异体骨髓间充质干细胞移植对大鼠子宫内膜异位症炎症反应的调节作用

Regulating Effect of Allogeneic Bone Marrow Mesenchymal Stem Cell Transplantation on Inflammatory Reaction of Endometriosis

为探讨异体移植大鼠骨髓源间充质干细胞对大鼠内异症炎症反应的调节及机制,采用贴壁法获取大鼠骨髓间充质干细胞(BMSCs),流式鉴定表型。选取25只健康SD雌性大鼠自体移植构建内异症模型,随机5只作为假手术组,4周后将选取其中成功建模的15只大鼠中的12只随机分成两组,每组6只,尾静脉分别给予生理盐水和1×10^(6)个BMSCs的悬液。细胞移植4周后处死大鼠,检测血液和腹腔液VEGF、TNF-α含量,并提取腹腔液中的巨噬细胞,流式检测CD86、CD206的表达,同时取下异位囊肿组织和假手术组正常内膜组织行HE染色,在病理组织水平观察各组的差异。发现通过贴壁法成功获取大鼠BMSCs,流式鉴定CD14、CD19、HLA-DR≤2%、CD90≥95%。对照组血清和腹腔液VEGF、TNF-α的含量显著高于假手术组(P<0.05)。BMSCs组TNF-α的含量显著低于对照组,但VEGF的浓度高于对照组(P<0.05)。巨噬细胞表型:BMSCs组的M1型巨噬细胞(CD86+)低于对照组,而M2型巨噬细胞(CD206+)高于对照组,二者差异具有统计学意义(P<0.05)。组织病理学:对照组与假手术组相比腺体数量减少伴有纤维化,血管分布较多,而BMSCs组腺体出现萎缩,与对照组相比腺体和血管明显减少。说明异体BMSCs移植可以诱导巨噬细胞的M2型极化,通过分泌消炎因子抑制异位症炎症进程,促进局部组织修复。

In order to explore the regulating effect of allogeneic bone marrow mesenchymal stem cell transplantation on inflammatory reaction of endometriosis,the research adopts adherent method to gain rat BMSCs and to make flow identification phenotype.25 healthy SD female rats are selected to make autotransplantation endometriosis model.5 rats are randomly divided into sham-operation group.After 4 weeks,12 in 15 rats which are under successful modeling are randomly divided into two groups,with 6 rats in each group.They are injected with normal saline and 1×10^(6) BMSCs suspension in caudal vein.Rats are executed 4 weeks after cell transplantation.Blood and peritoneal fluid VEGF and TNF-αcontent of control group are significantly higher than sham-operation group(P<0.05).TNF-αof BMSCs group is significantly lower than control group,but VEGF concentration is significantly higher than control group(P<0.05).M1 macrophage cell phenotype:macrophage(CD86+)of BMSCs group is lower than control group,but macrophage(CD206+),and M2 macrophage(CD206+)are higher than control group,with statistically significant difference(P<0.05).Histopathology:compared with control group,gland number reduces with fibrosis and more vascularity.The gland atrophies in BMSCs group,and the gland and blood vessel reduce significantly compared with control group.This indicates that allosome BMSCs transplantation can induce M2 polarization of macrophage,inhibit ectopia inflammation through the secretion of antiphlogosis,and promote local tissue repair.