發(fā)布時間：2018-06-27 01:26

  本文選題：茶黃素 + 骨關(guān)節(jié)炎�。� 參考：《浙江大學(xué)學(xué)報(農(nóng)業(yè)與生命科學(xué)版)》2017年04期

【摘要】：采用白介素-1β(interleukin-1β,IL-1β)對體外培養(yǎng)的正常大鼠軟骨細(xì)胞進(jìn)行誘導(dǎo),構(gòu)建骨關(guān)節(jié)炎(osteoarthritis,OA)炎性退變細(xì)胞模型,探討茶黃素(theaflavins,TFs)治療骨關(guān)節(jié)炎的可行性。通過甲苯胺藍(lán)及免疫熒光染色對細(xì)胞進(jìn)行鑒定,利用CCK-8細(xì)胞活力檢測篩選TFs藥物濃度;取第2代生長狀況良好的大鼠膝關(guān)節(jié)軟骨細(xì)胞,根據(jù)所加培養(yǎng)物的不同將實驗分為3組:空白組G0、模型組G1(10 ng/mL IL-1β刺激)、TFs組G2(不同濃度的TFs+10 ng/mL IL-1β刺激),通過實時熒光定量聚合酶鏈?zhǔn)椒磻?yīng)(quantitative real-time polymerase chain reaction,qRT-PCR)檢測各組Ⅱ型膠原(typeⅡcollagen,ColⅡ)、蛋白聚糖(aggrecan,ACAN)、基質(zhì)金屬蛋白酶-13(matrix metalloproteinases-13,MMP-13)、IL-1β及環(huán)氧合酶-2(cyclooxygenase-2,COX-2)mRNA的表達(dá)。結(jié)果表明,甲苯胺藍(lán)及免疫熒光染色結(jié)果呈陽性,證實體外分離培養(yǎng)的細(xì)胞為軟骨細(xì)胞。CCK-8檢測結(jié)果顯示:TFs質(zhì)量濃度在100μg/mL時,細(xì)胞活力較空白組明顯減弱(P0.05);TFs質(zhì)量濃度在0~75μg/mL時不影響軟骨細(xì)胞活力。qRT-PCR結(jié)果表明:與空白組相比,模型組中合成因子ColⅡ和ACAN mRNA表達(dá)量極顯著降低(P0.01),分解因子MMP-13、炎癥細(xì)胞因子IL-1β及炎癥誘導(dǎo)酶COX-2 mRNA表達(dá)量極顯著增加(P0.01);與模型組相比,TFs組能抑制IL-1β引起的炎癥相關(guān)因子mRNA的表達(dá),且呈濃度依賴性(P0.05)。綜上所述,茶黃素可通過增強軟骨細(xì)胞合成因子活性、減弱分解因子活性并抑制細(xì)胞炎癥反應(yīng),有效延緩大鼠軟骨細(xì)胞炎性退變進(jìn)程,對炎癥軟骨細(xì)胞有一定的保護(hù)作用。
[Abstract]:Normal rat chondrocytes were induced by interleukin-1 尾 (interleukin-1 尾). The inflammatory degeneration cell model of osteoarthritis (OA) was established, and the feasibility of theaflavinsTFs in the treatment of osteoarthritis was discussed. The cells were identified by toluidine blue and immunofluorescence staining, the concentration of TFs was screened by CCK-8 cell viability test, and the articular chondrocytes of the second generation of rat knee joint were obtained. The experiment was divided into three groups according to the different cultures: blank group G0, model group G1 (10 ng / mL IL-1 尾 stimulation) and TFs group G2 (different concentrations of TFs 10 ng / mL IL-1 尾 stimulation). Real time quantitative real-time polymerase chain quantitative polymerase chain reaction (QRT-PCR) was used to detect group 鈪，

本文編號：2072142