门静脉高压症小鼠模型构建
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摘要
目的构建小鼠胆总管结扎(BDL)及四氯化碳(CCl_4)诱导的两种门静脉高压症(PHT)模型。方法 24只C57BL/6小鼠,随机分成4组(BDL组及相应对照组,CCl_4诱导组及相应对照组),每组6只。BDL及CCl_4诱导法构建PHT小鼠模型后,经门静脉主干穿刺测取门静脉压,并通过血清谷氨酸转氨酶(ALT)及天冬氨酸转氨酶(AST)水平检测,肝脏切片苏木精-伊红(HE)及天狼星红染色,α-平滑肌肌动蛋白(SMA)免疫组化检查分别对模型肝功能、肝纤维化及肝星状细胞激活情况进行评价。结果两种建模方法均使小鼠门静脉压上升,CCl_4诱导组上升更为显著。两模型组小鼠与相应对照组相比,均呈现严重肝功能损伤、肝纤维化及肝星状细胞激活。结论 BDL及CCl_4诱导方法均能成功构建PHT小鼠模型,其门静脉压、血清学生化指标及肝脏病理学改变均符合PHT特点。
Objective To establish two types of portal hypertension(PHT) models in mice by using bile duct ligation(BDL) method and carbon tetrachloride(CCl_4) induction technique respectively. Methods A total of 24 C57BL/6 mice were randomly and equally divided into the following four groups with 6 mice in each group: group BDL, control group of BDL, group CCl_4, and control group of CCl_4. After the establishment of PHT, the main portal vein was punctured in all experimental mice to measure the portal vein pressure, and blood sampling was collected to test serum alanine aminotransferase(ALT) and aspartate aminotransferase(AST) levels. Using hematoxylin eosin(HE) and sirius red staining the liver tissues were pathologically examined. Immunohistochemical study of alpha smooth muscle actin(SMA) was adopted to evaluate the liver function, hepatic fibrosis and hepatic stellate cell activation status. Results Both modeling methods could make the portal vein pressure increased in experimental mice. The increasing of portal vein pressure in group CCl_4 was more obvious. Compared with their corresponding control groups, the degree of liver damage, hepatic fibrosis and hepatic stellate cell activation in group BDL and group CCl_4 were more serious. Conclusion Both BDL method and CCl_4 induction technique can successfully establish the mouse model of PHT. All the portal venous pressure, the serum biochemical indices and the changes of liver pathology of the mouse model are well in line with the characteristics of PHT in human.
Objective To establish two types of portal hypertension(PHT) models in mice by using bile duct ligation(BDL) method and carbon tetrachloride(CCl_4) induction technique respectively. Methods A total of 24 C57BL/6 mice were randomly and equally divided into the following four groups with 6 mice in each group: group BDL, control group of BDL, group CCl_4, and control group of CCl_4. After the establishment of PHT, the main portal vein was punctured in all experimental mice to measure the portal vein pressure, and blood sampling was collected to test serum alanine aminotransferase(ALT) and aspartate aminotransferase(AST) levels. Using hematoxylin eosin(HE) and sirius red staining the liver tissues were pathologically examined. Immunohistochemical study of alpha smooth muscle actin(SMA) was adopted to evaluate the liver function, hepatic fibrosis and hepatic stellate cell activation status. Results Both modeling methods could make the portal vein pressure increased in experimental mice. The increasing of portal vein pressure in group CCl_4 was more obvious. Compared with their corresponding control groups, the degree of liver damage, hepatic fibrosis and hepatic stellate cell activation in group BDL and group CCl_4 were more serious. Conclusion Both BDL method and CCl_4 induction technique can successfully establish the mouse model of PHT. All the portal venous pressure, the serum biochemical indices and the changes of liver pathology of the mouse model are well in line with the characteristics of PHT in human.
引文
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[12]薛挥,李培杰,李伟之,等.经颈静脉肝内门体分流术的并发症及其防治[J].临床肝胆病杂志,2016,32:238-241.
[13]曹青,彭小刚,洪葵.诺贝尔生理或医学奖:基因靶向技术和人类疾病小鼠模型[J].江西医学,2012,47:414-416.
[14]Novo E,Cannito S,Paternostro C,et al.Cellular and molecular mechanisms in liver fibrogenesis[J].Arch Biochem Biophys,2014,15:20-37.
[15]李生财,李彤.肝纤维化动物模型的造模原理及应用[J].中医药学刊,2016,24:2267-2268.
[16]Wynn TA,Ramalingam TR.Mechanisms of fibrosis:therapeutic translation for fibrotic disease[J].Nat Med,2012,18:1028-1040.
[17]Schwabl P,Payer BA,Grahovac J,et al.Pioglitazone decreases portosystemic shunting by modulating inflammation and angiogenesis in cirrhotic and non-cirrhotic portal hypertensive rats[J].J Hepatol,2014,60:1135-1142.
[2]原姗姗,韩国宏,樊代明.TIPS治疗门静脉高压并发症预后评价的现状[J].介入放射学杂志,2011,20:245-248.
[3]Vorobioff JD,Groszmann RJ.Prevention of portal hypertension:from variceal development to clinical decompensation[J].Hepatology,2015,61:375-381.
[4]Blomme B,Van Steenkiste C,Vanhuysse J,et al.Impact of elevation of total bilirubin level and etiology of the liver disease on serum N-glycosylation patterns in mice and humans[J].Am J Physiol Gastrointest Liver Physiol,2010,298:G615-G624.
[5]Mookerjee RP,Mehta G,Balasu bramaniyan V,et al.Hepatic dimethylarginine-dimethylaminohydrolase-1 is reduced in cirrhosis and is a target for therapy in portal hypertension[J].J Hepatol,2015,62:325-331.
[6]Van Steenkiste C,Ribera J,Geerts A,et al.Inhibition of placental growth factor activity reduces the severity of fibrosis,inflammation,and portal hypertension in cirrhotic mice[J].Hepatology,2011,53:1629-1640.
[7]Altrock E,Sens C,Wuerfel C,et al.Inhibition of fibronectin deposition improves experimental liver fibrosis[J].J Hepatol,2015,62:625-633.
[8]Iwakiri Y.Pathophysiology of portal hypertension[J].Clin Liver Dis,2014,18:281-291.
[9]Sanyal AJ,Bosch J,Blei A,et al.Portal hypertension and its complications[J].Gastroenterology,2008,134:1715-1728.
[10]陈皓,杨卫平,李宏为.肝移植时代传统外科手术在肝硬化门静脉高压症中的治疗地位[J].外科理论与实践,2016,21:372-374.
[11]徐克,韩国宏.正确认识经颈静脉肝内门体分流术治疗肝硬化门静脉高压并发症的地位[J].中华肝脏病杂志,2011,19:481-482.
[12]薛挥,李培杰,李伟之,等.经颈静脉肝内门体分流术的并发症及其防治[J].临床肝胆病杂志,2016,32:238-241.
[13]曹青,彭小刚,洪葵.诺贝尔生理或医学奖:基因靶向技术和人类疾病小鼠模型[J].江西医学,2012,47:414-416.
[14]Novo E,Cannito S,Paternostro C,et al.Cellular and molecular mechanisms in liver fibrogenesis[J].Arch Biochem Biophys,2014,15:20-37.
[15]李生财,李彤.肝纤维化动物模型的造模原理及应用[J].中医药学刊,2016,24:2267-2268.
[16]Wynn TA,Ramalingam TR.Mechanisms of fibrosis:therapeutic translation for fibrotic disease[J].Nat Med,2012,18:1028-1040.
[17]Schwabl P,Payer BA,Grahovac J,et al.Pioglitazone decreases portosystemic shunting by modulating inflammation and angiogenesis in cirrhotic and non-cirrhotic portal hypertensive rats[J].J Hepatol,2014,60:1135-1142.