地塞米松磷酸钠纳米制剂对IL-1β诱导大鼠原代软骨细胞炎症的影响OACSTPCD
Effect of dexamethasone sodium phosphate nanomaterials on IL-1β-induced inflammation in primary rat chondrocytes
目的:探讨地塞米松(Dex)磷酸钠纳米制剂(Nano-Dex)对白细胞介素(IL)-1β诱导的大鼠原代软骨细胞炎症的影响.方法:利用透射电镜观察Nano-Dex纳米粒的形态,马尔文粒径仪检测Nano-Dex的粒径和电位.培养SD大鼠软骨细胞,将细胞分为空白组、IL-1β组、IL-1β+Dex组和IL-1β+Nano-Dex组.用实时荧光定量PCR(RT-qPCR)法检测软骨细胞基质金属蛋白酶(MMP)13、MMP3、IL-1β、IL-6、Ⅱ型胶原α1(Col2a1)和蛋白聚糖(ACAN)基因表达水平,免疫荧光染色观察MMP13的表达,western blotting法检测软骨细胞MMP13、Col2a1、IL-6、IL-1β、P65和Caspase 3蛋白表达水平.结果:Nano-Dex形状圆润,大小均一,直径约在90 nm,带正电荷.与空白组比较,IL-1β组软骨修复基因Col2a1、ACAN表达下调,而炎症基因IL-1β、IL-6、MMP3、MMP13表达上调(均P<0.05);与IL-1β组比较,Nano-Dex+IL-1β组Col2a1基因及蛋白表达上调,ACAN基因表达上调,IL-1β、IL-6、MMP3、MMP13基因表达下调,MMP13、P65、IL-6、IL-1β和Caspase 3蛋白表达均下调(均P<0.05).免疫荧光结果显示,IL-1β+Nano-Dex组MMP13的荧光强度明显低于IL-1β组(P<0.05).结论:Nano-Dex能有效抑制IL-1β诱导的软骨细胞炎症.
Objective:To explore the effect of dexamethasone(Dex)sodium phosphate nanomaterials(Nano-Dex)on interleukin(IL)-1β-induced inflammation in primary rat chondrocytes.Methods:Transmission electron microscopy was used to observe the morphology of Nano-Dex nanoparticles;Malvern particle size meter was used to detect the particle size as well as the potential of Nano-Dex.SD rat chondrocytes were cultured,and the cells were randomly divided into blank group,IL-1β group,IL-1β+Dex group,and IL-1β+Nano-Dex group.The expression levels of matrix metalloproteinase(MMP)13,MMP3,IL-1β,IL-6,collagen type Ⅱ alpha 1(Col2a1)and proteoglycan(ACAN)genes in chondrocytes were detected by reverse transcription-quantitative PCR(RT-qP-CR).The expression of MMP13 was observed by immunofluorescence staining.The expression levels of MMP13,Col2a1,IL-6,IL-1β,P65 and Caspase 3 in chondrocytes were detected by western blotting.Results:Na-no-Dex had a rounded shape,uniform size,diameter of about 90 nm,and a positive charge.Compared with the blank group,the expression of cartilage repair genes Col2a1 and ACAN in the IL-1β group was down-regulated,while the expression of inflammatory genes IL-1β,IL-6,MMP3 and MMP13 was up-regulated(all P<0.05).Compared with the IL-1β group,the expression of Col2a1 gene and protein as well as the expression of ACAN gene in the Nano-Dex+IL-1β group were up-regulated,and the expression of IL-1β,IL-6,MMP3 and MMP13 genes as well as the expression of MMP13,P65,IL-6,IL-1β and Caspase 3 proteins were down-regulated(all P<0.05).Immunofluorescence results showed that the fluorescence intensity of MMP13 in the IL-1β +Nano-Dex group was significantly lower than that in the IL-1β group(P<0.05).Conclusion:Nano-Dex can effectively in-hibit IL-1β-induced chondrocyte inflammation.
何浩强;黄婵婷;黄权新;覃再嫩;刘刚
广西医科大学再生医学与医用生物资源开发应用省部共建协同创新中心,南宁 530021||广西组织器官修复医用生物材料工程技术研究中心,南宁 530021
临床医学
地塞米松纳米制剂抗炎骨关节炎
dexamethasonenanomaterialsanti-inflammationosteoarthritis
《广西医科大学学报》 2024 (004)
锌(II)介导的纳米化药物携载p65 siRNA调控自噬对骨关节炎的作用研究
508-515 / 8
国家自然科学基金资助项目(No.81960414)
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