透明质酸对人髓核细胞聚集蛋白多糖和ADAMTSs表达的影响  

Effects of Hyaluronic Acid on the Expression of Aggrecan and ADAMTSs in Human Nucleus Pulposus Cells

作　　者：张楚杭 钱胜龙 王华松 黄明 ZHANG Chuhang;QIAN Shenglong;WANG Huasong;HUANG Ming(Department of Medicine,Wuhan University of Science and Technology,Wuhan Hubei 430065,China)

机构地区：[1]武汉科技大学医学部医学院,湖北武汉430065 [2]中部战区总医院骨科

出　　处：《联勤军事医学》2024年第9期740-744,共5页Military Medicine of Joint Logistics

摘　　要：目的通过研究透明质酸(hyaluronic acid,HA)对人髓核细胞中聚集蛋白多糖及ADAMTSs(a disintegrin and metalloproteinase with thrombospondin motifs)表达的影响,探讨HA调节ADAMTSs减缓椎间盘退变(intervertebral disc degeneration,IDD)的作用机制。方法根据Pfirrmann分级获取正常及退变人椎间盘髓核,采用组织块法培养髓核细胞。将患者手术后的髓核组织标本分为正常组(正常椎间盘)、正常治疗组(正常椎间盘)、退变组(退变椎间盘)、退变治疗组(退变椎间盘),每组各10个标本。正常组和退变组髓核细胞不行HA干预;正常治疗组和退变治疗组在加压前1 h行HA 1 mg/ml干预。各组髓核组织细胞均构建IDD模型。采用逆转录聚合酶链反应(reverse transcription quantitative polymerase chain reaction,RT-PCR)检测各组髓核细胞中聚集蛋白多糖和ADAMTSs的mRNA表达;采用Western blot检测ADAMTSs蛋白的表达。结果经HA干预后,正常治疗组及退变治疗组的聚集蛋白多糖mRNA相对表达量均较干预前升高(P均<0.05),ADAMTS-4、ADAMTS-5的mRNA相对表达量均较干预前明显降低(P均<0.05),而ADAMTS-1 mRNA相对表达量较干预前无显著变化(P>0.05)。Western blot结果显示,正常治疗组和退变治疗组髓核细胞ADAMTS-4和ADAMTS-5蛋白水平较干预前明显降低(P均<0.05)。结论HA可能通过减少ADAMTSs的表达,降低其对聚集蛋白多糖的降解,从而减缓IDD。Objective To investigate the effect of hyaluronic acid(HA)on the expression of aggrecan and a disintegrin and metalloproteinase with thrombospondin motifs(ADAMTSs)in human nucleus pulposus cells,and to explore the mechanism of HA regulating ADAMTSs in alleviating intervertebral disc degeneration(IDD).Methods Normal and degenerated human intervertebral disc nucleus pulposus was obtained according to Pfirrmann classification,nucleus pulposus cells were cultured by the tissue block method.The nuclear pulposus tissue samples of patients after surgery were divided into normal group(normal intervertebral disc),normal treatment group(normal intervertebral disc),degenerative group(degenerative intervertebral disc)and degenerative treatment group(degenerative intervertebral disc),with 10 samples in each.Nucleus pulposus cells in normal and degenerative groups did not receive HA intervention,and the normal treatment group and the degenerative treatment group received HA 1 mg/ml intervention 1h before pressure.IDD models were constructed by nucleus pulposus tissue cells in each group.The mRNA expression of aggrecan and ADAMTSs in each group was detected by reverse transcription quantitative polymerase chain reaction(RT-PCR);the protein expression of ADAMTSs was detected by Western blot.Results After the treatment of HA,the mRNA expression of aggrecan in normal treatment group and degenerated group was higher than that before intervention(all P<0.05),and the mRNA expression of ADAMTS-4 and ADAMTS-5 was significantly reduced(all P<0.05),whereas the mRNA expression of ADAMTS-1 had no significant change compared with that before intervention(P>0.05).Western blot analysis result shows that protein levels of ADAMTS-4 and ADAMTS-5 were significantly reduced compared with that before intervention(all P<0.05).Conclusion Hyaluronic acid may retard IDD by reducing the expression of ADAMTSs to decrease their degradation of aggrecan.

关 键 词：髓核细胞 椎间盘退变 透明质酸 ADATMSs 聚集蛋白多糖 

分 类 号：R521.7[医药卫生—内科学]