阳明腑实证大鼠小肠神经-ICC网络形态学损伤和下法施治研究

Morphological changes of enteric nerve- interstitial cells of Cajal-smooth muscle cells network in Yang Ming Fu Shi syndrome and therapeutic mechanism of Xia-Fa in traditional chinese

关键词:  阳明腑实证  Cajal间质细胞  肠神经

Key words:  [Key words]　Yang Ming Fu Shi Sydrom  ICC  entric nerve

摘要:

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10.0 cm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。

Abstract:

[Abstract] Objective　To observe the morphological changes of enteric nerve and ICC of enteric nerve-ICC-smooth muscle cells network in rats with Yang Ming Fu Shi Syndrome, and to investigate the function of gastrointestinal motility and the therapeutic Mechanism of Xa-Fa in Traditional Chinese. 　Methods　One handred Wistar rats of both sexes were randomly divided into three groups, the contrpl group 20 rats, the model group 40 rats and the therap group 40 rats. The proximal 10.0 cm segment of jejunum beginning 2 cm distal to the pylorus from each group, was studied using c-Kit and vesicular acetylcholine transporter (VAChT)/ neuronal nitric oxide synthase (nNOS) immunohistochemical double-staining with whole-mount preparation technique and confocal laser scanning microscopy. 　Results　Change of enteric nerve-ICC net work with confocal laser scanning microscopy:Model group: Compare with control group, the numbers of ICC were significantly decreased (P<0.01), network was disrupted, fluorescence intensity of ICC decreased. Compare with control group, the numbers of vesicular acetylcholine transporter (VAChT) and neuronal nitric oxide synthase (nNOS) were significantly decreased (P<0.01), nerves network disordered, each connection decreased, fluorescence intensity of enteric nerve decreased(P<0.01).The connection of ICCs and enteric nerves were loosen or disappeared.ICC- enteric nerve network was significantly disrupted.Treat group: Compare with control group, the numbers of ICC were significantly increased (P<0.01), network was recovered, fluorescence intensity of ICC increased. the numbers of vesicular acetylcholine transporter (VAChT) and neuronal nitric oxide synthase (nNOS) were significantly increased (P<0.01), network was recovered, fluorescence intensity of nerves increased (P<0.01). The connection of ICCs and enteric nerves were approaching normal, ICC- enteric nerve network was significantly recovered. 　Conclusion Gastrointestinal tract distended in Yang Ming Fu Shi Sydrom rats. Function of gastrointestinal was significantly disordered; and recoveried after Da Cheng Qi decoction treated.In Yang Ming Fu Shi Sydrom rats, numbers of vesicular acetylcholine transporter (VAChT) and neuronal nitric oxide synthase (nNOS) were significantly decreased; and numbers of ICC were decreased too.entric nerve-ICCnetwork was disrupted; after treated, numbers of nerves and ICC were increased,and network was recoveried.