摘要
背景:研究证实,滑膜炎分泌的炎症因子通过加速关节软骨的分解代谢而成为骨性关节炎发展的主要原因,基质金属蛋白酶3和基质金属蛋白酶13在骨性关节炎中起关键作用。目的:探究富血小板血浆治疗骨性关节炎的有效作用机制。方法:分离并提取大鼠膝关节滑膜细胞和软骨细胞分别培养,腹主动脉取血制备富血小板血浆制剂。将滑膜细胞分为空白对照组和大肠杆菌脂多糖处理组,脂多糖处理组用脂多糖刺激滑膜细胞制造滑膜炎模型;再将滑膜炎细胞分为富血小板血浆处理组和未处理组(脂多糖组)。对照组、富血小板血浆处理组和未处理组3组细胞培养24 h后,用ELISA法检测培养基中炎性细胞因子白细胞介素1β、白细胞介素6和肿瘤坏死因子α的水平,另一部分培养基分别处理软骨细胞,培养48 h后,Western blot检测软骨细胞内Ⅰ型、Ⅱ型胶原蛋白、基质金属蛋白酶3和基质金属蛋白酶13蛋白水平的变化,荧光定量PCR检测基质金属蛋白酶3和基质金属蛋白酶13 mRNA表达水平的变化。结果与结论:①ELISA检测显示,脂多糖处理组滑膜细胞培养基中白细胞介素1β、白细胞介素6和肿瘤坏死因子α的水平显著高于对照组和富血小板血浆处理组(P<0.01);富血小板血浆处理组培养基中白细胞介素1β、白细胞介素6和肿瘤坏死因子α的水平高于对照组(P<0.05);②Western blot检测显示,富血小板血浆处理组软骨细胞中Ⅰ型、Ⅱ型胶原蛋白显著高于对照组和脂多糖处理组(P<0.01),脂多糖处理组低于对照组(P<0.05);③脂多糖组基质金属蛋白酶3和基质金属蛋白酶13蛋白显著高于富血小板血浆处理组和对照组(P<0.01);④PCR检测显示脂多糖组基质金属蛋白酶3,13 mRNA显著高于富血小板血浆处理组和对照组;⑤结果表明,富血小板血浆处理能够通过降低滑膜细胞炎性因子白细胞介素1β、白细胞介素6和肿瘤坏死因�
BACKGROUND:Studies have confirmed that inflammatory factors secreted by synovitis that accelerate the catabolism of articular cartilage have become the main cause of osteoarthritis.Matrix metalloproteinase 3 and matrix metalloproteinase 13 play a key role in osteoarthritis.OBJECTIVE:To study the effective mechanism of platelet-rich plasma(PRP)in the treatment of osteoarthritis.METHODS:The rat knee synovial cells and chondrocytes were isolated and extracted separately.The blood samples of rats were extracted to prepare PRP preparation.Then,synovial cells were divided into a control group and an E.coli lipopolysaccharide(LPS)treatment group,where synovial cells were stimulated with LPS to create a synovitis model.The synovitis cells were further divided into a PRP treatment group and an untreated group.The cells in each group were cultured for 24 hours.A portion of the medium was taken,in which the levels of interleukin-1β,interleukin-6 and tumor necrosis factor-α was detected using ELISA.The other part of the medium was used to treat chondrocytes.After 48 hours of culture,the changes of type I,II collagens,matrix metalloproteinase 3 and matrix metalloproteinase 13 protein levels in chondrocytes were detected by western blot.The expression levels of matrix metalloproteinase 3 and matrix metalloproteinase 13 mRNA were detected by real-time PCR.RESULTS AND CONCLUSION:Determined by the ELISA,the levels of interleukin-1β,interleukin-6 and tumor necrosis factor-αin the synovial cell culture medium of the LPS treatment group were significantly higher than those of the control group and the PRP treatment group(P<0.01).The levels of interleukin-1β,interleukin-6 and tumor necrosis factor-α in the medium of PRP treatment group were significantly higher than those in the control group(P<0.05).Western blot showed that the expression of type I and II collagens in the chondrocytes of the PRP treatment group were significantly higher than those in the control and LPS treatment groups(P<0.01);the expression of type I and II
作者
陈尉
张国如
何健东
霍少川
Chen Wei;Zhang Guoru;He Jiandong;Huo Shaochuan(Department of Joint Surgery,Third People’s Hospital of Hainan Province,Sanya 572000,Hainan Province,China;Department of Joint Surgery,Guangzhou Orthopedic Hospital,Guangzhou 510000,Guangdong Province,China;Shenzhen Hospital,Guangzhou University of Chinese Medicine,Shenzhen 518034,Guangdong Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2020年第29期4643-4649,共7页
Chinese Journal of Tissue Engineering Research
基金
中国博士后基金面上项目(2018M633088),项目负责人:霍少川。
关键词
富血小板血浆
滑膜
炎症
软骨
细胞
关节炎
platelet-rich plasma
synovial membrane
inflammation
cartilage
cells
arthritis
