文献类型：期刊文献

中文题名：四君子汤对脾虚大鼠海马生理功能的影响及其机制研究

英文题名：Effect and its mechanism of Sijunzi Decoction on physiological function of hippocampus in rats with spleen deficiency

作者：巩子汉[1,2];李楷[2];楚天云[3];杨晓轶[2];段永强[2];李能莲[2];王惠楠[4];付晓艳[2];成映霞[2];马骏[2];白敏[2]

第一作者：巩子汉

机构：[1]中国中医科学院中医基础理论研究所,北京100700;[2]甘肃中医药大学基础医学院,甘肃兰州730020;[3]河南中医药大学基础医学院,河南郑州450046;[4]甘肃省第二人民医院,甘肃兰州730000

第一机构：中国中医科学院中医基础理论研究所,北京100700

年份：2021

卷号：37

期号：13

起止页码：1692

中文期刊名：中国临床药理学杂志

外文期刊名：The Chinese Journal of Clinical Pharmacology

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；

基金：国家自然科学基金地区基金资助项目(81660758)。

语种：中文

中文关键词：四君子汤;海马;环磷酸腺苷;蛋白激酶A;环磷腺苷效应元件结合蛋白

外文关键词：Sijunzi Decoction;hippocampus;cyclic adenosine monophosphate;protein kinase A;camp-response element binding protein

摘要：目的研究四君子汤对脾气虚模型大鼠P物质(SP)、促胰液素(S)和海马组织环磷酸腺苷(cAMP)、蛋白激酶A(PKA)和环磷腺苷效应元件结合蛋白(CREB)基因和蛋白表达影响。方法按随机数字表法将造模成功大鼠分为5组:模型组、自然恢复对照组(均给予等量生理盐水)和大、中、小3个剂量实验组(给予24,12,6 g·kg^(-1)四君子汤药剂灌胃),每组12只。另取12只正常大鼠作为空白组(等量生理盐水),1次/天,连续21 d。用综合因素法复制大鼠脾虚模型。用酶联免疫吸附法检测各组大鼠海马组织中SP和血清促胰液素S含量,实时荧光定量-PCR法合蛋白质印迹法检测海马组织cAMP、PKA、CREB基因(2^(-△△)ct值)和蛋白(灰度值)表达情况。结果空白组、模型组、自然恢复对照组和大、中、小3个剂量实验组SP含量分别为(2.64±0.19),(2.35±0.16),(2.36±0.10),(2.60±0.17),(2.50±0.09)和(2.38±0.16)ng·mL^(-1);这6组的S含量分别为(0.58±0.01),(0.54±0.01),(0.54±0.02),(0.57±0.01),(0.56±0.01)和(0.54±0.01)ng·mL^(-1);这6组的cAMP蛋白表达量分别为0.76±0.12,0.47±0.04,0.48±0.05,0.68±0.08,0.50±0.05和0.48±0.04;这6组的PKA蛋白表达量分别为0.84±0.09,0.55±0.04,0.56±0.03,0.69±0.06,0.63±0.05和0.59±0.03;这6组的CREB蛋白表达量分别为1.02±0.22,0.57±0.04,0.60±0.04,0.89±0.14,0.67±0.07和0.62±0.05。基因结果的趋势与蛋白一致。上述指标:模型组与空白组比较,差异均有统计学意义(均P<0.01);高、中2个剂量实验组与模型组比较,差异均有统计学意义(P<0.05,P<0.01)。结论四君子汤可促进细胞因子SP和S的异常分泌,并可上调海马组织cAMP、PKA和CREB基因和蛋白异常表达,从而发挥保护脾虚大鼠海马的正常生理功能。
Objective To study the effect of Sijunzi Decoction on substance P(SP),serum tryptophan(S),cyclic adenosine monophosphate(cAMP),protein kinase A(PKA)and cAMP effector element binding protein(CREB)gene and proteins expression in hippocampus of spleen Qi deficiency rats.Methods Wistar rats were randomly divided into 5 groups according to random number table:model group,natural recovery control group(normal saline by gavage)and low,medium and high dose experimental groups(24,12 and 6 g·kg^(-1)Sijunzi Decoction by gavage),12 rats in each group;another 12 normal rats were taken as blank group(normal saline by gavage),1 times a dayfor 21 d.The rats were prepared to duplicate spleen deficiency model by comprehensive factor method.The contents of SP and S were detected by enzyme-linked immunosorbent assay.The expression of cAMP,PKA and CREB genes(2^(-△△)ctvalue)and proteins(gray value)were detected by real-time fluorescence quantitative-PCR and Western blot.Results The contents of SP in blank group,model group,natural recovery control group and large,medium and small dose experimental groups were(2.64±0.19),(2.35±0.16),(2.36±0.10),(2.60±0.17),(2.50±0.09)and(2.38±0.16)ng·mL^(-1),respectively;(0.58±0.01),(0.54±0.01),(0.54±0.02),(0.57±0.01),(0.56±0.01)和(0.54±0.01)ng·mL^(-1),respectively;the expression of cAMP protein expression in the six groups were 0.76±0.12,0.47±0.04,0.48±0.05,0.68±0.08,0.50±0.05 and 0.48±0.04,respectively;the expression of PKA protein expression in the six groups were 0.84±0.09,0.55±0.04,0.56±0.03,0.69±0.06,0.63±0.05 and 0.59±0.03,respectively;the expression of CREB protein expression in the six groups were 1.02±0.22,0.57±0.04,0.60±0.04,0.89±0.14,0.67±0.07 and 0.62±0.05.The trend of gene results is consistent with that of protein.The above indicators:comparison between model group and blank group,the difference was statistically significant(all P<0.01);and comparison between large and medium dose experimental groups and model group,the difference was statistically significant(P<0.05,P<0.01).Conclusion Sijunzi Decoction can promote the abnormal secretion of SP and S,and up regulate the expression of cAMP,PKA and CREB gene and protein in hippocampus,so as to protect the normal physiological function of hippocampus in spleen deficiency rats.