| Objective:In this experiment, we established model rats of surgical excision of the stomach and intervened the rats with Qi Huang Decoction by dropping it in the intestinal cavity. We studied the effects of Qi Huang Decoction on the number of molecules and gene expression in the process of intestinal lymphocyte homing. We revealed the truth that Qi Huang Decoction protected the function of intestinal mucosal barrier by the realization of lymphocyte homing. The studies would provide basic research for the prevention and treatment in intestinal barrier dysfunction and the related critical illness.Methods:We established the gastrectomy model and sham operation model, divided the rats into three groups randomly:(1) sham operation group:We only took rats’middle abdomen away without gastrectomy. After the operation, we did not give enteral nutrition or Chinese medicine, but let rats drink and eat freely.(2) Contrast group:After the operation, rats were put into the cages. The rats that were not given anything to eat could move about freely when they woke up. Until the first day after operation, the rats should be dropped6ml saline water before the time of giving enteral nutrition (Protein Enteral Nutrition Powder). The treatment lasted seven days.(3) Qi Huang Decoction group:The formulation was the same to the Contrast group which included heat、method、and treatment before dropped6ml Qi Huang decoction.After the end of the treatment, we executed rats、separated specimen and then observed changes of molecule expression in intestinal lymphocyte homing process.(1) Homing receptor(α4β7、L-selectin、LFA-1):Flow cytometry was used to test express-quantity of homing receptor and count positive rate of mononuclear cells in lymph fluid; We observed changes of molecule expression on homing receptor(α4β7、 L-selectin、LFA-1) in lymph fluid by RT-PCR.(2) Homing ligands (MAdCAM-1、ICAM-1):immunohistochemical method was used to measure average value of integrated optical density in Peyer’s patches’ positive (3) cells; we also observed changes of molecule expression on homing ligands (MAdCAM-1、ICAM-1) in lymph fluid by RT-PCR.Result(1)The changes of homing receptors(α4β7、L-selectin、LFA-1) in lymph fluid of every group:①he changes of positive rate of Mononuclear cells in lymph fluid:A week later, compared with that of the sham operation group, the positive rate of Mononuclear cells in Qi Huang Decoction group and contrast group decreased significantly (P<0.01). The results showed the fact that the immune barrier on intestinal mucosal suffered damage from the Surgery, The reason is that the positive rate of Mononuclear cells in lymph fluid decreased. Compared with that of the contrast group, the positive rate of Mononuclear cells in Qi Huang decoction group increased, But the result was not of significance in statistics (P<0.05). The fact was proved that we intervened the rats with Qi Huang decoction by dropping it in the intestinal cavity could increase the positive rate of Mononuclear cells. Qi Huang decoction inducted lymphocytes gather together. It was a good way to repair the damaged immune barrier on intestinal mucosal.(2) The changes of gene expression of homing receptors in lymph fluid: Compared with that of the sham operation group, the gene expression of Qi Huang decoction group and contrast group decreased significantly (P<0.01). Compared with that of the contrast group, Qi Huang decoction group increased significantly (P<0.05). We intervened the rats with Qi Huang decoction by dropping it in the intestinal cavity could increase the gene expression. Qi Huang decoction group would combine homing receptors with homing ligands better in the epithelium and lamina propria than other groups. Additionally, the number of LFA-1molecule expression was not only more than α4β7、L-selectin, but also the difference was significant (P<0.01). Because LFA-1Played a major role in the later period of lymph node homing. (2) The changes of homing ligands (MAdCAM-1、ICAM-1) in the Peyer’s patches in every group:①he number of positive expression in the Peyer’s patches:Compared with that of the sham operation group, the number of positive expression of Qi Huang Decoction group and contrast group in the Peyer’s patches decreased significantly (P<0.01). The result proved that we intervened the rats with Qi Huang Decoction and enteral nutrition could combine homing receptors with homing ligands better, they repaired the damaged immune barrier on intestinal mucosal quickly.②he changes of gene expression of homing ligands in the Peyer’s patches:A week later, compared with that of the sham operation group, the gene expression of Qi Huang decoction group and contrast group decreased significantly (P<0.01). The Phenomenon showed trauma could lead to destruction within mucosal epithelium and lamina propria and make gene expression of adhesion molecules decrease significantly. Compared with that of the contrast group, Qi Huang Decoction group increased significantly(P<0.01). Relative expression level of adhesion molecule (MADCAM-1、 ICAM-1) rose after Qi Huang Decoction was dropped. It restored mucosal immune barrier on epithelium and lamina propria and promoted adhesion molecules to swim out. Additionally, we found that the number of MADCAM-1molecule expression was much more than ICAM-1. The reason was that MADCAM-1could combine with the homing receptors of α4β7and also with L-selectin. The homing receptors of LFA-1combined with homing ligands of ICAM-1much later and less than the homing receptors of α4β7L-selectin.Conclusion:After gastric resection, we intervened the rats with Qi Huang Decoction by dropping it in the intestinal cavity. Qi Huang Decoction promoted higher mRNA molecule expression of homing receptor(α4β7L-selectin、LFA-1) in blood lymphocyte’s surface, caused naive lymphocyte to pass through high endothelial venule. The naive lymphocyte was generated in central lymphoid organs and intestinal sensed by induction site(Peyer’s patches). Besides, Qi Huang Decoction could increase higher mRNA expression of homing ligands (MAdCAM-1、ICAM-1) on the epithelium and lamina propria. The receptors and ligands which were expressed highly not only mook T cell in lymph and IgA+B lymphocyte mellow and multiple indirectly, but also improved mucosal immune barrier function after surgery. |
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