运脾消食颗粒对厌食模型幼龄大鼠下丘脑、胃窦及十二指肠β-EP、nesfatin-1影响研究

Effect of Yunpi Xiaoshi Granules(运脾消食颗粒)onβ-EP and Nesfatin-1 in Hypothalamus,Antrum and Duodenum of Anorexia Model in Young Rats

目的研究厌食模型幼龄大鼠下丘脑、胃窦、十二指肠中β-内啡肽(β-EP)、新型神经肽-1(nesfatin-1)的含量及蛋白表达情况,以及中药复方运脾消食颗粒对两者的干预作用。方法选取SPF级72只Wistar幼龄大鼠,雌雄各半,日龄30 d,体质量(60±10)g,随机分为空白组(12只)和造模组(60只),造模组以病因和症状模型法建立幼龄大鼠厌食症模型。造模成功后的动物随机分为模型组、阳性组(儿宝颗粒组)、运脾消食颗粒高剂量组、运脾消食颗粒中剂量组、运脾消食颗粒低剂量组,每组12只,空白组和模型组灌胃等量0.9%氯化钠溶液,其余4组分别给予等体积的儿宝颗粒混悬液(1.75 g/kg)、运脾消食颗粒高剂量混悬液(4.20 g/kg)、运脾消食颗粒中剂量混悬液(2.10 g/kg)、运脾消食颗粒低剂量混悬液(1.05 g/kg),连续给药14 d,最后一次给药结束禁食禁水24 h后,采集标本,用ELISA法和Western Blot法分别检测大鼠下丘脑、胃窦、十二指肠中β-EP、nesfatin-1的含量和蛋白表达。结果①模型组大鼠下丘脑、胃窦、十二指肠β-EP含量及蛋白表达降低,治疗后儿宝颗粒组大鼠下丘脑、胃窦、十二指肠β-EP含量及蛋白表达增加,运脾消食颗粒高、中剂量组大鼠下丘脑、胃窦、十二指肠β-EP含量及蛋白表达增加;②模型组大鼠下丘脑、胃窦、十二指肠nesfatin-1含量及蛋白表达增加,治疗后儿宝颗粒组大鼠下丘脑、胃窦、十二指肠nesfatin-1含量及蛋白表达降低,运脾消食颗粒高、中剂量组大鼠下丘脑、胃窦、十二指肠nesfatin-1含量及蛋白表达降低。结论运脾消食颗粒可显著降低下丘脑、胃窦、十二指肠中nesfatin-1含量及蛋白表达水平,而增加下丘脑、胃窦、十二指肠中β-EP含量及蛋白水平;可改善模型大鼠的胃肠功能,调节脑肠肽水平,增加食欲,这可能是运脾消食颗粒治疗小儿厌食症的重要机制之一。

Objective To study the content and protein expression of β-endorphin(β-EP)and new neuropeptide-1(nesfatin-1)in hypothalamus,antrum and duodenum of young anorexia model rats,as well as the intervention effect of Yunpi Xiaoshi Granules(运脾消食颗粒)on both.Methods Seventytwo young Wistar rats with SPF grade,male and female,30 days of age,weight(60±10)g,were randomly divided into blank group(12 rats)and model group(60 rats).The model group was used to establish anorexia model of young rats by etiology and symptom model.The animals were randomly divided into model group,positive group[Erbao Granules(儿宝颗粒)group],Yunpi Xiaoshi Granules high-dose group,Yunpi Xiaoshi Granules medium-dose group and Yunpi Xiaoshi Granules low-dose group with 12 animals in each group.The blank group and model group were given the same amount of 0.9%sodium chloride solution.The other four groups were given equal volume of Erbao Granules suspension(1.75 g/kg),Yunpi Xiaoshi Granules high-dose suspension(4.20 g/kg),Yunpi Xiaoshi Granules medium-dose suspension(2.10 g/kg)and Yunpi Xiaoshi Granules low-dose suspension(1.05 g/kg),respectively,for 14 consecutive days.24 hours after the last administration,samples were collected,and the contents and protein expressions of β-EP and nesfatin-1 in hypothalamus,antrum and duodenum of rats were detected by ELISA and Western Blot.Results①β-EP content and protein expression were decreased in hypothalamus,gastric antrum and duodenum of rats in model group;β-EP content and protein expression were increased in hypothalamus,gastric antrum and duodenum of rats in Erbao Granules group after treatment;β-EP content and protein expression were increased in hypothalamus,gastric antrum and duodenum of rats in high-dose and medium-dose groups of Yunpi Xiaoshi Granules;②Nesfatin-1 content and protein expression were increased in hypothalamus,antrum and duodenum of rats in model group,while nesfatin-1 content and protein expression were decreased in hypothalamus,antrum and duodenum of rats in Erbao Granules group after treatment.The content and protein expression of nesfatin-1 decreased in hypothalamus,gastric antrum and duodenum of rats in Yunpi Xiaoshi Granules high-dose and medium-dose groups.Conclusion Yunpi Xiaoxian Granules can significantly reduce nesfatin-1 content and protein expression level in hypothalamus,antrum and duodenum,and increase β-EP content and protein level in hypothalamus,antrum and duodenum.It can improve the gastrointestinal function of model rats,regulate the level of brain intestinal peptide and increase appetite,which may be one of the important mechanisms of Yunpi Xiaoshi Granules in the treatment of anorexia in children.