雷公藤红素对类风湿关节炎小鼠骨侵蚀的影响  

Effect of celastrol on bone erosion in mice with rheumatoid arthritis

作　　者：周伟 刘志刚 宋卫中[3] ZHOU Wei;LIU Zhi-gang;SONG Wei-zhong(School of Nursing and Health,Henan University,Kaifeng 475001,Henan Province,China;Department of Orthopedic,The First Affiliated Hospital of Henan University,Kaifeng 475001,Henan Province,China;Teaching and Research Office of Chinese Medicine,School of Pharmacy,Henan University,Kaifeng 475001,Henan Province,China)

机构地区：[1]河南大学护理与健康学院,河南开封475001 [2]河南大学第一附属医院骨科,河南开封475001 [3]河南大学药学院中药教研室,河南开封475001

出　　处：《中国临床药理学杂志》2021年第18期2476-2480,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：目的研究雷公藤红素(CSL)在胶原诱导性类风湿关节炎(CIA)小鼠骨侵蚀中的作用及其可能的作用机制。方法将24只DBA/1小鼠分为空白组、模型组(CIA小鼠模型)和实验组(腹腔注射2 mg·kg^(-1)CSL的CIA小鼠)。统计各组小鼠的关节炎指数评分,用苏木精-伊红(HE)染色法评估滑膜炎浸润及骨侵蚀情况,用TRAP染色检测骨组织破骨细胞(OC)数量。从小鼠股骨中分离诱导破骨细胞前体细胞(osteoclast precursors cells,OCPs),采用M-CSF+RNAKL诱导OC分化,分为OCPs组、诱导分化组和CSL低、中、高剂量组(分别用含有0.1,0.25,0.5μmol·L^(-1)CSL的诱导分化培养基培养)。用流式细胞仪检测OCPs的凋亡率;用TRAP染色观察OC数量;用蛋白质印迹(Western blot)法检测Fas/FasL死亡受体途径相关蛋白Fas、FasL、cleaved-caspase-8和cleaved-caspase-3的表达。结果CSL能够缓解CIA小鼠的关节炎症,抑制炎性细胞的浸润和骨侵蚀,减少OC的数量。OCPs组、诱导分化组和CSL低、中、高剂量组中细胞凋亡率分别为(1.82±0.24)%,(7.98±0.94)%,(8.41±0.84)%,(8.87±0.95)%,(12.57±1.53)%,OC细胞数分别为(0.8±0.7),(84.8±8.9),(70.0±6.6),(46.5±8.2),(32.5±4.6)个。与OCPs组相比,诱导分化组中细胞凋亡率和OC细胞增多(P<0.05)。与诱导分化组相比,CSL高剂量组细胞凋亡率升高,而OC细胞数量减少(P<0.05)。CSL高剂量组中Fas、FasL、cleaved-caspase-8和cleaved-caspase-3的表达水平显著高于诱导分化组(P<0.05)。结论CSL能够有效缓解CIA小鼠的骨侵蚀作用,其机制可能是CSL激活Fas/FasL死亡受体途径诱导OCPs的凋亡并抑制OC分化,抑制骨破坏的发生。Objective To investigate the effect of celastrol(CSL)on bone erosion in collagen-induced rheumatoid arthritis(CIA)mice and its possible mechanism.Methods A total of 24 DBA/1 mice were divided into blank group,model group(CIA mouse model),and experimental group(CIA mice treated by intraperitoneal injection of 2mg·kg^(-1)CSL).Arthritis index score of mice in each group was counted,synovitis infiltration and bone erosion were assessed by hematoxylin-eosin(HE)staining,and the amount of bone tissue osteoclast(OC)was detected by TRAP staining.Osteoclast precursors cells(OCPs)were isolated and induced from mouse femoral bone marrow,and differentiation of OC cells was induced by M-CSF+RNAKL.The cells were divided into the OCPs group,RANKL/OPCs group,CSL-L,CSL-M and CSL-H groups(cultured in induced differentiation medium containing 0.1,0.25 and0.5μmol·L^(-1)CSL,respectively).The apoptosis rate of OCPs was detected by flow cytometry;the number of OC was observed by TRAP staining.Expression of Fas/Fas L death receptor pathway related proteins:Fas,Fas L,cleavedcaspase-8 and cleaved-caspase-3 were detected by Western blot.Results CSL can alleviate joint inflammation,inhibit inflammatory cell infiltration and bone erosion,reduce the number of OC in CIA mice.The apoptosis rate in OCPs group,RANKL/OPCs group,CSL-L group,CSL-M group and CSL-H group were(1.82±0.24)%,(7.98±0.94)%,(8.41±0.84)%,(8.87±0.95)%,(12.57±1.53)%and the numbers of OC were(0.8±0.7),(84.8±8.9),(70.0±6.6),(46.5±8.2),(32.5±4.6),respectively.Compared with OCPs group,the apoptosis rate and OC cells were increased in RANKL/OCPs group(P<0.05).Compared with RANKL/OCPs group,the apoptosis was increased,while the OC cells were reduced in CSL-H group(P<0.05).The expression of Fas,Fas L,cleaved-caspase-8,and cleaved-caspase-3 in CSL-H group were significantly higher than those in RANKL/OCPs group(P<0.05).Conclusion CSL can effectively alleviate bone erosion in CIA mice,and its mechanism may be that CSL activates the Fas/Fas L death receptor pathway to induce

关 键 词：雷公藤红素 破骨前体细胞 破骨细胞 骨侵蚀 

分 类 号：R28[医药卫生—中药学]