复元胶囊含药血清促进骨关节炎软骨细胞合成Ⅱ型胶原、蛋白多糖的机制研究  

Mechanism of Serum Containing Fuyuan Capsule in Promoting Synthesis of Collagen Type Ⅱ and Aggrecan in Osteoarthritis Chondrocytes

作　　者：张玉笛[1] 周凌云[1] 钟玉[1] 李荣亨[1] 

机构地区：[1]重庆医科大学附属第一医院中西医结合科,重庆400016

出　　处：《中药新药与临床药理》2015年第5期639-644,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：渝卫中医([2008]38号2008-2-35)

摘　　要：目的观察复元胶囊是否通过调节smad 3磷酸化水平促进骨关节炎软骨细胞合成Ⅱ型胶原(COL2A1)及蛋白多糖(ACAN)。方法体外培养人软骨瘤细胞株(SW1353),随机分为3组:空白组、白介素1β(IL-1β)组、IL-1β+复元胶囊组。以最适浓度15%复元胶囊含药血清进行实验干预。CCK8法观察细胞的生长情况;流式细胞术检测细胞周期;实时荧光定量PCR(q RT-PCR)检测细胞COL2A1、ACAN m RNA合成水平;Western blot检测细胞smad 3磷酸化水平。结果 IL-1β组的细胞生长曲线,细胞增殖率(25.50±0.63),COL2A1(0.523±0.009)、ACAN(0.201±0.035)m RNA及磷酸化smad 3(p-smad 3)(1.026±0.003)蛋白表达水平均低于空白组(P<0.05);IL-1β复元胶囊组的细胞生长曲线,细胞增殖率(37.55±0.71),COL2A1(0.701±0.068)、ACAN(0.901±0.230)m RNA及p-smad 3(1.527±0.003)蛋白表达水平均高于IL-1β组(P<0.01)。结论复元胶囊可以通过上调smad 3磷酸化水平,促进骨关节炎软骨细胞COL2A1及ACAN的合成,起到保护软骨细胞的作用。Objective To investigate the effect of Fuyuan capsule （FC）on the synthesis of collagen typeⅡ （COL2A1） and aggrecan （ACAN） by activating the phosphorylation state of smad3 in osteoarthritis （OA） chondrocytes. Methods Human chondroblastoma cells （SW1353） were cultured in vitro and were randomly divided into blank control group, IL-1β group, and combination group （treated with IL-1β and serum containing FC）. The optimal intervention concentration of serum containing FC was set at 15 %. The cell growth curve was measured by CCK8 assays, and cell cycle was analyzed by flow cytometry. The levels of COL2A1 and ACAN mRNA expression were examined by quantitative reverse transcription polymerase chain reaction （qRT-PCR）. The phosphorylated smad 3 was tested by Western blot method. Results The cell growth curve, cell proliferation rate（25.50 ± 0.63 ）, gene levels of COL2A1 （0. 523 ± 0.009） and ACAN（0.201± 0.035）, protein level of p-smad3（ 1.026 ± 0.003） in IL-1β group were significantly lower than those of the blank control group （P 〈 0.05）. In the combination group, the cell growth curve, cell proliferation rate （37.55± 0.71 ）, gene levels of COL2A 1 （0.701 ± 0.068 ） and ACAN （0.901±0.230）, protein level of p-smad3（ 1.527 ±0.003） were high as compared with those in IL-1β group（P 〈 0.01 ）. Conclusion FC can enhance the synthesis of COL2A1 and ACAN by increasing the phosphorylated smad3 level in OA chondrocytes, thus to protect the cartilage from injury.

关 键 词：骨关节炎 复元胶囊 Ⅱ型胶原 蛋白多糖 SMAD 3 

分 类 号：R285.5[医药卫生—中药学]