摘要
目的:观察独活寄生汤血清对膝骨性关节炎退变软骨细胞Aggrecan和Collagen X mRNA表达的影响。方法:选择6例患者标本均来源于云南省中医医院骨科膝骨性关节炎住院患者,行手术截取的新鲜退变软骨组织,采用组织块法培养原代细胞;MTT法检测传代第一代细胞生长情况,并绘制生长曲线;制备独活寄生汤大鼠含药血清;均取传代第一代的退变软骨细胞,配制5%、10%、20%独活寄生汤含药血清及相应体积比浓度的生理盐水血清作为对照;MTT法检测不同浓度的独活寄生汤血清对退变软骨细胞增殖的影响;实时荧光定量PCR检测不同给药时间点各组退变软骨细胞Aggrecan和Collagen Ⅹ mRNA的表达。结果:退变软骨细胞传代第一代生长曲线见传代第8天时细胞增殖达到顶峰,故选择细胞传代第8天为给药干预时间点;与含同体积比的生理盐水血清组比较,不同浓度的独活寄生汤血清可对退变软骨细胞增殖产生促进作用(P<0.01),尤以10%独活寄生汤血清更为明显(P<0.05);与含同体积比生理盐水血清组比较,不同时间点的5%、10%、20%独活寄生汤血清干预后的退变软骨细胞Aggrecan mRNA的表达均上调(P<0.01),Collagen Ⅹ mRNA的表达均下调(P<0.01),且以10%独活寄生汤血清组的作用最为明显(P<0.05)。结论:独活寄生汤可能通过调节退变软骨组织的胶原及其蛋白多糖的表达而产生延缓膝骨性关节炎退变软骨组织的部分作用。
Objective： To observe Aggrecan and Collagen Ⅹ mRNA expression of the degeneration knee osteoarthritis chondrocytes by Duhuo Jisheng Decoction serum intervening. Methods： To select 6 cases of inpatients diagnosed knee osteoarthritis from Yunnan Province Hospital of Traditional Chinese Medicine to remove degeneration cartilage tissue. Small tissue block was used to culture primary chondrocytes, and rat serum containing Duhuo Jisheng Decoction was prepared. MTT method was used to detect proliferation rate of degenerative chondrocytes of the first passage generation and draw growth curve. 5%, 10%, 20% serum containing Duhuo Jisheng Decoction were made, and the corresponding concentration serum of saline was used as control group. MTT method was used to detect different concentrations of Duhuo Jisheng Decoction. Serum effecting on growth of degenerative knee osteoarthritis chondrocytes of the first passage generation were observed by real time quantitative PCR detected the Aggrecan and Collagen Ⅹ mRNA expressions. Results： The growth curve of the first passage generation chondrocytes of the degeneration knee osteoarthritis showed that chondrocytes proliferation peak was the 8 th day, so the 8 th day of cell passage was selected as the time point of drug administration intervention. Compared with the control groups, different concentrations of Duhuo Jisheng Decoction serum groups had promoted chondrocytes different degree proliferation（P0.01）, especially the 10% concentration was more obvious（P0.05）. At different time points, compared with control group, after intervention of 5%, 10%, 20% containing Duhuo Jisheng Decoction serum, Aggrecan mRNA expression in degeneration chondrocytes were increased（P0.01）, Collagen Ⅹ mRNA expression were reduced（P0.05）, especially the 10% concentration was more obvious（P0.05）. Conclusion： Duhuo Jisheng Decoction effectively prevents and treats degeneration knee osteoarthritis, which may probably regulate collagen and proteoglycan expression in degeneration cartilage tissue.
作者
李具宝
何瑞建
高焕焕
韦佳佳
彭文潘
侯刚
蒯永丽
李骥征
周柏健
张伟强
向俊宜
LI Ju-bao;HE Rui-jian;GAO Huan-huan;WEI Jia-jia;PENG Wen-pan;HOU Gang;KUAI Yong-li;LI Ji-zheng;ZHOU Bo-jian;ZHANG Wei-qiang;XIANG Jun-yi(yunnan Province Hospital of Traditional Chinese Medicine,Kunming 650021,China;yunnan University of Traditional Chinese Medicine,Kunming 650500,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2018年第11期5125-5129,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金地区科学基金项目(No.81460736)
云南省科技计划面上项目(No.2016FB141)
云南省医疗卫生单位内设研究机构科研项目(No.2016NS170
No.2016NS171)~~
