摘要
目的探讨鱼腥草素钠在体外对软骨细胞及其细胞外基质合成降解代谢的影响,从而尝试阐述其对骨关节炎的潜在作用。方法从小鼠软骨组织提取小鼠原代软骨细胞,用于在体外研究鱼腥草素钠对软骨细胞胞外基质合成降解的影响。通过增殖实验和活死染色实验研究低、中、高不同浓度的鱼腥草素钠对软骨细胞增殖能力的影响;通过荧光染色实验,检测COLⅡ和MMP13的表达,探究鱼腥草素钠对于软骨细胞基质合成降解的影响;通过实时荧光定量聚合酶链反应研究鱼腥草素钠对软骨细胞细胞基质合成降解基因表达水平的影响;通过蛋白印迹实验研究鱼腥草素钠对软骨细胞细胞基质合成降解蛋白表达水平的影响。结果细胞增殖及活死细胞染色表明:与对照组相比,白介素组细胞增殖能力明显下降,而鱼腥草素钠干预对细胞增殖无显著影响,即鱼腥草素钠干预对细胞无明显毒性;荧光染色、PCR及蛋白印迹实验结果表明:鱼腥草素钠显著降低IL-1β上调的小鼠软骨细胞中MMP13及ADAMTS5基因及蛋白的表达,同时增加COLⅡ和ACAN基因及蛋白的表达。结论鱼腥草素钠在体外能够促进软骨细胞外基质合成并抑制降解,维持细胞外基质稳态,且呈现一定浓度依赖性,中、高浓度鱼腥草素钠组的促进软骨细胞外基质合成抑制降解能力显著,因而鱼腥草素钠可能对骨关节炎的发生发展具有一定抑制作用,可能会是一种能够用于治疗早期骨关节炎的新型药物。
Objective To investigate the effect of Sodium Houttuynia(SH)on chondrocytes and extracellular matrix metabolism and to expound its potential protective effect on osteoarthritis in vitro.Methods Cartilage tissues used in the in-vitro research were obtained from shoulders and knees of mice to extract chondrocytes to investigate the effect of SH on chondrocytes and extracellular matrix metabolism.Then CCK-8 and staining of live and dead cells were used to observe cell proliferation and to get knowledge of effects of different concentrations of SH on chondrocytes and their proliferation.COLⅡand MMP13 immunofluorescence staining were performed to determine cartilage matrix synthesis and degradation.And then mRNA expression levels of cartilage matrix synthesis gene and degradation were determined by qRT-PCR,while protein expression levels of cartilage matrix synthesis and degradation were determined by western blot analysis.Results CCK-8 and staining of live and dead cells showed that cell proliferation ability of the group intervented with IL decreased significantly,compared with the control group's,while cell proliferation ability of the group intervented with SH was almost the same as that of the control group's, which meant that SH had no significant toxicity to cells. Immunofluorescence staining showed that in chondrocytes, protein level of COL Ⅱ increased in SH group compared with IL group, while protein level of MMP13 decreased in SH group. Subsequently, results of qRT-PCR and Western blot suggested that COL Ⅱ and Aggrecan expression levels increased in SH group compared with IL group, while MMP13 and ADAMTS5 expression levels decreased in SH group. Conclusion In vitro, SH promotes the synthesis of proteoglycan and type Ⅱ collagen secreted by chondrocytes, and suppresses the decomposition of cartilage extracellular matrix conductive to maintain the balance between the degradation and synthesis of the extracellular matrix. In particular, high and medium dosage SH can significantly decrease matrix synthesis and increase matrix degradation, which may inhibit osteoarthritis progression. This study suggests that SH can be used as a kind of potential drug for the treatment of early osteoarthritis.
作者
朱天峰
赵志坚
刘洋
张一健
陆英杰
朱雪松
ZHU Tian-feng;ZHAO Zhi-jian;LIU Yang;ZHANG Yi-jian;LU Ying-jie;ZHU Xue-song(Department of Orthopedics,First Affiliated Hospital of Soochow University,Suzhou,Jiangsu,215006,China)
出处
《中国血液流变学杂志》
CAS
2022年第1期10-15,F0002,共7页
Chinese Journal of Hemorheology
基金
国家自然科学基金资助项目(82072442),江苏省研究生科研创新计划(KYCX21_2971)。
关键词
骨关节炎
鱼腥草素钠
细胞外基质
基质合成和降解
osteoarthritis
Sodium Houttuynia
extracellular matrix
synthesis and degradation of extracellular matrix
