基于肠神经系统研究芪黄煎剂对胃切除术后大鼠肠运动功能的影响

Study on effects of Qihuang Decoction on intestinal motility after gastrectomy in rats based on enteric nervous system

目的:探讨芪黄煎剂对胃切除术后大鼠肠神经元和神经递质表达的影响。方法:将45只大鼠随机分为对照组、肠内营养组、芪黄煎剂组,每组15只,其中肠内营养组和芪黄煎剂组建立胃切除模型。通过HPLC构建指纹图谱控制不同批次芪黄煎剂的药物质量;干预7d后,采用酚红排空测定法,计算小肠推进率;将铂金丝电极插入小肠肌层,通过生物机能实验系统检测小肠电生理;通过激光扫描共聚焦显微镜,观察小肠肠神经系统(ENS)网络神经元中乙酰胆碱酯酶(AChE)、P物质(SP)、血管活性肠肽(VIP)、一氧化氮合成酶(NOS)的量判定阳性神经元的数量;通过Western Blot检测AChE、SP、VIP、NOS的蛋白表达情况。结果:与对照组比较,肠内营养组小肠推进率显著降低,小肠神经元中AChE、SP数量和蛋白表达量显著减少,而NOS、VIP数量和蛋白表达量显著增加(P<0.01);芪黄煎剂组较肠内营养组可显著提高小肠推进率,促进肠神经元中AChE、SP数量和蛋白表达量显著增加,而NOS、VIP数量和蛋白表达量显著减少(P<0.01);各组小肠电生理比较,芪黄煎剂组较肠内营养组小肠电生理波幅未见明显改善,但波形变得更加规整。结论:芪黄煎剂可以通过调控ENS中相应阳性神经元数量以及ACh E、SP、VIP、NOS蛋白表达,重建ENS网络结构,恢复小肠电生理活动,促进肠道功能恢复。

Objective: To explore the effects of Qihuang Decoction on the expression of enteric neurons and neurotransmitters in rats after gastrectomy. Methods: Forty-five rats were randomly divided into three groups of control group,enteral nutrition group, and Qihuang Decoction group, 15 rats in each group. The enteric nutrition group and Qihuang Decoction group were established into gastrectomy model. HPLC method was applied to determine the fingerprint of Qihuang Decoction quality in different batches. After 7 days of intervention, the small intestine propulsion rate was calculated by phenol red emptying method. The platinum wire electrode was inserted into the myenteron, and the electrophysiology of the small intestine was systematically detected by biological function experiment. The amount of AChE, SP, VIP and NOS in the ENS was observed by laser scanning confocal microscope to determine the number of positive neurons. The protein expression of AChE, SP, VIP and NOS were detected by Western Blot. Results: Compared with the control group, the small intestine propulsion rate of the enteral nutrition group was significantly reduced, the number and protein expression of AChE and SP in small intestine neurons were significantly reduced, but significantly increased the number and protein expression of NOS and VIP(P<0.01). Compared with the enteral nutrition group, the propulsion rate of the small intestine in Qihuang Decoction group was significantly increased,the number and protein expression of AChE, SP in enteric neurons were significantly increased, while the number and protein expression of NOS, VIP were significantly reduced(P<0.01). When comparing the small intestinal electrophysiology of each group, the small intestinal electrophysiological waveform amplitude was not significantly improved in the Qihuang Decoction group compared with the enteral nutrition group, but the waveform became more regular. Conclusion: These results indicated that Qihuang Decoction can regulate the number of corresponding positive neurons and the expression of AChE, SP, VIP and NOS in ENS, which is beneficial for reconstructing ENS network structure and the recovery of electrophysiological activity in small intestine, and ultimately contribute to promoted intestinal function.