背景:母猪的繁殖能力是影响养猪发展的主要因素。当归的草药提取物(Oliv。)Diels,蒙古黄芪,杜仲。,和甘草D.C.Eaton对母猪的繁殖有改善作用。
目的:为了研究这四种中药以1:1:1:1比例(ALAE)的复方对子宫内膜异位症的治疗作用机制,子宫内膜炎,子宫粘连,宫内发育迟缓,先兆子痫,及其对母猪繁殖效率的提高。
方法:采用超高效液相色谱-质谱分析和网络药理学方法对ALAE的活性成分进行鉴定。然后,我们使用结果构建了一个可视化网络。在母猪动物和猪子宫内膜上皮细胞(PEEC)中验证了ALAE参与母猪繁殖改善的关键靶标和途径。
结果:在ALAE中总共发现了62种活性化合物(在甘草D.C.Eaton中发现了41种,5在黄芪中,11在杜仲中。,5在当归中(Oliv。)Diels)与563个疾病相关靶标(例如caspase-3、EGFR、IL-6)参与EGFR酪氨酸激酶抑制剂耐药,PI3K-AKT,和其他信号通路。分子对接结果显示GC41(光甘草定),GC18(medicarpin),EGFR和CCND1可能是与母猪生殖改善相关的关键成分和靶蛋白。在PEEC中,EGFR表达在mRNA和蛋白质水平降低了三个剂量(160、320和640μg/mL)的ALAE。下游通路PI3K-AKT1的磷酸化增强。炎症因子(IL-6、IL-1β)的表达,ESR1和caspase-3通过多种途径降低。此外,抗炎因子(IL-10)的表达水平,血管生成相关因子(MMP9,PIGF,PPARγ,IgG),和胎盘连接相关因子(CTNNB1,occludin,和claudin1)增加。此外,仔猪出生总数,活的和健康的垃圾数量显着增加。母猪动物中通过ALAE处理减少了死胎的数量。
结论:服用ALAE可显著增加仔猪出生总数,通过改善胎盘结构,减少了活的和健康的窝的数量,减少了死胎的数量,减弱炎症反应,母猪胎盘血管生成和生长因子受体的调节。生殖能力的提高可能与PEECs中EGFR-PI3K-AKT1通路的激活有关。此外,ALAE可能参与雌激素受体的调节,凋亡因子,和细胞周期蛋白。
BACKGROUND: Reproductive ability of sows is a primary element influencing the development of pig farming. Herbal extracts of Angelica sinensis (Oliv.) Diels, Astragalus mongholicus Bunge, Eucommia ulmoides Oliv., and Polypodium glycyrrhiza D.C.Eaton showed effects on improvement of reproduction in sows.
OBJECTIVE: To investigate the mechanism of the treatment effects by a compound of these four Chinese herbs in a 1:1:1:1 ratio (ALAE) on endometriosis, endometritis, uterine adhesion, intrauterine growth retardation, pre-eclampsia, and its enhancement of reproductive efficiency in sows.
METHODS: Active components of ALAE were identified by using ultra-performance liquid chromatography-mass spectrometry analysis and network pharmacology. Then we used the results to construct a visualization network. Key targets and pathways of ALAE involved in
sow reproduction improvement were validated in
sow animals and porcine endometrial epithelial cells (PEECs).
RESULTS: A total of 62 active compounds were found in ALAE (41 in Polypodium glycyrrhiza D.C.Eaton, 5 in Astragalus mongholicus Bunge, 11 in Eucommia ulmoides Oliv., 5 in Angelica sinensis (Oliv.) Diels) with 563 disease-related targets (e.g. caspase-3, EGFR, IL-6) involved in EGFR tyrosine kinase inhibitor resistance, PI3K-AKT, and other signaling pathways. Molecular docking results indicated GC41 (glabridin), GC18 (medicarpin), EGFR and CCND1 are possible key components and target proteins related to reproductive improvement in sows. In PEECs, EGFR expression decreased at the mRNA and protein levels by three doses (160, 320, and 640 μg/mL) of ALAE. The phosphorylation of downstream pathway PI3K-AKT1 was enhanced. The expression of inflammatory factors (IL-6, IL-1β), ESR1 and caspase-3 decreased through multiple pathways. Additionally, the expression levels of an anti-inflammatory factor (IL-10), angiogenesis-related factors (MMP9, PIGF, PPARγ, IgG), and placental junction-related factors (CTNNB1, occludin, and claudin1) increased. Furthermore, the total born number of piglets, the number of live and healthy litters were significantly increased. The number of stillbirths decreased by ALAE treatment in
sow animals.
CONCLUSIONS: Dministration of ALAE significantly increased the total number of piglets born, the numbers of live and healthy litters and decreased the number of stillbirths through improving placental structure, attenuating inflammatory response, modulating placental angiogenesis and growth factor receptors in sows. The improvement of reproductive ability may be related to activation of the EGFR-PI3K-AKT1 pathway in PEECs. Moreover, ALAE maybe involved in modulation of estrogen receptors, apoptotic factors, and cell cycle proteins.