加味柴芍六君颗粒对TRL-NF-κB/SOCS串流介导UC免疫失衡的调节效应

Ulcerative colitis (UC) is a difficult self-limited inflammatory disease to genetic susceptibility people, which intestinal mucosal immune tolerance and immune activation is continuously stimulated by the intestinal flora product lipopolysaccharide (LPS) antigen and environmental factors. that resulting of immune disorders or imbalances is consistent with Chinese medicine imbalance pathogenesis of "yin and yang". Based on the TRL4-MyD88-NF-κB pathway mediating intestinal mucosal immune and inflammatory response and the TRL4-SOCS1 pathway negatively regulating NF-κB and JAK-STAT- mediated immune tolerance, and the two pathway "cross talking" out of control in the immune imbalance in UC , in order to explore the pathophysiological mechanisms of UC, in this study，we will replicate the UC rat model , use ELISA, RT-PCR to detect LPS and IFN-γ, TNF-alpha, IFN-beta, IL-1β, IL-10; Wstern blot, immunohistochemical method detects SOCS1, IKK, JAK , STAT and TLR4, NF-κB p65, MyD88 expression. So as to Explore the effect of the Chaishao Liujun particles regulating UC rats immune imbalance, and support scientific meaning of the Chinese medicine theory of" yin and yang" balance, and also provide an objective basis for Chinese medicine preventing UC.

溃疡性结肠炎(UC)是环境因素作用于遗传易感者，在肠道菌群产物脂多糖(LPS)等抗原的持续刺激或(和)免疫调节紊乱，致使肠粘膜免疫耐受与免疫激活失衡，而致难于自限的炎症反应，这与中医"阴阳失衡"病机相一致。本研究从TRL4-MyD88-NF-κB通路介导肠粘膜免疫炎症反应与TRL4-SOCS1负调控NF-κB和JAK-STAT介导免疫耐受及两者"串流"失控在UC免疫失衡中的关键关节出发，探讨UC病理生理机制，复制UC大鼠模型，运用ELISA、RT-PCR法检测LPS、IFN-γ、TNF-α、IFN-β、IL-1β、IL-10；Western blot、免疫组化检测SOCS1、IKK、JAK、STAT、TLR4、NF-κBp65、MyD88的变化。探讨加味柴芍六君颗粒对UC大鼠免疫失衡的调节效应，佐证中医"阴平阳秘"阴阳平衡理论的科学内涵，为中医药防治UC提供客观依据。

溃疡性结肠炎(UC)是肠道菌群产物脂多糖(LPS)等抗原的持续刺激致使肠粘膜免疫耐受与免疫激活失衡，而致难于自限的炎症反应。全国名老中医李桂贤教授运用加味柴芍六君颗粒治疗UC取得了较好的疗效，为进一步探讨加味柴芍六君颗粒治疗UC机制，本课题从TRL4-MyD88-NF-κB通路介导肠粘膜免疫炎症反应与TRL4-SOCS1负调控NF-κB和JAK-STAT介导免疫耐受出发，复制UC大鼠模型，运用免疫组化法检测TRL4、MyD88、NF-κBP65、SOCS1、JAK2、STAT3；ELISA法检测IL-1β、IL-10 、IFN-β、IFN-γ、LPS、TNF-α；RT-PCR法检测 TNF-α、IFN-γ、IL-1β、IL-10；Western blot法检测SOCS1、TRL4、NF-κBP65、MyD88的变化。结果：①加味柴芍六君颗粒大、中、小剂量组及SASP组与模型组比较LPS、IFN-γ、IL-1β（ELISA法）、SOCS1、NF-κBP65（Western blot法）、TRL4（免疫组化法）有明显统计学意义（P＜0.01）；②大、中、小剂量组与SASP组比较IL-1β（ELISA法）有统计学意义（P＜0.05）；③中剂量组与SASP组比较NF-κBP65（免疫组化法）有统计学意义（P＜0.05）。④大、中剂量组及SASP组与模型组比较STAT3、SOCS1、NF-κBP65（免疫组化法）、TNF-α（ELISA法）有统计学意义（P＜0.05或P＜0.01）；⑤大、中剂量组与小剂量组比较STAT3、NF-κBP65（免疫组化法）有统计学意义（P＜0.05）；⑥大剂量组及SASP组与小剂量组比较SOCS1、JAK2（免疫组化法）有统计学意义（P＜0.05）；⑦大剂量组与模型组比较MyD88、JAK2（免疫组化法）、IFN-γ（RT-PCR法）、IL-10（ELISA法、RT-PCR法）有统计学意义（P＜0.05或P＜0.01）；⑧中剂量组与模型组比较MyD88、TRL4（Western blot法）、IL-1β、TNF-α（RT-PCR法）有统计有统计学意义（P＜0.05或P＜0.01）。结论：加味柴芍六君颗粒可调节TRL4-MyD88-NF-κB通路与TRL4- SOCS1负调控NF-κB和JAK-STAT通路平衡，从而达到治疗UC疗效。