禹州漏芦总甾酮对缺乳模型大鼠的催乳作用及分子机制研究

目的：探讨催乳颗粒（国家药品标准WS3-413(Z-085)-2003(Z)）主要组分禹州漏芦中有效成分总甾酮（total sterone, TSR）的催乳效果，并研究其作用机制。方法：SD大鼠雌雄交配后，取产仔时间相差不超过24 h的60只雌鼠随机分为正常对照组、模型组、TSR低、高剂量组和催乳颗粒阳性药对照组，每组12只母鼠，每窝仔鼠为8只。除正常对照组外，各组母鼠自分娩第2日起腹腔注射左旋多巴2 mg/kg，每天1次，连续注射7 d。造模当日开始灌胃给药，每天1次，连续给药14 d，正常对照组母鼠生理盐水灌胃，每天1次。自给药开始每天测定母鼠单次泌乳量至第14天，然后处死各组母鼠。采用病理HE染色观察各组母鼠乳腺组织形态变化，采用ELISA检测血清催乳素（PRL）和5-羟色胺（5-HT）水平，采用免疫组化检测各组母鼠乳腺组织PRL分布变化，采用Real-time qPCR检测乳蛋白、乳脂相关基因β-casein、FAS、ACC表达和各组母鼠乳腺组织经典Wnt信号通路相关基因β-catenin、c-Myc、CCND1、SFRP4、DNMT1、MeCP2表达的变化。结果：TSR低、高剂量组都能显著提高缺乳模型母鼠单次泌乳量，改善模型组母鼠乳腺组织病理形态，提高PRL和5-HT血清水平。TSR增加模型组母鼠乳腺组织PRL分布，上调乳蛋白、乳脂相关基因β-casein、FAS、ACC和经典Wnt信号通路相关基因β-catenin、c-Myc、CCND1、SFRP4、DNMT1、MeCP2表达。 结论：TSR对缺乳模型母鼠具有明显的催乳作用，其机制可能与促进母鼠血清PRL及下丘脑5-HT释放，增加乳腺组织PRL分布，上调乳蛋白、乳脂相关基因和激活经典Wnt信号有关。

Prolactinic effects and molecular mechanisms of total sterone from Echinops latifolius Tausch on the milk deficient model rats

AIM: To investigate the effect of an effective component total sterone (TSR) of Echinops latifolius Tausch, the main component of a Chinese patent medicine Cuiru Keli (national drug standard WS3-413 (Z-085)-2003 (Z), on lactation and its possible mechanism. METHODS: After mating between male and female SD rats, 60 female rats were randomly divided into normal control group, model group, TSR low-dose and high-dose groups and prolactin granule positive control group, with 12 female rats in each group and 8 newborn rats in each nest. In addition to the normal control group, the rats in each group were intraperitoneally injected with levodopa 2 mg/kg once a day for 7 days from the second day of delivery. The rats in the normal control group were given normal saline by gavage once a day for 14 days. From the beginning of self-sufficiency, the single lactation of the female rats was measured every day until the 14th day, and then the female rats in each group were killed. Pathological HE staining was used to observe the morphological changes of mammary gland tissue in each group. ELISA was used to detect the levels of serum prolactin (PRL) and 5-hydroxytryptamine (5-HT). Immunohistochemistry was used to detect the distribution of PRL in mammary gland tissue of each group. Furthermore, Real-time qPCR was used to detect the expression of milk protein, milk fat related genes β-casein, FAS, ACC and the expression of canonical Wnt signaling pathway related genes β-catenin, c-Myc, CCND1, SFRP4, DNMT1, MeCP2 in mammary gland of each group. RESULTS: Both low and high dose TSR could significantly increase the single lactation volume, improve the pathological morphology of mammary gland, and increase the serum levels of PRL and 5-HT. TSR increased the distribution of PRL and up-regulated the expression of milk protein, milk fat related genes β-casein, FAS, ACC and canonical Wnt signaling pathway related genes β-catenin, c-Myc, CCND1, SFRP4, DNMT1, MeCP2.CONCLUSION: TSR can significantly promote lactation in lactation deficient rats, and its mechanism may be related to promoting the release of PRL and 5-HT in serum, increasing the distribution of PRL in mammary gland, up-regulating the milk protein and milk fat related genes and activating the canonical Wnt signal.