Protective effect of Yunkang oral liquid via regulating androgen receptor in polycystic ovary syndrome rats.

Objectives: This study aimed to explore the effect and mechanism of Yunkang oral liquid (YK) on polycystic ovary syndrome (PCOS). Methods: PCOS model rats were prepared by injecting exogenous androgen dehydroepiandrosterone, and YK was administered simultaneously for 28 days during modeling. The morphology of ovaries and uterus was observed using H&E staining, and serum levels of testosterone (T), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) were determined by radioimmunoassay. Additionally, serum lipids (TG, HDL-c), blood glucose (GLU), and aminotransferase (AST, ALT) levels were detected. The expression of androgen receptor (AR) protein was determined by Western blotting. Results: YK treatment resulted in reduced serum levels of T, LH and FSH, ameliorated ovarian polycystic-like pathological changes and uterine morphology in PCOS rats, and decreased serum TG, GLU, AST and ALT levels, elevated serum HDL-c levels, and improved abnormalities of glycolipid metabolism accompanying PCOS. Moreover, YK decreased the expression of ovarian AR in PCOS rats. Conclusions: This study indicates that YK may protect the ovaries by inhibiting the expression of AR, which could be a potential treatment for PCOS.

Yunkang Oral Liquid Promotes Restoration of Mice from Postpartum Kidney Deficiency via TGF-β/Smads Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Yunkang oral liquid on postpartum kidney deficiency in mice. MethodPostpartum mice were randomized into model and low-dose （6 mL·kg-1）， medium-dose （9 mL·kg-1）， and high-dose （12 mL·kg-1） Yunkang oral liquid groups. The mouse model of postpartum kidney deficiency was established by sleep deprivation combined with forced swimming. Another 9 female ICR mice were selected as the normal control group. The mice were administrated with Yunkang oral liquid during the period of modeling. The levels of estradiol （E2）， progesterone （P）， follicle-stimulating hormone （FSH）， and luteinizing hormone （LH） in the serum were measured by the enzyme-linked immunosorbent assay. The morphological changes of ovaries and uterus were observed by hematoxylin-eosin （HE） staining， and the expression of transforming growth factor （TGF）-β and Smad2/3 was determined by immunohistochemistry and Western blotting. ResultThe mice in the model group showed prolonged estrous cycle， reduced voluntary activity， dorsal temperature， grip strength， and bone strength， and whitening tongue coating. Compared with the model group， Yunkang oral liquid shortened the estrous cycle， increased the voluntary activity， dorsal temperature， grip strength， and bone strength， and alleviated the whitening of tongue coating. Moreover， it elevated the E2 and P levels and lowered the FSH and LH levels in the serum， decreased ovarian follicular atresia rate， promoted uterine repair， and down-regulated the expression of TGF-β and Smad2/3 in the ovarian and uterine tissues. ConclusionYunkang oral liquid can ameliorate postpartum kidney deficiency in mice by regulating the TGF-β/Smads signaling pathway.

[Effect of Yunkang Oral Liquid on preventing LPS-induced abortion and regulating immune tolerance in mice].

To investigate the effect of Yunkang Oral Liquid on preventing lipopolysaccharide(LPS)-induced abortion and regulating immune tolerance in mice,sixty normal ICR mice were randomly divided into normal group,model group,Yunkang Oral Liquid high,middle and low dose groups and progesterone group.Abortion model was established by tail vain injection of LPS(0.1µg/mouse)on the 7th day of pregnancy.Since the first day of pregnancy,the same volume of distilled water,Yunkang Oral Liquid at the dose of 36,18 and 9 m L·kg~(-1)·d~(-1),and progesterone at the dose of 0.038 g·kg~(-1)·d~(-1)were given in corresponding groups.The mice were sacrificed at the 9th day of pregnancy,and the embryo loss of each group was calculated.The levels of Th1 type cytokines(TNF-α,IFN-γ)and Th2 type cytokines(IL-4,IL-10)in uterus homogenate were detected by ELISA.HE staining was performed to examine the histopathological changes in the decidua.The expression levels of CD14,F4/80 in macrophages of uterus were detected by immunohistochemistry.Western blot was used to investigate the protein expression of TLR4,MyD88 and NF-κB in uterine decidua.In our study,all Yunkang Oral Liquid groups could significantly reduce the embryo absorption rate of mice,while high dose group can significantly increase the levels of IL-10 and IL-4;both medium and high dose groups can significantly decrease TNF-α,and IFN-γlevelsin the uterus of model mice,reduce the protein expression of NF-κB,MyD88 and TLR4 in uterine decidua tissue.Various treatment groups could reduce the counts of F4/80,CD14 macrophages and decrease expression area in uterine tissue.Our results showed that Yunkang Oral Liquid could prevent LPS-induced abortion,and the mechanism may be associated with inhibiting the TLR4/MyD88/NF-κB signaling pathway and regulating the balance of Th1/Th2 immune factors,which could improve the endometrial receptivity of mice,and promote the development of decidua and implantation of embryo.

Effect of Yunkang Oral Liquid on preventing LPS-induced abortion and regulating immune tolerance in mice / 中国中药杂志

To investigate the effect of Yunkang Oral Liquid on preventing lipopolysaccharide(LPS)-induced abortion and regulating immune tolerance in mice,sixty normal ICR mice were randomly divided into normal group,model group,Yunkang Oral Liquid high,middle and low dose groups and progesterone group.Abortion model was established by tail vain injection of LPS(0.1μg/mouse)on the 7th day of pregnancy.Since the first day of pregnancy,the same volume of distilled water,Yunkang Oral Liquid at the dose of 36,18 and 9 m L·kg~(-1)·d~(-1),and progesterone at the dose of 0.038 g·kg~(-1)·d~(-1)were given in corresponding groups.The mice were sacrificed at the 9th day of pregnancy,and the embryo loss of each group was calculated.The levels of Th1 type cytokines(TNF-α,IFN-γ)and Th2 type cytokines(IL-4,IL-10)in uterus homogenate were detected by ELISA.HE staining was performed to examine the histopathological changes in the decidua.The expression levels of CD14,F4/80 in macrophages of uterus were detected by immunohistochemistry.Western blot was used to investigate the protein expression of TLR4,MyD88 and NF-κB in uterine decidua.In our study,all Yunkang Oral Liquid groups could significantly reduce the embryo absorption rate of mice,while high dose group can significantly increase the levels of IL-10 and IL-4;both medium and high dose groups can significantly decrease TNF-α,and IFN-γlevelsin the uterus of model mice,reduce the protein expression of NF-κB,MyD88 and TLR4 in uterine decidua tissue.Various treatment groups could reduce the counts of F4/80,CD14 macrophages and decrease expression area in uterine tissue.Our results showed that Yunkang Oral Liquid could prevent LPS-induced abortion,and the mechanism may be associated with inhibiting the TLR4/MyD88/NF-κB signaling pathway and regulating the balance of Th1/Th2 immune factors,which could improve the endometrial receptivity of mice,and promote the development of decidua and implantation of embryo.

[Effect and mechanism of Yunkang oral liquid in regulating endocrine system and VEGF signaling pathway and reducing abortion rate in recurrent abortion mice].

This experiment focuses on the effect of Yunkang oral liquid on abortion rate, endocrine system and VEGF signal pathway in Clark classical recurrent abortion model mice. RSA mice were randomly divded into model group, low, middle and high-dose groups and progesterone group. The normal pregnancy mice were included into normal group. Since the first day of pregnancy, the normal group and the RSA model group were given the same dose of distilled water, while low, middle and high-dose groups were given Yunkang oral liquid at the dose of 9, 18, 36 mL·kg¹·d⁻¹; progesterone group were given progesterone by 0.039 g·kg¹·d⁻¹. The mice were put to deathat the 15th day of pregnancy, and the embryo loss rate of each group was observed. Serum estradiol (E2), progesterone (P), prolactin (PRL), luteinizing hormone (LH), follicle stimulating hormone (FSH) level were tested; the protein expressions of estrogen receptor(ER), progesterone receptor (PR), prolactin receptor (PRLR) in decidua and RAS, MAPK, VEGF, VEGFR-2 gene and protein expressions in deciduas were studied. The results showed that middle, high dose Yunkang and progesterone could significantly decrease the embryo loss rate of RSA mice. The levels of FSH, LH, PRL, P and E2 in serum in Yunkang and progesterone groups were increased, and the serum levels of FSH, LH, and E2 in Yunkang group were higher than those in progesterone group. Western blot analysis showed that Yunkang oral liquid and progesterone can significantly increase the expressions of PRLR, PR in the uterine decidua of RSA mice, and the expression of ER in Yunkang group was higher than that in progesterone group. Western blot and PCR showed that the Yunkang oral liquid and progesterone can significantly increase RAS, MAPK, VEGF, VEGFR-2 gene and protein expressions in the uterine decidua of RSA mice. The results showed that Yunkang oral liquid can effectively reduce the embryo loss rate of RSA model mice, increase the levels of FSH, LH, PRL, P and E2 in serum, promote the expressions of PRLR, PR, ER protein in decidua and the RAS, MAPK, VEGF, VEGFR-2 gene and protein expressions in the decidua, improve the vascular remodeling of fetal interface, the endometrial receptivty, the development of decidua and the blastocyst implantation.

Effect and mechanism of Yunkang oral liquid in regulating endocrine system and VEGF signaling pathway and reducing abortion rate in recurrent abortion mice / 中国中药杂志

This experiment focuses on the effect of Yunkang oral liquid on abortion rate, endocrine system and VEGF signal pathway in Clark classical recurrent abortion model mice. RSA mice were randomly divded into model group, low, middle and high-dose groups and progesterone group. The normal pregnancy mice were included into normal group. Since the first day of pregnancy, the normal group and the RSA model group were given the same dose of distilled water, while low, middle and high-dose groups were given Yunkang oral liquid at the dose of 9, 18, 36 mL·kg¹·d⁻¹; progesterone group were given progesterone by 0.039 g·kg¹·d⁻¹. The mice were put to deathat the 15th day of pregnancy, and the embryo loss rate of each group was observed. Serum estradiol (E₂), progesterone (P), prolactin (PRL), luteinizing hormone (LH), follicle stimulating hormone (FSH) level were tested; the protein expressions of estrogen receptor(ER), progesterone receptor (PR), prolactin receptor (PRLR) in decidua and RAS, MAPK, VEGF, VEGFR-2 gene and protein expressions in deciduas were studied. The results showed that middle, high dose Yunkang and progesterone could significantly decrease the embryo loss rate of RSA mice. The levels of FSH, LH, PRL, P and E₂ in serum in Yunkang and progesterone groups were increased, and the serum levels of FSH, LH, and E₂ in Yunkang group were higher than those in progesterone group. Western blot analysis showed that Yunkang oral liquid and progesterone can significantly increase the expressions of PRLR, PR in the uterine decidua of RSA mice, and the expression of ER in Yunkang group was higher than that in progesterone group. Western blot and PCR showed that the Yunkang oral liquid and progesterone can significantly increase RAS, MAPK, VEGF, VEGFR-2 gene and protein expressions in the uterine decidua of RSA mice. The results showed that Yunkang oral liquid can effectively reduce the embryo loss rate of RSA model mice, increase the levels of FSH, LH, PRL, P and E₂ in serum, promote the expressions of PRLR, PR, ER protein in decidua and the RAS, MAPK, VEGF, VEGFR-2 gene and protein expressions in the decidua, improve the vascular remodeling of fetal interface, the endometrial receptivty, the development of decidua and the blastocyst implantation.