脾切除对原发性胆汁性肝硬化小鼠肝组织纤维化影响研究
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摘要
目的探讨脾切除对原发性胆汁性肝硬化(PBC)小鼠的影响及其相关因素。方法 40只雌性C57BL/6小鼠被分为脾切除组(n=10)、假手术组(n=10)、模型组(n=10)和正常对照组(n=10)。在前三组,采用1%聚肌胞苷酸腹腔注射建立PBC模型,在正常对照组,给予等体积生理盐水腹腔注射。在实验20 w末,对动物分别行脾切除术,或在假手术组采取腹壁切开后仅轻微翻动脾脏,或在模型组和正常对照组不行任何处理。在实验32w,行血生化和肝组织病理学检查。采用RT-PCR法检测肝组织转化生长因子-β1(TGF-β1)及其Ⅰ型受体(TβRI)和Ⅱ型受体(TβRII)mRNA水平。取末端回肠组织,采用ELISA法检测其匀浆TNF-α水平。结果在32w时,脾切除组、假手术组和模型组动物肝指数均显著低于对照组[(41.5±5.2)、(39.6±7.5)、(39.1±1.0)对(53.2±2.1),P值均<0.05];脾切除组血清ALT、AST和ALP水平分别为(43.5±6.4) U/L、(157.7±20.9) U/L和(178.1±38.0)U/L,显著低于模型组[(52.0±9.0) U/L、(183.4±12.4) U/L和(195.3±62.6) U/L,P值均<0.05];脾切除组肝组织纤维化程度明显轻于假手术组和模型组;脾切除组肝组织TGF-β1、TβRI和TβRII mRNA水平分别为(0.5±0.09)、(0.5±0.05)和(0.5±0.09),显著低于模型组[(1.0±0.10)、(1.0±0.08)和(1.0±0.09),P值均<0.05];脾切除组回肠末端匀浆TNF-α水平为(50.0±8.6) ng/L,显著低于模型组[(106.8±19.8) ng/L,P<0.05]。结论脾切除可缓解PBC小鼠肝组织纤维化进程,可能与抑制肝脏TGF-β1表达和抑制回肠末端TNF-α分泌有关。
Objective To investigate the effects of splenectomy on hepatic fibrosis in mice with poly I:Cinduced primary biliary cholangitis(PBC). Methods Forty C57 BL/6 female mice were randomly divided into the splenectomy group(n =10),sham-operation group(n=10),model control group(n=10) and normal control group(n=10). Mice in the first three groups were injected with poly I:C to establish PBC models,while animals in the control were injected with equal volume of saline. At the end 20 weeks,splenectomy was performed in ten mice,and sham operations were performed in another ten mice. Liver index, liver function index and liver histology were performed at each group at the end of 32 w. The mRNA levels of TGF-β1,TβRI and TβRII in liver tissues were assayed by RT-PCR,and TNF-α levels in terminal ileum were detected by ELISA. Results At the end of 32 weeks,the liver index were(41.5±5.2),(39.6±7.5),(39.1±1.0) and(53.2±2.1),in splenectomy,sham-operation,model and control group,respectively(P<0.05);serum ALT,AST and ALP levels in splenectomy group were(43.5±6.4)U/L,(157.7±20.9) U/L and(178.1±38.0) U/L,significantly lower than those in model group [(52.0±9.0) U/L,(183.4±12.4) U/L and(195.3±62.6) U/L,P<0.05 for all];liver fibrosis staging in splenectomy group were significantly milder than those in sham-operation and model control group;hepatic TGF-β1,T(3 RI,TβRII mRNA levels were(0.5±0.09),(0.5±0.05) and(0.5±0.09) in splenectomy group,significantly lower than those in model group [(1.010.10),(1.0±0.08) and(1.0±0.09), P<0.05];the TNF-α level in terminal ileum homogenate was(50.0±8.6)ng/L in splenectomy group,obviously lower than [(106.8±19.8) ng/L,P<0.05] in model group. Conclusion Splenectomy could relieve the progression of liver fibrosis in PBC mice,which might be related to the inhibition of hepatic TGF-β1 and ileal TNF-α expression.
Objective To investigate the effects of splenectomy on hepatic fibrosis in mice with poly I:Cinduced primary biliary cholangitis(PBC). Methods Forty C57 BL/6 female mice were randomly divided into the splenectomy group(n =10),sham-operation group(n=10),model control group(n=10) and normal control group(n=10). Mice in the first three groups were injected with poly I:C to establish PBC models,while animals in the control were injected with equal volume of saline. At the end 20 weeks,splenectomy was performed in ten mice,and sham operations were performed in another ten mice. Liver index, liver function index and liver histology were performed at each group at the end of 32 w. The mRNA levels of TGF-β1,TβRI and TβRII in liver tissues were assayed by RT-PCR,and TNF-α levels in terminal ileum were detected by ELISA. Results At the end of 32 weeks,the liver index were(41.5±5.2),(39.6±7.5),(39.1±1.0) and(53.2±2.1),in splenectomy,sham-operation,model and control group,respectively(P<0.05);serum ALT,AST and ALP levels in splenectomy group were(43.5±6.4)U/L,(157.7±20.9) U/L and(178.1±38.0) U/L,significantly lower than those in model group [(52.0±9.0) U/L,(183.4±12.4) U/L and(195.3±62.6) U/L,P<0.05 for all];liver fibrosis staging in splenectomy group were significantly milder than those in sham-operation and model control group;hepatic TGF-β1,T(3 RI,TβRII mRNA levels were(0.5±0.09),(0.5±0.05) and(0.5±0.09) in splenectomy group,significantly lower than those in model group [(1.010.10),(1.0±0.08) and(1.0±0.09), P<0.05];the TNF-α level in terminal ileum homogenate was(50.0±8.6)ng/L in splenectomy group,obviously lower than [(106.8±19.8) ng/L,P<0.05] in model group. Conclusion Splenectomy could relieve the progression of liver fibrosis in PBC mice,which might be related to the inhibition of hepatic TGF-β1 and ileal TNF-α expression.
引文
[1]中华医学会肝病学分会,中华医学会消化病学分会,中华医学会感染病学分会等.胆汁淤积性肝病诊断和治疗共识(2015).实用肝脏病杂志,2016,19(6):771-781.
[2]程询,杨长青.检测门脉高压程度评估肝硬化严重程度研究进展.实用肝脏病杂志,2017,20(1):10-12.
[3]刘雪莲,杨见权.脾功能亢进症治疗进展.实用肝脏病杂志,2013,16(4):382-384.
[4] Yao YM,Liu QG,Yang W,et al. Effect of spleen on immune function of rats with liver cancer complicated by liver cirrhosis.Hepatobiliary Pancreat Dis Int,2003,2(2):242-246.
[5]郭晓霞,李良学,武玉鹏,等.聚肌胞苷酸诱导的早期原发性胆汁性肝硬化小鼠模型的建立.实用肝脏病杂志,2014,17(6):619-622.
[6] Irma M,Markku N. Human leukocyte antigen associations in Finnish liver transplantations due to primary sclerosing cholangitis and primary biliary cirrhosis. Open Med,2007,2(1):12-25.
[7] Chuang N, Gross RG, Odin JA. Update on the epidemiology of primary biliarycirrhosis.Expert Rev Gastroenterol Hepatol,2011,5(5):583-590.
[8] Liu H, Liu Y, Wang L, et al. Prevalence of primary biliary cirrhosis in adults referring hospital for annual health check-up in Southern China. BMC Gastroenterol, 2010,10(100):2-5.
[9] Chan CW,Carpenter JR,Rigamonti C,et al.Survival following the development of ascites and/or peripheral oedema in primary biliary cirrhosis:a staged prognostic model. Scand J Gastroenterol,2005,40(9):1081-1089.
[10]何方平,陈兰.原发性胆汁性肝硬化的流行病学及自然病程研究进展.实用肝脏病杂志,2013,16(2):97-98.
[11] Cartin-Ceba R,Krowda MJ.Portopulmonary hypertension.Clin Liver Dis,2014,18(2):421-438.
[12] Fed erica L, Caterina M. The diagnostic accuracy of biomarkers for diagnosis of primary biliary cholangitis(PBC)in anti-mitochondrial antibody(AM A)-negative PBC patients:a review of literature. Clin Chem Laborat Med,2017,56(1):25-31.
[13] Morinaga A,Ogata T,Kage M,et al. Comparison of liver regeneration after a splenectomy and splenic artery ligation in a dimethylnitrosamine-induced cirrhotic rat model. HPB,2010,12(1):22-30.
[14] Kaoru 0, Kanako Y. Familial occurrence of autoimmune liver disease with overlapping features of primary biliary cholangitis and autoimmune hepatitis in a mother and her daughter. Clin J Gastroenterol,2016,9(5):312-318.
[15] Okada Y,Tsuzuki Y,Hokari R,et al.Pressure loading and ethanol exposure differentially modulate rat hepatic stellate cell activation.J Cell Physiol,2008,215(2):472-480.
[16]陈柳莹,陆伦根.重视慢性肝病的病情评估.实用肝脏病杂志,2017,20(1):4-6.
[17] Mousa S, Carbone M. Novel therapeutics for primary biliary cholangitis:Toward a disease-stage-based approach. Autoimmun Rev,2016,15(9):870-876.
[18] Urrunaga N, Rockey D. Portal hypertensive gastropathy and colopathy. Clin Liver Dis, 2014,18(2):389-406.
[19] Norman K, Pirlich M. Gastrointestinal tract in liver disease:which organ is sick. Curr Opin Clin Nutr Metab Care,2008,11(5):613-619.
[20] Hua F, Wang L. Elevation of Vδ1 T cells in peripheral blood and livers of patients with primary biliary cholangitis. Clin Exp Immunol,2016,186(3):347-355.
[2]程询,杨长青.检测门脉高压程度评估肝硬化严重程度研究进展.实用肝脏病杂志,2017,20(1):10-12.
[3]刘雪莲,杨见权.脾功能亢进症治疗进展.实用肝脏病杂志,2013,16(4):382-384.
[4] Yao YM,Liu QG,Yang W,et al. Effect of spleen on immune function of rats with liver cancer complicated by liver cirrhosis.Hepatobiliary Pancreat Dis Int,2003,2(2):242-246.
[5]郭晓霞,李良学,武玉鹏,等.聚肌胞苷酸诱导的早期原发性胆汁性肝硬化小鼠模型的建立.实用肝脏病杂志,2014,17(6):619-622.
[6] Irma M,Markku N. Human leukocyte antigen associations in Finnish liver transplantations due to primary sclerosing cholangitis and primary biliary cirrhosis. Open Med,2007,2(1):12-25.
[7] Chuang N, Gross RG, Odin JA. Update on the epidemiology of primary biliarycirrhosis.Expert Rev Gastroenterol Hepatol,2011,5(5):583-590.
[8] Liu H, Liu Y, Wang L, et al. Prevalence of primary biliary cirrhosis in adults referring hospital for annual health check-up in Southern China. BMC Gastroenterol, 2010,10(100):2-5.
[9] Chan CW,Carpenter JR,Rigamonti C,et al.Survival following the development of ascites and/or peripheral oedema in primary biliary cirrhosis:a staged prognostic model. Scand J Gastroenterol,2005,40(9):1081-1089.
[10]何方平,陈兰.原发性胆汁性肝硬化的流行病学及自然病程研究进展.实用肝脏病杂志,2013,16(2):97-98.
[11] Cartin-Ceba R,Krowda MJ.Portopulmonary hypertension.Clin Liver Dis,2014,18(2):421-438.
[12] Fed erica L, Caterina M. The diagnostic accuracy of biomarkers for diagnosis of primary biliary cholangitis(PBC)in anti-mitochondrial antibody(AM A)-negative PBC patients:a review of literature. Clin Chem Laborat Med,2017,56(1):25-31.
[13] Morinaga A,Ogata T,Kage M,et al. Comparison of liver regeneration after a splenectomy and splenic artery ligation in a dimethylnitrosamine-induced cirrhotic rat model. HPB,2010,12(1):22-30.
[14] Kaoru 0, Kanako Y. Familial occurrence of autoimmune liver disease with overlapping features of primary biliary cholangitis and autoimmune hepatitis in a mother and her daughter. Clin J Gastroenterol,2016,9(5):312-318.
[15] Okada Y,Tsuzuki Y,Hokari R,et al.Pressure loading and ethanol exposure differentially modulate rat hepatic stellate cell activation.J Cell Physiol,2008,215(2):472-480.
[16]陈柳莹,陆伦根.重视慢性肝病的病情评估.实用肝脏病杂志,2017,20(1):4-6.
[17] Mousa S, Carbone M. Novel therapeutics for primary biliary cholangitis:Toward a disease-stage-based approach. Autoimmun Rev,2016,15(9):870-876.
[18] Urrunaga N, Rockey D. Portal hypertensive gastropathy and colopathy. Clin Liver Dis, 2014,18(2):389-406.
[19] Norman K, Pirlich M. Gastrointestinal tract in liver disease:which organ is sick. Curr Opin Clin Nutr Metab Care,2008,11(5):613-619.
[20] Hua F, Wang L. Elevation of Vδ1 T cells in peripheral blood and livers of patients with primary biliary cholangitis. Clin Exp Immunol,2016,186(3):347-355.