芪黄煎剂对大鼠胃切除后肠淋巴细胞归巢的影响

Surgery and other trauma can cause dysfunction of gut mucosal immunobarrier, leading to bacterial translocation, systemic inflammatory response syndrome，and multiple organ dysfunction syndrome, seriously affect the outcome of the disease. Maintaining of gut mucosal immunobarrier is related to intestinal lymphocyte homing.However, molecular basis of lymphocyte homing is on the mutual identification and combination of homing receptors α4β7, L-selectin, LFA-1, and homing ligands MAdCAM-1, ICAM-1.It is proved by previous clinical and animal experiments that gastrectomy can cause gut mucosal immunobarrier dysfunction and bacterial translocation, while Qihuang Decoction of tonifying spleen and catharsis has the function of protecting gut mucosal immunobarrier. But it is still unclear whether the function is achieved by regulating the process of intestinal lymphocyte homing to promote the expression of homing molecules..This project is planned on the basis of our preliminary studies through the estabilishment of gastrectomy surgical trauma model. After intestinal infusion with Qihuang Decoction, the following procedures will be done: 1. With 51Cr-labeled lymphocytes in vitro and vein injection in vivo, the number of lymphocyte homing is measured by γ counter method. 2. lymphocyte homing species will be detected with flow cytometry and immunohistochemistry. 3. Lymphocyte homing molecular expression number, protein content, gene transcription, and other indicators will be observed with reverse transcription polymerase chain reaction, immunohistochemistry and flow cytometry. The purpose of this study is to multi-channelly and multi-levelly observe the effect and molecular expression of Qihuang Decoction on gut lymphocyte homing after surgical stress, reveal the preoection mechanism of Qihuang Decoction on gut mucosal immunobarrier function, lay a foundation for protection of gut mucosal barrier, prevention and treatment of related critical diseases with traditional Chinese medicine.   .

手术等创伤引起肠黏膜免疫屏障功能障碍，导致细菌易位、SIRS和MODS发生，严重影响疾病转归和预后。肠淋巴细胞归巢是肠黏膜实现免疫屏障的先决条件；归巢受体和配体的互相辨认与结合，是归巢的分子基础。我们以往研究证实，健脾通里中药芪黄煎剂具有保护胃切除大鼠肠黏膜免疫屏障功能，但是否通过调节归巢过程以及促进归巢分子的表达而实现，尚不清楚。.本项目拟在我们前期研究基础上，通过建立胃切除手术大鼠模型，芪黄煎剂肠内滴注干预后：（1）淋巴细胞51Cr标记γ计数器计数法测定淋巴细胞归巢数量；（2）流式细胞、免疫组化法检测淋巴细胞归巢种类；（3）流式细胞、免疫组化、RT-PCR技术观察肠淋巴细胞归巢分子表达数量、蛋白含量和基因转录等指标。多途径、多层次观察芪黄煎剂对手术应激后肠淋巴细胞归巢的影响及其分子表达的干预，揭示其保护肠黏膜免疫屏障功能机制，为中药保护肠黏膜屏障功能及其危重病的防治奠定基础。

手术等创伤引起肠黏膜免疫屏障功能障碍，导致细菌易位、SIRS和MODS发生，严重影响疾病转归和预后。肠淋巴细胞归巢是肠黏膜实现免疫屏障的先决条件；归巢受体和配体的互相辨认与结合，是归巢的分子基础。我们以往研究证实，健脾通里中药芪黄煎剂具有保护胃切除大鼠肠黏膜免疫屏障功能，但是否通过调节归巢过程以及促进归巢分子的表达而实现，尚不清楚。本项目拟在我们前期研究基础上，通过建立胃切除手术大鼠模型，芪黄煎剂肠内滴注干预后：（1）淋巴细胞51Cr标记γ计数器计数法测定淋巴细胞归巢数量；（2）流式细胞、免疫组化法检测淋巴细胞归巢种类；（3）流式细胞、免疫组化、RT-PCR技术观察肠淋巴细胞归巢分子表达数量、蛋白含量和基因转录等指标。多途径、多层次观察芪黄煎剂对手术应激后肠淋巴细胞归巢的影响及其分子表达的干预。结果表明芪黄煎剂能够增加胃切除手术后肠淋巴细胞归巢数量、T细胞数量、B细胞数量，从而促进整个的归巢过程。同时芪黄煎剂能增加血液淋巴细胞表面归巢受体（α4β7、L-selectin、LFA-1）数量及mRNA 表达含量，促使在中枢淋巴器官（胸腺、骨髓等）和肠道自身生成的初始淋巴细胞通过诱导部位（PP结）致敏，再次穿越高内皮微静脉，回到血液循环，促进黏附分子的游出；同时也能增加肠黏膜效应部位的肠固有层或上皮内归巢配体（MAdCAM-1、ICAM-1）的数量及mRNA表达含量，使黏膜地址素更能捕获归巢受体，归巢受体与配体高表达结合后，激活淋巴组织中的T淋巴细胞的分化、成熟和增殖、并间接地促进淋巴组织中B细胞的增殖、活化。肠淋巴细胞归巢改善了胃切除术后肠黏膜免疫屏障功能，防治细菌移位发生，有助于减少手术创伤后肠源性感染以及SIRS、MODS等发生。本项目揭示其保护肠黏膜免疫屏障功能机制，为中药保护肠黏膜屏障功能及其危重病的防治奠定基础。