栀子苷抑制HSP70释放改善风湿热痹证胶原性关节炎大鼠血管新生的作用研究  

Inhibition of HSP70 release by geniposide improves angiogenesis in moist heat arthralgia spasm syndrome collagen induced arthritis rats

作　　者：舒寅 甘珮荣 王言 卜妍红 吴虹 SHU Yin;GAN Pei-rong;WANG Yan;BU Yan-hong;WU Hong(School of Pharmacy,Anhui University of Chinese Medicine,Key Lab of Xin’an Medicine,Ministry of Education,Anhui Province Key Lab of Chinese Medicinal Formula,Hefei 230012,China)

机构地区：[1]安徽中医药大学药学院,新安医学教育部重点实验室,中药复方安徽省重点实验室,安徽合肥230012

出　　处：《中国药理学通报》2024年第2期324-334,共11页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金面上资助项目(No 82374117,81874360);安徽省高校自然科学基金重点项目(No 2022AH050520)。

摘　　要：目的探讨栀子苷(geniposide,GE)改善风湿热痹证胶原性关节炎(collagen induced arthritis,CIA)大鼠血管新生的作用是否与调控热休克蛋白70(heat shock proteins 70,HSP70)的释放有关。方法在大鼠背部和尾根部多点皮内注射完全弗氏佐剂(complete Freund’s adjuvant,CFA)与鸡Ⅱ型胶原蛋白(chicken type II collagen,CCII)等体积混合物,构建CIA模型,在此基础上,给予风湿热刺激,建立风湿热痹证CIA大鼠(CIA rats with moist heat arthralgia spasm syndrome,CIA-S)模型。造模成功后随机分组,给药组分别用GE(60、120 mg·kg^(-1)·d^(-1))、MTX(0.5 mg·kg^(-1)·3d^(-1))灌胃两周,取材。观察大鼠体质量、关节炎指数(arthritis index,AI)等评价模型情况,关节表面温度和血流变指标评价热痹模型热属性,HE染色观察大鼠滑膜病理,多普勒彩超检测血流信号,免疫组化和Westren blot法检测滑膜组织HSP70、CD31、VEGF含量,ELISA法检测大鼠血清HSP70含量。EdU染色、细胞划痕和Transwell迁移实验以及成管实验探究HSP70对人脐静脉血管内皮细胞(human umbilic vein endothelial cells,HUVECs)的增殖、迁移、成管能力的影响。结果与空白组相比,CIA-S组关节肿胀,热属性明显,血管新生强烈,滑膜组织异常增生,炎性细胞浸润,可见微血管形成;CIA-S组与CIA组比较,关节红肿程度加重,炎症峰值提前、持续时间延长,消退缓慢,血管新生与HSP70表达更为强烈。GE给药组呈剂量依赖性,有效改善关节炎症状,减少HSP70含量。细胞实验结果表明,GE抑制HSP70的表达和释放,进而抑制HSP70诱导的HUVECs增殖、迁移和成管能力增强。结论GE有可能通过抑制HSP70的释放改善风湿热痹证CIA大鼠血管新生,进而改善关节炎。Aim To investigate the relation between the effect of geniposide(GE)in improving angiogenesis in arthralgia spasm syndrome collagen induced arthritis(CIA)rats and the modulation of heat shock proteins 70(HSP70)release.Methods A CIA model was constructed by multiple intradermal injections of complete Freund’s adjuvant(CFA)and an equal volume mixture of chicken type II collagen(CCII)into the dorsal and caudal root regions of rats,on the basis of which a rheumatic fever stimulus was given to build up a moist heat arthralgia spasm syndrome in CIA rats.After successful modeling,the groups were randomly grouped,and the administered groups were gavaged with GE(60,120 mg·kg^(-1)·d^(-1)),and MTX(0.5 mg·kg^(-1)·3d^(-1))for two weeks.Body mass and arthritis index of rats were observed to evaluate the model condition,joint surface temperature and blood rheology index to evaluate the thermal property of heat paralysis model,HE staining to observe the pathology of synovial membrane of rats,Doppler ultrasound to detect the blood flow signal,immunohistochemistry and Western blot to detect the content of HSP70,CD31,and VEGF in synovial membrane tissues,and ELISA to detect the content of HSP70 in the serum of rats.EdU staining,cell scratch and Transwell migration assays,and tube formation assays were performed to investigate the effects of HSP70 on the proliferation,migration,and tube formation of human umbilical vein vascular endo-thelial cells(HUVECs).Results Compared with the blank group,theCIA-S group had swollen joints,obvious thermal properties,intense neovascularization,abnormal proliferation of synovial tissue,inflammatory cell infiltration,and visible microvessel formation;the CIA-S group had increased redness and swelling in the joints compared with the CIA group,with the peak of inflammation advancing,prolonging the duration,and slowing down,and the expression of neovascularization and HSP70 was more intense.The GE administration group showed a dose-dependent effect,effectively improving the arthritis symptoms and

关 键 词：风湿热痹证CIA大鼠 HSP70 血管新生 栀子苷 人脐静脉血管内皮细胞 热应激 

分 类 号：R-332[医药卫生] R255.6R284.1R323.72R364.3R593.22