黄芩总黄酮对人骨关节炎软骨细胞增殖凋亡及COL10A1/SOX9信号通路的影响  

Effects of Total Flavonoids in Scutellaria Baicalensis Georgi on the Proliferation and Apoptosis of Human Osteoarthritis Chondrocytes and COL10A1/SOX9 Signaling Pathway

作　　者：王冠贤 李宝林 唐爱民 黄永湘 Wang Guanxian;Li Baolin;Tang Aimin;Huang Yongxiang(Department of Orthopedics,Danzhou People's Hospital,Hainan Danzhou 571700,China)

机构地区：[1]儋州市人民医院骨科,海南儋州571700

出　　处：《中国药师》2021年第4期693-697,共5页China Pharmacist

基　　金：海南省自然科学基金项目(编号:817395)。

摘　　要：目的:探讨黄芩总黄酮对人骨关节炎(OA)软骨细胞增殖凋亡及X型胶原蛋白α1链(COL10A1)/性别决定区Y框蛋白9(SOX9)信号通路的影响。方法:从5例行膝关节置换术的OA患者的关节软骨分离软骨细胞,用甲苯胺蓝染色进行鉴定。实验分为对照组、黄芩总黄酮低、中、高剂量组(25,50,100 mg·L-1)和塞来昔布组(9 mg·L-1)。噻唑蓝(MTT)法测定人OA软骨细胞增殖情况,流式细胞仪测定细胞凋亡水平,免疫印迹法检测人OA软骨细胞中COL10A1、SOX9、细胞外信号调节激酶1/2(ERK1/2)、B细胞淋巴瘤/白血病-2(Bcl-2)和Bcl-2相关X蛋白(Bax)蛋白水平。结果:人OA软骨细胞附着于培养皿后呈多边形或长条形,甲苯胺蓝染色可见OA软骨细胞胞浆以蓝紫色为主,异染颗粒散布于细胞间或细胞周围;与对照组比较,黄芩总黄酮低、中、高剂量组和塞来昔布组人OA软骨细胞增殖率、COL10A1、SOX9、Bcl-2蛋白和Bcl-2/Bax水平升高,凋亡率、ERK1/2和Bax蛋白水平降低(P<0.05);随着黄芩总黄酮浓度的增加,黄芩总黄酮各剂量组人OA软骨细胞增殖率、COL10A1、SOX9、Bcl-2蛋白和Bcl-2/Bax水平逐渐升高,凋亡率、ERK1/2和Bax蛋白水平逐渐降低(P<0.05);黄芩总黄酮高剂量组与塞来昔布组作用相似,上述指标差异无统计学意义(P>0.05)。结论:黄芩总黄酮可能通过激活COL10A1/SOX9信号通路,抑制人OA软骨细胞凋亡,促进OA软骨细胞增殖。Objective: To investigate the effect of total flavonoids in Scutellaria baicalensis Georgi on the proliferation and apoptosis of human osteoarthritis chondrocytes and collagen type X alpha 1 chain(COL10A1)/sex determining region Y-box protein 9(SOX9)signaling pathway. Methods: Chondrocytes were isolated from articular cartilage of 5 OA patients who underwent knee replacement and identified by toluidine blue staining. The experimental groups included control group,total flavonoids in Scutellaria baicalensis Georgi low,medium and high dose groups(25,50 and 100 mg·L-1) and celecoxib group(9 mg·L-1). The proliferation of human OA chondrocytes was measured by MTT,the cell apoptosis was measured by flow cytometry,the protein levels of COL10A1,SOX9,extracellular signal regulated kinase 1/2(ERK1/2),B-cell lymphoma/leukemia-2(Bcl-2) and Bcl-2 related X protein(Bax) were detected by Western blot. Results: After human OA chondrocytes attached to the culture dish,they were polygonal or strip-shaped. Toluidine blue staining showed that the cytoplasm of OA chondrocytes was mainly blue and purple,and heterochromate particles were scattered between the cells or around the cells. Compared with those in the control group,the proliferation rate and levels of COL10A1,SOX9,Bcl-2 protein and Bcl-2/Bax of human OA chondrocytes in total flavonoids in Scutellaria baicalensis Georgi low,medium and high dose groups and celecoxib group were increased,and the apoptosis rate and levels of ERK1/2 and Bax protein were decreased(P<0.05);the proliferation rate and levels of COL10A1,SOX9,Bcl-2 protein and Bcl-2/Bax of human OA chondrocytes increased gradually with the concentration increase of total flavonoids in Scutellaria baicalensis Georgi,while the apoptosis rate and the levels of ERK1/2 and Bax protein decreased gradually(P<0.05). The effect of high dose total flavonoids in Scutellaria baicalensis Georgi group was similar to that of celecoxib group,and the difference was not statistically significant(P>0.05). Conclusion: The total flavonoi

关 键 词：黄芩总黄酮 骨关节炎 细胞增殖 细胞凋亡 X型胶原蛋白α1链/性别决定区Y框蛋白9信号通路 

分 类 号：R285.5[医药卫生—中药学]