摘要
目的 观察槲皮素通过影响B细胞淋巴瘤/白血病-2相关X蛋白（bax）、B细胞淋巴瘤/白血病-2 （bcl-2）表达及其对诱导人类风湿性关节炎成纤维样滑膜细胞（RAFLSs）凋亡作用的影响.方法 收集类风湿性关节炎（RA）患者的滑膜组织标本,RA的诊断标准符合美国风湿病学协会修订的诊断标准.采用胰蛋白酶消化法分离滑膜细胞,用光学显微镜观察细胞的形态,流式细胞术分析体外培养的RAFLSs表面标记的白细胞分化抗原3（CD3）、CD19、CD14、以及CD90的表达.选用不同浓度的槲皮素（0、100、200、300 μmol/L）处理RAFLSs 48 h.利用Western blot法检测槲皮素处理后RAFLSs中促凋亡蛋白bax和抗凋亡蛋白bcl-2的灰度值,DNA片段化试验检测槲皮素对RAFLSs凋亡率,分析bcl-2/bax值对槲皮素处理后成纤维样滑膜细胞（FLSs）凋亡率的影响.结果 以槲皮素浓度为0 μmol/L时bcl-2/bax值（8.26±0.08）作为溶剂对照组,槲皮素浓度为100、200、300 μmol/L时处理48 h bcl-2/bax值（3.23±0.1l、0.26 ±0.05、0.12 ±0.03）,与对照组比较bcl-2/bax显著减小,差异有统计学意义（P＜0.01）.与槲皮素浓度为0tμmol/L对照组的RAFLSs的凋亡率[（6.5±0.9）％]比较,100 μmol/L槲皮素处理48 h RAFLSs的凋亡率[（19.9±3.8）％]增加（P＜0.05）,而200、300 μmol/L槲皮素处理48 h RALFSs的凋亡率[（34.3±6.7）％和（42.5±4.8）％]则显著增加,差异有统计学意义（P＜0.01）.结论 槲皮素能够通过调控bax、bcl-2的表达来影响RAFLSs的凋亡,且RAFLSs凋亡率与bcl-2/bax值成反比.
Objective To observe the effect of Quercetin regulating the expression levels of B cell lymphoma/leukemia-2 associated X protein （bax）,B cell lymphoma/leukemia-2 （bcl-2） on the apoptosis of human rheumatoid arthritis fibroblasts-like synoviocytes （RAFLSs）.Methods The synovial tissues were collected from patients with rheumatoid arthritis （RA）.The standard of diagnosis of RA was the criteria revised by American College of Rheumatology （ACR）.Fresh synovial tissues were chopped into fragments,washed in sterile phosphate-buffered saline （PBS）,and digested with collagenase to obtain synoviocytes.Human RAFLSs morphology was observed under light microscope.Flow cytometry was used to measure the expression of cluster of differentiation 3 （CD3）,CD14,CD19 and CD90 on the cell surface of RAFLSs.Human RAFLSs were treated with different concentrations of quercetin （0,100,200,and 300 μmol/L） for 24 h.Western blotting was used to detect the expression of anti-apoptotic protein bcl-2 and pro-apoptotic protein bax.The effect of quercetin through bcl-2/bax gray ratio on the apoptosis of fibroblast-like synoviocytes （FLSs） was analyzed.Results As compared with control group （the bcl-2/bax value was 8.26 ±0.08）,the bcl-2/bax in the quercetin-treated groups （100,200,and 300 μmol/L）（bcl-2/bax values were 3.23 ± 0.1 1,0.26 ± 0.05 and 0.12 ± 0.03 respectively） significantly decreased （P 〈0.01）.The apoptosis rate of RAFLSs in the control group （6.5% ± 0.9%） was significantly lower than in 100 μmo]/L quercetin-treated group （19.9% ± 3.8%） （P 〈 0.05）,and 200,300 μmol/L quercetin-treated groups （34.3% ± 6.7% and 42.5% ± 4.8% respectively） （P 〈 0.01）.Conclusion Quercetin can regulate the expression of bax and bcl-2 to influence RAFLSs apoptosis,and the apoptosis rate of RAFLSs is inversely proportional to the bcl-2/bax value.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2016年第10期2378-2380,共3页
Chinese Journal of Experimental Surgery
基金
河南省高等学校重点科研项目（14A310029）
