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目的:研究大鼠骨髓间充干细胞(r BMSC)抵抗人血清介导的异种体液性杀伤的作用及其主要机制。方法:分离培养SD大鼠BMSC,将第4代的r BMSC作为实验材料,以大鼠淋巴细胞(r LC)作为对照。采用流式细胞技术,检查两种细胞异种抗原α-Gal的表达情况,体积分数20%正常人血清对两种细胞的杀伤作用、两种细胞分别与正常人血清中天然抗体Ig M和Ig G的结合情况,以及与正常人血清作用后补体C3c、C4c、C5b-9在两种细胞上的沉积情况。结果:成功分离和培养r BMSC。相对于r LC,r BMSC对人血清介导的异种体液性杀伤具有明显的抵抗作用(P<0.01);r BMSC上α-Gal的表达显著低于r LC(P<0.05);r BMSC与人血清中Ig G和Ig M的结合量显著低于r LC(P<0.01);与正常人血清作用后,r LC可见显著的C3c、C4c和C5b-9沉积,但r BMSC仅有较少C3c和C4c沉积,两种细胞间比较,差异均有统计学意义(P<0.01)。结论:r BMSC能明显抵抗人血清介导的异种体液免疫杀伤作用,其机制可能与r BMSC低表达异种抗原α-Gal及抑制了补体攻膜复合物形成有关。
OBJECTIVE: To investigate the role of rBMSCs against human serum-mediated xenogeneic humoral killing and its underlying mechanisms. Methods: BMSC of SD rats were isolated and cultured. The 4th generation of rBMSC was used as experimental material and rat lymphocytes (rLC) as control. Flow cytometry was used to examine the expression of α-Gal, a heterologous antigen of two kinds of cells. The cytotoxicity of 20% normal human serum on two kinds of cells was compared with that of normal human serum Ig M and Ig G , And the deposition of complement C3c, C4c, C5b-9 on the two kinds of cells after being treated with normal serum. Results: r BMSCs were successfully isolated and cultured. Compared with rLC, rBMSC had a significant resistance to human serum-mediated xenogeneic humoral killing (P <0.01); α-Gal expression on r BMSC was significantly lower than r LC (P <0.05); r BMSC and The binding of Ig G to Ig M in human serum was significantly lower than that of r LC (P <0.01). Significant accumulation of C3c, C4c and C5b-9 was observed in r LC after exposure to normal human serum, but r BMSC was less C3c and C4c deposition, the difference between the two kinds of cells was statistically significant (P <0.01). CONCLUSION: rBMSC can significantly resist the human serum-mediated xenogeneic humoral immune-killing effect, which may be related to the low expression of α-Gal of rBMSC and the inhibition of the formation of complement membrane.