本文選題：BMSCs + 關(guān)節(jié)炎��； 參考：《山西醫(yī)科大學(xué)》2015年碩士論文

【摘要】：目的:觀察BMSCs移植對膠原誘導(dǎo)關(guān)節(jié)炎(CIA)大鼠血清中TNF-α、IL-6、IL-17、RANKL、OPG、RANTES、MCP-1、IP-10、VEGF的影響,探討B(tài)MSCs免疫調(diào)節(jié)作用的機制。方法:1.體外分離、培養(yǎng)Wistar大鼠BMSCs,流式細胞術(shù)檢測表面抗原CD44、CD105、CD29、CD34、CD45、CD31;BMSCs分化為成骨細胞、脂肪細胞的鑒定。2.實驗分組:CIA早期對照組、CIA晚期對照組、BMSCs早期干預(yù)組、BMSCs晚期干預(yù)組、MTX干預(yù)組、空白對照組,每組8只。3.給藥方法:按1×107/kg/鼠計算,將適量BMSCs注到大鼠體內(nèi),MTX為按0.9mg/kg給藥,1次/周。4.檢測指標:42d時,流式多因子檢測技術(shù)檢測外周血中TNF-α、IL-6、IL-17、RANKL、OPG、MCP-1、RANTES、IP-10、VEGF的表達水平,收集數(shù)據(jù)做統(tǒng)計分析。結(jié)果:1.經(jīng)流式細胞術(shù)檢測:CD44、CD105、CD29陽性,CD45、CD34、CD31陰性。BMSCs誘導(dǎo)成骨、成脂分化能力強。按照間充質(zhì)干細胞國際標準,證實獲得的細胞為BMSCs。2.CIA模型組大鼠血清中TNF-α、IL-6、RANKL、RANTES、MCP-1、IP-10、VEGF水平明顯較空白對照組升高,P0.05;IL-17a、OPG在CIA模型組與空白對照組間比較差異無統(tǒng)計學(xué)意,P0.05。3.與CIA模型組比較,MTX治療組大鼠血清中TNF-α、IL-6、RANKL、RANTES、MCP-1、IP-10水平明顯下降,P0.05。MTX治療組VEGF下降不明顯,與CIA模型組比較P0.05。4.早期BMSCs治療組較早期CIA模型組TNF-α、IL-6明顯降低,P0.05;晚期BMSCs治療組較晚期CIA模型組TNF-α、IL-6明顯下降,P0.05;早期BMSCs治療組與MTX治療組比較TNF-α、IL-6水平接近,差異無統(tǒng)計學(xué)意義P0.05;晚期BMSCs治療組TNF-α、IL-6水平也有下降趨勢,但較MTX治療組下降不明顯,P0.05。5.早期BMSCs治療組較早期CIA模型組RANTES、IP-10明顯下降,P0.05,但晚期BMSCs治療組與晚期CIA模型組比較RANTES、IP-10差別無統(tǒng)計學(xué)意義,P0.05;早期BMSCs治療組較晚期BMSCs治療組RANTES、IP-10下降更明顯,P0.05;MTX治療組與早期BMSCs治療組、晚期BMSCs治療組的RANTES、IP-10均無統(tǒng)計學(xué)差別,P0.05。6.早期BMSCs治療組較早期CIA模型組MCP-1明顯下降,P0.05,但晚期BMSCs治療組與CIA模型組比較RANTES差別無統(tǒng)計學(xué)意義,P0.05;早期BMSCs治療組較晚期BMSCs治療組MCP-1下降更明顯,差別有統(tǒng)計學(xué)意義P0.05;MTX治療組與早期BMSCs治療組MCP-1水平無統(tǒng)計學(xué)差別P0.05,晚期BMSCs治療組與MTX治療組比較MCP-1水平有統(tǒng)計學(xué)差異,P0.05。結(jié)論:1.CIA大鼠中存在免疫功能紊亂,血清中TNF-α、IL-6、RANKL、RANTES、MCP-1、IP-10、VEGF水平明顯升高。2.BMSCs可通過抑制血清中TNF-α、IL-6、RANTES、MCP-1、IP-10水平而發(fā)揮免疫調(diào)節(jié)作用。3.BMSCs干預(yù)CIA大鼠早期比晚期療效好。
[Abstract]:Aim: to observe the effect of BMSCs transplantation on VEGF in serum of rats with collagen induced arthritis (CIA-TNF- 偽) 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- 偽, 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- 偽 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- 偽 IL-6 decreased significantly compared with the early CIA model group, while in the late BMSCs treatment group, the TNF- 偽 IL-6 decreased significantly compared with the late CIA model group, and the level of TNF- 偽 IL-6 in the early BMSCs treatment group was similar to that in the MTX group, and the level of TNF- 偽 IL-6 in the BMSCs treatment group was similar to that in the MTX group. The level of TNF- 偽 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- 偽 IL-6 RANKLRANTES-1 / MCP-10 may play an immunomodulatory role by inhibiting the level of TNF- 偽 -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- 偽, IL-6, RANTES, MCP-1and IP-10.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R593.22

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