大黄不同萃取物对大鼠下丘脑和腺垂体超微结构及其GnRH、GnRH-R和FSHβ表达的影响  

Effects of different extracts of Rheum palmatum on ultrastructure and expression of GnRH,GnRH-R and FSHβ in hypothalamus and pituitary of rats

作　　者：赵盼盼[1] 佟继铭[1] 马淑月 高飞[1] 郭建恩[1] 张树峰[1] 

机构地区：[1]河北省中药研究与开发重点实验室/承德医学院中药研究所,承德067000 [2]华北理工大学,唐山063000

出　　处：《中药药理与临床》2017年第2期125-130,共6页Pharmacology and Clinics of Chinese Materia Medica

基　　金：河北省教育厅重大项目(2009301)

摘　　要：目的:比较大黄不同萃取物对成年雌性大鼠下丘脑、腺垂体超微结构及功能的影响,筛选大黄具有生殖毒性的主要萃取物。方法:采用极性梯度萃取法制备大黄水提物的氯仿萃取物、乙酸乙酯萃取物、正丁醇萃取物和水溶物。雌性成年大鼠随机分为正常对照组、大黄水提物组和各萃取物组,连续灌胃给药60天。采用透射电镜观察下丘脑、腺垂体超微结构病理变化;免疫组化法检测下丘脑弓状核中GnRH、腺垂体促性腺细胞中GnRH-R和FSHβ表达水平。结果:(1)超微结构:电镜下,与正常对照组相比,大黄水提物(生药4.0 g/kg)、氯仿(4.44 mg/kg)和乙酸乙酯萃取物(40.96 mg/kg)组均出现不同程度的病理变化,下丘脑弓状核神经元出现核仁增大,内质网、线粒体数目增多,同时伴有染色质边集和溶酶体增多;腺垂体促性腺细胞可见核膜断裂,线粒体肥大,内质网和高尔基体扩张,以及由粗面内质网组成的同心圆性膜性小体。正丁醇萃取物(79.43 mg/kg)和水溶物(835.72 mg/kg)组超微结构病变不明显。(2)大黄水提物(生药4.0 g/kg)组、氯仿(4.44 mg/kg)和乙酸乙酯萃取物(40.96 mg/kg)组GnRH和FSHβ表达明显高于正常对照组,大黄水提物(生药4.0 g/kg)组GnRH-R表达明显低于正常对照组。大黄水提物(生药4.0 g/kg)组GnRH和FSHβ表达明显高于正丁醇萃取物(79.43 mg/kg)和水溶物(835.72 mg/kg)组,乙酸乙酯萃取物(40.96 mg/kg)组GnRH和FSHβ表达高于正丁醇萃取物(79.43 mg/kg)组,其余各组间比较差异均无统计学意义。结论:氯仿(4.44 mg/kg)和乙酸乙酯萃取物(40.96 mg/kg)具有与大黄水提物相类的毒性作用,是大黄主要的生殖毒性萃取物。Objective： To compare the effect of different extracts of Rheum palmatum L. on the ultrastructure and function of hypothalamus and pituitary of adult female rats, and screen the main reproductive toxicity extract of Rheum palmatum. Methods： The chloroform, ethyl acetate and n-butanol extracts and water-soluble substance of Rheum palmatum were prepared by polarity gradient extraction method. Female adult rats were randomly divided into normal control group, rhubarb water extract group and different extracts groups. Medicine was continuously administered for 60 days. The ultrastructures of hypothalamus and pituitary were observed by transmission electron microscope. And the ex- pressions of GnRH in hypothalamic arcuate nucleus and GnRH-R and FSHβ in glandular pituitary gonadotropic cells were detected by immu- nohistochemistry method. Results： （ 1 ） Ultrastructure：The hypothalamus and pituitary of rats in groups of rhubarb water extract（4.0 g/kg for crude drug） , chloroform（4.44 mg/kg）and ethyl acetate extract （40.96 mg/kg）all appeared different degrees of pathological changes com- pared with normal control group. The hypothalamic arcuate nucleus neurons showed enlarged nucleolus, increased endoplasmic reticulum and mitochondria, as well as rolling chromatin and increased lysosome, glandular pituitary gonadotropic cells showed broken nuclear membrane, hypertrophied mitochondria, dilated endoplasmic reticulum and Golgi apparatus, as well as concentric membranous bodies composed of rough endoplasmic reticulum. The ultrastructural changes of n-butanol extract（79.43 mg/kg）and water soluble （835.72 mg/kg）groups were not ob- vious. （2） The expression of GnRH and FSHβ in rhubarb water extract（4.0 g/kg for crude drug） group , chloroform（4.44 mg/kg） and ethyl acetate extract（40.96 mg/kg） groups was higher than those in the normal control group （P 〈 0.05 or P 〈 0.01 ） , and the expression of GnRH-R in rhubarb water extract（4.0 g/kg for crude drug） group was lower

关 键 词：大黄 萃取物 生殖毒性 GNRH GnRH-R FSHΒ 

分 类 号：R285.5[医药卫生—中药学]