Objective To explore the effect of Siwu mixture on secretion level of E2 in ovarian granulose cell induced by cisplatin chemotherapy.Methods Ovarian granulosa cells from ten SD female rat of 22 to 24 days were taken to culture cells.48 healthy SD female rats from 22 to 24 days were randomly divided into 6 groups: normal group, model group, positive medicine group, and the high、medium and low dose of Siwu mixture group, 8 rats in each group.Rats were given corresponding drug by gavage for 3 days:Physiological saline (normal group and model group, 0.5 mL/d),Estradiol valerate tablets(positive medicine group, 0.4 mg/kg·d), Siwu mixture high group(13.5 mL/kg·d), Siwu mixture medium group(6.75 mL/kg·d), Siwu mixture low group(3.38 mL/kg·d).The ovarian granulosa cells were cultured by 20% pharmacological serum for 24 hours.MTT method was used to observe the effect of pharmacological serum on proliferation of primary granulosa cells after cisplatin injury.RIA was used to observe the intervention effect of pharmacological serum on E2 secretion.Mmunohistochemistry was used to detect intervention effect of serum pharmacological on CYP19a1 gene expression and activation of Smad2/3.Results MTT assay results showed that absorbance of positive medicine group and Siwu mixture groups were higher than the normal group, and the Siwu mixture high dose group was the highest(P<0.01), RIA method results showed that Siwu mixture had a auxo-action of E2, and high dose group was the most obvious(P<0.05).Immunohistochemical method results showed that Siwu mixture could improve the expression of CYP19a1 gene and Smad2/3 and Psmad2/3 protein, the effect of high dose group was the most obvious(P<0.01,P<0.01,P<0.01).Conclusion Siwu mixture has the effect of improvement on the secretion of E2.But it may not be able to play the role of completely repair, and need for further experimental observation.

This study was aimed to investigate the effects of Si-Wu mixture on the secretion of E2 and the expression of CYP19al gene in granulosa cells after cell injury.Ovarian granulosa cells of SD rats were treated with cisplatin (CDDP).And the expression levels of E2 and CYP19a1 were determined by different testing methods.The results showed that the level of E2 induced by radioimmunoassay of the Si-Wu group was significantly higher than that of the CDDP group.Meanwhile,the group which added TGF-β3 protein pathway blocker was lower than others.The results of immunohistochemistry and western blotting showed that the expression level of CYP 19a1 of the Si-Wu group was higher than that of the CDDP group.In the western blotting,the group which added blocker was significantly lower than the non-blocking group.It was concluded that the pharmacological serum of Si-Wu mixture can enhance the level of E2 in CDDP cells through TGF-β3 protein pathway.And the effect is accomplished by the intervention of CYP 19a1.

Objective To explore the effect of hydroxylsafflor yellow A (HSYA) on related protein of estrogenic effect. Methods The positive cells T47D and ER, and negative cells SK-BR-3 of estrogen receptor (ER) were cultured, and 1×10-7 mol/L× 10-9 mol/L of HSYA act on this two cells., negative cells SK-BR-3. The expression of protein ERα, ERβ, extracellular regu-lated protein kinases 1/2 (ERK1/2), p-ERK1/2, Progesterone Receptor (PR), P27 was detected by Western Blot. Results Com-pared with the blank control group, the high concentration HSYA promoted the expression of Erα, ERβ, ERK1/2, p-ERK1/2, PR, while inhibite the expression of P27 in T47D cells (P<0.01). HSYA showed significant inhibitory effects on ERK1/2, while had no obvious effects on p-ERK1/2 in SK-BR-3 cells (P>0.05). Conclusion HSYA has different influnces on related protein of estrogenic effect in the cells, which possiblly through regulating the expression of ER and activating the ERK sig-naling pathway.

Objective To observe the influences of Erxian decoction on the proliferation and cell cycle ovarian granulosa cells caused by CDDP. Methods Different doses of Erxian decoction pharmacological serum were injected in ovarian granulosa cells injured by CDDP,normal pharmacological serum was used as blank control,progynova pharmacological serum was used as positive control,and focus on the comparison of changes after joined LY294002 which was PI3K/AKT pathway inhibitor in each group at the same time. Each group of ovarian granulosa cells proliferation was detected by MTT method when pharmacological serum dosing for 24h,48h and 72h. Cell cycle and phase changes in each group were determined by flow cytometry. Results (1) Erxian decoction has proliferation promoting effect on CDDP injury ovarian granulosa cells,and has proliferation effect of ovarian granulosa cells after injured by CDDP and added LY294002 inhibitors. (2)Erxian decoction each dose group and progynova group can rise the proliferation index by enhancing the proportion of S phase granulosa cells and decreasing G0/G1 phase proportion. Conclusions Erxian decoction has significant proliferation promoting effect of CDDP injury ovarian granulosa cells. The possible mechanism on one hand is that it activates or enhances AKT signaling pathways and inhibits cell apoptosis. On the other hand,it perhaps promotes S phase proportion transformation from G0 phase therefore increases the ratio of S phase cells.

源自中药的植物雌激素是天然的杂环多酚类化合物,能够与雌激素核受体或膜受体结合发挥作用,被称为选择性雌激素受体调节剂.植物雌激素的药理作用体现在治疗骨质疏松、心血管疾病、恶性肿瘤及对抗艾滋病毒等方面.高水平、大规模地深入研究植物雌激素的作用机制应当成为未来重要的研究方向,以期为新药研发提供理论依据.

Objective:To observe the effect of serum containing Erxian decoction on regulating the expression of bcl-2and bax protein from rats' ovarian granulosa cells induced by cisplatin and to investigate the mechanism of Erxian Decoction serum on apoptosis of ovarian granulosa cells from the perspective of apoptosise.Methods:Thirty SD female rats for 22 to 24 days birth were randomly divided into three groups:the normal saline group,progynova group and the Erxian Decoction group with 10 rats in each group.After 4-day drug administration,rat hearts were taken to prepare drug serum.Ovarian granulosa cells were taken from SD rats and cultured to acquire adherent cells.Then,primary ovarian granulosa cells in rats were induced by cisplatin.After pharmacological action to serum,radio immunity method was used to detect estrogen and progestin and make analysis.Immunohistochemistry was applied to detect the expression of bcl-2 and bax protein in each group and then for image analysis.Results:Test results showed that E2 expression significantly changed in the model group and progynova group (P<0.05) with statistically significant difference,compared with the normal group.Compared with the model group,the expression of E2 increased significantly in the progynova group and the Erxian Decoction group.Compared with the normal group,P expression in the model group was somewhat changed (P<0.05),which showing statistically significant difference.Compared with the model group,P expression in Erxian Decoction group was slightly higher than that of the progynova group,but lower than that of the normal group.Immunohistochemistry results showed that brown granules of bcl-2 and bax protein were expressed in the cytoplasm of ovarian granulosa cells.Compared with the normal group,the expression of bcl-2 in each group under the intervention of cisplatin was decreased,and the model group was the most obvious (P<0.05).Compared with model group,the positive expression of bcl-2 in each drug group was increased,but it was weaker than that in the normal group (P<0.05).The intensity of bax and bcl-2's expression was opposite.Compared with the normal group,Bax expression in each group under the intervention of cisplatin were enhanced and that in the model group especially showed a strong expression (P<0.05).Compared with the model group,the positive expression of the drug groups was decreased,but was higher than that of the normal group (P<0.05) with statistically significant difference.Conclusion:Erxian Decoction can decrease the expression of Bax protein in follicular granulosa cells induced by cisplatin injury and increase the expression of bcl-2 protein.It can inhibit the apoptosis of ovarian granulosa cells and protect the ovarian function by adjusting the expression of bcl-2 and bax.

目的:观察顺铂干预大鼠卵巢颗粒细胞凋亡情况,从而探讨二仙汤含药血清经PI3k/Akt通路抑制卵巢颗粒细胞凋亡的作用机制.方法:SD雌性大鼠30只,随机分为3组,分别为生理盐水组、补佳乐组、二仙汤组.心脏取血,制备药物血清.SD大鼠卵巢颗粒细胞,经培养贴壁后,用顺铂损伤大鼠原代卵巢颗粒细胞.经各组药理血清作用后,应用Annexin V/PI双染流式细胞仪检测细胞凋亡,并对各实验组大鼠卵巢颗粒细胞的凋亡情况进行分析.应用免疫组化法,测定各分组PI3k、Akt、p-Akt蛋白的表达情况,并进行图像分析.结果:流式细胞凋亡结果显示,二仙汤可以降低顺铂损伤后的大鼠卵巢颗粒细胞的凋亡率.免疫组化结果显示,PI3k、p-Akt、Akt在细胞质内均有表达,可见棕黄色阳性物质;而二仙汤可以上调顺铂干预后大鼠卵巢颗粒细胞的PI3k、p-Akt、Akt的表达.结论:二仙汤含药血清上调了PI3k、p-Akt、Akt的表达,从而抑制了卵巢颗粒细胞凋亡,该过程可能通过调节PI3k/Akt通路实现,进而有效地保护卵巢功能.

The increasing incidence of female menopausal cardiovascular disease is closely related to the decline of the levels and hypofunction of estrogen.As an important steroid hormone,estrogen has widely distributed targets and complex biological effects.It enrolls in the regulation of the functions of multiple organs and systems.Estrogen or estrogenic compounds exert their biological effects through the non-genomic and genomic pathways via the typical estrogen receptor (ER)αand ERβor G protein coupled es-trogen receptor (GPER).The research of estrogen on cardiovascular system via ER pathway and its mechanisms will present as an important reference for the clinical treatment of female menopausal cardiovascular disease.This paper will discuss mainly on the role and its mechanisms of estrogen or estrogenic compounds on cardiovascular system.