短刺加电针法对兔膝骨关节炎模型软骨细胞肥大的影响

发布时间：2017-12-31 23:26

  本文关键词：短刺加电针法对兔膝骨关节炎模型软骨细胞肥大的影响　出处：《重庆医科大学》2017年硕士论文　论文类型：学位论文

  更多相关文章： 膝骨关节炎 短刺 核心结合蛋白因子2 Ⅹ型胶原

【摘要】：目的:观察短刺加电针法对兔膝骨关节炎的疗效,探索短刺加电针治疗兔膝骨关节炎的软骨细胞作用机制。方法:40只新西兰兔随机分为正常组(N组)10只、造模组30只,对造模组行Hulth-Telhag手术法复制膝骨关节炎模型,X线评估造模情况,造模成功后随机分为模型组(M组)、短刺组(CN组)和普通针刺组(NTN组)。术后6周开始治疗,M组不予任何处理,CN和NTN组取足三里、内外膝眼、梁丘和阴陵泉穴,CN组采用贴骨深刺的短刺加电针法,NTN组采用普通针刺加电针法,留针20min,每天1次,每个疗程5天,共治疗4个疗程。实验结束后采用HE染色观察软骨细胞的组织学改变,甲苯胺蓝染色观察软骨细胞形态,分别在治疗前和治疗结束后观察各组新西兰兔的行为学改变;免疫组化检测Runx2和ColⅩ蛋白的表达;Western-Blot法检测Runx2、Sox9和ColⅩ蛋白的表达;RT-PCR检测Sox9 mRNA的表达。结果:Lequesne MG评分、染色、Mankin评分、免疫组化、WB及RT-PCR结果一致。(1)行为学评分:N组Lequesne MG评分在0~1分,行为学正常,与N相比,治疗前M、CN、和NTN组Lequesne MG评分显著增高,有统计学差异(P0.01),M、CN、NTN组在治疗前Lequesne MG评分比较差异无统计学差异(P0.01),治疗结束后,与M组相比,CN和NTN组评分显著降低,其中CN组降低更加明显,有统计学差异(P0.01);(2)染色观察及Mankin评分:N组软骨表面光滑、软骨细胞分布均匀、排列整齐、甲苯胺蓝染色均匀无失染、潮线完整;M组软骨表层有裂隙、软骨细胞层次不清晰、排列无序、有成簇细胞出现、甲苯胺蓝染色深染、潮线不完整;CN组软骨表面不规则、软骨细胞数量较少、排列稍紊乱、轻度失染、潮线较完整;NTN组软骨表面粗糙、软骨细胞排列紊乱、出现成簇细胞、轻度失染、潮线不完整。CN组和NTN组与M组相比软骨细胞分布较均匀,软骨缺损程度减轻;CN组与NTN组相比细胞染色更均匀、细胞排列更整齐、层次清晰。Mankin评分显示:N组Mankin评分0~1分,M组Mankin评分7~9分,CN组Mankin评分3~5分,NTN组Mankin评分4~6分,四组评分有显著差异(P0.01)。(3)免疫组化和Western-Blot(WB)检测:与M组比较,N、CN、NTN组Runx2和ColⅩ蛋白表达量显著降低,比较差异有统计学意义(P0.01);CN组较NTN组显著降低,两组比较有差异(P0.01),说明短刺加电针法能有效下调Runx2和ColⅩ蛋白表达,且效果优于普通针刺组。(4)RT-PCR检测:与N组比较,M组、CN组和NTN组Sox9 mRNA表达量明显降低(P0.01);与M组比较,CN组和NTN组Sox9 mRNA表达量明显升高(P0.01);且CN组Sox9 mRNA表达量明显高于NTN组(P0.01)。结论:短刺加电针法较普通针刺法更能有效地治疗KOA,短刺加电针法能上调Sox9和下调Runx2、ColⅩ的表达,从而抑制兔膝骨关节软骨细胞肥大分化,这可能是其治疗KOA的机制之一。
[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，

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