细胞因子在骨性关节炎的发生及治疗方面的相关研究

发布时间：2018-05-07 15:14

本文选题：骨性关节炎 + 细胞因子　；参考：《武汉大学》2015年博士论文

【摘要】：目的：判断原发性膝关节骨性关节炎患者的血清及关节滑液中的IL-17浓度,评估IL-17浓度与骨性关节炎严重程度之间的相关性。观察bFGF对大鼠BMSCs体外增殖及其分化潜能的影响。方法： (1)收集原发性膝关节骨性关节炎患者的血清及关节滑液。收集年龄、性别相匹配的健康人群的血清样本。酶联免疫吸附试验法定量测定IL-17浓度。骨性关节炎的严重程度使用Lequesne指数评分标准和Kellgren-Lawrence放射学诊断标准评分分级进行评估。(2)体外培养大鼠BMSCs,分别设对照组和bFGF处理组,bFGF作用浓度分别为1、10、1001μg/L。作用72 h后,噻唑蓝(MTT)测吸光度(A)值反映细胞增殖,细胞免疫组织化学测细胞CD44的累积A值,逆转录-聚合酶链反应(RT-PCR)检测细胞CD44的mRNA相对表达量。结果：(1)实验组的血清IL-17浓度显著高于对照组(P0.05)。将患者根据KL评分分组后,KL评分4级的患者血清IL-17浓度显著高于对照组(P0.05)。KL评分2级、KL评分3级和对照组之间的血清IL-17浓度无显著性差异。KL评分3级、KL评分4级的关节滑液IL-17浓度显著高于KL评分2级(P0.05),KL评分4级的关节滑液IL-17浓度显著高于KL评分3级(P0.05)。关节滑液IL-17浓度与Lequesne指数具有相关性(r=0.6232,P0.05),但是血清IL-17浓度与Lequesne指数不具有相关性。(2)在1ug/LbFGF作用下其MTTA值为0.334±0.036,较对照组A值0.251±0.033明显增高(P0.05)。在1ug/LbFGF作用下其细胞免疫组织化学测得CD44的累积A值较对照组明显增高(P0.01),其RT-PCR测得CD44的mRNA相对表达量较对照组明显增高(P0.01)。结论：(1)原发性膝关节骨性关节炎患者关节滑液中的IL-17浓度与骨性关节炎的严重程度具有高度的正相关。IL-17可以作为一种有用的生化标记物来反应骨性关节炎的严重程度及进程。(2)低浓度的bFGF促进BMSCs快速增殖的同时,保证了BMSCs高增殖的同时又使其处于未分化状态。这种改良培养可获得高增殖率的BMSCs,并且保证了BMSCs较好的分化潜能。这种优化的扩增措施,在BMSCs应用于骨、软骨、脂肪等组织工程方面具有较好的应用前景。
[Abstract]:Aim: to determine the concentration of IL-17 in serum and synovial fluid of patients with primary knee osteoarthritis and to evaluate the correlation between IL-17 concentration and the severity of osteoarthritis. To observe the effect of bFGF on proliferation and differentiation potential of rat BMSCs in vitro. Methods: serum and synovial fluid were collected from patients with primary knee osteoarthritis. Collect serum samples from healthy people matched for age and sex. Enzyme linked immunosorbent assay (Elisa) was used to determine the concentration of IL-17. The severity of osteoarthritis was evaluated by Lequesne index score and Kellgren-Lawrence radiologic diagnostic criteria. BMSCs were cultured in vitro. The concentration of bFGF in control group and bFGF treated group was 110 渭 g / L and 100 1 渭 g / L, respectively. After 72 h exposure, the absorbance (A) value of thiazolyl blue was measured to reflect cell proliferation, the cumulative A value of cell CD44 was measured by cell immunohistochemistry, and the relative mRNA expression of CD44 was detected by reverse transcription-polymerase chain reaction (RT-PCR). Results the serum IL-17 concentration in the experimental group was significantly higher than that in the control group (P 0.05). The serum IL-17 concentration of the patients with KL grade 4 was significantly higher than that of the control group (P0.05U. KL grade 2, KL grade 3) and the control group. There was no significant difference in serum IL-17 concentration between the control group and the control group. There was no significant difference in the serum IL-17 concentration between the patients with KL score 4 and the control group with KL score 3 and KL grade 4. The concentration of IL-17 in synovial fluid was significantly higher than that in grade 2, grade 2 and grade 4, the concentration of IL-17 in synovial fluid was significantly higher than that in grade 3 of KL score. There was a correlation between IL-17 concentration and Lequesne index in synovial fluid, but there was no correlation between serum IL-17 concentration and Lequesne index. The MTTA value of synovial fluid was 0.334 卤0.036 when treated with 1ug/LbFGF, which was significantly higher than that of control group A (0.251 卤0.033). Under the action of 1ug/LbFGF, the cumulative A value of CD44 was significantly higher than that of the control group, and the relative mRNA expression of CD44 detected by RT-PCR was significantly higher than that of the control group. Conclusion (1) the concentration of IL-17 in synovial fluid of patients with primary knee osteoarthritis is positively correlated with the severity of osteoarthritis. IL-17 can be used as a useful biochemical marker to reflect the severity of osteoarthritis. Low concentration of bFGF promoted the rapid proliferation of BMSCs at the same time, The high proliferation of BMSCs and its undifferentiated state were ensured at the same time. BMSCs with high proliferation rate could be obtained by this improved culture, and the differentiation potential of BMSCs was guaranteed. The optimized amplification method has a good prospect in the application of BMSCs in bone, cartilage, fat and other tissue engineering.
【学位授予单位】：武汉大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R684.3

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