针刺足三针通过CO途径对大鼠结肠吻合术后ICCs修复的影响

发布时间：2018-04-27 03:02

  本文选题：CO + HO-1　； 参考：《广州中医药大学》2016年硕士论文

【摘要】：目的：腹部手术后,常出现术后胃肠功能障碍,导致一系列的临床症状。在促进术后胃肠功能恢复的临床研究中,大量的临床研究表明针刺能减轻术后胃肠功能障碍的程度,促进胃肠功能恢复,目前在临床上广泛应用。通过前期研究发现,术后胃肠功能的恢复可能与胃肠道ICCs修复再生存在密切的联系。CO气体信号通路是ICCs修复再生过程中一条重要的途径,CO是介导胃肠道平滑肌舒张的抑制性神经递质,当神经元受刺激时,HO合成CO并释放,介导神经传导。本课题通过观察针刺“靳三针”穴组中“足三针”对结肠吻合术后SD大鼠胃肠道CO气体信号通路的影响,探索针刺“足三针”对术后胃肠功能障碍SD大鼠胃肠功能恢复的作用机制。方法：将SD大鼠随机分为3组：正常组、模型组、针刺组,模型组及针刺组再按干预时间的不同分为3天组、5天组、10天组。模型组：行结肠吻合术造模。麻醉后沿腹白线于中下腹部切开腹部,与盲肠下端2cm处剪短,缝合结肠,依次缝合腹膜、肌层和皮肤,术后不予针刺。针刺组：造模同模型组,术后按干预时间不同予以针刺干预。小肠推进率的测量：SD大鼠取材前,用灌胃器抽取lml墨水对大鼠进行灌胃,灌胃30min后颈椎脱臼处死,用组织剪剪开腹部,充分暴露肠道,仔细剥离大肠、小肠,量取小肠全长及幽门至墨水远端的距离。参照公式：小肠推进率=(墨水行进长度／小肠总长度)×100%,计算每只大鼠的小肠推进率。CO、HO-1、HO-2含量的测定：将SD大鼠脱臼处死后,截取盲肠下端吻合口约lcm长,将肠组织清洗干净,沿纵轴分为三部分,分别用于检测H0-1、HO-2、C0和电镜观察；按照不同的标本处理方法将肠组织做前期处理。使用ELISA法测定HO-1、HO-2活性,生化法测定C0含量；使用电镜观察Cajal细胞的超微结构。结果：1.小肠推进率测定结果：比较各组大鼠小肠推进率,正常组与5天针刺组、10天针刺无统计学意义(P0.05),3天模型组与5天模型组无统计学差异(P0.05),3天针刺组与10天模型组无统计学差异(P0.05),5天模型组与10天模型组有统计学差异(P0.05),其余各组相比较均有显著统计学意义(P0.01)。2.HO-1活性测定结果：与3天正常组比较,3天模型组结果有显著统计学差异(P0.01),3天针刺组结果有统计学差异(P0.05)；与3天模型组比较,5天正常组、5天模型组、5天针刺组、10天正常组、10天模型组、10天针刺组结果有显著统计学差异(P0.01),3天针刺组结果有统计学差异(P0.05)；与3天针刺组比较,5天正常组、5天针刺组、10天正常组、10天模型组结果有统计学差异(P0.05)；余结果无统计学意义。3.HO-2活性的测定结果：与正常组比较,3天模型组、3天针刺组、5天模型组、5天针刺组、10天模型组、10天针刺组结果有统计学意义(P0.05)；与3天模型组比较,3天针刺组结果有统计学意义(P0.05),5天模型组、5天针刺组、10天模型组、10天针刺组结果有统计学意义(P0.01)；与3天针刺组比较,5天模型组、5天针刺组、10天模型组、10天针刺组有统计学意义(P0.05)；余无统计学意义。4.CO含量的测定结果：与正常组比较,3天模型组、3天针刺组、5天模型组结果有统计学差异(P0.01)；与3天模型组比较,3天针刺组、5天模型组5天针刺组、10天模型组、10天针刺组结果有统计学差异(P0.01)；与3天针刺组比较,5天针刺组、10天模型组、10天针刺组结果有统计学差异(P0.01)；与5天模型组比较,5天针刺组、10天模型组结果有统计学差异(P0.01)。余结果无统计学差异。5.电镜Cajal细胞结果：(1).正常组：Cajal细胞多成梭形,细胞核清晰,呈卵圆形,细胞内染色质均匀、清晰,细胞核周围胞质较少,胞内细胞器如：线粒体、内质网、高尔基体等发育良好；胞浆内中间丝清楚,数量较多,多分布在细胞核周围；基底膜完整；神经突触连接清楚。(2).模型组：①模型3天组：Cajal细胞凸起明显减少,胞浆少,胞浆内出现数量较多的空泡；细胞器数量明显减少,细胞器存在广泛病变,线粒体肿胀明显,线粒体嵴断裂；内质网扩张明显,粗面内质网脱颗粒；基底膜不完整。②模型5天组：Cajal细胞形状不规则,细胞核固缩,异染色质趋边,细胞内胞浆稍丰富,细胞结构稍清晰；线粒体肿胀,结构不清；中间丝数量稍多,基底膜不完整。③模型10天组：细胞内胞浆丰富,细胞器数量较多,部分结构不清晰；线粒体部分肿胀,可见少量溶酶体；部分粗面内质网脱颗粒,中间丝清晰,基底膜稍完整。(3)针刺组：①针刺3天组：细胞内胞浆丰富,细胞器数量较多,结构较清晰；线粒体部分肿胀；内质网扩张不明显；中间丝清晰,基底膜欠完整。②针刺5天组：Cajal细胞形状规则,多为梭形,细胞内染色质较均匀,细胞核周围胞质丰富,细胞器数量较多,结构清楚,线粒体数量较多,结构清晰；内质网无扩张；可见少量高尔基体、溶酶体；基底膜较完整。③针刺10天组：Cajal细胞多成梭形,细胞核清晰,呈卵圆形,细胞内染色质均匀、清晰,细胞核周围胞质较少,胞内细胞器发育良好；胞浆内中间丝清楚,数量较多,多分布在细胞核周围；基底膜完整；神经突触连接清楚。结论：1.针刺能改善胃肠功能障碍SD大鼠的小肠推进率,针刺组小肠蠕动恢复时间明显比模型组恢复时间缩短。2.针刺能降低术后应激增高的胃肠道HO-1、HO-2活性,使CO合成量减少,CO介导的胃肠道平滑肌舒张功能减弱,促进胃肠道蠕动功能的恢复。3.针刺对手术损伤带来的HO-2活性下降具有调节作用,能促进其回复正常。4.在POI中,高浓度CO与KIT表达下降并存,提示CO在高浓度下反而不利于ICC生存
[Abstract]:Objective : To study the effect of acupuncture on gastrointestinal function recovery in SD rats after abdominal surgery . It was used to detect HO - 1 , HO - 2 , C0 and electron microscope respectively .
The intestinal tissues were treated in advance according to different methods of specimen treatment . The contents of HO - 1 and HO - 2 were determined by ELISA .
Results : 1 . There was no significant difference between the 3 - day model group and the 10 - day model group ( P0.05 ) . There was no statistical difference between the three - day model group and the 10 - day model group ( P0.05 ) .
Compared with the 3 - day model group , 5 - day normal group , 5 - day model group , 5 - day acupuncture group , 10 - day normal group , 10 - day model group and 10 - day acupuncture group had significant statistical difference ( P0.01 ) .
Compared with the 3 - day acupuncture group , 5 - day normal group , 5 - day acupuncture group , 10 - day normal group and 10 - day model group had statistical difference ( P0.05 ) ;
The results of HO - 2 activity were as follows : compared with normal group , 3 - day model group , 3 - day acupuncture group , 5 - day model group , 5 - day acupuncture group , 10 - day model group and 10 - day acupuncture group had statistical significance ( P0.05 ) ;
Compared with the 3 - day model group , the results of the three - day acupuncture group had statistical significance ( P0.05 ) , 5 - day model group , 5 - day acupuncture group , 10 - day model group and 10 - day acupuncture group .
Compared with 3 - day acupuncture group , 5 - day model group , 5 - day acupuncture group , 10 - day model group and 10 - day acupuncture group had statistical significance ( P0.05 ) ;
No significant difference was found between the 3 - day model group , the 3 - day acupuncture group and the 5 - day model group ( P0.01 ) .
Compared with the 3 - day model group , the acupuncture group , 5 - day model group , 5 - day acupuncture group , 10 - day model group and 10 - day acupuncture group had statistical difference ( P0.01 ) .
Compared with the 3 - day acupuncture group , the acupuncture group , 10 - day model group and 10 - day acupuncture group had statistical difference ( P0.01 ) .
5 . The results of the 5 - day acupuncture group and the 10 - day model group were significantly different ( P0.01 ) .
The middle filament in the cytoplasm is clear , the quantity is more , the distribution is around the nucleus ;
the basement membrane is complete ;
Neurosynaptic connections are clear . ( 2 ) . The model group : 鈶，

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