加味蒌石汤对肾阴虚型PCOS模型小鼠卵泡抑素及激活素的影响

Effects of Modified Loushi Medicinal Broth on Follistatin and Activin in Polycystic Ovary Syndrome Model Mice with Chinese Medical Differentiation of Kidney-yin Deficiency

目的观察加味蒌石汤对肾阴虚多囊卵巢综合征(PCOS)模型小鼠卵泡抑素及激活素的影响。方法 ICR小鼠54只随机分为空白组,模型组,中药高、中、低剂量组,达英-35组六组,每组9只。采用"硫酸脱氢表雄酮(DHEA)联合甲状腺素及利血平"法制备肾阴虚型多囊卵巢综合征小鼠模型。除空白组、模型组外,其余各组予相应的药物灌胃40天,观察小鼠一般情况、体质量,酶联免疫吸附法测定小鼠血清性激素[血清睾酮(T)、雌二醇(E2)、促卵泡刺激素(FSH)和黄体生成素(LH)]、血清卵泡抑素(FS)、激活素(ACT)、环磷酸腺苷(cAMP)及环磷酸鸟苷(cGMP)。结果与空白组及各用药组比较,模型组小鼠出现不安定、多动、体毛稀疏、腹泻、消瘦等表现。实验前各组小鼠体质量差异无统计学意义(P>0.05),实验后模型组小鼠体质量低于空白组[(25.96±0.88)g比(28.36±0.77)g,P<0.05],中药高、中剂量组与达英-35组小鼠体质量均高于模型组和低剂量组[(28.25±0.55)g、(27.66±1.21)g、(28.08±0.50)g比(25.96±0.88)g、(26.71±0.74)g,P<0.05];模型组血清T、LH、FS、cAMP/cGMP高于空白组[(146.30±4.04)nmol/L比(134.36±2.30)nmol/L,(1772.66±122.37)ng/L比(1409.02±47.88)ng/L,(45.07±5.63)ng/L比(34.59±2.52)ng/L,(1.85±0.37)比(1.28±0.15),P<0.01],模型组ACT低于空白组[(6.12±0.63)ng/m L比(6.87±0.87)ng/m L,P<0.05],中药高、中、低剂量组与达英-35组血清FS均低于模型组[(36.27±3.05)ng/L、(38.33±2.47)ng/L、(39.27±4.09)ng/L、(36.64±2.55)ng/L比(45.07±5.63)ng/L,P<0.01],中药高、中、低剂量组与达英-35组ACT均高于模型组[(6.96±0.60)ng/m L、(6.90±0.81)ng/m L、(6.86±0.48)ng/m L、(7.14±0.73)ng/m L比(6.12±0.63)ng/m L,P<0.01或P<0.05],中药高、中、低剂量组与达英-35组cAMP/cGMP均低于模型组[(1.36±0.10)、(1.48±0.11)、(1.67±0.08)、(1.42±0.11)比(1.85±0.37),P<0.01]。结论加味蒌石汤能有效调节肾阴虚PCOS模型小鼠性激素水平,改善肾阴虚状态,降低FS含量和cAMP/cGMP比值,提高ACT含量,促进卵泡发育。

Objective To investigatetheeffect of modified Loushi medicinal broth（MLSMB） onfollistatin（FS） and activin（ACT） in polycystic ovary syndrome（PCOS） model mice with Chinese medical differentiation of deficiency of kidney-yin. Methods Fifty-four female ICR mice were randomly divided into 6 groups： normal control group,model group,MLSMB high-, medium-, low-dose groups and Diane-35 group, with 9 mice in each group. PCOS with deficiency of kidney-yin syndromewas induced by dehydroepiandrosterone-sulfate combined with thyroxin and reserpine in mice. Except normal control group, other groups were given the corresponding drugfor 40 days. All mice were weighted and their general condition was observed. Serum testosterone（T）, estradiol（E2）, follicle-stimulating hormone（FSH）, luteinizing hormone（LH）, FS, ACT, and cyclic adenosine monophosphate（cAMP）/cyclicguanosine monophosphate（cGMP） were detected by enzyme linked immunosorbent assay（ELISA）. Results The mice in model group showed signs of instability, hyperactivity, sparse body hair, diarrhea and emaciation. The body weight of model group was lower than that of normal control group（25.96±0.88 g vs 28.36±0.77 g, P0.05）, the body weight of MLSMB high dose, medium dose and Diane-35 groupswasheavier than that of model group and MLSMB low dose group（28.25±0.55 g, 27.66±1.21 g, 28.08±0.50 g vs 25.96±0.88, 26.71±0.74 g, P0.05）. Thelevels of serum T, LH, FS,cAMP/cGMPin model group were higher than those in normal control group（146.30 ±4.04 nmol/L vs 134.36 ±2.30 nmol/L, 1772.66±122.37 ng/L vs 1409.02±47.88 ng/L, 45.07±5.63 ng/L vs 34.59±2.52 ng/L, 1.85±0.37 vs 1.28±0.15;all P0.01）, while the level of serum ACT was lower（6.12±0.63 ng/m L vs 6.87±0.87, P0.01）. Thelevel ofserum FS in MLSMB high dose, medium dose, low dose and Diane-35 groupswas lower than that in model group（36.27 ±3.05 ng/L, 38.33 ±2.47 ng/L, 39.27 ±4.09 ng/L, 36.64 ±2.55 ng/L vs 45.07 ±5.63 ng/L, P 0.01）; ACT was higher（6.96 ±0.60 ng/m L, 6.90±0.81 ng/m L, 6.86±0.48 ng/m L, 7.14±0.73 ng/m L vs 6.12±0.63 ng/m L, P0.01 or P0.05）; cAMP/cGMP was lower（1.36±0.10, 1.48±0.11, 1.67±0.08, 1.42±0.11 vs 1.85±0.37, P0.01）. Conclusion MLSMB could regulate the levels of sex hormone, improve the state of Kidney-yin deficiency, reduce the level of FS and cAMP/cGMP, increase the level ACT, and promote follicular development.