Effect Of Yanghe Asthma-Easing Granules On Airway Inflammation And Treg\STAT6 In Asthmatic Rats With Injected Bone Marrow Mesenchymal Stem Cells

Objective:Observing the effect of Yang He Asthma-Easing Granules on airway inflammation in asthmatic rats after injected BMSCs,and further exploration its influence on CD4+CD25+Treg,STAT6.Methods:seventy SD rats of clean grade, were randomly divided into into normal group,model group, BMSCs group, the asthmatic group depleted of Treg and administrated with BMSCs, the asthmatic group depleted of Treg and administrated with both BMSCs and Yang He Asthma-Easing Granules,the BMSCs group administrated with Yang He Asthma-Easing Granules, he BMSCs group administrated with dexamethasone, each group of ten only, use ovalbumin(OVA) sensitized asthma,via the tail vein injection of anti-CD25+mab and BMSCs, and were given drug for treatment six weeks. The changes of lung tissue pathology morphology was measured by HE.Bronchoalveolar lavage fluid were meatured by Wright’s staining and observed under light microscope. The level of IL-4\5\13 in serum was measured by enzyme-linked immuno sorbent assay(ELISA).Flow cytometry(FCM) were detected in peripheral blood CD4+CD25+regulatory T cells level.The gene level of STAT6 in rats lung tissue was measured by real-time PCR.The protein level of STAT6 in rats lung tissue was measured by Western blot.Results:1 The morphology of BMSCs are similar to fibroblasts, such as spindle and polygon,which are observed under the microscope after be isolated and culture.And it could expanded and passaged.The expressions of CD44,CD29,CD45,CD11b/c were98.69%,99.67%,0.15% and 0.26%,and it could be induced into osteoblasts and adipocytes.2 Compared with normal group,model group rats with a large number of inflammatory cells infiltration in the lung tissue,mainly eosinophils, the levels of inflammatory cellsin bronchoalveolar lavage fluid and IL-4,IL-5,IL-13 in serum significantly increased(P<0.01).Compared with model group,the levels of inflammatory cells and IL-4,IL-5,IL-13 in serum decreased(P<0.05).The level of inflammatory cells in asthmatic group administrated with Yang He Asthma-Easing Granules or dexamethasone lower than that of normal group and BMSCs group(P<0.05).3 Compared with normal group,the level of Treg in peripheral blood significantly decreased(P<0.01),the gene and protein level of STAT6 in lung tissue significantly increased(P<0.01).The level of Treg in BMSCs group higher than model group,and the gene and protein level of STAT6 lower than that in model group(P<0.05).Compared with BMSCs group, the level of Tregs in asthmatic group administrated with Yang He Asthma-Easing Granules or dexamethasone higher,the gene and protein level of STAT6lower(P<0.01).Conclusions:1 The BMSCs can be isolated and expanded by using bone marrow adherent culture which is efficient and stable and were the adherent cells of similar fibroblasts morphology. After repeated passaging,BMSCs purity is high,and could be induced into osteoblasts or adipocytes.2 BMSCs can reduce the level of inflammatory cells in bronchoalveolar lavage fluid and IL-4,IL-5,IL-13 in serum and STAT6 in lung tissue, improve the level of CD4+CD25+Treg,and the airway inflammation.3 Yang He Asthma-Easing Granules can reduce the level of inflammatory cells in bronchoalveolar lavage fluid and IL-4,IL-5,IL-13 in serum in asthmatic rats with injected BMSCs.4 Yang He Asthma-Easing Granules can improve the airway inflammation by regulated the expression of CD4+CD25+Treg, STAT6.