黄精超微粉及水提物对自然围绝经期综合征模型大鼠的作用研究

Effect of superfine powder and aqueous extract of Polygonati Rhizoma on rats with natural perimenopausal syndrome

该实验将观察黄精超微粉及水提物对自然围绝经期综合征模型大鼠的作用,并探讨其可能的作用机制。将14~15月龄雌性SD大鼠经阴道涂片,筛选出60只动情周期紊乱的大鼠,随机分为6组,即模型对照组、苯甲酸雌二醇组(0.1 mg·kg-1)、黄精超微粉组(0.25、0.5 g·kg^(-1))、黄精水提物组(0.25、0.5 g·kg^(-1)),另设10只6~7月龄雌性SD大鼠为青年对照组,连续给药6周,测定体温、耳面部微循环血流量、眩晕时间、唾液分泌量、抓力、骨强度、旷场实验等围绝经期综合征症状相关指标;检测胸腺和脾脏湿重及系数、外周血T淋巴细胞及其亚群占比、血液学指标等免疫系统相关指标;检测动情周期、子宫、卵巢湿重及系数、卵巢组织形态学及细胞凋亡情况等卵巢相关指标;检测血清性激素水平、卵巢组织中细胞色素P450-11A1(CYP11A1)、芳香化酶(CYP19A1)、细胞色素P450-17A1(P45017A1)蛋白表达等下丘脑-垂体-卵巢轴(hypothalamic-pituitary-ovarian axis,HPO)相关指标。结果显示,黄精超微粉及水提物能显著降低围绝经期大鼠体温(肛、面、背温),耳部微循环血流量,眩晕时间,增加唾液分泌量、抓力、骨强度、旷场中水平运动总距离及总速度;显著升高胸腺和脾脏湿重及系数、淋巴细胞比率、CD3^(+)水平、CD4^(+)/CD8^(+)比值,减少中性粒细胞数量及比率;显著降低动情周期紊乱比率、血清促卵泡激素(follicle-stimulating hormone,FSH)、黄体生成素(luteinizing hormone,LH)含量,改善卵巢组织形态及细胞凋亡情况,增加子宫湿重及系数、卵巢湿重、血清抑制素B(inhibin B,INHB)、雌二醇(estradiol,E2)、抗缪勒管激素(anti-müllerian hormone,AMH)含量及卵巢CYP11A1、CYP19A1蛋白表达。提示黄精超微粉及水提物能够改善自然围绝经期综合征模型大鼠相关症状,并能改善卵巢功能和提高免疫功能,其作用机制可能与促进雌激素合成从而调节HPO轴功能有关。

This study aims to examine the effect of superfine powder and aqueous extract of Polygonati Rhizomaon on natural perimenopausal syndrome in rats and explore the underlying mechanism. To be specific, a total of 60 female SD rats(14-15 months old) with estrous cycle disorder were screened by the vaginal smear and randomized into model control group, β-estradiol 3-benzoate group(0.1 mg·kg^(-1)), superfine powder of Polygonati Rhizoma group(0.25, 0.5 g·kg^(-1)) and aqueous extract of Polygonati Rhizoma group(0.25, 0.5 g·kg^(-1)), and another 10 female SD rats(14-15 months old) were selected as the youth control group. The administration lasted 6 weeks. Then the perimenopausal syndrome-related indexes such as body temperature, microcirculatory blood flow of face and ear, vertigo period, salivary secretion, grip force, and bone strength were determined and open field test was conducted. The immune system-related indexes such as the wet weight and index of thymus and spleen, percentage of T lymphocytes and subgroups in peripheral blood, and hematological indexes were measured. In addition, the ovary-related indexes such as estrous cycle, the wet weight and index of uterus and ovary, ovarian tissue morphology, and cell apoptosis were determined. Moreover, hypothalamus-pituitary-ovary axis(HPO)-related indexes such as serum sex hormone levels, cytochrome P450 family 11 subfamily A member 1(CYP11A1), cytochrome P450 family 19 subfamily A member 1(CYP19A1), and cytochrome P450 family 17 subfamily A member 1(P450 17A1) in ovarian tissue were measured. The results showed that the superfine powder and aqueous extract of Polygonati Rhizoma significantly decreased body temperature(anal, facial and dorsal temperature), microcirculatory blood flow in the ear, and vertigo period, increased salivary secretion, grip force, bone strength, total distance and total speed in the open field test, wet weight and index of thymus and spleen, lymphocyte ratio, CD3^(+)level, and CD4^(+)/CD8^(+)ratio, reduced neutrophil number and ratio, estrous cycle disorder ratio, and number of ovarian apoptotic cells, raised wet weight and index of uterus, wet weight of ovary, levels of inhibin B(INHB), estradiol(E2), anti-müllerian hormone(AMH), and ovarian CYP11A1 and CYP19A1, decreased follicle-stimulating hormone(FSH) and luteinizing hormone(LH) content, and improved ovarian tissue morphology. It is suggested that the superfine powder and aqueous extract of Polygonati Rhizoma can improve the symptoms associated with natural perimenopausal syndrome in rats and enhance ovarian function and immune function. The mechanism is that they regulate HPO axis function by increasing estrogen synthesis.