Please wait a minute...
Journal of Integrative Agriculture  2023, Vol. 22 Issue (11): 3500-3516    DOI: 10.1016/j.jia.2023.04.010
Animal Science · Veterinary Medicine Advanced Online Publication | Current Issue | Archive | Adv Search |
miR-27b-5p regulates chicken liver disease via targeting IRS2 to suppress the PI3K/AKT signal pathway

ZHAO Jing1*, WU Ya-mei1*, ZHANG Yao1#, TANG Shu-yue1, HAN Shun-shun1, CUI Can1, TAN Bo2, YU Jie3, KANG Hou-yang4, CHEN Guang-deng5, MA Meng-gen5, ZHU Qing1#, YIN Hua-dong1#

1 Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu 611130, P.R.China
2 College of Forestry, Sichuan Agricultural University, Chengdu 611130, P.R.China 
3 Key Laboratory for Animal Disease Resistance Nutrition of China, Institute of Animal Nutrition, Sichuan Agricultural University, Chengdu 611130, P.R.China 
4 Triticeae Research Institute, Sichuan Agricultural University, Chengdu 611130, P.R.China
5 College of Resources, Sichuan Agricultural University, Chengdu 611130, P.R.China
Download:  PDF in ScienceDirect  
Export:  BibTeX | EndNote (RIS)      
摘要  

肝脏是鸡最重要的器官之一,具有储存肝糖原、合成蛋白质、解毒和脱氧等生理功能。肝脏的代谢和生长是一个复杂过程,受到环境、饮食和遗传等因素的调控。microRNAs (miRNAs) 作为转录后调控因子可参与多种生理过程。越来越多的证据表明,miR-27b-5p在不同物种的肝脏发育和代谢中发挥着至关重要的调控作用。但是miR-27b-5p在鸡肝脏中的潜在作用尚未被探索。在本次试验中,我们首先发现脂肪肝鸡血清中TGTC的含量显著高于正常鸡,而VLDL和血清激素(FSHLHE2)含量较正常组鸡显著降低。随后的研究发现miR-27b-5p在鸡脂肪肝中表达量较高,表明miR-27b-5p可能参与调节鸡肝脏的发育和代谢。接下来,我们探索了miR-27b-5p在鸡肝脏中的潜在功能,发现miR-27b-5p促进脂肪合成、增加细胞内氧化应激水平和炎症反应,并最终导致肝细胞发生凋亡。之后,我们又构建了miR-27b-5p与其靶基因间的调控机制,发现miR-27b-5p通过吸附IRS2来调控PI3K/AKT信号通路。此外,我们又探索了IRS2在鸡肝脏细胞中的效应,结果表明干扰IRS2与外源性过表达miR-27b-5p具有相同的作用。最终,我们的研究揭示miR-27b-5p通过结合IRS2来抑制PI3K/AKT信号通路,从而导致鸡肝脏脂肪变性、氧化应激,炎症反应和细胞凋亡。



Abstract  

The liver is a vital organ in chickens that performs a number of crucial physiological functions, including the storage of hepatic glycogen, protein synthesis, detoxification, and deoxidation.  The growth and metabolism of the liver are complex processes influenced by factors such as environment, diet, and genetics.  MicroRNAs (miRNAs), as post-transcriptional regulatory molecules, play a role in various biological processes.  There is growing evidence that miR-27b-5p plays a key role in the regulation of liver development and metabolism in various species.  However, its role in chicken livers has yet to be determined.  In our experiment, we found that chickens with fatty livers had significantly higher levels of serum triglyceride (TG) and total cholesterol (TC) compared to the normal chickens, while the control group had significantly higher levels of very low-density lipoprotein (VLDL) and serum hormones.  Further research showed that the mRNA of miR-27b-5p was highly expressed in fatty livers.  By exploring the function of miR-27b-5p in chicken livers, we discovered that it promotes lipogenesis, oxidative stress, and inflammatory responses, leading to hepatocyte apoptosis.  Our study also established the mechanism by which miR-27b-5p interacts with its target gene, and found that miR-27b-5p targets insulin receptor substrate 2 (IRS2) and modulates the PI3K/AKT signaling pathway.  Additionally, our investigation of IRS2 in chicken hepatocytes revealed that knocking down IRS2 has the same effects as overexpressing miR-27b-5p.  In conclusion, our study revealed that miR-27b-5p directly binds to IRS2, inhibiting the PI3K/AKT signaling pathway and causing steatosis, oxidative stress, inflammation, and apoptosis in chicken liver.

Keywords:  miR-27b-5p        IRS2        lipogenesis        oxidative stress        inflammation        apoptosis  
Received: 28 December 2022   Accepted: 07 March 2023
Fund: This experiment was financially supported by the National Key Research and Development Program of China (2021YFD1300600), the Sichuan Science and Technology Program, China (2021YFYZ0007 and 2022YFYZ0005), and the China Agriculture Research System of MOF and MARA (CARS-40-K06).
About author:  ZHAO Jing, E-mail: zhaojing@stu.sicau.edu.cn; WU Ya-mei, E-mail: wuyamei@stu.sicau.edu.cn; ZHANG Yao, E-mail: zhangyao@sicau.edu.cn; #Correspondence YIN Hua-dong, E-mail: yinhuadong@sicau.edu.cn; ZHU Qing, E-mail: zhuqing@sicau.edu.cn * These authors contributed equally to this study.

Cite this article: 

ZHAO Jing, WU Ya-mei, ZHANG Yao, TANG Shu-yue, HAN Shun-shun, CUI Can, TAN Bo, YU Jie, KANG Hou-yang, CHEN Guang-deng, MA Meng-gen, ZHU Qing, YIN Hua-dong. 2023. miR-27b-5p regulates chicken liver disease via targeting IRS2 to suppress the PI3K/AKT signal pathway. Journal of Integrative Agriculture, 22(11): 3500-3516.

Ayala I, Castillo A M, Adánez G, Fernández-Rufete A, Pérez B G, Castells M T. 2009. Hyperlipidemic chicken as a model of non-alcoholic steatohepatitis. Experimental Biology and Medicine234, 10–16.

Bannister D W, Evans A J, Whitehead C C. 1975. Evidence for a lesion in carbohydrate metabolism in fatty liver and kidney syndrome in chicks. Research in Veterinary Science18, 149–156.

Bao S, Wu Y L, Wang X, Han S, Cho S, Ao W, Nan J X. 2020. Agriophyllum oligosaccharides ameliorate hepatic injury in type 2 diabetic db/db mice targeting INS-R/IRS-2/PI3K/AKT/PPAR-γ/Glut4 signal pathway. Journal of Ethnopharmacology257, 112863.

Benito-Vicente A, Uribe K B, Rotllan N, Ramírez C M, Jebari-Benslaiman S, Goedeke L, Canfrán-Duque A, Galicia-García U, Saenz De Urturi D, Aspichueta P, Suárez Y, Fernández-Hernando C, Martín C. 2020. miR-27b Modulates insulin signaling in hepatocytes by regulating insulin receptor expression. International Journal of Molecular Sciences21, 8675.

Bensadoun A, Rothfeld A. 1972. The form of absorption of lipids in the chicken, Gallus domesticus. In: Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine, New York. pp. 814–817.

Cai Y, Song Z, Zhang X, Wang X, Jiao H, Lin H. 2009. Increased de novo lipogenesis in liver contributes to the augmented fat deposition in dexamethasone exposed broiler chickens (Gallus gallus domesticus). Comparative Biochemistry and Physiology (C: Toxicology & Pharmacology), 150, 164–169.

Castro R E, Ferreira D M, Afonso M B, Borralho P M, Machado M V, Cortez-Pinto H, Rodrigues C M. 2013. MiR-34a/SIRT1/p53 is suppressed by ursodeoxycholic acid in the rat liver and activated by disease severity in human non-alcoholic fatty liver disease. Journal of Hepatology58, 119–125.

Cherian G, Goeger M P. 2004. Hepatic lipid characteristics and histopathology of laying hens fed CLA or n-3 fatty acids. Lipids39, 31–36.

Choudhari S K, Chaudhary M, Gadbail A R, Sharma A, Tekade S. 2014. Oxidative and antioxidative mechanisms in oral cancer and precancer: A review. Oral Oncology50, 10–18.

Clarke S D, Jump D. 2018. Regulation of hepatic gene expression by dietary fats: A unique role for polyunsaturated fatty acids. In: Nutrition and Gene Expression. Taylor & Francis Group Country, London. pp. 227–246.

Day C P, James O F. 1998. Steatohepatitis: a tale of two “hits”? Gastroenterology114, 842–845.

Eckstein S S, Weigert C, Lehmann R. 2017. Divergent roles of IRS (Insulin Receptor Substrate) 1 and 2 in liver and skeletal muscle. Current Medicinal Chemistry24, 1827–1852.

Feng Z, Li Z, Zhu D, Ling W, Zheng L, Pu L, Kong L. 2017. Mir-24 regulates hepatocyte apoptosis via BIM during acute liver failure. American Journal of Translational Research9, 4925–4935.

Ferreira D M, Afonso M B, Rodrigues P M, Simão A L, Pereira D M, Borralho P M, Rodrigues C M, Castro R E. 2014. C-Jun N-terminal kinase 1/c-Jun activation of the p53/microRNA 34a/sirtuin 1 pathway contributes to apoptosis induced by deoxycholic acid in rat liver. Molecular and Cellular Biology34, 1100–1120.

Fraslin J M, Touquette L, Douaire M, Menezo Y, Guillemot J C, Mallard J. 1992. Isolation and long-term maintenance of differentiated adult chicken hepatocytes in primary culture. In Vitro Cellular & Developmental Biology28a, 615–620.

Giordano S, Columbano A. 2013. MicroRNAs: new tools for diagnosis, prognosis, and therapy in hepatocellular carcinoma? Hepatology57, 840–847.

He Y, Hwang S, Cai Y, Kim S J, Xu M, Yang D, Guillot A, Feng D, Seo W, Hou X, Gao B. 2019. MicroRNA-223 Ameliorates nonalcoholic steatohepatitis and cancer by targeting multiple inflammatory and oncogenic genes in hepatocytes. Hepatology70, 1150–1167.

Ke F, Ren C, Zhai Z, Gao X, Wei J, Zhu Y, Zhi Y. 2022. LINC01234 regulates microRNA-27b-5p to induce the migration, invasion and self-renewal of ovarian cancer stem cells through targeting SIRT5. Cell Cycle21, 1020–1033.

Kubota N, Kubota T, Itoh S, Kumagai H, Kozono H, Takamoto I, Mineyama T, Ogata H, Tokuyama K, Ohsugi M, Sasako T, Moroi M, Sugi K, Kakuta S, Iwakura Y, Noda T, Ohnishi S, Nagai R, Tobe K, Terauchi Y, Ueki K, Kadowaki T. 2008. Dynamic functional relay between insulin receptor substrate 1 and 2 in hepatic insulin signaling during fasting and feeding. Cell Metabolism8, 49–64.

Liu T, Yang T, Xu Z, Tan S, Pan T, Wan N, Li S. 2018. MicroRNA-193b-3p regulates hepatocyte apoptosis in selenium-deficient broilers by targeting MAML1. Journal of Inorganic Biochemistry186, 235–245.

Nassir F, Adewole O L, Brunt E M, Abumrad N A. 2013. CD36 deletion reduces VLDL secretion, modulates liver prostaglandins, and exacerbates hepatic steatosis in ob/ob mice. Journal of Lipid Research54, 2988–2997.

Pierantonelli I, Svegliati-Baroni G. 2019. Nonalcoholic fatty liver disease: Basic pathogenetic mechanisms in the progression from NAFLD to NASH. Transplantation103, e1–e13.

Shan W, Gao L, Zeng W, Hu Y, Wang G, Li M, Zhou J, Ma X, Tian X, Yao J. 2015. Activation of the SIRT1/p66shc antiapoptosis pathway via carnosic acid-induced inhibition of miR-34a protects rats against nonalcoholic fatty liver disease. Cell Death & Disease6, e1833.

Shini A, Shini S, Bryden W L. 2019. Fatty liver haemorrhagic syndrome occurrence in laying hens: impact of production system. Avian Pathology48, 25–34.

Singaravelu R, Chen R, Lyn R K, Jones D M, O’Hara S, Rouleau Y, Cheng J, Srinivasan P, Nasheri N, Russell R S, Tyrrell D L, Pezacki J P. 2014. Hepatitis C virus induced up-regulation of microRNA-27: A novel mechanism for hepatic steatosis. Hepatology59, 98–108.

Tachibana S, Sato K, Takahashi T, Akiba Y. 2002. Octanoate inhibits very low-density lipoprotein secretion in primary cultures of chicken hepatocytes. Comparative Biochemistry and Physiology132, 621–627.

Teimouri M, Hosseini H, Shabani M, Koushki M, Noorbakhsh F, Meshkani R. 2020. Inhibiting miR-27a and miR-142-5p attenuate nonalcoholic fatty liver disease by regulating Nrf2 signaling pathway. IUBMB Life72, 361–372.

Thirone A C, Huang C, Klip A. 2006. Tissue-specific roles of IRS proteins in insulin signaling and glucose transport. Trends in Endocrinology and Metabolism17, 72–78.

Tian W H, Wang Z, Yue Y X, Li H, Li Z J, Han R L, Tian Y D, Kang X T, Liu X J. 2019. MiR-34a-5p increases hepatic triglycerides and total cholesterol levels by regulating ACSL1 protein expression in laying hens. International Journal of Molecular Sciences20, 4420.

Trott K A, Giannitti F, Rimoldi G, Hill A, Woods L, Barr B, Anderson M, Mete A. 2014. Fatty liver hemorrhagic syndrome in the backyard chicken: A retrospective histopathologic case series. Veterinary Pathology51, 787–795.

Verma S, Mishra R, Malik A, Chaudhary P, Malhotra S S, Panda A K, Gupta S K. 2021. MiR-27b-5p inhibits BeWo cells fusion by regulating WNT2B and enzyme involved in progesterone synthesis. American Journal of Reproductive Immunology86, e13409.

Vickers K C, Shoucri B M, Levin M G, Wu H, Pearson D S, Osei-Hwedieh D, Collins F S, Remaley A T, Sethupathy P. 2013. MicroRNA-27b is a regulatory hub in lipid metabolism and is altered in dyslipidemia. Hepatology57, 533–542.

Videla L A, Rodrigo R, Orellana M, Fernandez V, Tapia G, Quiñones L, Varela N, Contreras J, Lazarte R, Csendes A, Rojas J, Maluenda F, Burdiles P, Diaz J C, Smok G, Thielemann L, Poniachik J. 2004. Oxidative stress-related parameters in the liver of non-alcoholic fatty liver disease patients. Clinical Science106, 261–268.

Wang X, He Y, Mackowiak B, Gao B. 2021. MicroRNAs as regulators, biomarkers and therapeutic targets in liver diseases. Gut70, 784–795.

Xu H, Zhou Y, Liu Y, Ping J, Shou Q, Chen F, Ruo R. 2016. Metformin improves hepatic IRS2/PI3K/Akt signaling in insulin-resistant rats of NASH and cirrhosis. The Journal of Endocrinology229, 133–144.

Xu Y, Li Z, Zhang S, Zhang H, Teng X. 2020. MiR-187-5p/apaf-1 axis was involved in oxidative stress-mediated apoptosis caused by ammonia via mitochondrial pathway in chicken livers. Toxicology and Applied Pharmacology388, 114869.

Yang P, Liang Y, Luo Y, Li Z, Wen Y, Shen J, Li R, Zheng H, Gu H F, Xia N. 2019. Liraglutide ameliorates nonalcoholic fatty liver disease in diabetic mice via the IRS2/PI3K/Akt signaling pathway. DiabetesMetabolic Syndrome and Obesity (Targets and Therapy), 12, 1013–1021.

Yilmaz Y, Younossi Z M. 2014. Obesity-associated nonalcoholic fatty liver disease. Clinics in Liver Disease18, 19–31.

You X, Liu F, Zhang T, Li Y, Ye L, Zhang X. 2013. Hepatitis B virus X protein upregulates oncogene Rab18 to result in the dysregulation of lipogenesis and proliferation of hepatoma cells. Carcinogenesis34, 1644–1652.

Zhang Y, Liu Z, Liu R, Wang J, Zheng M, Li Q, Cui H, Zhao G, Wen J. 2018. Alteration of hepatic gene expression along with the inherited phenotype of acquired fatty liver in chicken. Genes9, 199.

Zhang Y, Wang C, Lu J, Jin Y, Xu C, Meng Q, Liu Q, Dong D, Ma X, Liu K, Sun H. 2020. Targeting of miR-96-5p by catalpol ameliorates oxidative stress and hepatic steatosis in LDLr–/– mice via p66shc/cytochrome C cascade. Aging12, 2049–2069.

Zhao J, Zhao X, Shen X, Zhang Y, Zhang Y, Ye L, Li D, Zhu Q, Yin H. 2022. CircCCDC91 regulates chicken skeletal muscle development by sponging miR-15 family via activating IGF1-PI3K/AKT signaling pathway. Poultry Science101, 101803.

[1] Saif ULLAH, Sheeraz MUSTAFA, Wael ENNAB, Muhammad JAN, Muhammad SHAFIQ, Ngekure M. X. KAVITA, Lü Zeng-peng, MAO Da-gan, SHI Fang-xiong. A protective role of resveratrol against the effects of immobilization stress in corpora lutea of mice in early pregnancy[J]. >Journal of Integrative Agriculture, 2020, 19(7): 1857-1866.
[2] LI Fu-hua, ZHENG Shao-jie, ZHAO Ji-chun, LIAO Xia, WU Su-rui, MING Jian. Phenolic extract of Morchella angusticeps peck inhibited the proliferation of HepG2 cells in vitro by inducing the signal transduction pathway of p38/MAPK[J]. >Journal of Integrative Agriculture, 2020, 19(11): 2829-2838.
[3] WANG Jun, SHI xin-jin, SUN Hai-wei, CHEN Hong-jun. Insights into African swine fever virus immunoevasion strategies[J]. >Journal of Integrative Agriculture, 2020, 19(1): 11-22.
[4] RAN Mao-liang, WENG Bo, CAO Rong, PENG Fu-zhi, LUO Hui, GAO Hu, CHEN Bin. miR-34c inhibits proliferation and enhances apoptosis in immature porcine Sertoli cells by targeting the SMAD7 gene[J]. >Journal of Integrative Agriculture, 2019, 18(2): 449-459.
[5] ZHANG Feng, GAO Peng, GE Xin-na, ZHOU Lei, GUO Xin, YANG Han-chun. Critical role of cytochrome c1 and its cleavage in porcine reproductive and respiratory syndrome virus nonstructural protein 4-induced cell apoptosis via interaction with nsp4[J]. >Journal of Integrative Agriculture, 2017, 16(11): 2573-2585.
[6] HU Ting-xi, ZHU Hua-bin, SUN Wei-jun, HAO Hai-sheng, ZHAO Xue-ming, DU Wei-hua, WANG Zong-li. Sperm pretreatment with glutathione improves IVF embryos development through increasing the viability and antioxidative capacity of sex-sorted and unsorted bull semen[J]. >Journal of Integrative Agriculture, 2016, 15(10): 2326-2335.
No Suggested Reading articles found!