软骨细胞生物钟在大鼠骨关节炎发病中的作用  

The Role of Chondrocyte Clock in the Pathogenesis of Osteoarthritis in Rats

作　　者：陈颖颖 李冬霞 张丽宁 胡琪 周丽 罗莎 周同亮[2] 杜梦梦 陶庆文[1] CHEN Ying-ying;LI Dong-xia;ZHANG Li-ning;HU Qi;ZHOU Li;LUO Sha;ZHOU Tong-liang;DU Meng-meng;TAO Qing-wen(Department of Traditional Chinese Rheumatology,China-Japan Friendship Hospital,Beijing,100029;Clinical Research Institute of China-Japan Friendship Hospital,Beijing,100029)

机构地区：[1]中日友好医院中医风湿病科,北京100029 [2]中日友好医院临床研究所,北京100029

出　　处：《中国中西医结合杂志》2020年第4期476-479,共4页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金青年科学基金项目(No.81704050);国家自然科学基金面上项目(No.81673941)。

摘　　要：目的探索软骨细胞生物钟基因及蛋白表达在骨关节炎(OA)发病中的作用,为OA临床时辰医疗提供实验依据。方法选用雄性SPF级SD大鼠27只,随机分为正常组、模型组、盐酸氨基葡萄糖组3组,每组9只。其中正常组不干预,模型组采用Hulth法手术干预,盐酸氨基葡萄糖组采用Hulth法手术造模,术后每天使用盐酸氨基葡萄糖25 mg/kg灌胃,连续灌胃28天。造模成功后采用HE染色及Mankin′s评分评价软骨情况;采用RT-PCR检测bmal1、per1、cry1 mRNA水平;Western Blot法检测BMAL1、PER1、CRY1蛋白表达水平。结果通过HE染色及Mankin′s评分可知,模型组采用Hulth法造模之后,其表现符合OA标准,在采用盐酸氨基葡萄糖治疗之后,OA状态有所改善;RT-PCR及Western Blot结果显示,与正常组比较,模型组大鼠bmal1 mRNA及BMAL1蛋白表达量降低,per1、cry1 mRNA及PER1、CRY1蛋白表达量升高(P<0.05)。与模型组比较,盐酸氨基葡萄糖组大鼠bmal1 mRNA及BMAL1蛋白表达量升高,per1、cry1 mRNA及PER1、CRY1蛋白表达量降低(P<0.05)。结论软骨细胞生物钟基因及蛋白变化贯穿于OA发病与向愈的整个过程,其中OA的发生伴随着软骨细胞生物钟的紊乱,OA的恢复伴随着软骨细胞生物钟的恢复。Objective To observe the roles of cartilage cell clock genes and protein expressions in the pathogenesis of osteoarthritis(OA), providing experimental basis for clinical OA treatment. MethodsTwenty-seven male SPF-grade SD rats were randomly divided into three groups, a normal group, a model group, and glucosamine hydrochloride group, 9 rats in each group. Rats in the normal group received no intervention. Rats in the model group received Hulth surgical intervention. And those in glucosamine hydrochloride group received Hulth surgical modeling and glucosamine hydrochloride(25 mg/kg) by gastrogavage for 28 successive days. HE staining and Mankin’s score were used to evaluate the cartilage after successful modeling. The expression levels of bmal1, cry1, and per1 genes and proteins were detected by RT-PCR and Western Blot. Results Results of HE staining and Mankin’s score showed that the performance of the model group met OA standard after modelled with Hulth method, and the OA status was improved after treatment with glucosamine hydrochloride. Results of RT-PCR and Western Blot showed that, as compared with the normal group, the expression levels of bmal1 gene and protein decreased and the expression levels of cry1 and per1 gene and protein increased in the model group(P<0.05). Compared with the model group, the expressions of bmal1 gene and protein increased, while the expressions of cry1 and per1 gene and protein decreased in glucosamine hydrochloride group(P<0.05). ConclusionChanges in chondrocyteclock genes and proteins run through the whole process of OA onset and healing, in which the occurrence of OA is accompanied by the disorder of chondrocyte clock, and the recovery of OA is accompanied by the recovery of chondrocyte clock.

关 键 词：软骨细胞 骨关节炎 动物模型 生物钟 

分 类 号：R684.3[医药卫生—骨科学]