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
AIM To investigate the distribution and function of interstitialcells of Cajal(ICCs) and platelet-derived growth factor receptor-α positive(PDGFRα+) cells in the proximal and distal colon.METHODS The comparison of c...AIM To investigate the distribution and function of interstitialcells of Cajal(ICCs) and platelet-derived growth factor receptor-α positive(PDGFRα+) cells in the proximal and distal colon.METHODS The comparison of colonic transit in the proximal and distal ends was performed by colonic migrating motor complexes(CMMCs). The tension of the colonic smooth muscle was examined by smooth muscle spontaneous contractile experiments with both ends of the smooth muscle strip tied with a silk thread. Intracellular recordings were used to assess electrical field stimulation(EFS)-induced inhibitory junction potentials(IJP) on the colonic smooth muscle. Western blot analysis was used to examine the expression levels of ICCs and PDGFRα in the colonic smooth muscle.RESULTS Treatment with NG-nitro-L-arginine methyl ester hydrochloride(L-NAME) significantly increased the CMMC frequency and spontaneous contractions, especially in the proximal colon, while treatment with MRS2500 increased only distal CMMC activity and smooth muscle contractions. Both CMMCs and spontaneous contractions were markedly inhibited by NPPB, especially in the proximal colon. Accordingly, CyPPA sharply inhibited the distal contraction of both CMMCs and spontaneous contractions. Additionally, the amplitude of stimulationinduced nitric oxide(NO)/ICC-dependent slow IJPs(sIJPs) by intracellular recordings from the smooth muscles in the proximal colon was larger than that in the distal colon, while the amplitude of electric field stimulationinduced purinergic/PDGFRα-dependent fast IJPs(fIJPs) in the distal colon was larger than that in the proximal colon. Consistently, protein expression levels of c-Kit and anoctamin-1(ANO1) in the proximal colon were much higher, while protein expression levels of PDGFRα and small conductance calcium-activated potassium channel 3(SK3) in the distal colon were much higher.CONCLUSION The ICCs are mainly distributed in the proximal colon and there are more PDGFRα+ cells are in the distal colon, which generates a pressure gradient between the two ends of the colon to propel the feces to the anus.展开更多
[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
目的:探索前列腺Cajal间质细胞(interstitial cells of Cajal,ICCs)与交感神经及平滑肌细胞之间的形态及功能学联系。方法:(1)制作豚鼠前列腺组织冰冻切片并进行双重免疫荧光染色。使用酪氨酸激酶受体(c-Kit)作为ICCs的标记物,酪氨酸羟...目的:探索前列腺Cajal间质细胞(interstitial cells of Cajal,ICCs)与交感神经及平滑肌细胞之间的形态及功能学联系。方法:(1)制作豚鼠前列腺组织冰冻切片并进行双重免疫荧光染色。使用酪氨酸激酶受体(c-Kit)作为ICCs的标记物,酪氨酸羟化酶(tyrosine hydroxylase,TH)和多巴胺β羟化酶(dopamineβ-hydroxylase,DβH)作为交感神经纤维的标记物,α肌动蛋白(α-actin)作为平滑肌细胞的标记物。另外,分别对α1-肾上腺素受体与c-Kit以及缝隙连接蛋白Connexin43(Cx43)与c-Kit进行免疫荧光双重标记。(2)将前列腺肌条固定于灌流槽中,记录去甲肾上腺素(norepinephrine,NE)诱发的肌条收缩幅度和频率,以及加入Glivec后肌条收缩幅度和频率的变化。结果:(1)免疫荧光组织化学标记结果显示:ICCs与交感神经纤维、平滑肌细胞平行分布,ICCs边缘与交感神经、平滑肌细胞边缘密切接触,c-Kit标记的ICCs上存在α1-肾上腺素受体和缝隙连接蛋白Cx43的共表达。(2)在去甲肾上腺素的刺激下前列腺平滑肌肌条收缩幅度为(0.98±0.16)g,收缩频率为(2.54±0.28)次/min。使用Glivec后肌条收缩幅度显著下降,50μmol/L组为(0.56±0.07)g(P<0.01),200μmol/L组为(0.25±0.05)g(P<0.01),收缩频率未见明显改变,50μmol/L组为(2.67±0.32)次/min(P>0.05),200μmol/L组为(2.75±0.31)次/min(P>0.05)。结论:豚鼠前列腺ICCs具有介导前列腺交感神经信号,支配平滑肌活动的形态和功能学基础。展开更多
目的:研究白细胞介素6(IL-6)对豚鼠近端结肠平滑肌的影响及其机制.方法:观察IL-6对结肠收缩的影响;用河豚毒素(TTX)阻断肠神经后观察不同浓度IL-6对结肠平滑肌收缩的影响;损伤Cajal间质细胞(ICC)后观察IL-6对结肠平滑肌收缩的影响.结果...目的:研究白细胞介素6(IL-6)对豚鼠近端结肠平滑肌的影响及其机制.方法:观察IL-6对结肠收缩的影响;用河豚毒素(TTX)阻断肠神经后观察不同浓度IL-6对结肠平滑肌收缩的影响;损伤Cajal间质细胞(ICC)后观察IL-6对结肠平滑肌收缩的影响.结果:在带有ICC的近端结肠纵行肌加入IL-6后,结肠平滑肌的收缩振幅增加和频率加快,呈浓度依赖性:加入TTX后,收缩的幅度和频率,同拮抗前相比分别降低和减慢(0.206±0.027 g vs 0.300±0.039 g;9.770±1.711 s vs 8.483±1.113 s:P<0.01,P<0.05):TTX阻断后加入IL-6(80μg/L),振幅增加和频率加快(P<0.01,P<0.05):破坏结肠ICC,收缩幅度和频率分别与损伤ICC后加入IL-6无显著性差异(80μg/L),而与正常结肠收缩振幅和频率有显著性差异(P<0.01).结论:IL-6对豚鼠近端结肠平滑肌的收缩活动有兴奋作用,其兴奋效应主要是通过肠神经元介导.ICC是IL-6对平滑肌兴奋途径的一个不可缺少的中间环节.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.
基金Supported by The National Natural Science Foundation of China,No.31671192 and No.31571180Foundation of Xin Hua Hospital,No.JZPI201708
文摘AIM To investigate the distribution and function of interstitialcells of Cajal(ICCs) and platelet-derived growth factor receptor-α positive(PDGFRα+) cells in the proximal and distal colon.METHODS The comparison of colonic transit in the proximal and distal ends was performed by colonic migrating motor complexes(CMMCs). The tension of the colonic smooth muscle was examined by smooth muscle spontaneous contractile experiments with both ends of the smooth muscle strip tied with a silk thread. Intracellular recordings were used to assess electrical field stimulation(EFS)-induced inhibitory junction potentials(IJP) on the colonic smooth muscle. Western blot analysis was used to examine the expression levels of ICCs and PDGFRα in the colonic smooth muscle.RESULTS Treatment with NG-nitro-L-arginine methyl ester hydrochloride(L-NAME) significantly increased the CMMC frequency and spontaneous contractions, especially in the proximal colon, while treatment with MRS2500 increased only distal CMMC activity and smooth muscle contractions. Both CMMCs and spontaneous contractions were markedly inhibited by NPPB, especially in the proximal colon. Accordingly, CyPPA sharply inhibited the distal contraction of both CMMCs and spontaneous contractions. Additionally, the amplitude of stimulationinduced nitric oxide(NO)/ICC-dependent slow IJPs(sIJPs) by intracellular recordings from the smooth muscles in the proximal colon was larger than that in the distal colon, while the amplitude of electric field stimulationinduced purinergic/PDGFRα-dependent fast IJPs(fIJPs) in the distal colon was larger than that in the proximal colon. Consistently, protein expression levels of c-Kit and anoctamin-1(ANO1) in the proximal colon were much higher, while protein expression levels of PDGFRα and small conductance calcium-activated potassium channel 3(SK3) in the distal colon were much higher.CONCLUSION The ICCs are mainly distributed in the proximal colon and there are more PDGFRα+ cells are in the distal colon, which generates a pressure gradient between the two ends of the colon to propel the feces to the anus.
文摘目的:探索前列腺Cajal间质细胞(interstitial cells of Cajal,ICCs)与交感神经及平滑肌细胞之间的形态及功能学联系。方法:(1)制作豚鼠前列腺组织冰冻切片并进行双重免疫荧光染色。使用酪氨酸激酶受体(c-Kit)作为ICCs的标记物,酪氨酸羟化酶(tyrosine hydroxylase,TH)和多巴胺β羟化酶(dopamineβ-hydroxylase,DβH)作为交感神经纤维的标记物,α肌动蛋白(α-actin)作为平滑肌细胞的标记物。另外,分别对α1-肾上腺素受体与c-Kit以及缝隙连接蛋白Connexin43(Cx43)与c-Kit进行免疫荧光双重标记。(2)将前列腺肌条固定于灌流槽中,记录去甲肾上腺素(norepinephrine,NE)诱发的肌条收缩幅度和频率,以及加入Glivec后肌条收缩幅度和频率的变化。结果:(1)免疫荧光组织化学标记结果显示:ICCs与交感神经纤维、平滑肌细胞平行分布,ICCs边缘与交感神经、平滑肌细胞边缘密切接触,c-Kit标记的ICCs上存在α1-肾上腺素受体和缝隙连接蛋白Cx43的共表达。(2)在去甲肾上腺素的刺激下前列腺平滑肌肌条收缩幅度为(0.98±0.16)g,收缩频率为(2.54±0.28)次/min。使用Glivec后肌条收缩幅度显著下降,50μmol/L组为(0.56±0.07)g(P<0.01),200μmol/L组为(0.25±0.05)g(P<0.01),收缩频率未见明显改变,50μmol/L组为(2.67±0.32)次/min(P>0.05),200μmol/L组为(2.75±0.31)次/min(P>0.05)。结论:豚鼠前列腺ICCs具有介导前列腺交感神经信号,支配平滑肌活动的形态和功能学基础。
文摘目的:研究白细胞介素6(IL-6)对豚鼠近端结肠平滑肌的影响及其机制.方法:观察IL-6对结肠收缩的影响;用河豚毒素(TTX)阻断肠神经后观察不同浓度IL-6对结肠平滑肌收缩的影响;损伤Cajal间质细胞(ICC)后观察IL-6对结肠平滑肌收缩的影响.结果:在带有ICC的近端结肠纵行肌加入IL-6后,结肠平滑肌的收缩振幅增加和频率加快,呈浓度依赖性:加入TTX后,收缩的幅度和频率,同拮抗前相比分别降低和减慢(0.206±0.027 g vs 0.300±0.039 g;9.770±1.711 s vs 8.483±1.113 s:P<0.01,P<0.05):TTX阻断后加入IL-6(80μg/L),振幅增加和频率加快(P<0.01,P<0.05):破坏结肠ICC,收缩幅度和频率分别与损伤ICC后加入IL-6无显著性差异(80μg/L),而与正常结肠收缩振幅和频率有显著性差异(P<0.01).结论:IL-6对豚鼠近端结肠平滑肌的收缩活动有兴奋作用,其兴奋效应主要是通过肠神经元介导.ICC是IL-6对平滑肌兴奋途径的一个不可缺少的中间环节.
