温针灸对消化不良性腹泻大鼠结肠黏膜水通道蛋白3、4表达的影响研究

Effects of Needle-Warming Moxibustion on Expressions of Colonic Mucosal AQP3 and AQP4 in Rats with Chronic Diarrhea of Dyspepsia

目的:探讨温针灸对消化不良性腹泻大鼠结肠黏膜组织水通道蛋白3(AQP3)、水通道蛋白4(AQP4)以及炎性因子表达的影响,同时比较不同疗程温针灸的效果差异。方法:建立乳糖诱导的消化不良慢性腹泻大鼠模型,将大鼠随机分为A、B、C、D 4组。A组:空白组(5只);B组:模型组(5只);C组:温针灸1个疗程组(5只);D组:温针灸2个疗程组(5只)。C组给予为期5 d的温针灸,针灸部位:主穴有天枢、关元和中脘,配穴足三里,1次/d;D组给予5 d温针灸后休息2 d,再行温针灸5 d,针灸穴位与C组相同。观察大鼠腹泻情况,并于治疗结束后取对应各组大鼠结肠黏膜组织,WB检测AQP3和AQP4蛋白的表达,qPCR检测AQP3 mRNA和AQP4 mRNA的表达,ELISA法检测炎性因子TNF-α、IL-6、IL-8和IL-10的表达,免疫组化检测结肠黏膜组织AQP3、AQP4阳性表达情况。结果:温针灸1个疗程,腹泻率由100%下降至12.5%,2个疗程后腹泻率为0%;与B组相比,C组和D组AQP3和AQP4蛋白相对表达量均上升,A组、C组和D组AQP3 mRNA和AQP4 mRNA相对表达量均升高,D组AQP3 mRNA相对表达水平高于A组,差异均有统计学意义(P<0.01或P<0.05)。与B组相比,C组和D组TNF-α、IL-6和IL-8表达降低,IL-10表达升高,除C组IL-8以外,其余差异均有统计学意义(P<0.01或P<0.05)。同时,D组IL-6的表达低于A组,IL-10的表达高于A组,差异有统计学意义(P<0.05)。结论:温针灸可显著上调腹泻大鼠结肠黏膜AQP3、AQP4蛋白及其mRNA表达水平,同时显著减少腹泻大鼠结肠黏膜炎性因子TNF-α、IL-6和IL-8的表达,增加IL-10的表达。温针灸可改善大鼠消化不良性腹泻,对机体起到治疗和保健作用,2个疗程温针灸对于部分炎性因子的改善效果较1个疗程温针灸更加明显。

Objective:To explore the effects of needle-warming moxibustion on the expressions of aquaporin-3(AQP3)and aquaporin-4(AQP4)and inflammatory factors in colonic mucosa of rats with chronic diarrhea of dyspepsia,and to compare the effects of different courses of needle-warming moxibustion.Methods:The rat model of chronic diarrhea of dyspepsia was induced by lactose,and the rats were randomly divided into the blank group(group A),the model group(group B),the needle-warming moxibustion groups of one course(group C)and two courses(group D),with 5 rats in each group.Group C was given needle-warming moxibustion on Tianshu(ST25),Guanyuan(CV4),Zhongwan(CV12),and Zusanli(ST36)for 5 days;group D was given needle-warming moxibustion on the same above points for two courses of 10 days,with 2 days interval between each course.The diarrhea condition was observed,and the colonic mucosa samples were collected at the end of the treatment in each group.The expressions of AQP3 and AQP4 were detected by WB,the expressions of AQP3 mRNA and AQP4 mRNA were detected by qPCR,the expressions of inflammatory factors of TNF-α,IL-6,IL-8 and IL-10 were detected by ELISA,and the positive expressions of AQP3 and AQP4 in colonic mucosa were detected by immunohistochemistry.Results:The diarrhea rate decreased from 100%to 12.5%after one course of the treatment and the rate decreased to 0%after two courses of the treatment.Compared to those in group B,the relative expressions of AQP3 and AQP4 increased significantly in group C and group D,the relative expressions of AQP3 mRNA and AQP4 mRNA increased significantly in group A,group C and group D,and the relative expression of AQP3 mRNA in group D was significantly higher than that in group A(P<0.01,P<0.05).Compared to those in group B,the expressions of TNF-α,IL-6 and IL-8 decreased,whereas the expression of IL-10 increased in group C and group D;of which the differences were statistically significant(P<0.01,P<0.05),except IL-8 in group C;the expression of IL-6 was lower in group D than that in group A,whereas the expression of IL-10 was higher in group D than that in group A(P<0.05).Conclusion:Needle-warming moxibustion can significantly up-regulate the protein and the mRNA expressions of AQP3 and AQP4 in colonic mucosa of diarrhea rats,it can significantly reduce the expressions of TNF-α,IL-6 and IL-8,and increase the expression of IL-10.Needle-warming moxibustion can improve diarrhea of dyspepsia and play a treating and health-caring role in the patients.The effects of two courses of needle-warming moxibustion on some inflammatory factors are more obvious than those of one course of needle-warming moxibustion.