本文選題：骨性關(guān)節(jié)炎 + 細(xì)胞因子��； 參考：《武漢大學(xué)》2015年博士論文

【摘要】：目的：判斷原發(fā)性膝關(guān)節(jié)骨性關(guān)節(jié)炎患者的血清及關(guān)節(jié)滑液中的IL-17濃度,評(píng)估IL-17濃度與骨性關(guān)節(jié)炎嚴(yán)重程度之間的相關(guān)性。觀察bFGF對(duì)大鼠BMSCs體外增殖及其分化潛能的影響。方法： (1)收集原發(fā)性膝關(guān)節(jié)骨性關(guān)節(jié)炎患者的血清及關(guān)節(jié)滑液。收集年齡、性別相匹配的健康人群的血清樣本。酶聯(lián)免疫吸附試驗(yàn)法定量測(cè)定IL-17濃度。骨性關(guān)節(jié)炎的嚴(yán)重程度使用Lequesne指數(shù)評(píng)分標(biāo)準(zhǔn)和Kellgren-Lawrence放射學(xué)診斷標(biāo)準(zhǔn)評(píng)分分級(jí)進(jìn)行評(píng)估。(2)體外培養(yǎng)大鼠BMSCs,分別設(shè)對(duì)照組和bFGF處理組,bFGF作用濃度分別為1、10、1001μg/L。作用72 h后,噻唑藍(lán)(MTT)測(cè)吸光度(A)值反映細(xì)胞增殖,細(xì)胞免疫組織化學(xué)測(cè)細(xì)胞CD44的累積A值,逆轉(zhuǎn)錄-聚合酶鏈反應(yīng)(RT-PCR)檢測(cè)細(xì)胞CD44的mRNA相對(duì)表達(dá)量。結(jié)果：(1)實(shí)驗(yàn)組的血清IL-17濃度顯著高于對(duì)照組(P0.05)。將患者根據(jù)KL評(píng)分分組后,KL評(píng)分4級(jí)的患者血清IL-17濃度顯著高于對(duì)照組(P0.05)。KL評(píng)分2級(jí)、KL評(píng)分3級(jí)和對(duì)照組之間的血清IL-17濃度無顯著性差異。KL評(píng)分3級(jí)、KL評(píng)分4級(jí)的關(guān)節(jié)滑液IL-17濃度顯著高于KL評(píng)分2級(jí)(P0.05),KL評(píng)分4級(jí)的關(guān)節(jié)滑液IL-17濃度顯著高于KL評(píng)分3級(jí)(P0.05)。關(guān)節(jié)滑液IL-17濃度與Lequesne指數(shù)具有相關(guān)性(r=0.6232,P0.05),但是血清IL-17濃度與Lequesne指數(shù)不具有相關(guān)性。(2)在1ug/LbFGF作用下其MTTA值為0.334±0.036,較對(duì)照組A值0.251±0.033明顯增高(P0.05)。在1ug/LbFGF作用下其細(xì)胞免疫組織化學(xué)測(cè)得CD44的累積A值較對(duì)照組明顯增高(P0.01),其RT-PCR測(cè)得CD44的mRNA相對(duì)表達(dá)量較對(duì)照組明顯增高(P0.01)。結(jié)論：(1)原發(fā)性膝關(guān)節(jié)骨性關(guān)節(jié)炎患者關(guān)節(jié)滑液中的IL-17濃度與骨性關(guān)節(jié)炎的嚴(yán)重程度具有高度的正相關(guān)。IL-17可以作為一種有用的生化標(biāo)記物來反應(yīng)骨性關(guān)節(jié)炎的嚴(yán)重程度及進(jìn)程。(2)低濃度的bFGF促進(jìn)BMSCs快速增殖的同時(shí),保證了BMSCs高增殖的同時(shí)又使其處于未分化狀態(tài)。這種改良培養(yǎng)可獲得高增殖率的BMSCs,并且保證了BMSCs較好的分化潛能。這種優(yōu)化的擴(kuò)增措施,在BMSCs應(yīng)用于骨、軟骨、脂肪等組織工程方面具有較好的應(yīng)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 鹵0.036 when treated with 1ug/LbFGF, which was significantly higher than that of control group A (0.251 鹵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.
【學(xué)位授予單位】：武漢大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類號(hào)】：R684.3

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