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AIM:To investigate the neuroprotective effect of 17β-estradiol(E2)on retina light damage in BALB/c and C57BL/6 mice and provide experimental data for the successful construction of a research model for E2 against retinal light damage.METHODS:Totally 40～45 adult female BALB/c or C57BL/6 mice were divided into six groups, 6 for each group: normal control, ovariectomized control, ovariectomized light(mice were stimulated with continuous white light at 10000 lx for 4, 8, 12, 16, and 24h after 14d of ovariectomy), intravitreal administration sham operation, saline and E2 pre-treatment groups(2μL saline or 10-5mol/L E2 were intravitreal injected respectively after 14d of ovariectomy operation and 24h of dark adaptation). The morphological and functional changes of the retina were detected by paraffin section HE staining, TUNEL staining and electroretinogram.RESULTS:In the ovariectomized light group, the thickness of the inner/outer nuclear layer decreased significantly from the 4h stimulation of 10000 lx white light group. Intravitreal administration of E2 significantly inhibited the apoptosis of retinal cells in the two strains of mice(P<0.01)and the decrease of amplitudes of a- and b-waves in max-ERG of C57BL/6 mice(P<0.05).CONCLUSION:The light loss sensitivity of two strains of mice was different under the same light stimulation. E2 had a protective effect on both morphology and function of the retina in BALB/c mice, and had a significant protective effect on retina function in C57BL/6 mice.
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OBJECTIVE@#To investigate the influence of chronic masseter hyperalgesia induced by 17β-estradiol (E2) and experimental occlusal interference (EOI) on underlying mechanism in hippocampus of ovariectomized (OVX) rats.@*METHODS@#In the study, 32 OVX rats were randomly divided into 4 groups (8 rats/group): The control group was OVX group, and 0 μg/d E2 (vehicle) injection was started 7 d after OVX without EOI; in the experimental group (1) OVX + E2 group, 80 μg/d E2 injection was started 7 d after OVX without EOI; in the experimental group (2) OVX + EOI group, vehicle injection was started 7 d after OVX and EOI was applied 17 d after OVX; in the experimental group (3) OVX + E2 + EOI group, 80 μg/d E2 injection was started 7 d after OVX and EOI was applied 17 d after OVX. Bilateral masseter muscle mechanical withdrawal thresholds were measured before OVX, 7 days after OVX (before E2 injection), 17 days after OVX (10 days after E2 injection and before EOI) and 24 days after OVX (7 days after EOI). Immunofluorescence staining was used to reveal phospho-extracellular signal regulated kinase 1/2 (p-ERK1/2)-positive neurons in CA3 of hippocampus. The protein expression of p-ERK1/2 in hippocampus was detected using Western Blot.@*RESULTS@#Compared with the control group [left side: (135.3±8.5) g, right side: (135.4±10.8) g], bilateral masseter muscle mechanical withdrawal thresholds of OVX+E2 group [left side: (113.3±5.6) g, right side: (112.5 ± 5.6) g] and OVX+EOI group [left side: (93.3±5.4) g, right side: 90.8±5.5) g] were decreased (P < 0.01). Bilateral masseter muscle mechanical withdrawal thresholds were significantly lower in OVX+E2+EOI group [left side: (81.2±6.2) g, right side: 79.8±7.7) g] than in the control, OVX+E2 and OVX+EOI groups (P < 0.05). The proportion of p-ERK1/2 positive neurons in the CA3 region of the hippocampus was increased in the control, OVX+E2, OVX+EOI and OVX+E2+EOI groups in turn, and the difference between the groups was statistically significant (P < 0.05). p-ERK1/2 protein expression was increased in the control, OVX+E2 and OVX+EOI groups in turn, but the difference was not statistically significant (P>0.05). p-ERK1/2 expression was significantly higher in OVX+E2+EOI group than in the other three groups (P < 0.05).@*CONCLUSION@#High concentration of E2 could exacerbated EOI-induced chronic masseter hyperalgesia in ovariectomized rats, and its central mechanism may be related to the upregulation of the phosphorylation of ERK1/2 in hippocampus.
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Animals , Female , Humans , Rats , Estradiol , Hippocampus , Hyperalgesia/chemically induced , Masseter Muscle , Ovariectomy , Rats, Sprague-DawleyABSTRACT
Aim To study the regulatory effect of daidzein on osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) expression in MG-63 cells and its mechanism. Methods RT-PCR, Western blot and siRNA were used to study the regulatory effect of daidzein on OPG and RANKL expression in human osteoblast-like MG-63 cells. Results Daidzein could promote the expression of OPG mRNA and protein in MG-63 cells and inhibit the expression of RANKL mRNA and protein, which could be blocked by ICI 182780. It was confirmed that ERa and ER0 mediated not only the promoting effect of daidzein on OPG expression of MG-63 cells but also the inhibition of RANKL. Conclusions Daidzein promotes OPG gene expression in MG-63 cells and inhibits the expression of RANK gene expression through ERa and ERβ pathways.
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OBJECTIVES@#To study the effects of 17β-estradiol (E2) on the regulation of the proliferation of condylar chondrocytes and provide a preliminary discussion on the role of phosphorylate-mammalian target of rapamycin (p-mTOR) in this regulatory process.@*METHODS@#Condylar chondrocytes were isolated from 6-week-old female rats for primary culture. Drug treatment with different concentrations of E2 and/or rapamycin (RAPA) was carried out on second-generation cells. Cell Counting Kit 8 was used to measure the cell viability of condylar chondrocytes after culture for 24, 48, or 72 h, and reverse transcription-polymerase chain reaction (RT-PCR) was applied to detect the relative gene expression of estrogen receptor alpha (ERα), estrogen receptor beta (ERβ), collagen type Ⅱ (COLⅡ), autophagy-related gene 6 (Beclin-1), and autophagy-related gene 5 (ATG-5). Western blot was employed to determine the relative protein expression of ERα, ERβ, Beclin-1, lipid-modified light chain 3B (LC3-Ⅱ), and p-mTOR.@*RESULTS@#E2 could significantly promote the proliferation of chondrocytes cultured @*CONCLUSIONS@#At a concentration of 10
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Animals , Female , Rats , Autophagy , Cell Proliferation , Chondrocytes , Estradiol , Estrogen Receptor alpha/metabolism , Estrogen Receptor beta , PhosphorylationABSTRACT
Objective: To observe the effect of 17β-estradiol (17β-E2) on the calcium (Ca2) channels during the osteogenic differentiation of rat bone marrow mesenchymal stem cells (MSCs), and to elucidate the mechanism of 17β-E2 in the osteogenic differentiation of MSCs. Methods: The MSCs were separated by density gradient centrifugation and adherent screening, and passaged for 3 times continuously to induce osteoblast differentiation. The MSCs were divided into control group [cultivated in osteoblast culture medium alone (OBM)] and different doses of 17β-E2 groups (added with 0. 1, 1.0, 10. 0, and 100. 0 pmol · L-1 17β-E2 in OBM, respectively). On the 14th day of osteogenic induction, the cells in each group were stained with Fluo-3/AM, and the Ca2 levels were determinated by laser scanning confocal microscope; the mean fluorescence intensity (MFD was used to respresent the level of Ca2. Whole-cell Ca2 currents were recorded using whole-cell patch clamp technique under different conditions. Results: The MSCs with fibroblast-like cells, oval nuclei and visible nucleoli were successfully isolated by density gradient centrifugation and adherent screening. The subcultured MSCs grew vigorously and maintained the morphological characteristics of primary cells. Following the increase of 17β-E2 concentration, the Fluo-3 fluorescence staining intensity of Ca2 in each group was also gradually increased, especially in 100. 0 pmol · L-1 17β-E2 group. Compared with control group, the MFI of Ca' and the current peak values of Ca' in 10. 0 and 100. 0 pmol · L-1 17β-E2 groups were increased (P0. 05). Conclusion: The cells isolated by density gradient and adherent screening method are the rat MScs. 17β-E2 plays a role in promoting osteogenesis by enhancing the opening of Ca' channels in the MSCs and the inward current of calcium ions in a dose-dependent manner.
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To investigate the mechanism of 17β-estradiol (E2) induced tumor angiogenesis by VEGFα in the microenvironment of lung adenocarcinoma in A549 cells. Methods A549 cells were divided into three groups; PBS group, 10 nmol L"1 E2 group and 10 nmol L"1 E2 +5 (xmol L"1 estrogen receptor antagonist (ICI) group. Western blot was used to detect the expression of VEGFα in A549 cells, and ELISA was used to detect the expression of VEGFα in culture medium. The proliferation of human umbilical vein endothelial cells (HUVECs) was detected by MTT, the migration ability of HUVECs by scratch assay and the tubulogenesis in vitro by tube formation assay. Results Compared with control group, the expression of VEGFα in A549 cells in E2 group was higher, but the expression of VEGFα in A549 cells was significantly inhibited with the addition of ICI. Meanwhile, E2 induced the expression of VEGFα in A549 cells, promoting proliferation, migration and tubulogenesis of HUVEC, while ICI had the opposite effects. Conclusions E2 stimulates the expression of VEGFα by binding to estrogen receptor in A549 cells, promoting proliferation, migration and tube formation of HUVECs, contributing to lung adenocarcinoma angiogenesis.
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A rapid decline in egg production of laying hens begins after 480 d of age. Such a rapid decrease results predominantly from the ovarian aging, accompanied by endocrine changes, decreased yolk synthesis and accumulation, and the reduction in follicles selected into the preovulatory hierarchy. In this study, hens at 90, 150, 280, and 580 d old (D90, D150, D280, and D580, respectively) were compared for yolk precursor formation in the liver to elucidate effects of aging on laying performance. The results showed that liver lipid synthesis increased remarkably in hens from D90 to D150, but decreased sharply at D580 as indicated by the changes in triglyceride (TG) levels. This result was consistent with the age-related changes of the laying performance. The levels of liver antioxidants and total antioxidant capacity decreased significantly in D580 hens and the methane dicarboxylic aldehyde in D580 hens was much higher than that at other stages. The serum 17β-estradiol level increased from D90 to D280, but decreased at D580 (P<0.05). The expression of estrogen receptor α and β mRNAs in the liver displayed similar changes to the serum 17β-estradiol in D580 hens. Expressions of the genes related to yolk precursor formation and enzymes responsible for fat acid synthesis were all decreased in D580 hens. These results indicated that decreased yolk precursor formation in the liver of the aged hens resulted from concomitant decreases of serum 17β-estradiol level, transcription levels of estrogen receptors and critical genes involved in yolk precursor synthesis, and liver antioxidant status.
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Animals , Female , Age Factors , Antioxidants , Metabolism , Chickens , Egg Yolk , Metabolism , Estradiol , Blood , Lipids , Liver , Metabolism , Oviposition , Receptors, Estrogen , GeneticsABSTRACT
Objectives: To explore the effect of 17β-estradiol (E2) on hypoxic pulmonary hypertension (HPH) and explore if the effects were mediated through suppressing pulmonary artery smooth muscle cells (PASMCs) proliferation by targeting miRNA-21 (miR-21). Methods: Animal experiment: A total of 32 healthy female SD rats with castrated surgery were randomly divided into 4 groups: normoxia group, normoxia+E2 group, hypoxia group, hypoxia+E2 group (n=8 each). The rats in normoxia+E2 group and hypoxia+E2 group received subcutaneous injection of E2 20 μg/kg/d, and the rest groups received subcutaneous injection of equal volume saline. The hypoxic groups were placed in the hypoxic chamber (24 hours per day for 8 weeks) to establish HPH model and normoxic groups were kept in the room air. The pulmonary artery remodeling, mean pulmonary artery pressure (mPAP), right ventricle hypertrophy index (RVHI) were observed. Real-time PCR and Western blot were used to detect the levels of proliferation cell nuclear antibody (PCNA) and miR-21 expression in pulmonary artery. In vitro: human pulmonary artery smooth muscle cells (hPASMCs) were randomly divided into 3 groups: normoxia group, hypoxia group, hypoxia+E2 group. The levels of cell proliferation in each group were tested by MTT after 24 hours. Real-time PCR and Western blot were used to detect the levels of PCNA and miR-21 in cells. Results: Animal experiment: compared with normoxia group, the hypoxia group showed obviously thickened pulmonary artery wall, increased mPAP and RVHI, and significantly increased expression of miR-21 and PCNA (P<0.01);above changed were significantly attenuated in hypoxia+E2 group (P<0.01). In vitro: compared with normoxia group, the hypoxia group showed obvious proliferation and significantly increased expression of miR-21 and PCNA (P<0.01);compared with hypoxia group, the proliferation of hPASMCs and expression of miR-21 and PCNA were obviously reduced in hypoxia+E2 group (P<0.01). Conclusions: E2 could effectively reduce mPAP, attenuate the degree of right heart hypertrophy and pulmonary vascular remodeling, the protective effect may be mediated through downregulating miR-21 and PCNA expression, and subsequently inhibiting the proliferation of hPASMCs.
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Purpose To investigate the effects and mechanism of 17β-estradiol on the apoptosis and inflammation of renal tubular cells in rats with renal ischemia/reperfusion injury. Methods All the female Sprague-Dawley rats were ovariectomized and randomly divided into four groups: Control group, Sham group, I/R group and estrogen plus I/R (E2 + I/R) group (n = 8). Right kidney of the rat was excised and artery of the left kidney was blockaded for 45 min.24 h after the reperfusion, we collected the blood and nephridial tissue of each group. An automatic biochemical analyzer was used to measure the expression level of BUN and Cr in blood. Hematoxylin-eosin (HE) staining was used to observe the pathological changes and the degree of inflammatory reaction of the ischemia/reperfusion injury kidney. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) method was used to detect the apoptosis of renal tubular cells. The expression levels of Cleaved-Caspase-3 protein were measured by Western blot, while the numbers of CD4+ T lymphocyte infiltration in each group were tested by immunofluorescence (IF). Results Compared with the Sham group, expression level of BUN, Cr and Cleaved-Caspase-3 in I/R group significantly increased (P＜0.05) as well as the number of apoptotic cells (P＜0.05). In the meantime, inflammatory reaction significantly aggravated (P＜0.05) and the number of CD4 + T lymphocytes increased remarkably (P＜0.05). However, expression level of BUN, Cr and Gleaved-Caspase-3 in E2 + I/R group decreased significantly (P＜0.05) and the pathological damage in the kidney was alleviated (P＜0.05) compared with I/R group, furthermore, the number of apoptotic cells decreased (P＜0.05) compared with I/R group. The inflammatory reaction significantly blunted (P＜0.05) and the infiltration of CD4 + T lymphocytes decreased remarkably (P＜0.05) compared with I/R group. Conclusion Estrogen can inhibit the expression of Cleaved-Caspase-3 in renal tissue during ischemia/reperfusion injury and reduce the apoptosis of renal tubular cells. It can also reduce the infiltration of CD4 + T lymphocytes, thus playing a protective role on renal ischemia/reperfusion injury.
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In this paper, Fe3O4/MnO2doped graphene molecularly imprinted hybrid material ( Fe3O4/MnO2-MIP@ RGO ) was successfully synthesized via reversible addition fragmentation chain transfer ( RAFT ) molecularly imprinting technique by using methacrylic acid as functional monomer, divinylbenzene as cross-linker, Fe3O4/MnO2@ RGO as carrier, and 17β-estradiol ( 17β-E2 ) as template molecule. A novel molecularly imprinting electrochemical sensor by using Fe3O4/MnO2-MIP@ RGO modified electrode was constructed to specifically detect trace 17β-E2 in water. The experimental results showed that the Fe3O4/MnO2-MIP@ RGO electrochemical sensor exhibited rapid and linear current response to 17β-E2 in water samples with a linear range of 4 nmol/L to 0. 8 μmol/L ( R=0. 9852) , the detection limit was 47. 2 pmol/L (3σ) and the relative standard deviation (RSD) was from 2. 1% to 2. 5% . This study provides a simple and efficient, economical and reliable method for the monitoring of 17β-estradiol in the complex water environment.
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To investigate the effect of estrogen receptor (ER) agonist 17β-estradiol and G protein-coupled estrogen receptor 1 (GPER) agonist fulvestrant on masangial cell fibrogenesis under protein kinase C (PKC),we quantified type Ⅳ collagen (COL4A1),fibronectin (FN1),connective tissue growth factor (CTGF) and transforming growth factor-β1 (TGFβ1) gene transcription and semi-quantified phosphorylation of Akt signal upon Phorbol 12-myristate 13-acetate stimulation (which increased COL4A1,FN1,CTGF and TGFβ1 gene transcription to 2.5-0.5,1.4 ±0.2,26 ± 11 and 1.9 ±0.3 times compared with baseline,P ＜0.05) when incubated with the two drugs.It was found that 17β-estradiol and fulvestrant down-regulated COL4A1,FN1,CTGF and TGFβ1 genes transcription (P ＜0.05) and Akt signaling under PKC activation via ER and GPER.ER and GPER agonists are beneficial in protecting the mesangial cells from fibrogenic stimuli by inhibiting PKC signaling and excessive extracellular matrix production.
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Objective To investigate whether 17β-estradiol (E2) can stimulate the proliferation,migration,and secretion of trefoil factor family 1 (TFF1) in papillary thyroid cancer K-1 cells and explore the molecular mechanisms.Methods ELISA was used to detect the content of TFF1 in the supernatant of K-1 cells after the treatment of E2,propylpyrazoletriol (PPT,ERα agonist) or diarylpropionitrile (DPN,ERβ agonist).The expression of ERα and ERβ in the untreated cells was measured by Western blotting.ERα siRNA and ERβ siRNA by RNA interference were designed and synthesized,and the change of TFF1 was measured by ELISA again after the transfection.The interaction between TFF1 promoter and ER was evaluated by chromatin immunoprecipitation analysis (CHiP).The proliferation and migration were detected in the K-1 cells after E2 treatment by MTT assay and Transwell chamber test respectively.Festults After E2 treatment,the TFF1 content in the supernatant of K-1 cells was increased gradually,reached peak at 24 h,and then declined slowly.PPT treatment enhanced the secretion of TFF1 but DPN decreased it in the K-1 cells.Transfection of ERα siRNA obliterated the inductive effect of E2 on the secretion of TFF1,but that of ERβ siRNA increased the inductive effect in the K-1 cells.Western blotting showed that the expression level of ERα was higher than that of ERβ in the K-1 cells.ChIP results confirmed that ERα protein was bound to the promoter of TFF1 gene in K-1 cells.E2 treatment promoted cell proliferation and improved cell migration in the K-1 cells.Conclusion E2 induces the expression and secretion of TFF1 in K-1 cells through ERα-dependent manner,and thus promotes the proliferation and migration of the cells.
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Objective To investigate the protective effects of 17β-estradiol on human lens epithelial (HLE) cells in oxidative damage induced by H2 O2 and its involved mechanisms.Methods HLE cells cultured in vitro were collected and divided into 4 groups;cells in negative control group were cultured with normal medium,cells with 80％ fusion in n2O2 damage group was treated with 100 μmol · L-1 H2O2 for 12 h,cells in 17β-estradiol low dose group were incubated with 10 μmol · L-1 17β-estradiol for 24 h then subjected to 100 μmol · L-1 H2O2 for 12 h and ceEs in 17β-estradiol high dose group were cultured in 100 μmol · L-1 17β-estradiol for 24 h then treated with 100 μrnol · L-1 H2O2 for 12h.Next,cell viability was tested by CCK-8 colorimetric assay,while apoptotic rate was detected by flow cytometry,and intracellular reactive oxygen species (ROS) level was detected by H2 DCFDA fluorescence probe labeling method,as well as the contents of catalase (CAT),superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were determined by 721 D spectrophotometer.Results When compared with the negative control group,the survival rate of HLE ceils in the H2 O2 damage group was significantly decreased,and there was significantly different between both groups (P ＜0.05).Moreover,the survival rate of the 17β-estradiol low dose group (67.44％) and high dose group (78.52％) was obviously higher than that of the H2O2 damage group (59.34％),and the difference was statistically significant (P ＜ 0.05).After H2O2-induced injury,there was significant difference in the apoptotic rate of HLE cells between the H2O2 damage group (41.30 ±3.21)％ and the negative control group (1.67+0.32)％.In addition,the apoptotic rates of the 17β-estradiol low dose group and high dose group was (20.97 + 1.13)％ and (14.27 + 0.90)％ respectively,which was statistically different from the H2 O2 damage group (all P ＜ 0.05).H2 DCFDA fluorescent labeling test results showed that ROS fluorescence signal intensity gradually weakened after treated with 17β-estradiol.Besides,17β-estradiol significantly increased the expression of CAT、SOD and GSH-Px in HLE cells.Conclusion 17β-estradiol has obvious protective effects on HLE cells from the damage induced by H2O2.
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Objective To investigate the protective effects of 17β-estradiol on human lens epithelial (HLE) cells in oxidative damage induced by H2 O2 and its involved mechanisms.Methods HLE cells cultured in vitro were collected and divided into 4 groups;cells in negative control group were cultured with normal medium,cells with 80％ fusion in n2O2 damage group was treated with 100 μmol · L-1 H2O2 for 12 h,cells in 17β-estradiol low dose group were incubated with 10 μmol · L-1 17β-estradiol for 24 h then subjected to 100 μmol · L-1 H2O2 for 12 h and ceEs in 17β-estradiol high dose group were cultured in 100 μmol · L-1 17β-estradiol for 24 h then treated with 100 μrnol · L-1 H2O2 for 12h.Next,cell viability was tested by CCK-8 colorimetric assay,while apoptotic rate was detected by flow cytometry,and intracellular reactive oxygen species (ROS) level was detected by H2 DCFDA fluorescence probe labeling method,as well as the contents of catalase (CAT),superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were determined by 721 D spectrophotometer.Results When compared with the negative control group,the survival rate of HLE ceils in the H2 O2 damage group was significantly decreased,and there was significantly different between both groups (P ＜0.05).Moreover,the survival rate of the 17β-estradiol low dose group (67.44％) and high dose group (78.52％) was obviously higher than that of the H2O2 damage group (59.34％),and the difference was statistically significant (P ＜ 0.05).After H2O2-induced injury,there was significant difference in the apoptotic rate of HLE cells between the H2O2 damage group (41.30 ±3.21)％ and the negative control group (1.67+0.32)％.In addition,the apoptotic rates of the 17β-estradiol low dose group and high dose group was (20.97 + 1.13)％ and (14.27 + 0.90)％ respectively,which was statistically different from the H2 O2 damage group (all P ＜ 0.05).H2 DCFDA fluorescent labeling test results showed that ROS fluorescence signal intensity gradually weakened after treated with 17β-estradiol.Besides,17β-estradiol significantly increased the expression of CAT、SOD and GSH-Px in HLE cells.Conclusion 17β-estradiol has obvious protective effects on HLE cells from the damage induced by H2O2.
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AIM:To investigate the effects of estrogen on the expression of matrix metalloproteinases-2(MMP-2), tissue inhibitor of metalloproteinases-2(TIMP-2) and transforming growth factor-β1(TGF-β1) in cultured human corneal stromal cells.METHODS: Inflammatory environments of human corneal stromal cells were simulated by using 1.5ng/mL IL-1β.The cells were then treated with or without different concentrations of estrogen(0, 1×10-4, 1×10-6, 1×10-8, 1×10-10mol/L estradiol)in vitro.Cell viability was evaluated by MTT.Expression levels of MMP-2, TIMP-2 and TGF-β1 proteins were measured by enzyme-linked immunosorbent assay(ELISA).RESULTS:Estrogen did not affect the viability of human corneal stromal cells.Compared with the control group, expression levels of MMP-2 and TGF-β1 proteins in E2 treatment group significantly decreased after being treated with estrogen, while the expression level of TIMP-2 significantly increased.CONCLUSION: Estrogen could, to some extent, down-regulate the expression of MMP-2 and TGF-β1 and up-regulate the expression of TIMP-2, which might contribute to protecting human cornea.
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AIM: To discuss the protective effects and possible mechanisms of 17β-estradiol on human retinal pigment epithelial ( RPE) cells induced by high glucose. ·METHODS: RPE cells were cultured and divided into four groups according to randomized controlled method:blank control group:the cells were treated with 5. 5mmol/L routine glucose medium for processing; high glucose group: cells were treated with 100mmol/L glucose for 12h;17β-estradiol low concentration group: after treated with 10 μmol/L 17β-estradiol, cells were treated with 100mmol/L glucose for 12h; 17β-estradiol high concentration group: after treated with 100 μmol/L 17β-estradiol, cells were treated with 100mmol/L glucose for 12h. Cell viability were tested by MTT colorimetric detection. Cells apoptosis were detected by Hochest33258 staining. Intracellular reactive oxygen species( ROS) level were detected by H2 DCFDA staining. Expression of CAT, SOD and MDA were tested by colorimetric detection. · RESULTS: RPE cell activity decreased with the concentration of glucose increased; 17β-estradiol inhibited high glucose-induced cell viability decrease in RPE cells, decreased the apoptosis rate of RPE cells and intracellular ROS generation; besides, 17β-estradiol significantly increased the expression of CAT, SOD and decreased the expression of MDA in RPE cells. ·CONCLUSION: The 17β-estradiol effectively inhibited high glucose -induced RPE cells damage, which provide reliable experimental basis for the treatment of injuries in RPE cells.
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Objective To evaluate the role of phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2) in 17β estradiol-induced inhibition of propofol-caused neuroapoptosis in the hippocampus of newborn rats.Methods Seventy-eight male Sprague-Dawley rats,aged 7 days,weighing ll-18 g,were divided into 6 groups (n =13 each) using a random number table:dimethyl sulfoxide (DMSO) group,fat emulsion group (group F),17β estradiol group (group E),propofol group (group P),propofol plus 17β estradiol group (group PE) and propofol plusl7β estradiol plus mitogen-activated protein kinase kinase 1/2 inhibitor U0126 group (group PEU).17β estradiol 600 μg/kg was injected subcutaneously every 24 h for 7 consecutive days in group E,and the equal volume of DMSO was given instead in group DMSO.Propofol 75 mg/kg was injected intraperitoneally every 24 h for 7 consecutive days in group P,and the equal volume of fat emulsion was injected instead in group F.Propofol 75 mg/kg was injected intraperitoneally,and 17β estradiol 600 μg/kg was injected subcutaneously every 24 h for 7 consecutive days in group PE.Propofol 75 mg/kg was injected intraperitoneally,17β estradiol 600 μg/kg was injected subcutaneously,and U0126 10 mg/kg was injected intraperitoneally every 24 h for 7 consecutive days in group PEU.At 15 min after the last injection,3 rats in each group were randomly selected,and arterial blood samples from the cardiac apex were collected for determination of arterial oxygen partial pressure.The animals were sacrificed at 24 h after the last injection for determination of the expression of activated caspase-3 (by immunohistochemistry) and p-ERK1/2 (by Western blot).Results There was no significant difference in arterial oxygen partial pressure between the six groups (P＞0.05).Compared with group F,the expression of activated caspase-3 was significantly up-regulated,and the expression of p-ERK1/2 was downregulated in group P (P＜0.05).Compared with group P,the expression of activated caspase-3 was significantly down-regulated,and the expression of p-ERK1/2 was up-regulated in group PE (P＜0.05).Compared with group PE,the expression of activated caspase-3 was significantly up-regulated,and the expression of p-ERK1/2 was down-regulated in group PEU (P＜0.05).Conclusion The mechanism by which 17β estradiol inhibits propofol-caused neuroapoptosis in the hippocampus is related to up-regulation of the expression of p-ERK1/2 in newborn rats.
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Objective To examine the expression of stromal cell-derived factor-1 (SDF-1) in a rat model of retinal ischemia-reperfusion injury (RIRI),and investigate the protective effect of 17β-Estradiol (E2) on RIRI and explore the mechanism.Methods The RIRI model was established in Sprague-Dawley rats by increasing the intraocular pressure.Relative expression levels of SDF-1 mRNA and protein in the retina at 6 hours,12 hours and 24 hours following reperfusion was determined by RT-PCR and Western blot,respectively.E2 was administered to investigate the effects of estrogen on SDF-1 expression,and the estrogen receptor antagonist ICI 182-780 was administered to investigate the effect of estrogen receptor on the expression of SDF-1.Results SDF-1 expression in RIRI 6 hours group,12 hours group and 24 hours group was increased compared with normal control group (all P ＜ 0.05),with maximum expression at RIRI 12 hours group.As expected,pretreatment of RIRI rats with E2 had a protection on RIRI retina;SDF-1 expression was increased in RIRI + E2 group compared with IR control group and RIRI + vehicle group (all P ＜ 0.05).RIRI + E2 + ICI 182-780 group could decrease SDF-1 expression compared with RIRI + E2 group(all P ＜ 0.05).Conclusion E2 offers protection against RIRI by inducing an up-regulation in SDF-1 expression through activation of the estrogen receptor.
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Objective To study the regulation of serum FSH and E2 in the treatment of menopausal syndrome with 17 beta estradiol combined with DRSP. Methods A total of 90 female patients with menopausal syndrome treated in our hospital from November 2013 to November 2016 were selected. The patients were randomly divided into three groups: 17β group (30 patients, 17β-estradiol alone), DRSP group (30 patients treated with DRSP alone) and 30 patients (17β-estradiol combined with DRSP). Kupperman's menopausal syndrome was used to evaluate the physical symptoms of the patients and to compare the patients after the treatment. The patients' treatment efficiency was observed. The items examined before treatment included liver and kidney function, hematuria routine , Fasting blood glucose, FSH, E2, LDL, blood TC, electrocardiogram, intracavitary Doppler ultrasound, breast ultrasound, HDL, Kupperman evaluation table in accordance with the evaluation before the end of each treatment measure again, And performs the above-mentioned inspection. Results In the combined group, there was no significant difference in the scores of other symptoms except the differences between the ants on the skin and the treatment before treatment. There were significant differences in the scores of pain, headache, body sensation, agitation, palpitation, Pain, fatigue, insomnia and hot flashes were significantly different (P<0.01). In the DRSP group, pain, depression and insomnia were improved after treatment (P<0.05), other symptoms were (P<0.05), other symptoms were not improved;the total effective rate of combined group was 90.0%, DRSP (P<0.05), and the total effective rate of the group was significantly higher than that of the control group (P<0.05) .There was no significant difference The total effective rate was 73.33% in the group and 70.0% in the 17β group. There was significant difference between the combined group and the other groups (P<0.05). The difference of TC, E2 and FSH in the combined group before and after treatment (P<0.05). There was no difference in FSH between the two groups before and after treatment. There was no difference between the two groups before and after treatment. Conclusion 17β-estradiol combined with DRSP in the treatment of menopausal syndrome patients achieved good clinical effect, the serum levels of FSH and E2 in the body is significant.
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Objective To compare the ameliorating effect of collagen peptide chelated calcium (CPCC) and estrogen on the bone quality in ovariectomized rats in order to serve the development of safe drugs for prevention of osteoporosis (OP).Methods Bilateral ovariectomized rats were divided into ovariectomized group (OVX),sham group,17β-estradiol injection group (OVX+E2) and CPCC gavage group (OVX+CCCP).Bone and serum indices of these groups were assessed and compared at 9 weeks after treatment.Results Bone density of the OVX group was significantly lower than the sham group (P0.05),while the body weight gain of the E2 group at weeks 8 and 9 was significantly lower than those of the sham group (P<0.01).As regarding the prevention of bone loss,the Mg and Ca levels of the E2 group were significantly lower than those of the moderate and high dose CPCC groups.The Cu level was not significantly different compared with the sham group,while those in the moderate and low dose CPCC groups were significantly higher than the sham group.The Mn,Zn and hydroxyproline levels of the E2 group were significantly lower than those of the sham group,while the CPCC group maintained levels similar to that of the sham group.In regarding to the inhibiting effect on the increased blood BGP and StrACP,the E2 group was still maintained at levels similar to that of the OVX group,while those of the CPCC group were significantly lower than the OVX group.As regarding the decreased blood Ca,the E2 group was not significantly different with that of the OVX group,while that of the CPCC group was significantly higher than the OVX group.Conclusions CPCC is more effective than estrogen in ameliorating the bone quality of ovariectomized rats.