原发性胆汁性胆管炎小鼠模型肝内趋化因子CXCL13的表达

发布时间：2018-05-05 14:28

本文选题：原发性胆汁性胆管炎 + Poly　；参考：《医学研究生学报》2017年04期

【摘要】：目的原发性胆汁性胆管炎(PBC)患者肝内趋化因子CXCL13表达显著增加,但其来源和机制尚不明确。文章旨在通过建立PBC小鼠模型,探索PBC小鼠肝内趋化因子CXCL13的表达及其主要来源细胞。方法采用随机数字表法将C57BL/6小鼠分为实验组[n=20,腹腔注射聚肌苷酸-聚胞苷酸(Poly I:C,2次/周,共12周)]和对照组(n=10,腹腔注射,系同实验组等体积磷酸盐缓冲液)。酶联免疫吸附试验(ELISA)测定血清AMA水平,HE染色观察肝组织炎症细胞浸润情况。原位灌注酶消化结合磁珠分选法分离小鼠肝内Kupffer细胞、肝血窦内皮细胞以及肝内淋巴细胞,荧光定量PCR检测肝组织以及上述细胞亚群CXCL13 mRNA表达水平。结果实验组小鼠血清AMA水平随建模时间延长逐渐上升,首次注射Poly I:C后第4、8、12周阳性率分别为5.9%、52.9%以及76.5%,而对照组小鼠血清AMA水平在整个建模过程中均呈现较低水平,仅在第12周时有2只小鼠AMA水平略高于阳性界值。实验组和对照组小鼠肝组织HE染色结果表明,实验组小鼠肝内汇管区大量炎症细胞浸润,而对照组小鼠肝内未见炎症细胞浸润。流式细胞术检测实验组小鼠Kupffer细胞、肝血窦内皮细胞的分离纯度分别为76%~80%、68%~72%。与Kupffer细胞CXCL13 mRNA表达[2.34(0.22-8.64)]比较,肝血窦内皮细胞、肝内淋巴细胞表达下降[0.27(0.03-1.64)、0.05(0-0.22),P0.05]。结论 Poly I:C诱导建立的PBC小鼠模型中肝内升高的趋化因子CXCL13主要来源于Kupffer细胞。
[Abstract]:Objective the expression of chemokine CXCL13 was significantly increased in patients with primary biliary cholangitis, but its origin and mechanism were unclear. The aim of this study was to explore the expression of chemokine CXCL13 and its main cells in liver of PBC mice by establishing PBC mouse model. Methods C57BL/6 mice were randomly divided into two groups: the experimental group (n = 20) and the control group (n = 10) and the control group (n = 10, n = 10). The mice were treated with the same volume of phosphate buffer solution as the experimental group (n = 20, intraperitoneal injection) and the control group (n = 10, n = 10). The level of serum AMA was determined by Elisa and HE staining was used to observe the infiltration of inflammatory cells in liver tissue. Kupffer cells were isolated from mouse liver by in situ perfusion enzyme digestion combined with magnetic bead sorting method. Hepatic sinusoidal endothelial cells and intrahepatic lymphocytes were isolated. The expression of CXCL13 mRNA in liver tissue and its subsets were detected by fluorescence quantitative PCR. Results the serum AMA level of the experimental group increased gradually with the prolongation of modeling time. The positive rates of serum AMA in the experimental group were 5.92and 76.5respectively at the 8th week after the first injection of Poly I: C, while the serum AMA level in the control group was lower than that in the control group during the whole modeling process. Only at the 12th week, the AMA level of 2 mice was slightly higher than the positive limit. The results of HE staining in the liver tissue of the experimental group and the control group showed that a large number of inflammatory cells were infiltrated in the intrahepatic catchment area of the experimental group, but no infiltration of the inflammatory cells was found in the liver of the control group. Flow cytometry was used to detect Kupffer cells in experimental group and the purity of hepatic sinusoidal endothelial cells was 76% and 80% respectively. Compared with the expression of CXCL13 mRNA in Kupffer cells [2.34 ~ 0.22-8.64], the expression of hepatic lymphocytes in hepatic sinusoidal endothelial cells was decreased [0.27o0.03-1.640.55 (0-0.22) (P0.05)]. Conclusion the increased chemokine CXCL13 in the liver of PBC mice induced by Poly I: C is mainly derived from Kupffer cells.
【作者单位】：器官衰竭防治国家重点实验室广东省病毒性肝炎研究重点实验室南方医科大学南方医院感染内科暨肝病中心;北京大学深圳医院感染内科;
【基金】：国家自然科学基金(81470836)
【分类号】：R575.7;R-332

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