CAJ | 학술논문

目的：评价白介素-10（IL-10）和转化生长因子-β（TGF-β）抑制退变椎间盘细胞释放炎症因子的有效性，探索细胞因子抑制退变椎间盘细胞释放炎症因子的潜在生物学机制。方法建立6条1岁龄比格犬针刺退变模型，2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，未治疗组；并以正常髓核细胞作为正常对照组。应用流式细胞仪于治疗后12、24、48 h时检测髓核细胞肿瘤坏死因子-α（TNF-α）的表达量。结果治疗后12 h，正常髓核细胞组TNF-α的MFI值为7.92±0.32，IL-10治疗组为14.57±3.37，IL-10+TGF-β治疗组为11.9±2.91，均显著低于未治疗组（31.47±4.38，P<0.01）。治疗后24 h，IL-10治疗组为21.23±2.85，TGF-β治疗组为20.27±2.76， IL-10+TGF-β治疗组为12.9±2.91，均显著低于未治疗组（30.8±2.86，P<0.01），IL-10+TGF-β治疗组较IL-10治疗组及TGF-β治疗组显著降低（P<0.01）。而正常对照组与空白对照组差异无统计学意义。治疗后48 h，TGF-β治疗组与IL-10+TGF-β治疗组TNF-α阳性表达的细胞数达到最低水平，正常对照组TNF-α的MFI值为9.07±0.54，TGF-β治疗组为16.07±2.46，IL-10+TGF-β治疗组为12.3±4.42，均显著低于未治疗组（29.3±2.84，P<0.01）。结论 IL-10和TGF-β能够稳定地抑制TNF-α的表达，IL-10和TGF-β可能用于椎间盘退行性疾病的生物治疗。
목적：평개백개소-10（IL-10）화전화생장인자-β（TGF-β）억제퇴변추간반세포석방염증인자적유효성，탐색세포인자억제퇴변추간반세포석방염증인자적잠재생물학궤제。방법건립6조1세령비격견침자퇴변모형，2주후처사동물취출퇴변추간반수핵조직，체외분리배양。장배양적퇴변수핵세포분위4조：20 ng/ml IL-10치료조，20 ng/ml TGF-β치료조，20 ng/ml IL-10연합20 ng/ml TGF-β치료조，미치료조；병이정상수핵세포작위정상대조조。응용류식세포의우치료후12、24、48 h시검측수핵세포종류배사인자-α（TNF-α）적표체량。결과치료후12 h，정상수핵세포조TNF-α적MFI치위7.92±0.32，IL-10치료조위14.57±3.37，IL-10+TGF-β치료조위11.9±2.91，균현저저우미치료조（31.47±4.38，P<0.01）。치료후24 h，IL-10치료조위21.23±2.85，TGF-β치료조위20.27±2.76， IL-10+TGF-β치료조위12.9±2.91，균현저저우미치료조（30.8±2.86，P<0.01），IL-10+TGF-β치료조교IL-10치료조급TGF-β치료조현저강저（P<0.01）。이정상대조조여공백대조조차이무통계학의의。치료후48 h，TGF-β치료조여IL-10+TGF-β치료조TNF-α양성표체적세포수체도최저수평，정상대조조TNF-α적MFI치위9.07±0.54，TGF-β치료조위16.07±2.46，IL-10+TGF-β치료조위12.3±4.42，균현저저우미치료조（29.3±2.84，P<0.01）。결론 IL-10화TGF-β능구은정지억제TNF-α적표체，IL-10화TGF-β가능용우추간반퇴행성질병적생물치료。
Objective To investigate the biological mechanisms underlying the suppression of the release of in-flammatory cytokines from degenerative intervertebral disc cells and to evaluate the effectiveness of interleukin-10 (IL-10) and transforming growth factor-β (TGF-β) for suppressing the release of inflammatory cytokines from degenerative cells. Methods Six healthy 1 year old male beagle dogs were used for this study. A beagle model of intervertebral disc degen-eration (IDD) was established using an anular stab, and then the intervertebral discs were incised, and nucleus pulposus (NP) cells were isolated and cultured. The cultured degenerative NP cells were divided into four groups: the untreated de-generative NP cells, the degenerative NP cells treated with 20 ng/ml IL-10, the degenerative NP cells treated with 20 ng/ml TGF-β, and the degenerative NP cells treated with 20 ng/ml IL-10 and 20 ng/ml TGF-β. Fluorescence-activated cell sort-ing and tumor necrosis factor -α (TNF-α) monoclonal antibodies were used to determine TNF-α expression in NP cells. Results IL-10 and TGF-β blocked the cascade of inflammatory cytokines produced by degenerative intervertebral disc cells and limited the development of inflammatory responses. Higher expression of inflammatory cytokines was observed in the untreated degenerative NP cells. Conclusion Treatment with IL-10 and TGF-β can significantly and stably suppress the expression of TNF-α. IL-10 and TGF-βmay be suitable biological therapeutic agents for treating IDD.