温经活血外治法通过抑制NGF/TrKA信号通路改善膝骨关节炎外周痛敏的机制研究
Experimental Study on the Improvement of Peripheral Hyperalgesia in KOA through Inhibition of NGF/TrKA Signaling Pathway by Warming Channels and Activating Blood Circulation External Treatment
李明超 1张力 2时孝晴 3廖太阳 3揭立士 3王培民 3黄桂成 3黄正泉3
作者信息
- 1. 南京中医药大学附属医院/江苏省中医院,江苏 南京 210029;昆山市第三人民医院骨科,江苏 苏州 215300
- 2. 东南大学附属中大医院,江苏 南京 210009
- 3. 南京中医药大学附属医院/江苏省中医院,江苏 南京 210029
- 折叠
摘要
目的 基于NGF/TrKA信号通路探讨温经活血外治法减轻膝骨关节炎(Knee osteoarthritis,KOA)外周痛敏的效应及机制.方法 30 只SD大鼠随机分为正常组、KOA组和易层组,通过前交叉韧带切断术(ACLT)进行KOA造模,造模后 14 d,易层组大鼠予以易层贴敷治疗.不同时间点测定大鼠外周痛敏阈值,对大鼠软骨切片进行HE、Aggrecan、Ⅱ型胶原染色;滑膜切片进行HE、天狼星红、镀银染色及免疫染色;Western blot法测定滑膜及复合体经神经末梢逆转运至背根神经节(Dorsal root ganglia,DRG)组织中NGF/TrKA信号通路的关键分子NGF、TrKA,炎症指标IL-1β,疼痛介质TRPV1,泛神经标记物PGP9.5、S100 等蛋白表达,qPCR测定相应基因表达,ELISA法测定大鼠外周血中NGF、SP含量.结果 与KOA组相比较,易层组大鼠冷痛敏和机械痛敏阈值增加(P<0.05,P<0.01);滑膜组织中NGF、TrKA、TRPV1、IL-1β、PGP9.5 等蛋白和基因表达下降(P<0.05,P<0.01);DRG组织中TRPV1、PGP9.5、S100 等蛋白和基因表达下调(P<0.05,P<0.01).结论 温经活血外治法可抑制NGF/TrKA信号通路,下调NGF、TrKA、TRPV1、IL-1β、PGP9.5 的基因和蛋白表达,并可能抑制感觉神经纤维萌发,改善大鼠KOA外周痛敏状态.
Abstract
OBJECTIVE To explore the effect and mechanism of warming channels and activating blood circulation external treat-ment to alleviate peripheral hyperalgesia in knee osteoarthritis(KOA)based on NGF/TrKA signaling pathway.METHODS 30 SD rats were randomly divided into normal group,KOA group and Yiceng group.KOA model was established by anterior cruciate ligament transection(ACLT).14 days after model establishment,rats in Yiceng group were treated with Yiceng patch.The peripheral pain threshold of rats was measured at different time points.The cartilage sections were stained with HE,Aggrecan and type II collagen.The synovial sections were stained with HE,Sirius red,silver and performed with immunostaining.The protein expression of key molecules NGF and TrKA of NGF/TrKA signaling pathway,inflammatory index IL-1β,pain mediator TRPV1,pan-neural mark-ers PGP9.5 and S100 in synovium and complexes transported to dorsal root ganglia(DRG)tissues via nerve endings was determined by Western Blot.The corresponding gene expression was determined by qPCR.The levels of NGF and SP in peripheral blood of rats were determined by ELISA.RESULTS Compared with the KOA group,the cold allodynia and mechanical allodynia thresholds of the rats in the Yiceng group increased(P<0.05,P<0.01);the protein and gene expression of NGF,TrKA,TRPV1,IL-1β,PGP9.5 in the synovial tissue decreased(P<0.05,P<0.01);the protein and gene expression levels of TRPV1,PGP9.5,S100 in the DRG tissue were downregulated(P<0.05,P<0.01).CONCLUSION The warming channels and activating blood circulation external treatment can inhibit the NGF/TrKA signaling pathway,downregulate the gene and protein expressions of NGF,TrKA,TRPV1,IL-1β,PGP9.5,and may inhibit the sprouting of sensory nerve fibers and improve the peripheral hyperalgesia state of rats with KOA.
关键词
膝骨关节炎/滑膜炎症/温经活血外治法/外周痛敏/神经生长因子Key words
knee osteoarthritis/synovial inflammation/warming channels and activating blood circulation external treatment/pe-ripheral hyperalgesia/nerve growth factor引用本文复制引用
基金项目
国家自然科学基金面上项目(82074460)
国家自然科学基金面上项目(82274545)
黄桂成全国名老中医药专家传承工作室建设项目(国中医药人教函[2022]75号)
苏州市科技计划项目(SYS2020059)
苏州市科技计划项目(SKJYD2021024)
昆山市科技计划项目(KS2239)
昆山市科技计划项目(KS2336)
出版年
2024
NETL
NSTL
万方数据