摘要
目的观察GATA4对椎间盘退变的影响,并探究其促进髓核细胞发生炎症和衰老的可能机制。方法采用免疫组化、荧光、RT-qPCR和Western blot方法检测组织标本中GATA4在不同程度退变的椎间盘中的表达水平;Lenti-GATA4慢病毒转染髓核细胞,构建稳定表达GATA4的髓核细胞;通过Western blotting和β-Gal染色阳性计数,比较GATA4诱导髓核细胞发生炎症与叔丁基过氧化物(tert-butyl hydroperoxide,TBHP)诱导髓核细胞发生炎症的效果差异;加入NF-κB通路抑制剂,研究GATA4促进髓核细胞发生炎症和衰老的机制;构建大鼠椎间盘针刺退变模型,体内验证GATA4在促进椎间盘退变的的机制。结果严重退变的椎间盘中髓核细胞表达更多的GATA4,而且体外实验证实,过表达GATA4后,髓核细胞更易发生炎症和衰老。抑制NF-κB通路后,GATA4在促进髓核细胞发生炎症和衰老的作用被明显抑制,体内实验也证明椎间盘退变后髓细胞中GATA4和炎症因子IL-6表达都明显增加。结论GATA4可以活化NF-κB信号通路,增加促炎细胞因子IL-6和TNF-α表达,加重椎间盘退变的进展。
This study was designed to observe the effect of GATA4 on intervertebral disc degeneration(IVDD and to explore its possible mechanism of promoting inflammation and senescence in nucleus pulposus cells(NPCs).The expression level of GATA4 was detected in the tissue samples of different degrees of IVDD.Lenti-GATA4 lentivirus was used to transfect NPCs for constructing the NPCs stably expressing GATA4.By Western blotting andβ-Gal staining positive counting,the effect of inducing NPCs inflammation by GATA4 and TBHP were evaluated and compared.NF-κB pathway inhibitor was introduced to study the mechanism of GATA4 promoting the inflammation and senescence of NPCs.Rat intervertebral disc degeneration model was constructed to verify the mechanism of GATA4 in promoting intervertebral disc degeneration in vivo.Data indicated that NPCs with severe IVDD expressed more GATA4 compared with mild IVDD NPCs,and in vitro experiments confirmed that after overexpression of GATA4,NPCs showed more inflammation and senescence.After inhibiting NF-κB pathway,the effect of GATA4 in promoting NPCs inflammation and senescence were significantly inhibited.In vivo experiments also showed that the expression of GATA4 and inflammatory factor IL-6 were significantly increased after IVDD.In conclusion,GATA4 can activate the NF-κBpathway,increase the expression of pro-inflammatorycytokines IL-6 and TNF-α,thus aggravate theprogression of intervertebral disc degeneration.
作者
罗力文
简秀英
周跃
LUO Liwen;JIAN Xiuying;ZHOU Yue(Department of Orthopaedics,Xinqiao Hospital,Army Medical University,Chongqing 400038,China;Department of Infectious Diseases,Second Affiliated Hospital of Chongqing Medical University,Chongqing 400010,China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2021年第7期553-559,共7页
Immunological Journal
基金
国家自然科学基金(81874028)。
