风湿病与关节炎
風濕病與關節炎
풍습병여관절염
Rheumatism and Arthritis
2015年
4期
5-9
,共5页
林飞太%林煜%张怡元%夏春%张兵%肖莉莉%冯尔宥%王武炼
林飛太%林煜%張怡元%夏春%張兵%肖莉莉%馮爾宥%王武煉
림비태%림욱%장이원%하춘%장병%초리리%풍이유%왕무련
骨关节炎%独活寄生汤%软骨细胞%细胞凋亡%信号通路
骨關節炎%獨活寄生湯%軟骨細胞%細胞凋亡%信號通路
골관절염%독활기생탕%연골세포%세포조망%신호통로
osteoarthritis%Duhuo Jisheng Decoction%chondrocyte%apoptosis%signal pathway
目的：观察独活寄生汤含药血清对兔退变软骨细胞“caveolin-p38MAPK”信号通路的调控作用，探讨独活寄生汤治疗骨关节炎的作用机制。方法：将20只3月龄新西兰兔随机分为生理盐水组（空白血清组）和独活寄生汤组（含药血清组），每组10只。分别在24 h、36 h、48 h不同采血时间点采集独活寄生汤含药血清和空白血清，将5%、10%、15%、20%不同浓度两种血清作用于体外培养第2代软骨细胞，确定含药血清最佳干预条件；建立体外退变软骨细胞模型，分别给予独活寄生汤含药血清（含药血清组）和空白血清（空白血清组）干预36 h，收集软骨细胞，运用Western Blot法检测血清干预后退变软骨细胞caveolin-1、p38、p-p38蛋白表达，RT-PCR法检测血清干预后退变软骨细胞白细胞介素（IL）-1β、肿瘤坏死因子（TNF）-α、基质金属蛋白酶（MMP）-3、MMP-13、caveolin-1 mRNA表达。结果：浓度为15%的36 h采血时间点含药血清的促增殖作用最明显；退变软骨细胞中存在“caveolin-p38MAPK”信号通路的激活，独活寄生汤含药血清可抑制caveolin-1、p-p38蛋白表达及IL-1β、TNF-α、MMP-3、MMP-13、caveolin-1 mRNA的表达，差异有统计学意义（P<0.05）。结论：独活寄生汤能通过抑制“caveolin-p38MAPK”信号通路的激活及其下游效应分子，从而有效抑制软骨细胞凋亡。
目的：觀察獨活寄生湯含藥血清對兔退變軟骨細胞“caveolin-p38MAPK”信號通路的調控作用，探討獨活寄生湯治療骨關節炎的作用機製。方法：將20隻3月齡新西蘭兔隨機分為生理鹽水組（空白血清組）和獨活寄生湯組（含藥血清組），每組10隻。分彆在24 h、36 h、48 h不同採血時間點採集獨活寄生湯含藥血清和空白血清，將5%、10%、15%、20%不同濃度兩種血清作用于體外培養第2代軟骨細胞，確定含藥血清最佳榦預條件；建立體外退變軟骨細胞模型，分彆給予獨活寄生湯含藥血清（含藥血清組）和空白血清（空白血清組）榦預36 h，收集軟骨細胞，運用Western Blot法檢測血清榦預後退變軟骨細胞caveolin-1、p38、p-p38蛋白錶達，RT-PCR法檢測血清榦預後退變軟骨細胞白細胞介素（IL）-1β、腫瘤壞死因子（TNF）-α、基質金屬蛋白酶（MMP）-3、MMP-13、caveolin-1 mRNA錶達。結果：濃度為15%的36 h採血時間點含藥血清的促增殖作用最明顯；退變軟骨細胞中存在“caveolin-p38MAPK”信號通路的激活，獨活寄生湯含藥血清可抑製caveolin-1、p-p38蛋白錶達及IL-1β、TNF-α、MMP-3、MMP-13、caveolin-1 mRNA的錶達，差異有統計學意義（P<0.05）。結論：獨活寄生湯能通過抑製“caveolin-p38MAPK”信號通路的激活及其下遊效應分子，從而有效抑製軟骨細胞凋亡。
목적：관찰독활기생탕함약혈청대토퇴변연골세포“caveolin-p38MAPK”신호통로적조공작용，탐토독활기생탕치료골관절염적작용궤제。방법：장20지3월령신서란토수궤분위생리염수조（공백혈청조）화독활기생탕조（함약혈청조），매조10지。분별재24 h、36 h、48 h불동채혈시간점채집독활기생탕함약혈청화공백혈청，장5%、10%、15%、20%불동농도량충혈청작용우체외배양제2대연골세포，학정함약혈청최가간예조건；건입체외퇴변연골세포모형，분별급여독활기생탕함약혈청（함약혈청조）화공백혈청（공백혈청조）간예36 h，수집연골세포，운용Western Blot법검측혈청간예후퇴변연골세포caveolin-1、p38、p-p38단백표체，RT-PCR법검측혈청간예후퇴변연골세포백세포개소（IL）-1β、종류배사인자（TNF）-α、기질금속단백매（MMP）-3、MMP-13、caveolin-1 mRNA표체。결과：농도위15%적36 h채혈시간점함약혈청적촉증식작용최명현；퇴변연골세포중존재“caveolin-p38MAPK”신호통로적격활，독활기생탕함약혈청가억제caveolin-1、p-p38단백표체급IL-1β、TNF-α、MMP-3、MMP-13、caveolin-1 mRNA적표체，차이유통계학의의（P<0.05）。결론：독활기생탕능통과억제“caveolin-p38MAPK”신호통로적격활급기하유효응분자，종이유효억제연골세포조망。
Objective:To observe the effects of serum medicated with Duhuo Jisheng Decoction on the induced degeneration of articular chondrocytes in vitro culture based upon“caveolin-p38MAPK”signal path-way and probe the mechanism of Duhuo Jisheng Decoction in the treatment of osteoarthritis.Methods:20 March old New Zealand rabbits were randomly divided into normal saline group(blank serum group)and Duhuo Jisheng Decoction group (drug containing serum group),10 rats in each group.The second generation of carti-lage cells was cultured in vitro using different time points of sample collection such as 24 h,36 h and 48 h,serums medicated with 5%,10%,15%and 20%concentrations of Duhuo Jisheng Decoction and blank serums.The best intervention time of medicated serum was determined,which was then used to deal with the degenerating chon-drocytes.The degenerated cartilage cell models in vitro were established,which were intervened by the serum medicated with Duhuo Jisheng Decoction(drug containing serum group)and the blank serum(blank serum group)after 36 h,collecting chondrocytes. The Western Blot method was used to detect the expression of degenerated cartilage cells caveolin-1 and p-p38 protein after serum intervention.The RT-PCR method was used to detect the expression of degenerated cartilage cells IL-1β,TNF-α,MMP-3,MMP-13 and caveolin-1 mRNA after serum intervention.Results:The concentration of 15%and 36 h sampling time point serum on the prolifera-tion;the activation of"caveolin-p38MAPK"signaling pathway in the degeneration of cartilage cell,Duhuo Jisheng decoction containing serum can inhibit the expression of caveolin-1,p-p38 protein expression and IL-1β,TNF-α, MMP-3,MMP-13,caveolin-1,mRNA,the difference was statistically signiifcant(P<0.05).Conclusion:Duhuo Jisheng decoction can inhibit the"caveolin-p38MAPK"signal pathway and its downstream effector molecules, which can effectively inhibit the apoptosis of cartilage cells.
