本文關(guān)鍵詞：短刺加電針法對兔膝骨關(guān)節(jié)炎模型軟骨細胞肥大的影響　出處：《重慶醫(yī)科大學》2017年碩士論文　論文類型：學位論文

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【摘要】：目的:觀察短刺加電針法對兔膝骨關(guān)節(jié)炎的療效,探索短刺加電針治療兔膝骨關(guān)節(jié)炎的軟骨細胞作用機制。方法:40只新西蘭兔隨機分為正常組(N組)10只、造模組30只,對造模組行Hulth-Telhag手術(shù)法復制膝骨關(guān)節(jié)炎模型,X線評估造模情況,造模成功后隨機分為模型組(M組)、短刺組(CN組)和普通針刺組(NTN組)。術(shù)后6周開始治療,M組不予任何處理,CN和NTN組取足三里、內(nèi)外膝眼、梁丘和陰陵泉穴,CN組采用貼骨深刺的短刺加電針法,NTN組采用普通針刺加電針法,留針20min,每天1次,每個療程5天,共治療4個療程。實驗結(jié)束后采用HE染色觀察軟骨細胞的組織學改變,甲苯胺藍染色觀察軟骨細胞形態(tài),分別在治療前和治療結(jié)束后觀察各組新西蘭兔的行為學改變;免疫組化檢測Runx2和ColⅩ蛋白的表達;Western-Blot法檢測Runx2、Sox9和ColⅩ蛋白的表達;RT-PCR檢測Sox9 mRNA的表達。結(jié)果:Lequesne MG評分、染色、Mankin評分、免疫組化、WB及RT-PCR結(jié)果一致。(1)行為學評分:N組Lequesne MG評分在0~1分,行為學正常,與N相比,治療前M、CN、和NTN組Lequesne MG評分顯著增高,有統(tǒng)計學差異(P0.01),M、CN、NTN組在治療前Lequesne MG評分比較差異無統(tǒng)計學差異(P0.01),治療結(jié)束后,與M組相比,CN和NTN組評分顯著降低,其中CN組降低更加明顯,有統(tǒng)計學差異(P0.01);(2)染色觀察及Mankin評分:N組軟骨表面光滑、軟骨細胞分布均勻、排列整齊、甲苯胺藍染色均勻無失染、潮線完整;M組軟骨表層有裂隙、軟骨細胞層次不清晰、排列無序、有成簇細胞出現(xiàn)、甲苯胺藍染色深染、潮線不完整;CN組軟骨表面不規(guī)則、軟骨細胞數(shù)量較少、排列稍紊亂、輕度失染、潮線較完整;NTN組軟骨表面粗糙、軟骨細胞排列紊亂、出現(xiàn)成簇細胞、輕度失染、潮線不完整。CN組和NTN組與M組相比軟骨細胞分布較均勻,軟骨缺損程度減輕;CN組與NTN組相比細胞染色更均勻、細胞排列更整齊、層次清晰。Mankin評分顯示:N組Mankin評分0~1分,M組Mankin評分7~9分,CN組Mankin評分3~5分,NTN組Mankin評分4~6分,四組評分有顯著差異(P0.01)。(3)免疫組化和Western-Blot(WB)檢測:與M組比較,N、CN、NTN組Runx2和ColⅩ蛋白表達量顯著降低,比較差異有統(tǒng)計學意義(P0.01);CN組較NTN組顯著降低,兩組比較有差異(P0.01),說明短刺加電針法能有效下調(diào)Runx2和ColⅩ蛋白表達,且效果優(yōu)于普通針刺組。(4)RT-PCR檢測:與N組比較,M組、CN組和NTN組Sox9 mRNA表達量明顯降低(P0.01);與M組比較,CN組和NTN組Sox9 mRNA表達量明顯升高(P0.01);且CN組Sox9 mRNA表達量明顯高于NTN組(P0.01)。結(jié)論:短刺加電針法較普通針刺法更能有效地治療KOA,短刺加電針法能上調(diào)Sox9和下調(diào)Runx2、ColⅩ的表達,從而抑制兔膝骨關(guān)節(jié)軟骨細胞肥大分化,這可能是其治療KOA的機制之一。
[Abstract]:Objective: to observe the effect of short needling plus electric acupuncture on knee osteoarthritis in rabbits. Methods 40 New Zealand rabbits were randomly divided into normal group (n = 10) and model group (n = 30). The model group was established by Hulth-Telhag operation. The model of knee osteoarthritis was established by X-ray. After the model was successfully made, the model group was randomly divided into two groups: group M (model group). From 6 weeks after operation, no treatment was given in group N and NTN for Zusanli, internal and external knee eyes, Liangqiu and Yinlingquan points. CN group was treated with short needling plus electric acupuncture with deep bone sticking and NTN group with common acupuncture plus electric acupuncture for 20 minutes, once a day for 5 days. The histological changes of chondrocytes were observed by HE staining and the morphology of chondrocytes were observed by toluidine blue staining. The behavioral changes of New Zealand rabbits were observed before and after treatment. The expression of Runx2 and Col 鈪，

本文編號：1361950