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目的:通过研究肿瘤坏死因子-a(TNF-α)及Toll样受体4/核转录因子κB(TLR4/NF-κB)信号通路相关基因在原发性胆汁性胆管炎(PBC)模型小鼠回肠组织中的表达情况,探讨中药调肝理脾方(TGLP)联合熊去氧胆酸(UDCA)干预PBC小鼠的作用机制。方法:将100只C57BL/6雌性小鼠随机分为正常组、模型组、阳性组(UDCA 0.1g·kg~(-1)·d~(-1))、中药组(TGLP 5.4g·kg~(-1)·d~(-1))、联合组(TGLP 5.4g·kg~(-1)·d~(-1)+UDCA 0.1g·kg~(-1)·d~(-1)),每组20只;采用聚肌胞苷酸(poly l:C)5mg/kg复制PBC小鼠模型;给药8w、16w后颈脱位法处死小鼠,采集末端回肠,采用酶联免疫吸附试验(ELISA)法及实时荧光定量PCR(RT-PCR)法检测回肠组织中TLR4、TNF-α的表达水平,RTPCR法测定回肠组织中TLR4、NF-κB mRNA的相对表达水平。结果:与模型组比较,正常组小鼠回肠组织中NF-κB、TNF-α表达水平较低,差异有显著性意义(P<0.01);经治疗后3个治疗组小鼠TLR4、NF-κB、TNF-α表达水平均较模型组降低,差异有显著性意义(P<0.01或P<0.05),且联合组小鼠TLR4、NF-κB、TNF-α表达水平下降最显著;与联合组相比,中药组小鼠NF-κB表达水平较高,差异有显著性意义(P<0.01或P<0.05)。结论:负性调节小鼠回肠组织中TLR4/NF-κB信号通路并抑制TNF-α因子的释放可能是TGLP联合UDCA干预PBC模型小鼠的作用机制之一。
OBJECTIVE: To investigate the role of tumor necrosis factor-α (TNF-α) and TLR4 / NF-κB signaling pathway-related genes in primary biliary cholangitis (PBC) Ileum tissue, to explore the mechanism of action of traditional Chinese medicine Tiaoganlipi Fang (UDCA) in intervention of PBC mice. Methods: 100 C57BL / 6 female mice were randomly divided into normal group, model group, positive group (UDCA 0.1g · kg -1 · d -1), Chinese herb group (TGLP 5.4g · kg ~ (-1) · d ~ (-1)) in the combined group (TGG 5.4g · kg -1 · d -1 + UDCA 0.1g · kg -1 · d ~ (-1) 1), 20 in each group. The mice were sacrificed by 8 weeks and 16 weeks after cervical dislocation. The terminal ileum was harvested, The expressions of TLR4 and TNF-α in ileum were detected by ELISA and real-time fluorescence quantitative PCR (RT-PCR). The relative expression levels of TLR4 and NF-κB mRNA in ileal tissue were detected by RTPCR. Results: Compared with the model group, the expression of NF-κB and TNF-α in the ileum of normal mice was significantly lower than that of the model group (P <0.01). After treatment, the expressions of TLR4 and NF- (P <0.01 or P <0.05), and the expression of TLR4, NF-κB and TNF-α decreased significantly in the combination group; Compared with the model group, the expression of NF-κB and TNF- Compared with the control group, the expression of NF-κB in Chinese medicine group was significantly higher than that in the control group (P <0.01 or P <0.05). CONCLUSION: The negative regulation of TLR4 / NF-κB signaling pathway and the release of TNF-α in mouse ileum may be one of the mechanisms of TGLP and UDCA intervention in PBC model mice.