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目的:筛选大黄产生生殖毒性的主要萃取物,为生殖毒性成分的研究奠定基础。方法:依次用氯仿、乙酸乙酯、正丁醇对大黄水煎液进行萃取,得到相应的萃取物和水溶物。60只SD雌性大鼠随机分为正常对照组、大黄水煎液组、氯仿萃取物组、乙酸乙酯萃取物组、正丁醇萃取物组、水溶物组,各组用药剂量均相当于大黄生药4.0 g/kg,连续灌胃给药60天,末次给药后24h取血清、子宫、卵巢,HE染色,光镜下观察子宫腺体、卵巢中各级卵泡的结构变化;ELISA法检测大鼠血清中P、T、E2激素水平。结果:(1)组织结构:与正常对照组相比,各给药组子宫均出现不同程度的腺体扩张,上皮细胞呈扁平状,部分呈空泡状或高柱状,部分增生,数量增多,核增大。血管数量增多,扩张呈囊状。间质疏松、水肿。氯仿萃取物组表现最明显,乙酸乙酯萃取物组次之;各给药组卵巢中各级卵泡数均减少,黄体数、闭锁卵泡数增多,氯仿萃取物组卵巢中颗粒细胞减少,卵泡内细胞排列紊乱。(2)血清激素水平:与正常对照组比较大黄各萃取物组及大黄水煎液组大鼠血清中P、T激素水平均降低,E2激素水平均升高。其中氯仿萃取物组激素水平变化最为显著。结论:初步确定氯仿萃取物为大黄产生生殖毒性的主要萃取物。
OBJECTIVE: To screen the main extracts of reproductive toxicity caused by rhubarb, and lay the foundation for the study of reproductive toxic components. Methods: Rhubarb decoction was extracted with chloroform, ethyl acetate and n-butanol to obtain the corresponding extracts and water-soluble substances. Sixty SD female rats were randomly divided into normal control group, rhubarb decoction group, chloroform extract group, ethyl acetate extract group, n-butanol extract group, water soluble group, the dosage of each group were equivalent to rhubarb 4.0 g / kg of crude drug was administrated for 60 days. The serum, uterus, ovary and HE were stained by hematoxylin and eosin (HE) 24 h after the last administration. The changes of follicles in uterus gland and ovary were observed under light microscope. Serum P, T, E2 hormone levels. Results: (1) Tissue structure: Compared with the normal control group, the uterus of each administration group showed different degrees of glandular dilatation, the epithelial cells were flattened, vacuolar or tall columnar part, hyperplasia and number increased, Nuclear enlargement. Increased number of blood vessels, dilated cystic. Interstitial loose, edema. Chloroform extract group showed the most obvious, followed by ethyl acetate extract group; the number of ovarian follicles at all levels in each administration were reduced, the number of luteal phase, the number of atresia follicles increased, the chloroform extract group of ovarian granulosa cells, follicular Disorders of cells. (2) serum hormone levels: compared with the normal control group rhubarb extract group and rhubarb decoction serum P, T hormone levels were reduced, E2 hormone levels were increased. Chloroform extract of which the most significant changes in hormone levels. Conclusion: The main extracts of rhubarb with reproductive toxicity were initially identified by chloroform extract.