摘要
目的探索利用细胞因子抑制退变椎间盘细胞释放炎症因子的生物学机制及作用,评价IL-10和TGF-β抑制退变椎间盘细胞释放炎症因子的有效性。方法建立6条1岁龄比格犬针刺退变模型,6周后处死动物取出退变椎间盘髓核组织,体外分离培养,将培养的退变髓核细胞分为4组:20 ng/ml IL-10治疗组,20 ng/ml TGF-β治疗组,20 ng/ml IL-10联合20 ng/ml TGF-β治疗组及未治疗组,正常髓核细胞作为对照组。采用ELISA(enzyme-linked immunosorbent assay)和real-time PCR检测IL-10和TGF-β对细胞炎症因子蛋白水平与基因水平表达量的影响。结果检测未治疗组髓核细胞高表达炎症因子,而采用IL-10和TGF-β的组别炎症因子的表达受到了明显的抑制。IL-10和TGF-β能够阻断退行性椎间盘细胞产生的炎性细胞因子的级联反应和椎间盘细胞炎症反应的发展。结论 IL-10和TGF-β能够稳定地抑制椎间盘退变(intervertebral disc degeneration,IDD)中IL-1β和TNF-α的表达,有介导治疗IDD的应用前景。
Objective To explore the biological function and mechanism of underlying suppression of the release of inflammatory cytokines from degenerative disc cells and to evaluate the effectiveness of interleukin（ IL）-10 and transforming growth factor（ TGF）-β in suppressing the release of inflammatory cytokines from degenerative disc cells. Methods Six 1-year-old male beagle dogs were used. A beagle model of intervertebral disc degeneration（ IDD） was established using an anular stab. The intervertebral discs were incised when the dogs were sacrifi ced 6 weeks later, and nucleus pulposus（ NP） cells were isolated and cultured in vitro. The cultured degenerative NP cells were divided into 4 groups, including untreated degenerative NP cells, degenerative NP cells treated with 20 ng / ml IL-10, degenerative NP cells treated with 20 ng / ml TGF-β and degenerative NP cells treated with 20 ng / ml IL-10 and 20 ng / ml TGF-β. An enzyme-linked immunosorbent assay（ ELISA） and real-time polymerase chain reaction（ PCR） were performed to determine the effects of IL-10 and TGF-β on the protein and gene expressions of inflammatory cytokines. Results Higher expressions of inflammatory cytokines were observed in untreated degenerative NP cells. IL-10 and TGF-β blocked the cascade of inflammatory cytokines produced by degenerative disc cells and limited the development of infl ammatory responses. Conclusions The expressions of IL-1β and tumor necrosis factor（ TNF）-α in degenerative disc cells can be signifi cantly and stably suppressed by IL-10 and TGF-β. Thus, IL-10 and TGF-β may be suitable biological therapeutic agents for IDD.
出处
《中国骨与关节杂志》
CAS
2015年第3期230-234,共5页
Chinese Journal of Bone and Joint
基金
国家自然科学基金面上项目(81272044)
国家自然科学基金青年项目(81301033)
关键词
椎间盘退行性变
动物实验
白细胞介素10
转化生长因子Β
Intervertebral disc degeneration
Animal experimentation
Interleukin-10
Transforming growth factor beta
