摘要
目的:探讨加味当归四逆汤通过PI3K/Akt/mTOR信号通路对膝骨关节炎(KOA)模型大鼠关节软骨退变及细胞自噬相关因子表达的影响。方法:将96只SD大鼠饲养2周后,随机分为正常对照组、模型空白组、硫酸氨基葡萄糖组、加味当归四逆汤高、中和低剂量组,每组16只,采用改良的Hulth法造模。造模成功后24 h~4周各组分批次分别给予生理盐水、硫酸氨基葡萄糖、加味当归四逆汤高、中、低剂量灌服。各批次大鼠均在灌胃4周后观察软骨组织形态学变化和Mankin病理评分;实时PCR检测IL-1β、TNF-α、MMP-3、MMP-13、COL2A1表达;Western Blot法检测PI3K、p-Akt、p-mTOR和Beclin-1、LC3-Ⅰ/Ⅱ表达。结果:模型空白组关节软骨严重退变,软骨细胞分布紊乱,出现了严重萎缩、变性的情况;加味当归四逆汤低、中、高剂量各组,药物剂量越低,关节软骨细胞退变、萎缩程度越接近模型空白组,药物剂量越高,关节软骨细胞退变、萎缩程度越接近正常对照组;与模型空白组相比,硫酸氨基葡萄糖组、加味当归四逆汤高、中、低三种剂量组大鼠膝关节病理Mankin评分显著降低,差异有统计学意义(P<0.05),硫酸氨基葡萄糖组、加味当归四逆汤高、中剂量组大鼠膝关节IL-1β、MMP-3、MMP-13、PI3K、p-Akt、p-mTOR蛋白表达显著降低,差异有统计学意义(P<0.05),COL2A1、Aggrecan、Beclin-1、LC3-Ⅰ/Ⅱ蛋白表达明显上升,差异有统计学意义(P<0.05),且加味当归四逆汤高剂量组与硫酸氨基葡萄糖组各蛋白表达结果间差异无统计学意义(P>0.05)。结论:加味当归四逆汤能通过下调PI3K、p-Akt、p-mTOR蛋白表达,抑制PI3K/Akt/mTOR通路,诱导自噬基因Beclin1、LC3表达,提升膝骨关节炎软骨细胞自噬水平,以缓解膝骨关节炎关节软骨退变。
Objective:To investigate the efficacy of modified Danggui Sini decoction on articular degeration and the expression of autophagy related factors in knee osteoarthritis(KOA)rats through PI3 K/Akt/mTOR signaling pathway.Methods:96 SD rats were randomly divided into normal control group,blank model group,glucosamine sulfate group,modified Danggui Sini decoction high-dose group,middle-dose group and low-dose group with 16 rats in each group after feeding for 2 weeks.The modified Hulth method was used to build the model.Each group was administered in batches of physiological saline,glucosamine sulfate,and high,middle and low doses of modified Danggui Sini decoction 24 h to 4 weeks after the successful model building.The morphological changes of cartilage tissue were observed in each batch of rats after intragastric administration for 4 weeks;the expression of IL-1β,TNF-α,MMP-3,MMP-13,and COL2 A1 were detected by RT-PCR;the expression of PI3 K,p-Akt,p-mTOR,Beclin-1 and LC3-Ⅰ/Ⅱwere detected by Western Blot.Results:There were severe degeneration of articular cartilage,disorder of chondrocyte distribution,severe atrophy and degeneration in blank group.In the low-dose,middle-dose and high-dose groups of modified Danggui Sini decoction,with the increasing of drug dosage,the degree of degeneration and atrophy of articular chondrocytes was closer to that of the normal control group.Compared with model blank group,the pathological Mankin score of knee joint of rats in glucosamine sulfate group,modified Danggui Sini decoction high-dose,medium-dose and low-dose groups was significantly decreased(P<0.05).The protein expressions of IL-1β,MMP-3,MMP-13,PI3 K,p-Akt and p-mTOR in knee joint of rats in glucosamine sulfate group and high-dose and medium-dose modified Danggui Sini decoction groups were significantly decreased(P<0.05).The protein expressions of COL2 A1,Aggrecan,Beclin-1 and LC3-Ⅰ/Ⅱwere significantly increased(P<0.05),and there was no statistical significance between the high-dose modified Danggui Sini decoction
作者
王永涛
谢一舟
樊效鸿
余洋
WANG Yongtao;XIE Yizhou;FAN Xiaohong;YU Yang(Department of Orthopedics,Affiliated Hospital of Chengdu University of Traditional Chinese Medicine,Chengdu 610072,China)
出处
《中国中医骨伤科杂志》
CAS
2022年第11期7-12,共6页
Chinese Journal of Traditional Medical Traumatology & Orthopedics
基金
四川省中医药管理局课题(CKY2021127)。
关键词
膝骨关节炎
当归四逆汤
软骨细胞
自噬
knee osteoarthritis
Danggui Sini decoction
chondrocyte
autophagy
