摘要
目的探讨LncRNA HAND2-AS1对白细胞介素-1β(IL-1β)诱导的软骨细胞损伤的影响及其可能作用机制。方法采用qRT-PCR法检测对照组、骨关节炎组血浆中HAND2-AS1、miR-106b-5p的表达量;采用IL-1β诱导人正常软骨细胞C28/I2建立细胞损伤模型,随机分为con组、IL-1β组、IL-1β+pcDNA组、IL-1β+pcDNA-HAND2-AS1组、IL-1β+anti-miR-NC组、IL-1β+miR-106b-5p Inhibitor组、IL-1β+pcDNA-HAND2-AS1+miR-NC组、IL-1β+pcDNA-HAND2-AS1+miR-106b-5p mimic组;MTT法与流式细胞术分别检测细胞增殖及凋亡率;ELISA法检测IL-6、TNF-α的水平;双荧光素酶报告实验检测HAND2-AS1与miR-106b-5p的靶向关系。结果与对照组比较,骨关节炎组患者血浆中HAND2-AS1的表达量降低(P<0.05),miR-106b-5p的表达量升高(P<0.05);与con组比较,IL-1β组细胞增殖抑制率和细胞凋亡率升高(P<0.05),IL-6、TNF-α的水平升高(P<0.05);与IL-1β+pcDNA组、IL-1β组比较,IL-1β+pcDNA-HAND2-AS1组细胞IL-6、TNF-α的水平降低(P<0.05),细胞增殖抑制率和凋亡率降低(P<0.05);HAND2-AS1可靶向调控miR-106b-5p的表达;与IL-1β+anti-miR-NC组比较,IL-1β+miR-106b-5p Inhibitor组细胞增殖抑制率和细胞凋亡率降低(P<0.05),IL-6、TNF-α的水平降低(P<0.05);与IL-1β+pcDNA-HAND2-AS1+miR-NC组比较,IL-1β+pcDNA-HAND2-AS1+miR-106b-5p mimic组细胞增殖抑制率和细胞凋亡率升高(P<0.05),IL-6、TNF-α的水平升高(P<0.05)。结论HAND2-AS1过表达可通过靶向调控miR-106b-5p的表达而促进细胞增殖及抑制细胞凋亡、炎性反应从而减轻IL-1β诱导的软骨细胞损伤。
Objective To investigate the effects of LncRNA HAND2-AS1 on interleukin-1β(IL-1β)-induced chondrocyte damage and its possible mechanism.Methods The qRT-PCR method was used to detect the expression levels of HAND2-AS1 and miR-106b-5p in the plasma in control group and osteoarthritis group.Human normal chondrocyte C28/I2 was induced by IL-1βto establish cell injury models,which were randomly divided into con group,IL-1βgroup,IL-1β+pcDNA group,IL-1β+pcDNA-HAND2-AS1 group,IL-1β+anti-miR-NC group,IL-1β+miR-106b-5p inhibitor group,IL-1β+pcDNA-HAND2-AS1+miR-NC group,IL-1β+pcDNA-HAND2-AS1+miR-106b-5p mimic group.MTT method and flow cytometry were used to detect the cell proliferation and apoptosis rates;ELISA was used to detect the serum levels of IL-6 and TNF-α;dual luciferase reporter experiment was used to detect the targeting correlation between HAND2-AS1 and miR-106b-5p.Results Compared with those in control group,the expression levels of HAND2-AS1 in osteoarthritis group were significantly decreased(P<0.05),however,the expression levels of miR-106b-5p were significantly increased(P<0.05).Compared with those in control group,the cell proliferation inhibition rate and apoptosis rate as well as the levels of IL-6 and TNF-αin IL-1βgroup were significantly increased(P<0.05).Compared with those in IL-1β+pcDNA group and IL-1βgroup,the levels of IL-6 and TNF-αas well as cell proliferation inhibition rate and apoptosis rate in IL-1β+pcDNA-HAND2-AS1 group were significantly decreased(P<0.05).HAND2-AS1 could targetedlt regulate the expression of miR-106b-5p.Compared with those in IL-1β+anti-miR-NC group,the cell proliferation inhibition rate and apoptosis rate as well as the levels of IL-6 and TNF-αin IL-1β+miR-106b-5p inhibitor group were significantly decreased(P<0.05).Compared with those in IL-1β+pcDNA-HAND2-AS1+miR-NC group,the cell proliferation inhibition rate and apoptosis rate in IL-1β+pcDNA-HAND2-AS1+miR-106b-5p mimic group were significantly decreased(P<0.05).however,thelevels of IL-6 level and
作者
徐立
张旭然
张淼
XU Li;ZHANG Xuran;ZHANG Miao(Department of Orthopedics,Fuxin Central Hospital,Liaoning,Fuxin 123000,China)
出处
《河北医药》
CAS
2022年第19期2916-2919,2924,共5页
Hebei Medical Journal
