补骨脂-肉豆蔻醇提物对脾虚泄泻小鼠水通道蛋白及结肠中细胞因子的影响

孙祥瑞; 高家荣; 汪庆飞; 王婷

doi:10.13898/j.cnki.issn.1000-2200.2018.02.004

补骨脂-肉豆蔻醇提物对脾虚泄泻小鼠水通道蛋白及结肠中细胞因子的影响

Effect of ethanol extract of psoralen-nutmeg on aquaporin and cytokines in spleen deficiency diarrhea mice

摘要

摘要: 目的：探讨补骨脂-肉豆蔻醇提物对脾虚泄泻小鼠水通道蛋白（AQP）及结肠中细胞因子的作用。方法：昆明小鼠60只，随机分正常组、模型组、参苓白术散组和补骨脂-肉豆蔻醇提取物（醇提物）低、中、高剂量组，各10只。模型组、醇提物组和参苓白术散组均采用大黄诱导脾虚泄泻小鼠模型，正常组小鼠每天给予等量蒸馏水灌胃。建模成功后，醇提物低、中、高剂量组分别给予补骨脂-肉豆蔻醇提取物7.02、14.04、21.06 g/kg灌胃，参苓白术散组给予参苓白术散2.34 g/kg灌胃，正常组和模型组给予等量蒸馏水，每天1次，连续给药2周。比较各组小鼠体质量和胸腺指数、脾脏指数；免疫组织化学法检测AQP3、AQP4蛋白表达；ELISA法检测结肠白细胞介素（IL）-1β、IL-2、肿瘤坏死因子（TNF）水平。结果：各组小鼠体质量间差异有统计学意义（P<0.01），其中模型组小鼠体质量明显低于正常组（P<0.01），醇提物中、高剂量组小鼠体质量均高于模型组（P<0.05）。模型组小鼠脾脏指数和胸腺指数均低于正常组（P<0.05和P<0.01），而醇提物中、高剂量组小鼠脾脏指数和醇提物高剂量组小鼠胸腺指数均高于模型组（P<0.05）。免疫组织化学结果显示，模型组和醇提物各组小鼠AQP3、AQP4蛋白表达均低于正常组（P<0.05~P<0.01）；醇提物中、高剂量组和参苓白术散组小鼠AQP3、AQP4蛋白表达均明显高于模型组和醇提物低剂量组（P<0.01）。与正常组比较，模型组小鼠结肠中IL-1β、IL-2和TNF-α水平均明显升高（P<0.01）；与模型组和醇提物低、中剂量组比较，醇提物高剂量组和参苓白术散组小鼠结肠中IL-1β、IL-2和TNF-α水平均明显降低（P<0.01）。结论：补骨脂-肉豆蔻醇提物可上调AQP表达，降低肠道的炎性因子水平，对脾虚泄泻小鼠胃肠道功能有一定的改善作用。

Abstract: Objective: To explore the effects of ethanol extract of psoralen-nutmeg on aquaporin(AQP) and cytokines in spleen deficiency diarrhea mice.Methods: Sixty Kunming mice were randomly divided into the normal group,model group,Shenling Baizhu powder group,and low,medium and high dose of ethanol extract of psoralen-nutmeg group(10 mice each group).The spleen deficiency diarrhea mice in model group,Shenling Baizhu powder group and ethanol extract group were established using rhubarb,and the normal group was gavaged using the same volume distilled water.After successful modeling,the low,medium and high dose of ethanol extract group were gavaged with 7.02,14.04 and 21.06 g/kg of ethanol extract of psoralen-nutmeg,respectively.The Shenling Baizhu powder group was gavaged with Shenling Baizhu powder at the dose of 2.34 g/kg,and the normal group and model group were gavaged with the same volume of distilled water,once a week,for 2 weeks.The body mass,thymus index and spleen index between all groups were compared.The protein expression levels of AQP3 and AQP4 were detected by immunohistochemistry,and the levels of IL-1β、IL-2、TNF-α of colon were measured using ELISA.Results: The difference of the body mass between all groups was statistically significant(P<0.01),which in model group was significantly lower than that in normal group(P<0.01),and which in medium and high dose of ethanol extract group was higher than that in model group(P<0.05).The thymus index and spleen index in model group were significantly lower than those in normal group(P<0.05 and P<0.01),and which in middle and high dose of ethanol extract group were higher than those in model group(P<0.05).The results of immunohistochemistry showed that the protein expression levels of AQP3 and AQP4 in normal group and ethanol extract group were lower than those in normal group(P<0.05 to P<0.01),and the protein expression levels of AQP3 and AQP4 in medium and high dose of ethanol extract group and Shenling Baizhu powder group were significantly higher than those in model group and low dose of ethanol extract group(P<0.01).Compared with normal group,the levels of IL-1β,IL-2 and TNF-α in colon tissue in model group significantly increased(P<0.01).Compared with model group and low and medium dose of ethanol extract group,the levels of IL-1β,IL-2 and TNF-α in colon tissue in high dose of ethanol extract group and Shenling Baizhu powder group significantly decreased(P<0.01).Conclusions: Ethanol extract of psoralen-nutmeg can obviously enhance the expression of aquaporin,reduce the levels of intestinal inflammatory factors,and improve the gastrointestinal function of spleen deficiency diarrhea mice.

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