本文選題：原發(fā)性膽汁性膽管炎 + Poly�。� 參考：《醫(yī)學(xué)研究生學(xué)報(bào)》2017年04期

【摘要】：目的原發(fā)性膽汁性膽管炎(PBC)患者肝內(nèi)趨化因子CXCL13表達(dá)顯著增加,但其來(lái)源和機(jī)制尚不明確。文章旨在通過(guò)建立PBC小鼠模型,探索PBC小鼠肝內(nèi)趨化因子CXCL13的表達(dá)及其主要來(lái)源細(xì)胞。方法采用隨機(jī)數(shù)字表法將C57BL/6小鼠分為實(shí)驗(yàn)組[n=20,腹腔注射聚肌苷酸-聚胞苷酸(Poly I:C,2次/周,共12周)]和對(duì)照組(n=10,腹腔注射,系同實(shí)驗(yàn)組等體積磷酸鹽緩沖液)。酶聯(lián)免疫吸附試驗(yàn)(ELISA)測(cè)定血清AMA水平,HE染色觀察肝組織炎癥細(xì)胞浸潤(rùn)情況。原位灌注酶消化結(jié)合磁珠分選法分離小鼠肝內(nèi)Kupffer細(xì)胞、肝血竇內(nèi)皮細(xì)胞以及肝內(nèi)淋巴細(xì)胞,熒光定量PCR檢測(cè)肝組織以及上述細(xì)胞亞群CXCL13 mRNA表達(dá)水平。結(jié)果實(shí)驗(yàn)組小鼠血清AMA水平隨建模時(shí)間延長(zhǎng)逐漸上升,首次注射Poly I:C后第4、8、12周陽(yáng)性率分別為5.9%、52.9%以及76.5%,而對(duì)照組小鼠血清AMA水平在整個(gè)建模過(guò)程中均呈現(xiàn)較低水平,僅在第12周時(shí)有2只小鼠AMA水平略高于陽(yáng)性界值。實(shí)驗(yàn)組和對(duì)照組小鼠肝組織HE染色結(jié)果表明,實(shí)驗(yàn)組小鼠肝內(nèi)匯管區(qū)大量炎癥細(xì)胞浸潤(rùn),而對(duì)照組小鼠肝內(nèi)未見(jiàn)炎癥細(xì)胞浸潤(rùn)。流式細(xì)胞術(shù)檢測(cè)實(shí)驗(yàn)組小鼠Kupffer細(xì)胞、肝血竇內(nèi)皮細(xì)胞的分離純度分別為76%~80%、68%~72%。與Kupffer細(xì)胞CXCL13 mRNA表達(dá)[2.34(0.22-8.64)]比較,肝血竇內(nèi)皮細(xì)胞、肝內(nèi)淋巴細(xì)胞表達(dá)下降[0.27(0.03-1.64)、0.05(0-0.22),P0.05]。結(jié)論 Poly I:C誘導(dǎo)建立的PBC小鼠模型中肝內(nèi)升高的趨化因子CXCL13主要來(lái)源于Kupffer細(xì)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.
【作者單位】： 器官衰竭防治國(guó)家重點(diǎn)實(shí)驗(yàn)室 廣東省病毒性肝炎研究重點(diǎn)實(shí)驗(yàn)室 南方醫(yī)科大學(xué)南方醫(yī)院感染內(nèi)科暨肝病中心;北京大學(xué)深圳醫(yī)院感染內(nèi)科;
【基金】：國(guó)家自然科學(xué)基金(81470836)
【分類號(hào)】：R575.7;R-332

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