxygen-glucose deprivation by targeting eukaryotic translation elongation factor 1A2. PLoS One. 2016;11(2):e0149965. doi:10.1371/journal.pone.0149965 ] [
32. Chen ML, Lin K, Lin SK. NLRP3 inflammasome signaling as an early molecular response is negatively controlled by miR-186 in CFA-induced prosopalgia mice. Braz J Med Biol Res. 2018;51(9):e7602. doi:10.1590/1414-431x20187602 ] [
33. Schroder K, Tschopp J. The inflammasomes. Cell. 2010;140(6):821–832. doi:10.1016/j.cell.2010.01.040 [] [[PubMed]
34. Martinon F, Tschopp J. Inflammatory caspases and inflammasomes: master switches of inflammation. Cell Death Differ. 2007;14(1):10. doi:10.1038/sj.cdd.4402038 [] [[PubMed]
35. Eldridge Matthew JG, Sanchez-Garrido J, Hoben GF, Goddard PJ, Shenoy AR. The atypical ubiquitin E2 conjugase UBE2L3 is an indirect caspase-1 target and controls IL-1β secretion by inflammasomes. Cell Rep. 2017;18(5):1285. doi:10.1016/j.celrep.2017.01.015 ] [
36. Jia Y, Yao Z, Tianlin S, et al. Effect of Yinchen Shaogan decoction on NLRP3 inflammasome of endotoxin-induced acute liver injury’s mice. Chin Arch Trad Chin Med. 2015;11:2734–2737. [PubMed]
37. Park JK, Jeong DH, Park HY, et al. Hepatoprotective effect of Arazyme on CCl4-induced acute hepatic injury in SMP30 knock-out mice. Toxicology. 2008;246(2–3):132–142. doi:10.1016/j.tox.2008.01.006 [] [[PubMed]