探索肾阳虚证骨关节炎大鼠软骨细胞生物钟的变化

Changes of Biological Clock in Chondrocytes of Rats with Osteoarthritis Induced by Kidney Yang Deficiency

目的:探索肾阳虚证骨关节炎大鼠软骨细胞生物钟的表达情况,为肾阳虚证骨关节炎的病机提供新的理论支持。方法:选用6周龄SD大鼠36只,随机分为正常组(不干预)、模型组(hulth法干预)、去势组(hulth法+去势干预)、腺嘌呤组(hulth法+腺嘌呤灌胃干预),每组9只。造模成功后取材,以RT-PCR方法检测软骨细胞生物钟相关基因表达情况,包括pi3k、akt、bmal1、per1、cry1基因;以放射免疫法和化学发光法检测血清肾阳虚指标表达量,包括皮质酮(Corticosterone, CORT)、雌二醇(Estradiol,E2)、睾酮(Testosterone,T)。结果:生物钟表达情况可见,较正常组相比,模型组、去势组和腺嘌呤组均可见bmal1基因的表达量降低,去势组per1基因表达量升高,腺嘌呤组per1、cry1基因表达量升高(P<0.05),pi3k、akt基因表达未见明显差异(P>0.05);3组组间两两比较,bmal1基因表达量可见,腺嘌呤组低于去势组,去势组低于模型组(P<0.05);per1、cry1基因表达量3组间未见明显差异(P>0.05)。肾阳虚指标可见:较正常组相比,去势组、腺嘌呤组大鼠E2升高,T降低,CORT升高(P<0.05);组间两两比较可见腺嘌呤组E2及CORT较去势组表达更高(P<0.05),T表达未见明显差异(P>0.05)。结论:骨关节炎肾阳虚证大鼠软骨细胞生物钟会发生紊乱,bmal1基因表达水平降低,per1、cry1基因表达水平升高,且肾阳虚程度不同,生物钟紊乱水平也不同。

Objective:To explore the expression of biological clock in chondrocytes of rats with osteoarthritis of kidney Yang deficiency to provide new theoretical support for pathogenesis of osteoarthritis of kidney Yang deficiency.Method:Thirty-six SD rats of 6 weeks old were randomly divided into normal group(no intervention),model group(Hulth method intervention),castration group(Hulth method+castration intervention),adenine group(Hulth+adenine gavage intervention),9 rats in each group.After successful modeling,the expression of biological clock related genes in chondrocytes was detected by RT-PCR,including pi3 k,akt,bmal1,per1 and cry1 gene.The expression level of indexes of kidney Yang deficiency rats was detected by radioimmunoassay and chemiluminescence,including corticosterone(corticosterone,CORT),estradiol(Estradiol,E2)and testosterone(Testosterone,T).Result:Compared with the normal group,the expression of the bmal1 gene in the model group,the castrated group and the adenine group decreased,the expression of per1 gene in the castrated group increased,the expression of per1 and cry1 genes in the adenine group increased(P<0.05),and the expression of pi3 k and akt genes was not significantly different(P>0.05).In the three groups,the expression of bmal1 gene was visible,that in the adenine group was lower than that of the ovariectomized group,and the castration group’s expression was lower than that of the model group(P<0.05),and there was no significant difference among the three groups’per1 and cry1 gene expression(P>0.05).Compared with the normal group,the E2 in the ovariectomized group and the adenine group increased,the T decreasedand the CORT increased(P<0.05).The E2 and CORT in the adenine group were higher than those of the castrated group(P<0.05)and the expression of T was not significantly different(P>0.05)between the two groups.Conclusion:The biological clock of chondrocytes in rats with osteoarthritis of kidney Yang deficiency syndrome would be disturbed.The expression level of bmal1 gene is reduced,the expression level of per1 and cry1 is higher,and the degree of kidney Yang deficiency is different,and the level of the biological clock disorder is different.