摘要
目的:探讨荣筋拈痛方对白细胞介素-1β(IL-1β)诱导退变软骨细胞增殖的作用机制。方法:CCK8法筛选荣筋拈痛方对退变软骨细胞最佳干预条件后,将软骨细胞随机分为正常组、模型组、抑制剂组和荣筋拈痛方组,分组干预后采用激光共聚焦观察各组Ⅱ型胶原蛋白表达;流式细胞术检测各组周期分布情况;RTPCR与Western Blot检测各组G1/S检测点相关调节因子p16、Cyclin D1、CDK4、pRB基因和蛋白表达。结果:荣筋拈痛方100μg/mL干预退变软骨细胞48h可显著增强软骨细胞活力(P<0.01)。与正常组比较,模型组和抑制剂组Ⅱ型胶原蛋白表达量显著降低(P<0.01);与模型组和抑制剂组比较,荣筋拈痛方组Ⅱ型胶原蛋白表达量显著增加(P<0.01)。与正常组比较,模型组和抑制剂组G0/G1期比例升高,G2/M期、S期比例显著降低(P<0.01);与模型组和抑制剂组比较,荣筋拈痛方组G0/G1期比例显著降低,G2/M期、S期比例显著升高(P<0.01)。与正常组比较,模型组和抑制剂组Cyclin D1、CDK4、pRB基因和蛋白表达量显著降低,p16基因和蛋白表达量显著增加(P<0.01,P<0.05);与模型组和抑制剂组比较,荣筋拈痛方组Cyclin D1、CDK4、pRB基因和蛋白表达量显著增加,p16基因和蛋白表达量显著降低(P<0.01)。结论:荣筋拈痛方可调控软骨细胞G1/S期的转换,促进Ⅱ型胶原蛋白表达和软骨细胞增殖。
Objective:To investigate the mechanism of Rongjin Niantong Formula on the proliferation of degenerative chondrocytes induced by interleukin-1β(IL-1β).Methods:After screening the best intervention conditions of Rongjin Niantong Formula on degenerative chondrocytes by CCK8 method,chondrocytes were randomly divided into normal group,model group,inhibitor group and Rongjin Niantong Formula group.After group intervention,the expression of type II collagen in each group was observed by laser confocal method;The cycle distribution of each group was detected by flow cytometry;RTPCR and Western Blot were used to detect the gene and protein expressions of regulatory factors p16,Cyclin D1,CDK4,and pRB at G1/S detection points.Results:Rongjin Niantong Formula 100μg/mL for 48 h could significantly enhance the activity of degenerative chondrocytes(P<0.01).Compared with the normal group,the expression of type II collagen in the model group and the inhibitor group was reduced(P<0.01);Compared with the model group and the inhibitor group,the expression of type II collagen in the Rongjin Niantong Formula group increased(P<0.01).Compared with the normal group,the proportion of G0/G1 phase was increased in the model group and the inhibitor group,while the proportion of G2/M phase and S phase was decreased(P<0.01).Compared with model group and inhibitor group,the proportion of G0/G1 phase in Rongjin Niantong Formula group was decreased,while the proportion of G2/M phase and S phase was increased(P<0.01).Compared with normal group,the expression levels of Cyclin D1,CDK4,pRB gene and protein in model group and inhibitor group were decreased,while the expression levels of p16 gene and protein were increased(P<0.01,P<0.05).Compared with model group and inhibitor group,the expression levels of Cyclin D1,CDK4,pRB gene and protein in Rongjin Niantong Formula group were increased,while the expression levels of p16 gene and protein were decreased(P<0.01).Conclusion:Rongjin Niantong Formula can regulate the transition of chondrocytes in G1/S phase,promote the expression of type II collagen and the proliferation of chondrocytes.
作者
黄艳峰
陈俊
林洁
赵忠胜
叶锦霞
李西海
吴广文
HUANG Yan-feng;CHEN Jun;LIN Jie;ZHAO Zhong-sheng;YE Jin-xia;LI Xi-hai;WU Guang-wen(Academy of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;Fujian Key Laboratory of Integrative Medicine on Geriatrics,Fuzhou 350122,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2021年第4期2077-2082,共6页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金面上项目(No.82074465)
福建省自然科学基金面上项目(No.2020J01750)
陈可冀中西医结合发展基金(No.CKJ2020005)。
关键词
荣筋拈痛方
膝骨性关节炎
本痿标痹
白细胞介素-1Β
退变软骨细胞
细胞周期
增殖
大鼠
Rongjin Niantong Formula
Knee osteoarthritis
Deficiency in origin and excess in superficidity
Interleukin-1β
Degenerative chondrocytes
Cell cycle
Proliferation
Rats
