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目的用聚肌苷酸胞苷酸(polyinosinic polycytidylic acid,PolyI:C)注射 C57BL/6小鼠以研究建立原发性胆汁性肝硬化(PBC)动物模型,探讨 CD_(40)配体(CD_(40)L)在外周血 T 淋巴细胞表面的表达,和在 PBC 中 T 淋巴细胞的活化情况。方法 20只 C57BL/6小鼠随机分对照组和模型组。将polyI:C 以5mg/kg 的剂量腹腔注射模型组小鼠,对照组注射 PBS,每周2次,120 d 后处死,留取外周血和肝组织,间接免疫荧光法检测血清抗线粒体抗体(AMA),HE 染色观察胆管病理变化,生化分析仪测定血清碱性磷酸酶(ALP)含量,流式细胞术分析外周血 CD_4~+、CD_8~+淋巴细胞所占比例及 CD_(40)L在 T 淋巴细胞表面的表达情况。结果 PolyI:C 注射120 d 后模型组小鼠均出现 AMA 阳性,肝组织小胆管周围有不同程度的炎性细胞浸润,血清 ALP 明显高于对照组(P<0.001);对照组胆管及血清ALP 均未发生明显变化;模型组小鼠 CD_(40)L~+/CD_4~+、CD_(40)L~+/CD_8~+T 淋巴细胞[(4.35±0.34)%,(1.42±0.10)%)显著高于对照组[(0.78±0.10)%,(0.43±0.04)%,P<0.01];而模型组小鼠 CD_4~+、CD_8~+T 淋巴细胞所占比例[(25.83±1.80)%,(2.4.84±2.70)%]与对照组[(20.51±3.46)%,(17.84±1.53)%]比较,差异无统计学意义(P>0.05)。结论 PolyI:C 腹腔注射 C57BL/6小鼠120d 可引起 PBC 病变,活化的 CD_4~+、CD_8~+T 淋巴细胞在 PBC 发病中起重要作用。
Objective To investigate the effects of CD_ (40) ligand (CD_ (40)) on the development of primary biliary cirrhosis (PBC) animal model by injecting polyinosinic polycytidylic acid (PolyI: 40) L) on the surface of peripheral blood T lymphocytes, and the activation of T lymphocytes in PBC. Methods Twenty Twenty C57BL / 6 mice were randomly divided into control group and model group. The mice in the model group were injected intraperitoneally with 5 mg / kg polyI: C and the control group was injected with PBS twice a week for 120 days. Peripheral blood and liver tissues were collected for the detection of serum anti-mitochondrial antibodies AMA). Pathological changes of bile duct were observed by HE staining. Serum ALP levels were measured by biochemical analyzer. Flow cytometry was used to analyze the proportion of CD 4+, CD 8+ lymphocytes and CD 40 L in peripheral blood T lymphocyte surface expression. Results After 120 days of Polyl: C injection, AMA positive mice showed positive AMA in the model group, with varying degrees of inflammatory cell infiltration around the small bile ducts, the serum ALP levels were significantly higher in the model group than in the control group (P <0.001) (40.3 ± 0.34)%, (1.42 ± 0.10)% of the CD_ (40) L ~ + / CD_4 ~ + and CD_ (40) L ~ + / CD_8 ~ + T lymphocytes in the model group ) Was significantly higher than that of the control group [(0.78 ± 0.10)%, (0.43 ± 0.04)%, P <0.01], while the percentage of CD_4 ~ + and CD_8 ~ + T lymphocytes in the model group [(25.83 ± 1.80) %, (2.4.84 ± 2.70)%] compared with the control group [(20.51 ± 3.46)%, (17.84 ± 1.53)%], the difference was not statistically significant (P> 0.05). Conclusion Intraperitoneal injection of PolyI: C into C57BL / 6 mice for 120 days can cause PBC lesions. Activated CD_4 ~ + and CD_8 ~ + T lymphocytes play an important role in the pathogenesis of PBC.