独活寄生汤调控PI3K/Akt/mTOR信号通路介导细胞焦亡对膝骨关节炎模型兔的影响  

Effect of Duhuo Jisheng Decoction on knee osteoarthritis model rabbits through regulation of cell pyroptosis mediated by PI3K/Akt/mTOR signaling pathway

作　　者：贺琳钦 李鹏飞[1] 李小冬[1] 陈启鹏 唐宗瀚 宋雨鑫 宋寒冰[1] HE Lin-qin;LI Peng-fei;LI Xiao-dong;CHEN Qi-peng;TANG Zong-han;SONG Yu-xin;SONG Han-bing(Heilongjiang University of Chinese Medicine,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学,黑龙江哈尔滨150040

出　　处：《中国中药杂志》2025年第1期187-196,共10页China Journal of Chinese Materia Medica

基　　金：黑龙江省自然科学基金优秀青年科学基金项目(YQ2020H028)。

摘　　要：探究独活寄生汤防治膝骨关节炎(KOA)的可能作用机制。40只SPF级新西兰大兔采用SPSS 26.0软件随机分为空白组、模型组、独活寄生汤低剂量组、独活寄生汤高剂量组、独活寄生汤高剂量+磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路激活剂组(独活寄生汤高剂量+740Y-P组),每组8只。除空白组外,其余各组大兔采用木瓜蛋白酶膝关节腔注射+膝关节强迫屈曲位法构建KOA模型。造模完成后次日,空白组与模型组予以10 mL·kg^(-1)生理盐水灌胃,独活寄生汤低、高剂量组分别予以8.8、35.2 g·kg^(-1)独活寄生汤灌胃,独活寄生汤高剂量+740Y-P组予以35.2 g·kg^(-1)独活寄生汤灌胃+耳缘静脉注射0.15μmol·kg^(-1)740Y-P,各组均持续给药4周。造模及干预完成后对各组大兔进行行为学观察,干预完成后对各组大兔膝关节行影像学观察,取材后肉眼观察各组大兔膝关节软骨大体形态变化、苏木素-伊红(HE)染色对软骨组织行病理学观察,同时上述观察结果分别通过Lequesne MG评分、Kellgren-Lawrence(K-L)分级、Pelletier评分及Mankin评分进行量化评估,ELISA法检测软骨组织白细胞介素-1β(IL-1β)、白细胞介素-18(IL-18)、基质金属蛋白酶13(MMP13)含量,real-time RT-PCR法检测软骨组织PI3K、Akt、mTOR、Nod样受体蛋白3(NLRP3)、半胱氨酸蛋白酶1(caspase-1)、焦孔素D(GSDMD)mRNA表达,Western blot法检测软骨组织PI3K、Akt、mTOR、NLRP3、caspase-1、GSDMD蛋白表达。结果显示,与空白组比较,模型组膝关节退变明显;Lequesne MG评分、K-L分级、Pelletier评分及Mankin评分升高;软骨组织中IL-1β、IL-18、MMP13含量升高,PI3K、Akt、mTOR磷酸化激活且mRNA表达升高,NLRP3、caspase-1、GSDMD蛋白及mRNA表达升高。与模型组比较,独活寄生汤低、高剂量组上述指标皆逆转,且独活寄生汤高剂量组上述指标下降程度高于独活寄生汤低剂量组。与独活�This study aimed to investigate the underlying mechanisms of Duhuo Jisheng Decoction(DJD)in the prevention and treatment of knee osteoarthritis(KOA).Forty SPF New Zealand rabbits were randomly divided using SPSS 26.0 software into five groups:blank group,model group,low-dose DJD group,high-dose DJD group,and high-dose DJD+phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR)signaling pathway activator group(high-dose DJD+740Y-P group),with eight rabbits in each group.Except for the blank group,the KOA model was established in the other groups using papain injection into the knee joint cavity combined with forced flexion of the knee joint.The day after modeling,the blank group and model group were given normal saline at 10 mL·kg^(-1)by gavage,the low-dose DJD group received DJD at 8.8 g·kg^(-1)by gavage,the high-dose DJD group received DJD at 35.2 g·kg^(-1)by gavage,and the high-dose DJD+740Y-P group received DJD at 35.2 g·kg^(-1)by gavage along with 740Y-P at 0.15μmoL·kg^(-1)injected via the auricular vein.All groups received treatment continuously for four weeks.After modeling and intervention,behavioral observations were performed for all groups,and after the intervention,imaging assessments of the knee joints were conducted.Cartilage from the knee joints was collected,and gross morphological changes were observed.Pathological changes in cartilage tissue were examined using hematoxylin-eosin(HE)staining.The results of these observations were quantitatively evaluated using the Lequesne MG score,Kellgren-Lawrence(K-L)grading,Pelletier score,and Mankin score.ELISA was used to measure the levels of interleukin-1β(IL-1β),interleukin-18(IL-18),and matrix metalloproteinase 13(MMP13)in cartilage tissue.Real-time RT-PCR was used to detect the mRNA expression levels of PI3K,Akt,mTOR,Nod-like receptor protein 3(NLRP3),cysteine protease 1(caspase-1),and gasdermin D(GSDMD)in cartilage tissue.Western blot was employed to measure the protein expression levels of PI3K,Akt,mTOR,NL

关 键 词：PI3K/Akt/mTOR信号通路 NLRP3/caspase-1/GSDMD信号通路 细胞焦亡 独活寄生汤 膝骨关节炎 

分 类 号：R285.5[医药卫生—中药学] R-332[医药卫生—中医学]