Effect of sodium glycosaminodeoxycholate on apoptosis of hepatic stem cells in cholestatic microenvironment

doi:10.16098/j.issn.0529-1356.2019.06.008

Abstract

Abstract:

Objective To investigate the effects of hepatic stem cells survival in the pathological microenvironment of cholestatic cirrhosis, and the effects of sodium glycosaminodeoxycholate (GCDC) on apoptosis of hepatic stem cells in vitro. Methods Balb/c mice were subjected to bile duct ligation (BDL)to simulate the pathological environment of cholestatic cirrhosis；Liver stem cells HP14-19 were transplanted back into liver by the splenic vein and survival of cell colonization was detected；Effects of sodium glycosaminodeoxycholate on the viability of HP14-19 cells at different concentrations by cell counting kit-8(CCK-8) and crystal violet staining, flow cytometry was used to detect the apoptosis of HP14-19 cells treated with 600 μmol/L GCDC 24 hours later, and the expression of Bax，Bcl-2，Capase-3，phosphorylated adenosine 5’-monophosphate-activated protein kinase α(p-AMPKα) and adenosine 5’-monophosphate-activated protein kinase α(AMPKα) were detected by Western blotting. Results The results of CCK-8 and crystal violet staining showed that the proliferation of HP14-19 cells was inhibited with the increase of the concentration of sodium glycosaminodeoxycholate. Flow cytometry showed that the apoptosis rate of GCDC treated group was higher than that of control group. Compared with the control group, the expression of Bax, Capase-3 was up-regulated and the expression of Bcl-2 protein was down-regulated in the experimental group. The results showed that GCDC could induce apoptosis of HP14-19 (P<0.05) AMPK was activated. Conclusion Microenvironment of cholestatic cirrhosis induced apoptosis of liver stem cells.

Key words: Cholestatic cirrhosis, Sodium glycosaminodeoxycholate, Liver stem cell, Western blotting, Mouse

LI Zhi-peng WANG Jian XIAO Cheng KANG Quan LUO Qing. Effect of sodium glycosaminodeoxycholate on apoptosis of hepatic stem cells in cholestatic microenvironment[J]. Acta Anatomica Sinica, 2019, 50(6): 747-753.

References

［1］ Xiang B，Xie XL. Sequential treatment of “Kasai surgery-liver transplantation” for biliary atresia ［J］.Journal of Clinical Pediatric Surgery, 2018 17(11): 805-808. (in Chinese)

向波, 谢小龙. 胆道闭锁的“Kasai手术-肝移植”序贯治疗［J］. 临床小儿外科杂志, 2018,17(11):805-808.

［2］ LeeVan E, Matsuoka L, Cao S, et al. Biliary-enteric drainage vs primary liver transplant as initial treatment for children with biliary atresia［J］. JAMA Surg, 2019,154(1):26-32.

［3］ Zhang YJ, Li YY, Hao XN,et al.Vein transplantation of human umbilical cord-derived mesenchymal stem cells for treatment of hepatic fibrosis in rats［J］.Chinese Journal of Tissue Engineering Research, 2014, 18(28): 4485-4490. (in Chinese)

张英杰, 李玉云, 郝晓娜, 等. 人脐带源间充质干细胞静脉移植治疗大鼠肝纤维化［J］. 中国组织工程研究, 2014,18(28):4485-4490.

［4］ Guo CC, Lan L, Liu R,et al. Co-transplantation of endothelial progenitor cells and hepatocyte stem cells launches a counterattack against liver fibrosis in rats ［J］.Chinese Journal of Tissue Engineering Research, 2018, 22(5): 704-709. (in Chinese)

郭灿灿, 兰玲, 刘冉, 等. 内皮祖细胞和肝干细胞联合移植对大鼠肝纤维化的逆转作用［J］. 中国组织工程研究, 2018,22(05):704-709.

［5］ Bria A, Marda J, Zhou J, et al. Hepatic progenitor cell activation in liver repair［J］. Liver Res, 2017,1(2):81-87.

［6］ Zhao MX，Li BW，Wang DSh, et al. Role of miR-122 in bone marrow derived stem cell therapy for acute liver injury in rats［J］.Journal of Jilin University(Medicine Edition), 2015,41(6):1150-1153. (in Chinese)

赵民学, 李柏文, 王德盛, 等. miR-122在骨髓间充质干细胞治疗大鼠急性肝损伤中的作用［J］. 吉林大学学报（医学版）, 2015,41(6):1150-1153.

［7］ Bi Y, He Y, Huang J, et al. Functional characteristics of reversibly immortalized hepatic progenitor cells derived from mouse embryonic liver［J］. Cell Physiol Biochem, 2014,34(4):1318-1338.

［8］ éboli L, Tannuri AC, Gibelli N, et al. Comparison of the results of living donor liver transplantation due to acute liver failure and biliary atresia in a quaternary center［J］. Transplant Proc, 2017,49(4):832-835.

［9］ Namdaroglu S, Kaya AH, Batgi H, et al. Impacts of post-transplantation cyclophosphamide treatment after allogeneic hematopoietic stem cell transplantation in acute myeloid leukemia［J］. Sci Rep, 2019,9(1):2046.

［10］ Warren CFA, Wong-Brown MW, Bowden NA. BCL-2 family isoforms in apoptosis and cancer［J］. Cell Death Dis, 2019,10(3):177.

［11］ Tan KO, Fu NY, Sukumaran SK, et al. MAP-1 is a mitochondrial effector of Bax［J］. Proc Natl Acad Sci USA, 2005,102(41):14623-14628.

［12］ Shi Y. Mechanisms of caspase activation and inhibition during apoptosis［J］. Mol Cell, 2002,9(3):459-470.

［13］ Zhang JF,Zhao ZhH,Hao JH,et al.Effects of microＲNA-25 on cerebral ischemia/reperfusion injury-induced cell apoptosis［J］. Acta Anatomica Sinica, 2018,49(5):584-590. (in Chinese)

张军峰, 赵朝华, 郝佳晖, 等. MicroRNA-25在脑缺血/再灌注损伤诱导的细胞凋亡过程中的作用［J］. 解剖学报, 2018,49(5):584-590.

［14］ Xiao J，Yin SM，Xie ShF,et al.Quercetin modulate AMPK activity and induce apoptosis and autophagy in HL-60 cells［J］.Journal of Sun Yat-Sen University（Medical Sciences）, 2018,39(4):501-509. (in Chinese)

肖洁, 尹松梅, 谢双锋, 等. 槲皮素调控AMPK活性诱导HL-60细胞自噬与凋亡［J］. 中山大学学报（医学科学版）, 2018,39(4):501-509.

［15］ Zhu Y, Wang Q, Tang X, et al. Mesenchymal stem cells enhance autophagy of human intrahepatic biliary epithelial cells in vitro［J］. Cell Biochem Funct, 2018,36(5):280-287.

［16］ Gao L, Lv G, Guo X, et al. Activation of autophagy protects against cholestasis-induced hepatic injury［J］. Cell Biosci, 2014,4:47.

［17］ Yang B, Luo Q, Kang Q,et al. Tumor necrosis factor-α and transforming growth factor-β1 balance liver stem cell differentiation in cholestatic cirrhosis［J］ Journal of Southern Medical University, 2018, 38(4): 375-383. (in Chinese)

杨博, 罗庆, 康权, 等. TNF-α、TGF-β-1在胆汁淤积性肝硬化中平衡调控肝脏干细胞分化［J］. 南方医科大学学报, 2018,38(4):375-383.

［18］ Sun X, Zhang W. Silencing of Gal-7 inhibits TGF-β1-induced apoptosis of human airway epithelial cells through JNK signaling pathway［J］. Exp Cell Res, 2019,375(2):100-1010.

［19］ Park S, In Hwang S, Kim J, et al. The therapeutic potential of induced hepatocyte-like cells generated by direct reprogramming on hepatic fibrosis［J］. Stem Cell Res Ther, 2019,10(1):21.

［20］ Hu C, Zhao L, Duan J, et al. Strategies to improve the efficiency of mesenchymal stem cell transplantation for reversal of liver fibrosis［J］. J Cell Mol Med, 2019,23(3):1657-1670.

［21］ Hohenester S, Vennegeerts T, Wagner M, et al. Physiological hypoxia prevents bile salt-induced apoptosis in human and rat hepatocytes［J］. Liver Int, 2014,34(8):1224-1231.

[1]	HONG Xiao-min LI San-qiang CUI Qin-yi ZHENG Run-yue YANG Meng-li LUO Ren-li LI Qian-hui. Nuclear factor-κB signaling pathway and gender differences in alcoholic liver fibrosis [J]. Acta Anatomica Sinica, 2024, 55(1): 55-61.
[2]	WEI Xi-xi LI Chao-hong ZHAO Chen-lu TANG Jia-ping LIU Yu-zhen. Characterization of group Ⅰ metabotropic glutamate receptors in rat superior cervical ganglion and their changes following chronic intermittent hypoxia [J]. Acta Anatomica Sinica, 2024, 55(1): 3-9.
[3]	YUAN Lei YANG Zhi-wei YANG Hui LIU Zheng-hai HE Jie WAN Wei. Panax notoginseng saponin relieving the inflammatory pain caused by complete Freund’s adjuvant by inhibiting the activation of astrocytes in mice [J]. Acta Anatomica Sinica, 2024, 55(1): 25-31.
[4]	MA Ting-ting CHEN Jian-hong LIU Ai-cui LI Hai-ning. Role of inhibiting lncRNA TUG1 to down-regulate nucleotide binding oligomerization domain like receptor protein 1 inflammasome in delaying the progression of Alzheimer’s disease [J]. Acta Anatomica Sinica, 2024, 55(1): 32-42.
[5]	ZOU Cheng-gong FENG Hao CHEN Bing TANG Hui SHAO Chuan SUN Mou YANG Rong HE Jia-quan . Research on the dynamic changes of neurological dysfunction and cognitive function impairment in traumatic brain injury [J]. Acta Anatomica Sinica, 2024, 55(1): 43-48.
[6]	LÜ Hai-yan SHEN Xi-ya ZHAO Fu-sheng ZHU Mei . Effects of tricholoma matsutake polysaccharides on 1-methy-4-pehnyl-pyridine ion-induced PC12 cell damage [J]. Acta Anatomica Sinica, 2024, 55(1): 49-54.
[7]	ZHENG Li-ping LIU Xia DU Xiao-hua WU Ya-juan LIU Shan-shan . Expression and distribution of brain-derived neurotrophic factor in different cerebrum regions of yak and cattle [J]. Acta Anatomica Sinica, 2024, 55(1): 10-16.
[8]	HAO Yong-ming GUO Lei ZHOU Cheng-hui PEI Han-jun LI Li ZHAO Zhe . Establishment of a rat model of myocardial hypertrophy by a modified abdominal aortic coarctation method [J]. Acta Anatomica Sinica, 2024, 55(1): 120-124.
[9]	XU Ya-ping YU Yue-xin CHEN Jin-fu WANG Ke-ke GUO Zhikun . Structural distribution and mechanism of elastic fibers and collagen fibers in ventricular interstitium of aged rats [J]. Acta Anatomica Sinica, 2023, 54(6): 716-721.
[10]	WANG Wen-juan DING Yu-tong SU Hao REN Jie SUN Yan-rong WANG Han-fei LU Jia-li ZHANG Lin-qian BAI Yu QIN Li-hua. Changes of Nogo-A expression in hippocampus and striatum of rats under low estrogen condition [J]. Acta Anatomica Sinica, 2023, 54(6): 620-627.
[11]	ZHANG Qin YANG Jun-xia JIANG Qing-song. Effect of fosinopril on angiotensin converting enzyme 2 and vascular endothelial growth factor in diabetic retinopathy mice [J]. Acta Anatomica Sinica, 2023, 54(6): 628-634.
[12]	CHEN Zi-xuan SI Li-na SHU Wei-han ZHANG Xin WEI Chen-yang CHENG Lu-yang YANG Song-he QIAO Yue-bing. Mechanism of melatonin regulation of PI3K/Akt/mTOR signaling in hypothalamus delaying the initiation of puberty in female mice mice [J]. Acta Anatomica Sinica, 2023, 54(6): 644-651.
[13]	LIANG Pan-pan YU Ai-mei DU Jing KOU Wen-hui WANG Huan-huan SONG Ai-xia. Inhibitory effect of sodium ferulate on inflammatory response in migraine rats based on c-Jun N-terminal kinase/p38 mitogen-activated protein kinase signaling pathway [J]. Acta Anatomica Sinica, 2023, 54(6): 652-659.
[14]	XU Ying LIU Bin ZHENG Xing HE Zong-zhao. Role and mechanism of complement C3a receptor in the cognitive impairment of septic rats induced by cecal ligation and perforation [J]. Acta Anatomica Sinica, 2023, 54(6): 668-675.
[15]	LIU Zhe-chuan LI Kun MA Shuai-nan MENG Jia-qi WANG Yan-qin. Effects of liraglutide on inflammation and mitochondrial fusion/division in Parkinson’s disease model of mice induced by paraquat [J]. Acta Anatomica Sinica, 2023, 54(6): 676-681.