[Effect of SIRT1 on delay of D-gal-induced premature ovarian failure in mice with ginsenoside Rg_1].

To explore the effect of slient mating type information regulation 2 homolog 1(SIRT1) on the delay of D-galactose(D-gal) induced premature ovarian failure in mice with ginsenoside Rg_1(Rg_1). Fifty-four female SPF BALB/c mice were randomly divided into PBS group, D-gal group, and Rg_1 group. In the D-gal group, D-galactose(200 mg·kg~(-1)·d~(-1)) was injected subcutaneously into the neck and back for 42 days. In the PBS group, the equal amount of phosphate buffered saline(PBS) was injected into the neck and back for 42 days. In addition to the therapy of D-gal group, Rg_1 group was given Rg_1(20 mg·kg~(-1)·d~(-1)) through the intraperitoneal injection on the 15 th day for 28 days. At the same time, the D-gal group and the PBS group were also given an equal amount of PBS through hintraperitoneal injection on the 15 th day for 28 days. The changes in body weight and ovarian weight coefficient of mice were detected. Expressions of estradiol 2(E_2), luteinizing hormone(LH), superoxide dismutase(SOD), catalase(CAT) and follicle-stimulating hormone(FSH) in peripheral blood were detected. Morphological changes of ovaries were detected by HE staining. Changes in expression of aging regulator SIRT1 were detected by fluorescent quantitative PCR and Western blot. The results showed that compared with the PBS group, the body weight growth rate of the D-gal group was significantly slowed, the ovarian weight coefficient was decreased, the serum levels of E_2, LH, SOD, CAT were significantly reduced, and FSH was significantly increased. After the administration with Rg_1, the body weight growth rate, ovarian weight coefficient, serum levels of E_2, LH, SOD, and CAT in the mice were higher than those in the D-gal group, while FSH was lower than those in the D-gal group. HE staining showed that the follicular morphology and structure of the PBS group were normal; the number of follicles in the D-gal group was reduced, the corpus luteum was vacuolated, and the number of atretic follicles was increased. Compared with the D-gal group, the number of follicles in the Rg_1 group was increased, whereas the number of corpus luteum was decreased. Compared with the PBS group, SIRT1 expression was down-regulated in the D-gal group, while SIRT1 expression was up-regulated in the Rg_1 group compared with the D-gal group. The results suggested that Rg_1 could delay D-gal-induced premature ovarian failure in a mouse model of premature ovarian failure, and SIRT1 played an important role in this.

Effect of ginsenoside Rg_1 in delaying premature ovarian failure induced by D-gal in mice through PI3K/Akt/mTOR autophagy pathway / 中国中药杂志

The aim of this paper was to study the role of phosphoinositide 3-kinase(PI3 K), protein kinase B(Akt) and mamma-lian target of rapamycin(mTOR) in the inhibition of premature ovarian failure induced by D-galactose(D-gal) in mice model by ginsenoside Rg_1(Rg_1). Fifty-four female SPF BALB/c mice were randomly divided into PBS group, D-gal group, and Rg_1 group. In the D-gal group, D-galactose(200 mg·kg~(-1)·d~(-1)) was injected subcutaneously into the neck and back for 42 days. In the PBS group, an equal amount of phosphate buffered saline(PBS) was injected into the neck and back for 42 days. In addition to the therapy of D-gal group, Rg_1 group was given Rg_1(20 mg·kg~(-1)·d~(-1)) through intraperitoneal injection since the 15 th day for 28 days, at the same time, the D-gal group and the PBS group were also given an equal amount of PBS through intraperitoneal injection since the 15 th day for 28 days. After the treatment, the estrous cycle changes of the mice were detected, and the ovarian SA-β-Gal staining was used to detect the changes of ovarian aging. Western blot was used to detect the changes in protein expressions of PI3 K, Akt, mTOR, S6 k, LC3-Ⅱ and P16~(INK4 a). Fluorescence quantitative PCR was used to detect the changes in mRNA expressions of PI3 K, Akt, mTOR, S6 k, LC3-Ⅱ and P16~(INK4 a). According to the findings, compared with the PBS group, the D-gal group began to show estrous cycle disorder in the 3 rd week,the ovarian SA-β-Gal staining positive granulosa cells increased in the D-gal group, the expression of senescence marker P16~(INK4 a) increased, while the expression of autophagy signaling molecule LC3-Ⅱ decreased. After treatment with Rg_1, the positive rate of ovarian SA-β-Gal staining in Rg_1 group decreased, the expression level of autophagy signaling molecule LC3-Ⅱ in Rg_1 group was higher than that in D-gal group, while the expression level of senescence marker P16~(INK4 a) was lower than that in D-gal group. Compared with the PBS group, the protein and mRNA expressions of PI3 K, Akt, mTOR and S6 k in the D-gal group were up-regulated, the protein expressions of Akt, mTOR and S6 k in the Rg_1 group were up-regulated, and the mRNA expressions of PI3 K and mTOR were up-regulated. After treatment with Rg_1, the protein expressions of PI3 K, Akt, mTOR and S6 k in the Rg_1 group were lower than those in the D-gal group, while the mRNA expressions of Akt, mTOR and S6 k in the Rg_1 group were lower than those in the D-gal group. The finding ssuggested that Rg_1 has the effect in delaying ovarian premature failure in D-gal-induced mouse models, and PI3 K/Akt/mTOR autophagy signaling pathways play an important role.

A study on retinal angiomatous proliferation and its mechanism in apolipoprotein E-deficient mouse with dyslipidemia / 中华实验眼科杂志

Background Retinal angiomatous proliferation (RAP) is a subtype of neovascular age-related macular degeneration (AMD).There are currently very few studies on RAP.Objective This study was to explore the pathogenic mechanism of RAP in apolipoprotein E-deficient (ApoE-/-) mice with dyslipidemia.Methods Twenty-four 2-month-old SPF ApoE-/-mice were randomly divided into the high fat diet group and the normal diet group,and twelve 2-month-old C57BL/6 mice received the normal diet as controls.A diet with a higher content of fat was given for 4 consecutive months in the high fat diet group,and normal diet was given in the same way in the mice of the normal diet group.The mice were sacrificed at 6 months of age.The expression of vascular endothelial growth factor (VEGF) in the retinal pigment epithelial (RPE) cells,the expression of vascular endothelial growth factor receptor-2 (VEGFR-2) in the outer plexiform layer (OPL),microvascular density (MVD) and microvascular area (MVA) in the OPL were examined by immunohistochemistry and semi-quantitatively by histopathology with the Mias 2000 Imaging Analyzer System.The expression of VEGF protein in the retina was examined by Western blot.Results The MVD in the retinal OPL were (20.67±3.20) and (19.50± 1.87),respectively,in the ApoE-/-mice of the high fat diet group and the normal diet group,which were significantly higher than that (12.50±1.87) of the C57BL/6 normal diet group (all at P＜0.01).MVA in the retinal OPL were (626.49± 120.99) μm2 and (514.06±88.83) μm2 in the ApoE-/-mice of the high fat diet group and the normal diet group,respectively,showing a significant increase in comparison with the (336.52±84.96) μm2 of the C57BL/6 normal diet group (P＜0.01).The staining area of VEGF in RPE cells was (21 048±1849) μm2 in the ApoE-/-mice of the high fat diet group,showing a significant increase in comparison with the (17 116±2023) μm2 of the C57BL/6 normal diet group.However,no significant difference was found in the staining area of VEGF between the ApoE-/-mice with normal diet group and the C57BL/6 normal diet group ([17 854±2967] μm2 vs.[17 116±2023] μm2) (P＞0.05).Significant elevation was also seen in the staining area of VEGFR-2 in the retinal OPL of the ApoE-/-mice of the high fat diet group (12 193±3806)μm2 and the ApoE-/-mice of the normal diet group (11 969± 3616)xm2 compared with C57BL/6 mice of the normal diet group (5387±2225)μm2(all at P＜0.01).The relative expression values (VEGF/β-actin) of VEGF in the retinas were (1.51 ±0.32) and (1.17±0.39) in the ApoE-/-mice of the high fat diet group and the normal diet group,respectively,showing a significant increase in comparison with (0.28±0.14) of the C57BL/6 normal diet group (P＜0.01).Conclusions The expression of VEGF and VEGFR-2 in the retinas increases in the ApoE-/-mouse,which leads to the enlargement of MVD and MVA in the retinal OPL and subsequent RAP occurrence.