[目的]观察大鼠小肠神经-Cajal间质细胞(interstitial cells of Cajal,ICC)-平滑肌细胞网络的形态学结构,为临床提供理论依据。[方法]健康成年Wistar大鼠20只,上段小肠组织制作肠肌层全厚组织标本,进行c-Kit和囊泡装乙酰胆碱转运体(VACh...[目的]观察大鼠小肠神经-Cajal间质细胞(interstitial cells of Cajal,ICC)-平滑肌细胞网络的形态学结构,为临床提供理论依据。[方法]健康成年Wistar大鼠20只,上段小肠组织制作肠肌层全厚组织标本,进行c-Kit和囊泡装乙酰胆碱转运体(VAChT)/神经型一氧化氮合酶(nNOS)免疫荧光双标记染色后用于激光扫描共聚焦显微镜检测、分析;制作电镜标本用于透射电镜观测。[结果]ICC在环纵行肌间神经丛通过长的突触彼此相互连接形成密集的网络样完整结构。胆碱能/氮能神经彼此相接形成网状结构,ICC网络围绕着胆碱能/氮能神经网络,并与神经纤维和平滑肌纤维紧密连接,ICC相互之间、与平滑肌细胞间可见缝隙连接(gap junctions)。ICC与神经纤维末梢形成突触样连接(synaptic-like contacts);ICC突起与平滑肌细胞形成缝隙连接。形成完整的神经-ICC-平滑肌网络形态学基础。[结论]肠神经、ICC和平滑肌间形成相互连接的超微结构,三者细胞网络形态学基础的完整性是胃肠运动功能的基本保证。展开更多
Objective： To observe the effects of Dachengqi Decoction （大承气汤, DCQD） on morphological changes in the network of enteric nerve-interstitial cells of Cajal （ICCs）-smooth muscle cells （SMC） of enteric deep mu...Objective： To observe the effects of Dachengqi Decoction （大承气汤, DCQD） on morphological changes in the network of enteric nerve-interstitial cells of Cajal （ICCs）-smooth muscle cells （SMC） of enteric deep muscular plexuses （DMP） in the rats with multiple organ dysfunction syndrome （MODS）. Methods： One hundred Wistar rats of both sexes weighing 200 to 250 g were randomly divided into the control group, MODS group, and DCQD group. The morphologic changes of enteric nerve-ICC-SMC network in the DMP of intestine was observed using c-Kit and vesicular acetylcholine transporter/neuronal nitric oxide synthase immunohistochemical double-staining with whole-mount preparation technique, confocal laser scanning microscopy, and electron microscopy. Results： Compared with the control group, the distribution and densities of cholinergic/nitrergic nerves and ICC in the DMP （ICC-DMP） of intestine in the MODS group were significantly decreased （P0.01）, and the network of cholinergic nerve-ICC-SMC was disrupted; and the ultrastructural features of ICC-DMP, enteric nerve, and SMC were severely damaged. After treatment with DCQD, the damage in the network of enteric nerve-ICC-SMC was significantly recovered. Compared with the MODS group, the distribution and densities of cholinergic/nitrergic nerves and ICC-DMP in the DCQD group were significantly increased （P0.01）; and the ultrastructural features of ICC-DMP, enteric nerve, smooth muscle cells were significantly recovered. Conclusions： DCQD can improve the gastrointestinal motility in MODS. The mechanism may be related to the effect of repairing the damages in the network of enteric nerve-ICC-SMC.展开更多
目的观察阳明腑实证大鼠小肠神经-Cajal间质细胞(interstitial cells of Cajal,ICC)—平滑肌细胞网络的形态学变化,探讨阳明腑实证胃肠运动障碍的产生机制及下法的作用机制。方法成年Wistar大鼠100只,雌雄不限,随机分成3组,对照组20只;...目的观察阳明腑实证大鼠小肠神经-Cajal间质细胞(interstitial cells of Cajal,ICC)—平滑肌细胞网络的形态学变化,探讨阳明腑实证胃肠运动障碍的产生机制及下法的作用机制。方法成年Wistar大鼠100只,雌雄不限,随机分成3组,对照组20只;阳明腑实证模型组40只;大承气汤治疗组40只。存活大鼠取上段小肠10.0cm组织制作肠肌层全厚组织标本,进行c-Kit和乙酰胆碱转运体(VAChT)/一氧化氮合酶(nNOS)免疫荧光双标记染色后用于激光扫描共聚焦显微镜检测、分析。结果 (1)共聚焦显微镜检测ICC网络:与对照组相比,模型组大鼠小肠ICC数量明显减少(P<0.01),ICC突触的数量减少,相互间的连接不连续,完整的网络样结构消失,细胞荧光强度减弱(P<0.01)。(2)共聚焦显微镜检测肠神经-ICC网络:与对照组相比,模型组胆碱能/氮能神经数目明显减少(P<0.01/P<0.01),神经网结构紊乱,彼此间的连接减少,神经网络的荧光强度减弱(P<0.01/P<0.01);ICC与肠神经纤维两者间失去紧密连接,肠神经-ICC网络的完整结构受到破坏。(3)治疗组较模型组ICC数量明显恢复(P<0.01),网络样结构的完整性有所恢复,与模型组比较细胞荧光强度明显增强(P<0.01)。胆碱能神经/氮能神经纤维数目明显增加(P<0.01),神经网络结构再次好转,彼此间的网络连接的完整性恢复明显,神经网络的荧光强度增加(P<0.01),ICC与肠神经纤维间的连接接近正常,ICC-肠神经网络结构恢复明显。结论阳明腑实证大鼠小肠ICC和神经网络受到破坏。大承气汤治疗后小肠胆碱能/氮能神经和ICC数量恢复,肠神经-ICC网络结构得到改善。展开更多
基金Supported by the National Natural Science Foundation of China(No. 30772860 and No. 30800382)
文摘Objective： To observe the effects of Dachengqi Decoction （大承气汤, DCQD） on morphological changes in the network of enteric nerve-interstitial cells of Cajal （ICCs）-smooth muscle cells （SMC） of enteric deep muscular plexuses （DMP） in the rats with multiple organ dysfunction syndrome （MODS）. Methods： One hundred Wistar rats of both sexes weighing 200 to 250 g were randomly divided into the control group, MODS group, and DCQD group. The morphologic changes of enteric nerve-ICC-SMC network in the DMP of intestine was observed using c-Kit and vesicular acetylcholine transporter/neuronal nitric oxide synthase immunohistochemical double-staining with whole-mount preparation technique, confocal laser scanning microscopy, and electron microscopy. Results： Compared with the control group, the distribution and densities of cholinergic/nitrergic nerves and ICC in the DMP （ICC-DMP） of intestine in the MODS group were significantly decreased （P0.01）, and the network of cholinergic nerve-ICC-SMC was disrupted; and the ultrastructural features of ICC-DMP, enteric nerve, and SMC were severely damaged. After treatment with DCQD, the damage in the network of enteric nerve-ICC-SMC was significantly recovered. Compared with the MODS group, the distribution and densities of cholinergic/nitrergic nerves and ICC-DMP in the DCQD group were significantly increased （P0.01）; and the ultrastructural features of ICC-DMP, enteric nerve, smooth muscle cells were significantly recovered. Conclusions： DCQD can improve the gastrointestinal motility in MODS. The mechanism may be related to the effect of repairing the damages in the network of enteric nerve-ICC-SMC.
文摘目的观察阳明腑实证大鼠小肠神经-Cajal间质细胞(interstitial cells of Cajal,ICC)—平滑肌细胞网络的形态学变化,探讨阳明腑实证胃肠运动障碍的产生机制及下法的作用机制。方法成年Wistar大鼠100只,雌雄不限,随机分成3组,对照组20只;阳明腑实证模型组40只;大承气汤治疗组40只。存活大鼠取上段小肠10.0cm组织制作肠肌层全厚组织标本,进行c-Kit和乙酰胆碱转运体(VAChT)/一氧化氮合酶(nNOS)免疫荧光双标记染色后用于激光扫描共聚焦显微镜检测、分析。结果 (1)共聚焦显微镜检测ICC网络:与对照组相比,模型组大鼠小肠ICC数量明显减少(P<0.01),ICC突触的数量减少,相互间的连接不连续,完整的网络样结构消失,细胞荧光强度减弱(P<0.01)。(2)共聚焦显微镜检测肠神经-ICC网络:与对照组相比,模型组胆碱能/氮能神经数目明显减少(P<0.01/P<0.01),神经网结构紊乱,彼此间的连接减少,神经网络的荧光强度减弱(P<0.01/P<0.01);ICC与肠神经纤维两者间失去紧密连接,肠神经-ICC网络的完整结构受到破坏。(3)治疗组较模型组ICC数量明显恢复(P<0.01),网络样结构的完整性有所恢复,与模型组比较细胞荧光强度明显增强(P<0.01)。胆碱能神经/氮能神经纤维数目明显增加(P<0.01),神经网络结构再次好转,彼此间的网络连接的完整性恢复明显,神经网络的荧光强度增加(P<0.01),ICC与肠神经纤维间的连接接近正常,ICC-肠神经网络结构恢复明显。结论阳明腑实证大鼠小肠ICC和神经网络受到破坏。大承气汤治疗后小肠胆碱能/氮能神经和ICC数量恢复,肠神经-ICC网络结构得到改善。
