软骨细胞增殖与胰岛素和氢化可的松联合应用的关系(英文)

Effect of insulin, hydrocortisone and their combined application on the proliferation of chondrocytes

背景:关节腔内激素注射治疗关节炎能迅速缓解关节疼痛,但可发生进行性软骨损害。加用具有生长因子样作用的胰岛素,是否可产生对关节软骨细胞的保护作用?目的:观察胰岛素和氢化可的松联合作用对软骨细胞生长的影响。设计:分组对比观察,1种药物单独作用采用单因素方差分析,2种药物复合作用采用多因素实验设计资料方差分析的析因分析。单位:广州市创伤外科研究所。材料:4～6周龄的新西兰大白兔膝关节软骨。方法:实验于2000-02/2001-05在广州市创伤外科研究所完成。将软骨碎片分别用透明质酸酶,胰酶及Ⅱ型胶原酶消化,获得软骨单细胞悬液,予以不同作用及其不同剂量的药物干预。根据应用干预药物剂量的不同将其分为4组及相应亚组。对照组(不加氢化可的松和胰岛素),胰岛素(0.035,0.35,3.5,35mg/L)组,氢化可的松(1,5,10,50,100mg/L)组,胰岛素(0.35mg/L)+氢化可的松(50mg/L)组,应用四氮甲基唑蓝比色分析法观察其对软骨细胞增殖的影响。主要观察指标:①胰岛素对软骨细胞增殖的影响。②氢化可的松对软骨细胞增殖的影响。③氢化可的松与胰岛素联合使用对软骨细胞增殖的影响。结果:①胰岛素可促进软骨细胞增殖:单独使用0.035mg/L即可发挥效应,当浓度增至0.35mg/L时,其促进作用达到最大值。②氢化可的松对软骨细胞增殖的抑制作用:单独使用10mg/L时即可发挥效应,随着浓度增大抑制作用显著,当浓度增大至100mg/L,软骨细胞几乎全部死亡。③0.35mg/L胰岛素与50mg/L氢化可的松联合使用:存在着负协同作用(F=164.9,P<0.01)。结论:低浓度胰岛素对软骨细胞的增殖有促进作用。但氢化可的松对软骨细胞的增殖有抑制作用。二者联合使用时,氢化可的松可拮抗胰岛素的作用。

BACKGROUND:Intra-articular injection of hormone has been applied in the treatment of arthritis because it can alleviate arthralgia rapidly, which is accompanied commonly by progressive cartilage impairments. It is not clear if supplement of growth factor like insulin effect can play a protective role in articular chondrocytes. OBJECTIVE:To observe the effects of insulin or hydrocortisone alone and the combination on the proliferation of chondrocytes. DESIGN:Grouping comparative study, the effect of one medicine was analyzed by using one-factor analysis of variance, while the combined effect was analyzed with multi-factor analysis of variance. SETTING:Guangdong Institute of Trauma Sugery. MATERIALS:Articular cartilage from the knees of New Zealand white rabbits of 4-6 weeks old. METHODS:This study was carried out at Guangzhou Traumatic Research Institute from Feberary 2000 to May 2001.Chondrocytes were isolated from the knee joints of New Zealand white rabbits,digested with hyaluronidase, pancreatin and type Ⅱcollagenase and exposed to insulin, hydrocortisone or the combination of insulin and hydrocortisone of different dosage. They were divided into four groups:Control group ( without adding insulin and hydrocortisone), insulin group (0.035,0.35,3.5,35 mg/L subgroups), hydrocortisone group(1,5,10,50,100 mg/L subgroups)and insulin(0.35 mg/L)combined with hydrocortisone(50 mg/L)group.Their influence on chondrocytes proliferation was observed with methyl thiazolyl tetrazolium(MTT) method. MAIN OUTCOME MEASURES:①The influence of insulin on the proliferation of chondrocytes.②The influence of hydrocortisone on the proliferation of chondrocytes.③The combined effect of hydrocortisone and insulin. RESULTS:①Insulin could enhance the proliferation of chondrocytes even at the concentration of 0.035 mg/L(P< 0.01), reaching the maximum at the concentration of 0.35 mg/L(P< 0.01). ②10 mg/L hydrocortisone could inhibit the proliferation of chondrocytes ( P< 0.05 ),which became significant with increasing concentration and no viable chondrocytes could be observed at the concentration of 100 mg/L.③When the chondrocytes were exposed to 0 .35 mg/L insulin combined with 50 mg/L hydrocortisone, the promoting effect of insulin was inhibited due to negative cooperation. CONCLUSION:Insulin at low concentration could enhance the proliferation of chondrocytes,but hydrocortisone displayed inhibiting effect on the growth of chondrocytes. The function of insulin was antagonized when combined with hydrocortisone.