龙须藤总黄酮对膝关节炎软骨细胞增殖、凋亡的影响研究  

Study on Effect of Total Flavones from Bauhinia Championii on Proliferation and Apoptosis of Chondrocytes in Gonarthritis

作　　者：任安龙 季向荣[1] 方斌 REN Anlong;JI Xiangrong;FANG Bin

机构地区：[1]义乌市中心医院骨伤科,浙江义乌322000

出　　处：《新中医》2020年第23期12-16,共5页New Chinese Medicine

摘　　要：目的:探讨龙须藤总黄酮对膝关节炎软骨细胞增殖、凋亡的影响及其对长链非编码核糖核酸(LncRNA)MEG3的调控作用。方法:分离培养大鼠软骨细胞,对照1组为正常培养的软骨细胞,对照2组在含10 ng/L白细胞介素-1β(IL-1β)的培养基培养24 h,药物1组在含1μg/mL龙须藤总黄酮及10 ng/L IL-1β的培养基培养24 h,药物2组在含2μg/mL龙须藤总黄酮及10 ng/L IL-1β的培养基培养24 h,药物3组在含3μg/mL龙须藤总黄酮及10 ng/L IL-1β的培养基培养24 h,药物3+si-NC组用si-NC转染入软骨细胞后加入含3μg/mL的龙须藤总黄酮与10 ng/L IL-1β的培养基共同处理24 h,药物3+si-MEG3组用si-MEG3转染入软骨细胞后加入含3μg/mL的龙须藤总黄酮与10 ng/L IL-1β的培养基共同处理24 h。分别将pcDNA3.1、pcDNA3.1-MEG3转染至软骨细胞,用含有10 ng/L IL-1β的培养基培养24 h,分别记为pcDNA3.1组、pcDNA3.1-MEG3组。检测各组细胞存活率、凋亡率及MEG3、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、P21、半胱氨酰天冬氨酸特异性蛋白酶-3(Caspase-3)、B淋巴细胞瘤-2相关X蛋白(Bax)表达情况。结果:与对照1组比较,对照2组细胞存活率及MEG3表达降低(P<0.05),凋亡率及P21、Caspase-3、Bax、IL-6、TNF-α表达升高(P<0.05);与对照2组比较,药物1组、药物2组、药物3组细胞存活率及MEG3表达升高(P<0.05),凋亡率及P21、Caspase-3、Bax、IL-6、TNF-α表达降低(P<0.05);与pcDNA3.1组比较,pcDNA3.1-MEG3组细胞存活率及MEG3表达升高(P<0.05),凋亡率及P21、Caspase-3、Bax、IL-6、TNF-α表达降低(P<0.05);与药物3+si-NC组比较,药物3+si-MEG3组细胞存活率及MEG3表达降低(P<0.05),凋亡率及P21、Caspase-3、Bax、IL-6、TNF-α表达升高(P<0.05)。结论:龙须藤总黄酮可能通过上调MEG3表达促进膝关节炎软骨细胞增殖及抑制细胞凋亡,同时抑制炎症反应从而保护软骨细胞。Objective:To discuss the effect of total flavones from Bauhinia championii on proliferation and apoptosis of chondrocytes in gonarthritis and its regulatory effect on long non-coding RNAs(LncRNAs)maternally expressed gene 3(MEG3).Methods:Chondrocytes of rats were separated and cultured;the control group 1 was normally cultured chondrocytes;the control group 2 was cultured for 24 hours in medium with 10 ng/L of interleukin-1β(IL-1β);the medicine group 1 was cultured for 24 hours in medium with 1μg/mL of total flavones from Bauhinia championii and 10 ng/L of IL-1β;the medicine group 2 was cultured for 24 hours in medium with 2μg/mL of total flavones from Bauhinia championii and 10 ng/L of IL-1β;the medicine group 3 was cultured for 24 hours in medium with 3μg/mL of total flavones from Bauhinia championii and 10 ng/L of IL-1β;in the medicine 3+si-NC group,the chondrocytes were transferred with si-NC,and then treated with 3μg/mL of total flavones from Bauhinia championii and 10 ng/L of IL-1βfor 24 hours;in the medicine 3+si-MEG3 group,the chondrocytes were transferred with si-MEG3,and then treated with 3μg/mL of total flavones from Bauhinia championii and 10 ng/L of IL-1βfor 24 hours.The chondrocytes were transferred with pcDNA3.1 and pcDNA3.1-MEG3 respectively,and then cultured with for 24 hours in medium with 10 ng/L of IL-1β,namely the pcDNA3.1 group and the pcDNA3.1-MEG3 group.The cell viability,apoptosis rate and the expressions of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),P21,Caspase-3 and BCL-2-associated X protein(Bax)in each group were detected.Results:Compared with those in the control group 1,the cell viability and the MEG3 expression in the control group 2 were decreased(P<0.05),and the apoptosis rate and the expressions of P21,Caspase-3,Bax,IL-6 and TNF-αwere increased(P<0.05).Compared with those in the control group 2,the cell viability and the MEG3 expression in the medicine group 1,2 and 3 were increased(P<0.05),and the apoptosis rate and the expressions of P21,Caspase-3,Bax,I

关 键 词：膝关节炎 龙须藤总黄酮 LncRNA MEG3 软骨细胞 增殖 凋亡 细胞实验 

分 类 号：R285.5[医药卫生—中药学]