芹菜素在静水压下对人体腰椎间盘髓核蛋白多糖核心蛋白基因表达的影响

Effects of apigenin on proteoglycan core protein's gene expression in the human lumbar disc nucleus pulposus in response to hydrostatic pressure

[目的]探讨中药提取物芹菜素在静水压下对体外培养的人体腰椎间盘髓核蛋白多糖核心蛋白基因表达的影响。[方法]32例后路腰椎间盘髓核摘除术的志愿患者中获取的32个新鲜椎间盘髓核样品,每1例髓核样品被切成1~2 mm3大小的碎块,与1 ml培养基DMEM一同装入2.5 ml的注射器中。培养基中分别加入芹菜素（Apigenin）、一氧化氮合成酶的竞争性抑制剂NG-Monomethyl-L-arginine（简称L-NMMA）及一氧化氮的供体Sodium Nitroprus-side（简称SNAP）,在静水压装置中培养2 h后,对每组中的髓核样品进行石蜡包埋、切片,通过原位杂交来研究芹菜素对椎间盘髓核蛋白多糖核心蛋白的基因转录水平的调节作用。[结果]芹菜素3 atm组和L-NMMA3 atm组蛋白多糖核心蛋白原位杂交结果显示阳性髓核细胞数最多,平均光密度值（MOD）最大。[结论]对于退变的椎间盘的髓核组织,芹菜素能够促进蛋白多糖核心蛋白mRNA的基因转录水平。

[Objective]To investigate the effects of apigenin on proteoglycan core protein gene expression in the human lumbar intervertebral disc nucleus pulposus in response to hydrostatic pressure. [Methods]Thirty-two lumbar intervertebral disc nucleus pulposus specimens were obtained from patients who had undergone posterior discectomy.The syringes were placed in a water filled pressure vessel kept at 37℃.Hydrostatic pressures of 1（control）,3 and 30 atmospheres（atm） were applied.After incubation of 2 hours,each nucleus pulposus specimen was embedded in paraffin,sliced for situ hybridization to study the effect of the apigenin of the intervertebral disc nucleus pulposus proteoglycan core protein gene＇s transcription.As a competitive inhibitor of NO synthases,NG-Monomethy-L-arginine（L-NMMA,500 μmol） and as an organic donor of NO,S-nitroso-N-acetylpenicillamine（SNAP,200 μmol） were used.[Results]The results of apigenin 3 atm group and the L-NMMA 3 atm group in situ hybridization to proteoglycan core protein showed the largest number of positive nucleus pulposus cells,with a largest average optical density.[Conclusion]For the degenerative intervertebral disc nucleus pulposus,apigenin plays an important role in the promotion of proteoglycan core protein mRNA gene＇s transcription levels.