基于SOCS1/IRAKs"交叉对话"探讨涤痰通瘀法对脑出血肝损害的保护机制

Hepatic injury following cerebral hemorrhage has high incidence and crypticity, which is the key factor that aggravates the primary disease. The liver is the core target organ of severe stress injury. SOCS1/IRAKs "cross-talk" negative regulation to the cascade reaction of LPS intracellular signal transduction plays a key role in protecting hepatic injury. Its potential clinical application value has been paying close attention to in recent years. But the mechanism of action about SOCS1/IRAKs "cross-talk "is not found out. There is no effective method in activating the regulation. This study intends to observe SOCS1/IRAK-1 2,4, M "cross-talk" of the liver tissue in the model rats suffering from acute cerebral hemorrhage with the syndrome of phlegm-heat obstructing the excessive bowels by co-immunoprecipitation on the basis of preliminary studies, and detect the expression of TLR4、IRAK-1, 2,4, M and their downstream inflammatory factors such as NF-κB and TNF-α level at different time points in donor liver tissue and liver kupffer cells. The purpose of the study is to clear the mechanism of SOCS1/IRAKs "cross-talk". Meanwhile, the method of dissipating phlegm and removing blood stasis method in Chinese medcine is applied to identify the regulatory action to SOCS1/IRAKs "cross-talk" so as to reveal part protection mechanism of dissipating phlegm and removing blood Stasis method to hepatic injury following cerebral hemorrhage.

脑出血继发的肝损害具有高发性和极强隐蔽性，是严重应激损伤的核心靶器官，是引起和加重原发病并致死的关键性因素。SOCS1/IRAKs"交叉对话" 对LPS细胞内信号转导炎性级联反应抑制效应在肝损害保护效应中发挥关键作用，近年来其潜在的临床应用价值引起人们关注。但SOCS1/IRAKs"交叉对话"机制未明，活化调控缺乏有效手段。 本课题在前期基础上，采用免疫共沉淀观察模型大鼠SOCS1/IRAK-1、2、4、M相互作用，测定不同时相点在体肝组织及体外培养肝Kupffer细胞中CD14、IRAK-1、2、4、M及其下游炎性因子NF-κB、TNF-α表达水平，阐明SOCS1/IRAKs"交叉对话"内在机制，同时明确涤痰通瘀法对SOCS1/IRAKs"交叉对话"的调控作用，揭示该法对脑出血肝损害的部分保护机制。

脑出血继发的肝损害具有高发性和极强隐蔽性，是加重原发病并致死的关键性因素。SOCS1/IRAKs“交叉对话”对LPS细胞内信号转导炎性级联反应的抑制效应，在肝损害保护效应中发挥关键作用，但该“交叉对话”的机制未明，其活化调控缺乏有效手段。.本课题在前期基础上，采用免疫共沉淀观察模型大鼠SOCS1/IRAK-1、2、4、M相互作用，测定不同时相点在体肝组织及体外培养肝Kupffer细胞中CD14、IRAK-1、2、4、M及其下游炎性因子NF-κB、TNF-α表达水平，阐明SOCS1/IRAKs“交叉对话”内在机制，同时明确涤痰通瘀法对SOCS1/IRAKs“交叉对话”的调控作用，揭示该法对脑出血肝损害的部分保护机制。.通过研究，我们发现：（1）模型组IRAK-M表达水平明显降低（P<0. 05）；中药组明显升高（P<0. 05），且两组IRAK-M与TNF-α呈负相关。（2）急性脑出血大鼠48h可见凋亡细胞达高峰，24、72h可见凋亡细胞减少；涤痰通瘀方可显著降低血清ALT、AST水平（P<0.05，P<0.01）。（3）脑出血模型组在各时间点IRAK-M mRNA表达水平较正常组明显降低（P<0.05）；中药治疗组的IRAK-M mRNA较脑出血模型组明显升高（P < 0.05）。（4）脑出血模型组可见较多凋亡细胞，涤痰通瘀组KCs凋亡率较脑出血模型组显著减少（P<0. 05）。涤痰通瘀组在各时间点NF-κB表达水平和TNF-α含量较脑出血模型组显著降低（P<0.05，P<0.01）。.根据上述研究数据，我们认为：（1）急性脑出血大鼠模型可导致应激性肝损害细胞凋亡；（2）IRAK-M蛋白表达水平升高可能是抑制脑出血后诱发的LPS介导胞内信号转导炎性级联反应的重要因素；（3）涤痰通瘀方有以下作用：①可加强IRAK-M激酶活性，从而抑制炎症因子的释放，减轻肝损伤的程度；②可有效抑制肝细胞凋亡，从而提高肝功能，是防治脑出血应激性肝损害的重要途径；③可以提高KCs内 IRAK-M mRNA的表达，对急性脑出血应激性肝损害具有一定的保护作用；④能够有效抑制KCs凋亡，阻滞NF-κB的活化、减少TNF-α释放，对急性脑出血应激性肝损害具有保护作用。