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[1]袁普衛(wèi),李小群,康武林,等.蠲痹膠囊對(duì)膝骨關(guān)節(jié)炎豚鼠關(guān)節(jié)軟骨組織形態(tài)及滑膜中Toll樣受體4、NF-κB p65及腫瘤壞死因子-α表達(dá)的影響[J].中醫(yī)正骨,2016,28(09):5-12.
　YUAN Puwei,LI Xiaoqun,KANG Wulin,et al.Impact of Juanbi Jiaonang(蠲痹膠囊,JBJN)on articular cartilage tissue form and expression of Toll-like receptor 4,NF-kB p65 and tumor necrosis factor-alpha in synovium in guinea pigs with knee osteoarthritis[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2016,28(09):5-12.
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蠲痹膠囊對(duì)膝骨關(guān)節(jié)炎豚鼠關(guān)節(jié)軟骨組織形態(tài)及滑膜中Toll樣受體4、NF-κB p65及腫瘤壞死因子-α表達(dá)的影響()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第28卷
期數(shù):
2016年09期
頁碼:
5-12
欄目:
膝骨關(guān)節(jié)炎
出版日期:
2016-09-20

文章信息/Info

Title:
Impact of Juanbi Jiaonang(蠲痹膠囊,JBJN)on articular cartilage tissue form and expression of Toll-like receptor 4,NF-kB p65 and tumor necrosis factor-alpha in synovium in guinea pigs with knee osteoarthritis
作者:
袁普衛(wèi)1; 李小群1; 康武林1; 董博1; 陳斌2; 張曉亮2; 劉德玉1
1.陜西中醫(yī)藥大學(xué)附屬醫(yī)院,陜西 咸陽 712000; 2.陜西中醫(yī)藥大學(xué),陜西 咸陽 712046
Author(s):
YUAN Puwei1; LI Xiaoqun1; KANG Wulin1; DONG Bo1; CHEN Bin2; ZHANG Xiaoliang2; LIU Deyu1
1.The Affiliated Hospital of Shanxi University of Traditional Chinese Medicine,Xianyang 712000,Shanxi,China2.Shanxi University of Traditional Chinese Medicine,Xianyang 712046,Shanxi,China
關(guān)鍵詞:
骨關(guān)節(jié)炎; 膝;  Toll樣受體4;  NF-κB;  腫瘤壞死因子α;  軟骨; 關(guān)節(jié);  滑膜;  蠲痹膠囊;  氨基葡萄糖;  豚鼠;  動(dòng)物實(shí)驗(yàn)
Keywords:
osteoarthritis; knee;  Toll-like receptor 4;  NF-kappa B;  tumor necrosis factor-alpha;  cartilage; articular;  synovial membrane;  Juanbi Jiaonang;  glucosamine;  guinea pigs;  animal experimentation
摘要:
目的:觀察蠲痹膠囊對(duì)膝骨關(guān)節(jié)炎(knee osteoarthritis,KOA)豚鼠關(guān)節(jié)軟骨組織形態(tài)及滑膜中Toll樣受體4(Toll-like receptor 4,TLR4)、NF-κB p65及腫瘤壞死因子-α(tumor necrosis factor-α,TNF-α)表達(dá)的影響,探討其治療KOA的可能作用機(jī)制。方法:將40只1月齡豚鼠隨機(jī)分為正常組、3月齡組、5月齡組、氨基葡萄糖組和蠲痹膠囊組,每組8只。分組后正常組豚鼠直接進(jìn)行標(biāo)本采集,其余4組飼養(yǎng)至3月齡建立自發(fā)性KOA模型。造模結(jié)束后,3月齡組豚鼠進(jìn)行標(biāo)本采集; 5月齡組、氨基葡萄糖組和蠲痹膠囊組分別以純水、鹽酸氨基葡萄糖膠囊和蠲痹膠囊灌胃,每天1次,連續(xù)干預(yù)2個(gè)月后進(jìn)行標(biāo)本采集。采集標(biāo)本為豚鼠左后下肢膝關(guān)節(jié)髕下韌帶表面的滑膜和脛骨內(nèi)側(cè)平臺(tái)軟骨,制成石蠟切片。軟骨組織切片以番紅固綠染色后光鏡下觀察軟骨組織形態(tài),采用Mankin's病理評(píng)分標(biāo)準(zhǔn)進(jìn)行評(píng)分; 滑膜組織切片進(jìn)行免疫組織化學(xué)染色(SP法),用Image-pro Plus 6.0圖像分析軟件對(duì)陽性染色的平均光密度值進(jìn)行半定量分析,測定指標(biāo)包括TLR4、NF-κB p65及TNF-α的表達(dá)水平。結(jié)果:5組豚鼠關(guān)節(jié)軟骨Mankin's評(píng)分比較,差異有統(tǒng)計(jì)學(xué)意義[(0.132±0.125)分,(5.011±0.180)分,(7.324±0.293)分,(4.250±0.250)分,(2.500±0.327)分,F=108.400,P=0.000]; 3月齡組評(píng)分高于正常組(P=0.000),5月齡組評(píng)分高于3月齡組(P=0.000),氨基葡萄糖組評(píng)分低于5月齡組(P=0.000),蠲痹膠囊組評(píng)分低于氨基葡萄糖組(P=0.000)。5組豚鼠關(guān)節(jié)滑膜中TLR4、NF-κB p65及TNF-α表達(dá)量比較,組間差異均有統(tǒng)計(jì)學(xué)意義(0.000±0.000,0.268±0.021,0.454±0.043,1.879±0.013,0.116±0.015,F=97.000,P=0.000; 0.023±0.019,0.251±0.142,0.525±0.103,0.217±0.087,0.211±0.019,F=40.293,P=0.000; 0.047±0.023,0.386±0.142,0.812±0.174,0.289±0.151,0.204±0.108,F=78.242,P=0.000); 3月齡組TLR4、NF-κB p65及TNF-α表達(dá)量均高于正常組(P=0.000,P=0.000,P=0.000),5月齡組TLR4、NF-κB p65及TNF-α表達(dá)量均高于3月齡組(P=0.000,P=0.000,P=0.000),氨基葡萄糖組TLR4、NF-κB p65及TNF-α表達(dá)量均低于5月齡組(P=0.000,P=0.000,P=0.000),蠲痹膠囊組TLR4、NF-κB p65及TNF-α表達(dá)量均低于氨基葡萄糖組(P=0.011,P=0.019,P=0.008)。結(jié)論:蠲痹膠囊和鹽酸氨基葡萄糖均可延緩KOA豚鼠關(guān)節(jié)軟骨退變,蠲痹膠囊的作用更加明顯; 其作用機(jī)制可能是通過激活TLR4/NF-κB信號(hào)轉(zhuǎn)導(dǎo)通路來抑制炎性因子TNF-α表達(dá),從而減輕組織炎癥反應(yīng)。
Abstract:
Objective:To observe the impact of Juanbi Jiaonang(蠲痹膠囊,JBJN)on articular cartilage tissue form and expression of Toll-like receptor 4(TLR4),NF-κB p65 and tumor necrosis factor-alpha(TNF-a)in synovium in guinea pigs with knee osteoarthritis(KOA),and to explore the possible mechanism of action of JBJN in the treatment of KOA.Methods:Forty 1-month-old guinea pigs were randomly divided into normal group,3-month-old group,5-month-old group,glucosamine group and JBJN group,8 cases in each group.After grouping,the specimens were collected directly in normal group,while the guinea pigs in the other 4 groups were bred continuously and spontaneous KOA models were built when they were 3 months old.After modeling,the specimens were collected in 3-month-old group,while the guinea pigs in 5-month-old group,glucosamine group and JBJN group were intragastric administrated with pure water,glucosamine hydrochloride capsules and JBJN respectively,once a day for consecutive 2 months,and then the specimens were collected.The superficial synovium of infrapatellar ligament and the medial tibial plateau cartilage were collected from the knee of left posterior-inferior extremity and were made into paraffin sections.The cartilage tissue sections were received safranin-fast green staining and the cartilage tissue forms were observed under optical microscope and were evaluated according to Mankin score.The synovium tissue sections were received immunohistochemical staining(SP method),and the semiquantitative analysis were carried out on the average optical density value of the positive staining by using Image-pro Plus 6.0 image analysis software,and the main measuring indexes included the expression level of TLR4,NF-kB p65 and TNF-a.Results:There was statistical differences in Mankin scores of articular cartilage between the 5 groups(0.132+/-0.125,5.011+/-0.180,7.324+/-0.293,4.250+/-0.250,2.500+/-0.327 points,F=108.400,P=0.000),and 3-month-old group surpassed normal group(P=0.000), and 5-month-old group surpassed 3-month-old group(P=0.000)and glucosamine group(P=0.000),and glucosamine group surpassed JBJN group(P=0.000).There was statistical differences in the expression level of TLR4,NF-kB p65 and TNF-a in the articular synovium between the 5 groups(0.000+/-0.000,0.268+/-0.021,0.454+/-0.043,1.879+/-0.013,0.116+/-0.015,F=97.000,P=0.000; 0.023+/-0.019,0.251+/-0.142,0.525+/-0.103,0.217+/-0.087,0.211+/-0.019,F=40.293,P=0.000; 0.047+/-0.023,0.386+/-0.142,0.812+/-0.174,0.289+/-0.151,0.204+/-0.108,F=78.242,P=0.000),and 3-month-old group surpassed normal group(P=0.000,P=0.000,P=0.000),and 5-month-old group surpassed 3-month-old group(P=0.000,P=0.000,P=0.000)and glucosamine group(P=0.000,P=0.000,P=0.000),and glucosamine group surpassed JBJN group(P=0.011,P=0.019,P=0.008).Conclusion:Both JBJN and glucosamine hydrochloride can delay the articular cartilage degeneration in guinea pigs with KOA and the former has more significant effect.JBJN can inhibit the expression of inflammatory factor TNF-a through activating LR4/NF-kB signaling pathway,which may be the mechanism of action.

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備注/Memo:
2016-03-07收稿 2016-07-18修回
基金項(xiàng)目:國家自然科學(xué)基金項(xiàng)目(81574006); 陜西省重點(diǎn)科技創(chuàng)新團(tuán)隊(duì)項(xiàng)目(2013KCT-26); 陜西省自然科學(xué)基礎(chǔ)研究計(jì)劃項(xiàng)目(2010JM4002); 陜西省科學(xué)技術(shù)研究發(fā)展計(jì)劃項(xiàng)目(2011kjxx33); 咸陽市科學(xué)技術(shù)研究項(xiàng)目[2010K15-02(9)]; 全國名老中醫(yī)藥專家李堪印傳承工作室建設(shè)項(xiàng)目
袁普衛(wèi) E-mail:[email protected]

更新日期/Last Update: 1900-01-01