异基因骨髓间充质干细胞移植对胶原诱导关节炎大鼠血清多种细胞因子的影响

发布时间：2018-06-06 04:02

  本文选题：BMSCs + 关节炎　； 参考：《山西医科大学》2015年硕士论文

【摘要】：目的:观察BMSCs移植对胶原诱导关节炎(CIA)大鼠血清中TNF-α、IL-6、IL-17、RANKL、OPG、RANTES、MCP-1、IP-10、VEGF的影响,探讨BMSCs免疫调节作用的机制。方法:1.体外分离、培养Wistar大鼠BMSCs,流式细胞术检测表面抗原CD44、CD105、CD29、CD34、CD45、CD31;BMSCs分化为成骨细胞、脂肪细胞的鉴定。2.实验分组:CIA早期对照组、CIA晚期对照组、BMSCs早期干预组、BMSCs晚期干预组、MTX干预组、空白对照组,每组8只。3.给药方法:按1×107/kg/鼠计算,将适量BMSCs注到大鼠体内,MTX为按0.9mg/kg给药,1次/周。4.检测指标:42d时,流式多因子检测技术检测外周血中TNF-α、IL-6、IL-17、RANKL、OPG、MCP-1、RANTES、IP-10、VEGF的表达水平,收集数据做统计分析。结果:1.经流式细胞术检测:CD44、CD105、CD29阳性,CD45、CD34、CD31阴性。BMSCs诱导成骨、成脂分化能力强。按照间充质干细胞国际标准,证实获得的细胞为BMSCs。2.CIA模型组大鼠血清中TNF-α、IL-6、RANKL、RANTES、MCP-1、IP-10、VEGF水平明显较空白对照组升高,P0.05;IL-17a、OPG在CIA模型组与空白对照组间比较差异无统计学意,P0.05。3.与CIA模型组比较,MTX治疗组大鼠血清中TNF-α、IL-6、RANKL、RANTES、MCP-1、IP-10水平明显下降,P0.05。MTX治疗组VEGF下降不明显,与CIA模型组比较P0.05。4.早期BMSCs治疗组较早期CIA模型组TNF-α、IL-6明显降低,P0.05;晚期BMSCs治疗组较晚期CIA模型组TNF-α、IL-6明显下降,P0.05;早期BMSCs治疗组与MTX治疗组比较TNF-α、IL-6水平接近,差异无统计学意义P0.05;晚期BMSCs治疗组TNF-α、IL-6水平也有下降趋势,但较MTX治疗组下降不明显,P0.05。5.早期BMSCs治疗组较早期CIA模型组RANTES、IP-10明显下降,P0.05,但晚期BMSCs治疗组与晚期CIA模型组比较RANTES、IP-10差别无统计学意义,P0.05;早期BMSCs治疗组较晚期BMSCs治疗组RANTES、IP-10下降更明显,P0.05;MTX治疗组与早期BMSCs治疗组、晚期BMSCs治疗组的RANTES、IP-10均无统计学差别,P0.05。6.早期BMSCs治疗组较早期CIA模型组MCP-1明显下降,P0.05,但晚期BMSCs治疗组与CIA模型组比较RANTES差别无统计学意义,P0.05;早期BMSCs治疗组较晚期BMSCs治疗组MCP-1下降更明显,差别有统计学意义P0.05;MTX治疗组与早期BMSCs治疗组MCP-1水平无统计学差别P0.05,晚期BMSCs治疗组与MTX治疗组比较MCP-1水平有统计学差异,P0.05。结论:1.CIA大鼠中存在免疫功能紊乱,血清中TNF-α、IL-6、RANKL、RANTES、MCP-1、IP-10、VEGF水平明显升高。2.BMSCs可通过抑制血清中TNF-α、IL-6、RANTES、MCP-1、IP-10水平而发挥免疫调节作用。3.BMSCs干预CIA大鼠早期比晚期疗效好。
[Abstract]:Aim: to observe the effect of BMSCs transplantation on VEGF in serum of rats with collagen induced arthritis (CIA-TNF- 伪) and investigate the mechanism of immunomodulatory effect of BMSCs. Method 1: 1. BMSCs of Wistar rats were isolated and cultured in vitro, and the surface antigen CD44-CD105- CD105- CD29CD34-CD4- CD45-CD31- BMSCs differentiated into osteoblasts and adipocytes were identified by flow cytometry (FCM). The results showed that BMSCs were differentiated into osteoblasts and adipocytes were identified by flow cytometry (FCM). The experimental group was divided into two groups: the early control group (control group) and the late control group (BMSCs group), the late intervention group (MTX group) and the blank control group (8 rats in each group). Methods: according to 1 脳 107/kg/ mice, appropriate amount of BMSCs was injected into rats as 0.9mg/kg once a week. 4. Flow multifactor technique was used to detect the expression of VEGF in peripheral blood of TNF- 伪, IL-6, IL-17, RANKL, OPG, MCP-1 and RANTESU IP-10, and the data were collected for statistical analysis. The result is 1: 1. Flow cytometry was used to detect CD44 + CD105 + CD29 + CD45 + CD34 + CD31 negative. BMSCs were able to induce osteogenesis and the ability of adipogenic differentiation was strong. According to the international standard of mesenchymal stem cells, it was confirmed that the level of VEGF in serum of BMSCs.2. CIA model group was significantly higher than that of blank control group. There was no significant difference between CIA model group and control group (P 0.05.3). Compared with the model group of CIA, the serum levels of TNF- 伪 IL-6 and RANKLRANTESS-MCP-1 / IP-10 in the MTX group were significantly decreased. The VEGF level in the treatment group was not significantly decreased compared with that in the CIA model group, but it was not significantly decreased in the MTX group compared with that in the CIA model group (P0.05.4). In the early BMSCs treatment group, TNF- 伪 IL-6 decreased significantly compared with the early CIA model group, while in the late BMSCs treatment group, the TNF- 伪 IL-6 decreased significantly compared with the late CIA model group, and the level of TNF- 伪 IL-6 in the early BMSCs treatment group was similar to that in the MTX group, and the level of TNF- 伪 IL-6 in the BMSCs treatment group was similar to that in the MTX group. The level of TNF- 伪 and IL-6 in late BMSCs group also decreased, but it was not significantly decreased compared with MTX treatment group (P0.05.5). In the early BMSCs treatment group, RANTESlIP-10 decreased significantly than that in the early CIA model group, but there was no significant difference between the advanced BMSCs treatment group and the advanced CIA model group (P 0.05), while in the early BMSCs treatment group compared with the late BMSCs treatment group, the decrease of RANTESn IP-10 was significantly lower than that in the late BMSCs treatment group and the early BMSCs treatment group, and the P0.05MTX group and the early BMSCs treatment group were significantly lower than those in the late BMSCs treatment group. There was no significant difference in RNTES- IP-10 in advanced BMSCs treatment group (P0.05.6). MCP-1 decreased significantly in the early BMSCs treatment group compared with the early CIA model group, but there was no significant difference in RANTES between the late BMSCs treatment group and the CIA model group, and the decrease of MCP-1 in the early BMSCs treatment group was more significant than that in the late BMSCs treatment group, but in the late BMSCs treatment group, there was no significant difference in RANTES between the early BMSCs treatment group and the CIA model group. There was no significant difference in MCP-1 level between the BMSCs treatment group and the early BMSCs treatment group (P 0.05), while the MCP-1 level in the late BMSCs treatment group was significantly higher than that in the MTX treatment group (P 0.05). Conclusion: 1. There is immune dysfunction in CIA rats. The level of VEGF in serum TNF- 伪 IL-6 RANKLRANTES-1 / MCP-10 may play an immunomodulatory role by inhibiting the level of TNF- 伪 -IL-6, RANTES+ MCP-1and IP-10 in CIA rats. 3. BMSCs have a better therapeutic effect in the early stage than in the late stage in the intervention of CIA rats with the level of TNF- 伪, IL-6, RANTES, MCP-1and IP-10.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R593.22

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