疏肝止泻方经VIPR2/PLC/PKC通路改善肝郁脾虚型IBS-D大鼠肠道通透性的机制研究OA
Mechanism of Shugan Zhixie Formula(疏肝止泻方)in improving intestinal permeability in IBS-D rats with liver depression and spleen deficiency syndrome via the VIPR2/PLC/PKC pathway
目的 基于肠道通透性探讨疏肝健脾法通过VIPR2/PLC/PKC信号通路发挥对肝郁脾虚型腹泻型肠易激综合征(IBS-D)模型大鼠的干预效应及机制.同时,优化肝郁脾虚型IBS-D大鼠模型造模方法,以期建立更加满足IBS-D典型症状及情绪应激障碍的病证结合要求的大鼠模型.方法 50只SPF级Wistar大鼠,7只纳入空白组,将剩余大鼠模型复制成功后,分为模型组、匹维溴铵组,以及疏肝止泻方低、中、高剂量组.连续干预14天后取血液及结肠组织进行相关指标检测及分析.结果 与空白组相比,其余各组ELISA法检测血浆D-乳酸、血清PIP-2和DAG含量均有不同程度的增加(P<0.001),检测发现VIPR2、PLC、PKC蛋白及mRNA表达水平显著升高(P<0.001),PI3K蛋白及mRNA表达水平降低(P<0.001),而Zo-1、occludin蛋白及mRNA表达水平显著降低(P<0.01).另外,同模型组相比,L-SGZXF组、M-SGZXF组及H-SGZXF组中的各类蛋白及mRNA表达水平均具有显著性差异(P<0.05).最后,免疫组化法检测显示各组大鼠下丘脑中VIP、VIPR2蛋白的平均荧光强度相较于空白组升高(P<0.01,P<0.05).且参照相关造模方法,我们优化后的大鼠模型造模成功.结论 通过肠道灌注乙酸+饥饱失常+束缚夹尾应激的方案可以建立肝郁脾虚型IBS-D大鼠模型,且疏肝健脾法通过调节VIPR2/PLC/PKC信号通路上的蛋白表达水平,改善肠道通透性,发挥干预效应.
Objective To examine,using intestinal permeability and the VIPR2/PLC/PKC signaling pathway,the intervention impact and mechanism of the liver-soothing and spleen-strengthening technique on diarrhea-predominant irritable bowel syndrome(IBS-D)with liver depression and spleen deficiency in rat models.In order to create a rat model that better satisfies the needs of integrating normal IBS-D symptoms with emotional stress disorders,it is also necessary to refine the modeling approach for IBS-D rats with liver depression and spleen deficiency.Methods Seven of the fifty SPF-grade Wistar rats were placed in the blank group.The remaining rats were split into the model group,pinaverium bromide group,and low-,medium-,and high-dose Shugan Zhixie Formula(疏肝止泻方,SGZXF)groups following successful model replication.Blood and colon tissues were obtained for associated indicator testing and analy-sis following a 14-day continuous intervention.Results According to ELISA,the other groups'plasma D-lactate,serum PIP-2,and DAG levels increased to differing degrees when compared to the blank group(P<0.001).PI3K protein and mRNA expression declined(P<0.001),whereas VIPR2,PLC,and PKC protein and mRNA expression levels were dramatically increased(P<0.001).Zo-1 and occludin mRNA and protein expression were dramatically decreased(P<0.01).Furthermore,the protein and mRNA expression levels of the low-,medium-,and high-dose SGZXF groups differed significantly from the model group(P<0.05).Lastly,immunohis-tochemistry(IHC)revealed that each group's hypothalamus had higher average fluorescence intensity of VIP and VIPR2 proteins than the blank group(P<0.01,P<0.05).Furthermore,the rat model was effectively constructed using the optimal modeling approach.Conclusion Intestinal acetic acid perfusion,erratic feeding,and restraint-tail clamping stress can be used to create a rat model of IBS-D with liver depression and spleen deficiency.By controlling the expression of proteins in the VIPR2/PLC/PKC signaling pathway,the Liver-soothing and Spleen-strengthening approach improves intestinal permeability.
刘俊宏;李明;张娟;王淼蕾;李亚静;付兆媛;毛兰芳;郭欣
甘肃中医药大学附属医院,甘肃 兰州 730000西安市高陵区医院,陕西 西安 710000甘肃中医药大学,甘肃 兰州 730000甘肃中医药大学附属医院,甘肃 兰州 730000甘肃中医药大学附属医院,甘肃 兰州 730000甘肃中医药大学附属医院,甘肃 兰州 730000甘肃中医药大学附属医院,甘肃 兰州 730000甘肃中医药大学,甘肃 兰州 730000
医药卫生
腹泻型肠易激综合征肝郁脾虚证VIPR2/PLC/PKC信号通路疏肝健脾法肠道通透性疏肝止泻方
Diarrhea-predominant irritable bowel syndrome(IBS-D)Liver depression and spleen deficiency syndromeVIPR2/PLC/PKC signaling pathwayLiver-soothing and spleen-fortifying methodIntestinal permeabilityShugan Zhixie Formula(疏肝止泻方)
《时珍国医国药》 2026 (8)
1431-1438,8
国家中医药管理局青年岐黄学者基金项目(国中医药人教函[2022]256号)甘肃省临床医学研究中心建设项目(21JR7RA682)甘肃省科技厅联合科研项目基金(24JRRA874)甘肃省科技厅重点研发计划(社会发展类)(22YF7FA100)甘肃省卫生健康行业科研计划项目(GSWSQN2022-01)
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