【作者】 范李；

【导师】 杨述华；

【作者基本信息】 华中科技大学 ， 外科学， 2010， 博士

【摘要】 目的：观察辛伐他汀对兔骨性关节炎(OA)模型中软骨组织形态学的影响方法：健康新西兰5周龄大白兔30只,平均体重2.6kg,将动物随机分为A、B、C、D、E五组,每组6只动物,用戊巴比妥钠(40 mg/kg剂量)麻醉。除A组外其余4组按照Hulth法造模。术后C、D、E组动物分别给予患膝立即关节腔注射0.01mg/ml、0.1mg/ml和0.5mg/ml辛伐他汀稀释液,剂量为0.2ml/kg,每周1次,连续6周。B组则患膝关节腔注射等量生理盐水,每周1次,连续6周。A组为空白对照组,动物不做任何处理。所有动物不做固定,分笼饲养。术后6周处死动物。取患膝全层关节软骨,石蜡包埋切片,HE染色及番红O染色,中性树脂封片,利用解剖显微镜进行关节软骨大体观察,利用光学显微镜观察染色后切片并进行Mankin评分。结果：B组软骨关节面较其余各组破坏严重,且可见明显溃疡和裂隙,C、D、E组软骨结构、形态及软骨细胞基质对番红O异染性均优于B组。A、C、D、E组软骨Mankin评分均低于B组,其中A组、D组和E组较B组差异有显著性(P<0.05)。结论：①本实验采用Hulth法建立OA模型成功；②OA早期关节腔内注射辛伐他汀可以防止软骨退变,促进软骨基质合成,对软骨有保护作用。目的：研究辛伐他汀关节腔内注射对OA模型关节液及软骨中IL-1及MMP-13和Ⅱ型胶原表达的影响。方法：健康新西兰5周龄大白兔30只,平均体重2.6kg,将动物随机分为A、B、C、D、E五组,每组6只动物,用戊巴比妥钠(40 mg/kg剂量)麻醉。除A组外其余4组按照Hulth法造模。术后C、D、E组动物分别给予患膝立即关节腔注射0.01mg/ml、0.1mg/ml和0.5mg/ml辛伐他汀稀释液,剂量为0.2ml/kg,每周1次,连续6周。B组则患膝关节腔注射等量生理盐水,每周1次,连续6周。A组为空白对照组,动物不做任何处理。所有动物不做固定,分笼饲养。术后6周处死动物。取动物患膝关节液和关节软骨,利用RT-PCR技术检测软骨细胞中MMP-13和Ⅱ型胶原mRNA表达,用ELISA检测关节液IL-1的含量,运用免疫组织化学SABC法检测软骨组织内MMP-13的表达情况。结果：A、C、D、E组IL-1含量均明显低于B组,差异有显著性(P<0.05)；A、D、E组MMP-13 mRNA表达量均明显低于B组,且差异有显著性(P<0.05)；A、D、E组Ⅱ型胶原mRNA表达均明显高于B组,差异有显著性(P<0.05)。结论：在OA早期关节腔内注射辛伐他汀能够减轻关节软骨炎症反应,增强胶原合成代谢,减轻骨关节炎软骨的退变。目的：观察辛伐他汀对兔骨性关节炎(OA)模型中软骨细胞凋亡率的影响。方法：健康新西兰5周龄大白兔30只,平均体重2.6kg,将动物随机分为A、B、C、D、E五组,每组6只动物,用戊巴比妥钠(40 mg/kg剂量)麻醉。除A组外其余4组按照Hulth法造模。术后C、D、E组动物分别给予患膝立即关节腔注射0.01mg/ml、0.1mg/ml和0.5mg/ml辛伐他汀稀释液,剂量为0.2ml/kg,每周1次,连续6周。B组则患膝关节腔注射等量生理盐水,每周1次,连续6周。A组为空白对照组,动物不做任何处理。所有动物不做固定,分笼饲养。术后6周处死动物。取动物患膝关节软骨,利用流式细胞仪检测软骨细胞凋亡率；利用RT-PCR技术检测软骨细胞中Bcl-1mRNA表达。结果：C、D、E组软骨细胞凋亡率较B组下降,其中D组和E组软骨细胞凋亡率较B组显著下降,差异有显著性(P<0.05)；C、D、E组软骨细胞Bcl-1mRNA相对表达量均较B组下降,其中D组和E组Bcl-1mRNA相对表达量较B组显著下降,差异有显著性(P<0.05)。结论辛伐他汀可以呈剂量依赖性降低OA软骨细胞凋亡率,其机制可能与辛伐他汀下调Bcl-1mRNA表达有关。目的：观察辛伐他汀对兔骨性关节炎(OA)软骨细胞凋亡的影响。方法：试验分为5组,取5周龄兔原代关节软骨细胞进行体外培养,分别给予10μg/L的IL-1β以及不同浓度辛伐他汀进行预处理,然后分别检测不同组中软骨细胞的凋亡率、iNOSmRNA的表达、线粒体膜电位水平以及NO的含量。结果：经不同浓度辛伐他汀干预后,软骨细胞凋亡率明显下降,NO含量及iNOS mRNA的表达显著减少,线粒体膜电位明显上升。结论辛伐他汀对IL-1β诱导的关节软骨细胞凋亡有抑制作用,其机制可能与辛伐他汀改善软骨细胞线粒体功能,调节NO的代谢有关。更多还原

【Abstract】 Objective:To study the effect of Simvastatin on histology in cartilage of rabbit osteoarthritis model. Method:Thirty New-Zealand Rabbits were randomly divided into 5 groups:Group A、B、C、D、E. The OA model was established in group B、C、D、E by Hulth Method. Group A was normal control and received nothing.Group B was received intraarticular injection with sterilized normal saline.Group C、group D and group E were received intraarticular injection with different dose of simvastatin. All rabbits were killed 6 weeks postoperation,the knee joints were assessed by morphological method; and then, cartilage tissue were obtained and sections were made and stained with HE/SOFG for Mankin score. Result:Cartilage impairment in group B was more serious than other group.Mankin score in group A、C、D、E was lower than group B.Conclusion:OA model was established successfully. Intraarticular injection with simvastatin can protect articular cartilage from impairment by promoting synthesis of proteoglycan. Objective:To observe the effect of Simvastatin on level of IL-1 in synovia and type II collagen in cartilage of osteoarthritis rabbits. Method:Thirty New-Zealand Rabbits were randomly divided into 5 groups:Group A、B、C、D、E. The OA model was established in group B、C、D、E by Hulth Method. Group A was normal control and received nothing.Group B was received intraarticular injection with sterilized normal saline.Group C、group D and group E were received intraarticular injection with different dose of simvastatin. All rabbits were killed 6 weeks postoperation. The expression of MMP-13 was detected by immunohistochemical method and reverse transcription-polymerase chain reaction (RT-PCR).Gene expression of typeⅡcollagen was detected by using RT-PCR. The level of IL-1 in synovia was deteccted by ELISA method. Result:The expression of mmp-13 mRNA was significantly decreased in group C、D、E compared with group B.The expression of typeⅡcollagen was significantly increased in group C、D、E compared with group B.Conclusion:Intra-articular administration of Simvastatin can lower the level of MMP-13 and IL-1 and upregulate expression of typeⅡcollagen,thus limiting cartilage degradation. Objective:To observe the effect of Simvastatin on apoptosis of cartilage of osteoarthritis rabbits. Method:Thirty New-Zealand Rabbits were randomly divided into 5 groups:Group A、B、C、D、E. The OA model was established in group B、C、D、E by Hulth Method. Group A was normal control and received nothing.Group B was received intraarticular injection with sterilized normal saline.Group C、group D and group E were received intraarticular injection with different dose of simvastatin. All rabbits were killed 6 weeks postoperation. Cartilage tissue were obtained, chondrocyte apoptosis was detected by flow cytometry. The expression of Bcl-2 mRNA was detected by RT-PCR Result:The expression of BCL-2 mRNA was significantly decreased in group A、C、D、E compared with group B.The ratio of apoptotic cells was significantly increased in group D、E compared with group B.Conclusion:Intra-articular administration of Simvastatin can lower the ratio of chondrocyte apoptosis by downregulating expression of BCL-2 mRNA. Objective:to study the effect of hyaluronic acid (HA) on chondrocyte apoptosis in a rat osteoarthritis in vitro model and explore its mechanism. Method:Primary rabbit chondrocytes were cultured and induced to apoptosis by 10ng/ml interleukin-1β(IL-1β).After treatment with various concentrations of Simvastatin(10,50,100umol/l), the apoptotic rate, mitochondrial function, nitric oxide production, and the levels of inducible nitric oxide synthase (iNOS) mRNA in IL-1β-induced chondrocytes were examined. Results:Simvastatin could inhibit chondrocyte apoptosis in a dose-dependent manner. Furthermore, it could partly restore the levels of mitochondrial membrane potential, decrease nitric oxide production by down-regulation of iNOS mRNA expression in chondrocytes induced by IL-1β. Conclusion:The inhibitory effects of Simvastatin on IL-1β-induced chondrocyte apoptosis were possibly due to the protection of mitochondrial function, the decline in the levels of nitric oxide.更多还原