手法对兔膝关节NO水平及软骨细胞凋亡影响实验研究.pdf

目 录 中文摘要·············································1 英文摘要 ·············································3 英文缩略词表 ·········································6 引 言 ·············································7 正 文 ·············································9 1．实验材料和方法 ····································9 1．1 实验动物饲养及条件 ·······························9 1．2 主要试剂和药品 ···································9 1．3 实验器材·········································9 1．4 动物模型建立和分组 ······························10 1．5 实验过程········································11 2．实验结果 ·········································16 3．讨论 ·············································17 4．结论 ·············································38 5．存在问题及展望 ···································38 参考文献 ············································40 附 图 ···········································47 综 述 ············································48 致 谢 ············································63 攻读学位期间发表的学术论文 ··························64 作者声明 ············································65 个人简历 ············································66 中文摘要 目 的：研究手法对实验兔膝关节液一氧化氮（NO ）含量和膝骨关 节炎(KOA)软骨细胞凋亡的影响及探讨其作用机制。 方 法：24 只新西兰兔随机分成三组，造模组20 只，正常组4 只。 造模组随机分为手法组 10 只，对照组 10 只。造模组动物通过手术结扎 右侧股静脉和臀下静脉方法造模，正常组不作任何处理。手法组从第二 周(造模后第8 天)开始进行手法治疗，共5 周。对照组和正常组不治疗。 于第8 周末处死所有动物，抽取右膝关节液采用硝酸还原酶法检测NO 含 量；并取右膝关节胫骨平台内侧软骨制片，切片采用脱氧核糖核苷酸末 端转移酶介导的缺口末端标记法（terminal deoxy- nucleotidyl transf- erase mediated nick -end -labeling ，TUNEL ）标记，在光镜下进行凋亡细胞形态 学观察。 结 果：手法组右膝关节液中 NO 含量为 54.62 ±15.8 μmol∕L， 对照组为 72.47 ±16.4 μmol∕L，正常组为 49.23 ±14.5 μmol∕L。NO 含量手法组与对照组比较 p ＜0.05，有显著差异。病理切片中正常组见 关节软骨表面光滑，软骨细胞层次结构清晰，表层细胞小而扁平，向下 细胞增大并呈柱状排列，少见细胞分裂。正常组从表层至辐射层各层细 胞形态清晰，各层细胞未见或少见细胞体积萎缩，细胞核固缩等细胞凋 亡的形态学改变。对照组见软骨表面不光滑，表层软骨变薄，粗糙，软 骨表面出现多个裂隙，可见裂隙从表层延伸向下达辐射层，裂隙中可见 稀疏结缔组织及松散的网状连接，软骨下骨增厚，整个软骨细胞排列紊 乱，层次结构不易辨认。裂隙周围可见表现为