摘要
目的本研究旨在评价超活化血小板裂解液(super-activated platelet lysate,sPL)对软骨细胞增殖及损伤修复的作用。方法通过外周血制备sPL。使用RTCA S16系统检测sPL对C28/I2细胞增殖的影响。用10 ng/m L人白介素-1β(human interleukin 1β,h IL-1β)诱导C28/I2细胞炎症损伤及凋亡,并采用流式细胞术检测不同浓度sPL对C28/I2细胞凋亡修复的作用。利用RT-qPCR和ELISA方法检测sPL对C28/I2细胞炎症的调控作用。RT-qPCR、免疫细胞化学及Western blot分析sPL修复后软骨细胞特异性基因的表达。结果5%sPL及10%sPL可促进C28/I2细胞增殖,且10%sPL对细胞增殖的促进作用更强。凋亡检测结果显示,10 ng/m L h IL-1β显著增加C28/I2细胞的凋亡,而5%sPL和10%sPL均可显著降低凋亡率。同时,5%sPL和10%sPL显著降低C28/I2细胞中白介素-6(interleukin 6,IL-6)基因表达,且10%sPL的作用更强。在C28/I2细胞炎症损伤修复过程中,sPL能够显著提高Collagen II的表达,并降低Collagen I的表达。结论sPL在体外能够促进软骨细胞增殖,减少炎症反应及软骨细胞凋亡,从而促进纤维软骨向透明软骨的转化。
Objective Objective This study aims to evaluate the effects of super-activated platelet lysate(sPL)on the proliferation of chondrocytes and the repair of damaged chondrocytes.Methods sPL was prepared from peripheral blood.The RTCA S16 system was used to assess the effect of sPL on the proliferation of C28/I2 cells.Inflammatory injury and apoptosis of C28/I2 cells were induced with 10 ng/mL human interleukin-1β(hIL-1β),and flow cytometry was employed to assess the repair effect of different concentrations of sPL on C28/I2 cell apoptosis.RT-qPCR and ELISA were used to analyze the inflammatory regulation of C28/I2 cells by sPL.RT-qPCR,immunocytochemistry,and Western blot were performed to evaluate the expression of chondrocyte-specific genes after sPL treatment.Results Both 5%and 10%sPL promoted the proliferation of C28/I2 cells,with 10%sPL showing a stronger effect on cell proliferation.Apoptosis assays revealed that 10 ng/mL hIL-1βsignificantly increased apoptosis in C28/I2 cells,while both 5%and 10%sPL significantly reduced apoptosis rates.Additionally,both 5%and 10%sPL significantly reduced the expression of interleukin-6(IL-6)in C28/I2 cells,with 10%sPL showing a stronger effect.During the repair of C28/I2 cell inflammatory damage,sPL significantly increased the expression of Collagen II and decreased the expression of Collagen I.Conclusion In vitro,sPL promotes the proliferation of chondrocytes,reduces inflammation and apoptosis,and facilitates the conversion of fibrous cartilage to hyaline cartilage.
作者
刘春香
张天琦
孟令旗
隋福革
张怡
Liu Chunxiang;Zhang Tianqi;Meng Lingqi;Sui Fuge;Zhang Yi(National and Local Joint Stem Cell Research&Engineering Center for Aging Diseases,Tian Qing Stem Cell Co.,Ltd.,Harbin 150028,China;Department of Orthopedics,Daqing Longnan Hospital,Daqing 163453,China)
出处
《中国组织化学与细胞化学杂志》
CSCD
2024年第6期555-562,共8页
Chinese Journal of Histochemistry and Cytochemistry
基金
哈尔滨市松北区工业信息科技局资金项目(HSGK201913)。
