2021 年第 10 期 第 16 卷

四物汤在大鼠脑和骨骼肌组织中发挥雌激素样效应的机制研究

Study on mechanism of Siwu decoction exerting estrogenic effects on rats brain and skeletal muscle tissues

作者：陈梦1石丹宁2张则业2刘佳2何悦双2颜倩2赵丕文2

英文作者：Chen Meng1 Shi Danning2 Zhang Zeye2 Liu Jia2 He Yueshuang2 Yan Qian2 Zhao Piwen2

单位：1北京中医药大学中医学院102488；2北京中医药大学生命科学学院102488

英文单位：1School of Traditional Chinese Medicine Beijing University of Chinese Medicine Beijing 102488 China; 2School of Life Sciences Beijing University of Chinese Medicine Beijing 102488 China

关键词：四物汤；脑组织；骨骼肌；雌激素样效应 

英文关键词：Siwudecoction;Braintissues;Skeletalmuscle;Estrogeniceffects

• 摘要：

• 目的  探讨四物汤在大鼠脑和骨骼肌组织中发挥雌激素样效应的机制。方法将20只未达性成熟雌性SD大鼠完全随机分为正常对照组、雌激素组、四物汤高剂量组和低剂量组，各5只，适应性喂养4 d后，正常对照组予相等剂量0.9%氯化钠溶液灌胃，雌激素组予戊酸雌二醇0.104 mg/（kg·d）灌胃，四物汤高剂量组予四物汤2.08 g/（kg·d）灌胃，四物汤低剂量组予四物汤0.52 g/（kg·d）灌胃。每日早晚各灌胃1次，连续4 d。各组大鼠最后一次灌胃后3 h称重，10%水合氯醛腹腔麻醉后腹主动脉快速取血处死。观察大鼠脑及腿部骨骼肌组织细胞形态，采用蛋白质印迹法检测雌激素受体β（ERβ）、G蛋白偶联雌激素受体（GPER）、磷脂酰肌醇-3-激酶（PI3K）及蛋白激酶B（Akt）表达情况。结果 4组大鼠脑和骨骼肌组织细胞形态均未出现明显变化。在脑组织中，四物汤低剂量组ERβ表达量显著低于正常对照组和雌激素组［（0.354±0.066）比（0.450±0.022）、（0.509±0.042）］；四物汤高剂量组和四物汤低剂量组GPER表达量显著低于雌激素组；四物汤低剂量组PI3K表达量显著低于正常对照组［（0.255±0.086）比（0.404±0.100）］；雌激素组和四物汤高剂量组Akt表达量显著低于正常对照组［（0.224±0.049）、（0.268±0.097）比（0.467±0.102）］（均P<0.05或P<0.01）。在骨骼肌组织中，雌激素组和四物汤低剂量组ERβ表达量显著低于正常对照组；四物汤低剂量组PI3K表达量显著高于正常对照组和雌激素组，Akt表达量显著低于正常对照组和雌激素组（均P<0.05或P<0.01）。结论 四物汤可不同程度抑制脑组织中ERβ、GPER、PI3K、Akt及骨骼肌中ERβ、Akt的表达，促进骨骼肌中PI3K的表达。推断四物汤可能通过ERβ和GPER介导的PI3K/Akt信号通路发挥雌激素样效应。

• Objective To explore the mechanism of estrogenic effects of Siwu decoction (SWD) on rats brain and skeletal muscle tissues. Methods Twenty immature female SD rats were randomly divided into normal control group, estrogen group, high-dose SWD group and low-dose SWD group, with 5 rats in each group. After adaptively feeding for 4 d, the normal control group was given equal dose of 0.9% sodium chloride solution via oral administration, the estrogen group was given estradiol valerate 0.104 mg/（kg·d） via oral administration, the high-dose SWD group was given SWD 2.08 g/（kg·d） via oral administration, and the low-dose SWD group was given SWD 0.52 g/ （kg·d） via oral administration, with once every morning and evening for 4 d. Rats in each group were weighed 3 h after the last oral administration, and were sacrificed by rapid blood sampling from the abdominal aorta after intraperitoneal anesthesia with 10% chloral hydrate. The brain and leg skeletal muscle tissues cell morphology were observed. Western blot was used to detected the expressions of estrogen receptor β(ERβ), G protein-coupled estrogen receptor (GPER), phosphatidylinositol 3-kinase (PI3K) and protein kinase B (Akt). Results There were no significant changes in the morphology of brain and skeletal muscle tissues among the four groups of rats. In brain tissues, the expression of ERβ of low-dose SWD group was significantly lower than that of normal control group and estrogen group［（0.354±0.066） vs （0.450±0.022）,（0.509±0.042）］; the expression of GPER of high-dose SWD group and low-dose SWD group was significantly lower than that of estrogen group; the expression of PI3K of low-dose SWD group was significantly lower than that of normal control group［（0.255±0.086） vs （0.404±0.100）］; the expression of Akt of estrogen group and high-dose SWD group was significantly lower than that of normal control group［（0.224±0.049）,（0.268±0.097） vs （0.467±0.102）］（all P<0.05 or P<0.01）. In skeletal muscle tissues, the expression of ERβ of estrogen group and low-dose SWD group was significantly lower than that of normal control group; the expression of PI3K of low-dose SWD group was significantly higher than that of normal control group and estrogen group; the expression of Akt of low-dose SWD group was significantly lower than that of normal control group（all P<0.05 or P<0.01）. Conclusions  The expression of ERβ, GPER, PI3K and Akt in brain tissues and the expression of ERβ and Akt in skeletal muscle are inhibited by SWD, while the expression of PI3K in skeletal muscle is promoted. It is possible that SWD showes estrogenic effects through ERβ and GPER and their mediated PI3K/Akt signaling pathway.