单味药鹿茸调节骨关节炎模型兔软骨细胞外基质主要成分ADAMTS-4/TIMP-3基因的表达  

Pilose antler regulates the expression of ADAMTS-4/TIMP-3 gene in the extracellular matrix in a rabbit model of osteoarthritis

作　　者：孙志涛[1] 冯华龙 何升华[1] 赖居易 黄飞强[2] 王业广[1] Sun Zhi-tao;Feng Hua-long;He Sheng-hua;Lai Ju-yi;Huang Fei-qiang;Wang Ye-guang(Shenzhen Traditional Chinese Medicine Hospital,Shenzhen 518033,Guangdong Province,China;Fourth Clinical Medical College,Guangzhou University of Chinese Medicine,Shenzhen 518033,Guangdong Province,China)

机构地区：[1]深圳市中医院,广东省深圳市518033 [2]广州中医药大学第四临床医学院,广东省深圳市518033

出　　处：《中国组织工程研究》2018年第32期5133-5138,共6页Chinese Journal of Tissue Engineering Research

基　　金：深圳市卫生计生系统科研项目(201505016)~~

摘　　要：背景:大量研究证实鹿茸对于治疗骨关节炎有效,但是其与软骨细胞外基质主要成分ADAMTS-4/TIMP-3基因的相关性却报道较少。目的:探讨鹿茸对兔骨关节炎模型ADAMTS-4/TIMP-3基因表达的影响,进一步阐明鹿茸防治骨关节炎的作用机制。方法:选择健康雌性新西兰大白兔42只,采用前交叉韧带离断法建立兔骨关节炎模型,随机分为3组:对照组(生理盐水)、低剂量鹿茸组(0.21 g/kg)和高剂量鹿茸组(0.84 g/kg)。干预9周后,检测双膝关节中的羟脯氨酸含量,采用RT-PCR检测各组软骨靶基因mRNA的表达,采用Westernblot测量蛋白聚糖和Ⅱ型胶原蛋白的表达。结果与结论:(1)低剂量鹿茸组和高剂量鹿茸组软骨羟脯氨酸含量显著高于对照组,且高剂量鹿茸组显著高于低剂量鹿茸组(P <0.05);(2)高剂量鹿茸组和低剂量鹿茸组ADAMTS-4 mRNA表达较对照组明显降低,且高剂量显著组低于低剂量鹿茸组(P <0.05),而TIMP-3、蛋白聚糖、Ⅱ型胶原蛋白mRNA表达较对照组明显增加,且高剂量鹿茸组显著高于低剂量鹿茸组(P <0.05);(3)高剂量鹿茸组和低剂量鹿茸组蛋白聚糖、Ⅱ型胶原蛋白表达较对照组明显增高,且高剂量鹿茸组显著高于低剂量鹿茸组(P <0.05);(4)结果提示,鹿茸通过抑制ADAMTS-4的分泌,促进保护因子TIMP-3的分泌,阻止蛋白聚糖、Ⅱ型胶原蛋白蛋白的降解,起到修复软骨的作用。BACKGROUND:Pilose antler has been shown to be effective for osteoarthritis,but its association with ADAMTS-4/TIMP-3 gene(main composition of extracellular matrix)is rarely reported.OBJECTIVE:To investigate the effect of pilose antler on the expression of ADAMTS-4/TIMP-3 gene in rabbit models of osteoarthritis,and to further elucidate the mechanism of pilose antler in the prevention and treatment of osteoarthritis.METHODS:Forty-two healthy female New Zealand white rabbits were selected and used for establishing the models of osteoarthritis by anterior cruciate ligament rupture.The rabbit models were then randomized into three groups:control group(normal saline),low-dose(0.21 g/kg)and high-dose(0.84 g/kg)pilose antler groups.After intervention for 9 weeks,the content of hydroxyproline in both knees was detected.The expression levels of mRNA of cartilage target genes in each group were detected by RT-PCR.Protein expression levels of aggrecan and collagen type II were determined by western blot assay.RESULTS AND CONCLUSION:Compared with the control group,the content of hydroxyproline in cartilage in the low-dose and high-dose pilose antler groups was significantly increased,and the content in the high-dose pilose antler group was significantly higher than that in the low-dose pilose antler group(P<0.05).The order of mRNA expression level of ADAMTS-4 was as follows:high-dose pilose antler group<low-dose pilose antler group<control group(P<0.05).The order of mRNA expression levels of metallopeptidase inhibitor 3,aggrecan and collagen type II was as follows:high-dose pilose antler group>low-dose pilose antler group>control group(P<0.05).The expression levels of aggrecan and collagen type II were the highest in the high-dose pilose antler group,followed by the low-dose pilose antler group,and the lowest in the control group(P<0.05).Our findings indicate that pilose antler can repair the injured cartilage by inhibiting the secretion of ADAMTS-4,increasing the secretion of metallopeptidase inhibitor 3 and preventing the de

关 键 词：鹿茸 兔模型 骨关节炎 ADAMTS-4 TIMP-3 羟脯胺酸 蛋白聚糖 Ⅱ型胶原蛋白 软骨修复 中草药 骨关节炎 细胞外基质 软骨 组织工程 

分 类 号：R318[医药卫生—生物医学工程]