透骨消痛胶囊含药血清对软骨细胞TGF-β通路中相关信号分子影响的实验研究  

Effect of Touguxiaotong Capsule on Related Molecules in TGF-β Signal Pathway of Chondrocytes

作　　者：刘伯龄[1] 梁珪清[1] 李照辉[1] 刘少强[1] 吴彬彬[1] 林鹏程[1] LIU Boling LIANG Guiqing LI Zhaohui LIU Shaoqiang WU Binbin LIN Pengcheng(Department of Orthopaedic, Second Hospital of Fuzhou, Affiliated to Xiamen University,Fuzhou 350007,China.)

机构地区：[1]厦门大学附属福州第二医院骨科,福州350007

出　　处：《中国中医骨伤科杂志》2016年第12期1-6,共6页Chinese Journal of Traditional Medical Traumatology & Orthopedics

基　　金：福建省福州市科技计划项目(2013-S-123-8)

摘　　要：目的:观察透骨消痛胶囊含药血清对硝普钠造模后软骨细胞TGF-β(转化生长因子-β)信号通路中ALK1,ALK5,ALK1/ALK5及MMP-13的表达影响,探讨其防治骨关节炎的作用机制。方法:2月龄清洁级雄性SD大鼠90只,随机分为空白组30只、壮骨关节丸组30只和透骨消痛胶囊组30只,制备含药血清。4周龄清洁级雄性SD大鼠膝关节软骨建立软骨细胞体外培养体系。1 mmol/L SNP干预第2代软骨细胞复制软骨细胞凋亡模型。分为1 mmol/L SNP组,1 mmol/L SNP+10%壮骨关节丸组,1 mmol/L SNP+5%、10%和15%透骨消痛胶囊组,分别培养3,6和9d,Real-time PCR和Western blot分别检测各组软骨细胞ALK1,ALK5及MMP-13基因及蛋白表达。结果:含药血清分别干预细胞3,6和9d后,ALK1,ALK1/ALK5及MMP-13基因及蛋白的表达,壮骨关节丸组和透骨消痛胶囊组均明显低于1 mmol/L SNP组(P<0.01,P<0.05);ALK5基因及蛋白的表达,壮骨关节丸组和透骨消痛胶囊组均明显高于1 mmol/L SNP组(P<0.01,P<0.05)。结论:透骨消痛胶囊通过调节TGF-β信号通路发挥抗软骨细胞凋亡作用。Objective：To investigate the effect of Touguxiaotong capsule medicated serum on the expression of ALK1,ALK5,ALK1/ALK5 and MMP-13 in model chondrocytes intervened by SNP,and to explore its mechanism of prevention and treatment of osteoarthritis.Methods：Ninety male SD rats（two months old）were randomly divided into blank group,Zhuangguguanjie pill group,and Touguxiaotong capsule group（n=30 in each group）,preparing for the medicated serum respectively.Chondrocytes cultured in vitro,which was collected from knee joint cartilage of 4-weeks-old male SD rats,was established.1 mmol/L SNP was chosen to mould the degenerated chondrocytes model in passage 2 chondrocytes,which were divided as follows：1 mmol/L SNP group,1 mmol/L SNP＋ 10% Zhuangguguanjie pill group,and 1 mmol/L SNP＋ 5%,10% and 15% Touguxiaotong capsule group,intervened by 3 d,6 dand 9 drespectively.The expression of ALK1,ALK5,MMP-13 and related protein in the chondrocytes of each group were detected by RT-PCR and Western blot respectively.Results：Compared with 1 mmol/L SNP group,the expression of ALK1,ALK1/ALK5,MMP-13 and related protein in Zhuangguguanjie pill group and Touguxiaotong capsule group were significantly lower after treated with medicated serum at 3th,6th,and 9th day respectively（P〈0.01,P〈0.05）,while the expression of ALK5 were significantly increased（P〈0.01,P〈0.05）.Conclusion：Touguxiaotong capsule plays an anti-chondrocyte apoptosis effect by regulating the TGF-βsignal pathway.

关 键 词：透骨消痛胶囊 含药血清 软骨细胞 TGF-Β信号通路 

分 类 号：R33[医药卫生—人体生理学]