OBJECTIVE: To investigate the effect of human adipose-derived mesenchymal stem cells (hASCs) exosomes on osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) extracted from osteoporotic mice, and to evaluate the effect of hASCs exosomes on preventing bone loss induced by estrogen deficiency. METHODS: hASCs exosomes were extracted by ultracentrifugation. The osteoporotic mice were established by bilateral ovariectomy (OVX). BMSCs were isolated from osteo-porotic mice and cultured for further analysis. In the experimental group, these BMSCs were exposed to an osteogenic induction medium supplemented with hASCs exosomes to evaluate their potential effects on osteogenesis. In contrast, the control group was treated with the same osteogenic induction medium, but without the addition of hASCs exosomes, to serve as a baseline comparison for the study. To comprehensively assess the osteogenic differentiation of BMSCs influenced by hASCs exosomes, alkaline phosphatase (ALP) staining, ALP activity quantitative analysis and quantitative reverse transcription polymerase chain reaction (qPCR) were performed. These evaluations provided critical insights into the role of hASCs exosomes in promoting osteoblast differentiation and bone formation in osteoporotic conditions. The fluorescence labeled hASCs exosomes were injected via the tail vein to observe the biodistribution of exosomes. Two weeks after OVX, the mice were divided into three groups: The experimental group consisted of estrogen-deficient mice receiving hASCs exosome injections; the negative control group consisted of estrogen-deficient mice receiving phosphate-buffered saline (PBS) injections; and the positive control group consisted of mice that underwent Sham surgery and received PBS injections.The injections were administered once every 3 days, for a total of 8 injections. Afterward, the femurs were collected from the mice, and micro-computed tomography (micro-CT) was performed to measure bone mineral density and conduct bone morphometric analysis. RESULTS: hASCs exosomes were successfully extracted using ultracentrifugation. After the induction by hASCs exosomes, ALP staining and ALP activity in the BMSCs extracted from osteoporotic mice were significantly enhanced, the expression of osteogenesis related genes in BMSCs were significantly up-regulated. More trabecular bone and higher bone mineral density were observed in estrogen-deficient mice injected with hASCs exosomes compared with estrogen-deficient mice injected with PBS, and there was no significant decrease in bone mineral density compared with the Sham operation group. CONCLUSION hASCs exosomes promoted the osteogenic differentiation of BMSCs extracted from osteoporotic mice. hASCs exosomes prevented bone loss induced by estrogen deficiency.
Animals ; Mesenchymal Stem Cells/cytology* ; Exosomes ; Estrogens/deficiency* ; Humans ; Osteogenesis ; Cell Differentiation ; Female ; Mice ; Osteoporosis/prevention & control* ; Ovariectomy ; Adipose Tissue/cytology* ; Cells, Cultured

Animals ; Brain Stem ; drug effects ; Cochlea ; drug effects ; Cochlear Nerve ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Estrogens ; deficiency ; Female ; Ovariectomy ; Rats ; Signal-To-Noise Ratio

OBJECTIVETo observe the effect of Modified Zuoguiwan (MZ) on the balance between helper T cell subsets 17 (Th17) and regulatory T cell subsets (Treg) in estrogen deficiency induced bone loss mice and to explore its mechanism.

METHODSTotally 50 BALB/c mice were divided into the sham-operation group, the ovariectomy model group, the low dose MZ group, the middle dose MZ group, and the high dose MZ group by random digit table, 10 in each group. Mice in the low, middle, and high dose MZ groups were respectively administered with MZ at the daily dose of 7.25, 14.50, and 29.00 g/kg by gastrogavage, 0.5 mL each time for 12 successive weeks. Meanwhile, mice in the sham-operation group and the ovariectomy model group were administered with equal volume by gastrogavage, 0.50 mL each time. The serum estradiol (E2) level was assessed by enzyme linked immunosorbent assay (ELISA). Bone mineral density (BMD) of thigh bone was measured with dual energy X ray absorptiometry. In addition, the population of Th17/Treg subsets in spleen mononuclear cells was analyzed by extracellular and intracellular staining method using flow cytometry. Moreover, the mRNA expression of IL-17A and TGF-β in the spleen mononuclear cells was detected by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSCompared with the sham-operation group, both E2 and BMD significantly decreased, the percentage of Th17 subset and Th17/Treg ratio both increased, the percentage of Treg subset obviously decreased, the expression of IL-17A mRNA significantly increased, and the expression of TGF-β mRNA significantly decreased in the ovariectomy model group (all P < 0.05). Compared with the model group, BMD obviously increased, the percentage of Th17 subset and Th17/Treg ratio both decreased, the percentage of Treg subset obviously increased, the expression of IL-17A mRNA significantly decreased, and the expression of TGF-β mRNA significantly increased in the middle dose MZ group and the high dose MZ group (all P < 0. 05). Correlation analyses showed that BMD was positively related to both the serum E2 level and the percentage of Treg subset (P < 0.05), but negatively related to the percentage of Th17 subset (P < 0.05). In addition, the serum E2 level was positively related to the percentage of Treg subset, but obviously negatively related to that of Th17 subset (P < 0.05).

CONCLUSIONSThere was correlation between Th17/Treg imbalance and E2 deficient bone loss. MZ could decrease the proportion of Th17 subset, but elevate the proportion of Treg subset in E2 deficient bone loss mice. It could achieve therapeutic effect through adjusting the balance of Th17/Treg in E2 deficient bone loss mice.

Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Estrogens ; deficiency ; metabolism ; Female ; Flow Cytometry ; Humans ; Interleukin-17 ; Mice ; Mice, Inbred BALB C ; Osteoporosis, Postmenopausal ; drug therapy ; RNA, Messenger ; Spleen ; T-Lymphocyte Subsets ; T-Lymphocytes, Helper-Inducer ; T-Lymphocytes, Regulatory ; Th17 Cells ; Transforming Growth Factor beta ; metabolism

Estrogen is implicated as playing an important role in aging and tumorigenesis of estrogen responsive tissues; however the mechanisms underlying the mitogenic actions of estrogen are not fully understood. Here we report that estrogen deficiency in mice caused by targeted disruption of the aromatase gene results in a significant inhibition of telomerase maintenance of telomeres in mouse ovaries in a tissue-specific manner. The inhibition entails a significant shortening of telomeres and compromised proliferation in the follicular granulosa cell compartment of ovary. Gene expression analysis showed decreased levels of proto-oncogene c-Myc and the telomerase catalytic subunit, telomerase reverse transcriptase (TERT), in response to estrogen deficiency. Estrogen replacement therapy led to increases in TERT gene expression, telomerase activity, telomere length and ovarian tissue growth, thereby reinstating ovary development to normal in four weeks. Our data demonstrate for the first time that telomere maintenance is the primary mechanism mediating the mitogenic effect of estrogen on ovarian granulosa cell proliferation by upregulating the genes of c-Myc and TERT in vivo. Estrogen deficiency or over-activity may cause ovarian tissue aging or tumorigenesis, respectively, through estrogen regulation of telomere remodeling.
46, XX Disorders of Sex Development ; drug therapy ; genetics ; metabolism ; Aging ; genetics ; metabolism ; Animals ; Aromatase ; deficiency ; genetics ; metabolism ; Cell Proliferation ; drug effects ; Estrogen Replacement Therapy ; Estrogens ; deficiency ; pharmacology ; Female ; Gene Expression ; Genes, myc ; genetics ; Granulosa Cells ; drug effects ; metabolism ; pathology ; Gynecomastia ; drug therapy ; genetics ; metabolism ; Humans ; Infertility, Male ; drug therapy ; genetics ; metabolism ; Metabolism, Inborn Errors ; drug therapy ; genetics ; metabolism ; Mice ; Mice, Knockout ; Telomerase ; genetics ; metabolism ; Telomere ; chemistry ; metabolism ; pathology

INTRODUCTIONCardiovascular disease is the leading cause of death and morbidity among postmenopausal women, and oestrogen deficiency may be an important factor in its development. The role of oestrogen replacement in preventing cardiovascular disease is controversial. The aim of this descriptive review is to analyse the available data and to recommend evidence-based practice guidelines pertaining to hormone therapy in the context of cardiovascular and cerebrovascular health.

MATERIALS AND METHODSRelevant clinical trials were identified by computerised literature search. The collated data were presented to fellow gynaecologists for review, analysis of results and discussion in a series of meetings dedicated to finding the best evidence in menopause management. The evidence was used to formulate clinical practice guidelines for the management of women with significant cardiovascular risk factors.

RESULTSEvidence from animal studies and observational trials supported a cardio-protective effect of postmenopausal hormone therapy. More recent randomised clinical trial data have shown no significant reduction of coronary heart disease, and have confirmed a higher incidence of stroke and venous thromboembolism.

CONCLUSIONSThe evidence is widely divergent regarding postmenopausal hormone therapy and cardiovascular risk. More consistent data are available reporting an increased risk in the incidence of venous thromboembolism and stroke. It is important to be clear about the indications of hormone use and to utilise alternative modalities to promote cardiovascular health in the postmenopausal population.

Aged ; Cardiovascular Diseases ; prevention & control ; Cerebrovascular Disorders ; prevention & control ; Estrogens ; deficiency ; metabolism ; Female ; Hormone Replacement Therapy ; Humans ; Menopause ; Middle Aged ; Practice Guidelines as Topic

OBJECTIVETo explore the effect of Yijing Huoxue Cuyun Decoction (YHCD) in adjusting hypoestrogenemic response induced by clomiphene.

METHODSInfertile patients caused by ovulation disturbance were randomly divided into 2 groups. The 60 patients in the observed group were treated with clomiphine plus YHCD, and the 58 patients in the control group were given clomiphine plus estradiol valerate.

RESULTSBy scoring on the cervical relaxation and improvement of cervical mucus, 38 patients (63.3%) in the observed group had Insler score of more than 8 points, while that in the control group was only 25 (43.1%), comparison between the two groups showed significant difference (P < 0.05). The endometrium thickening in the observed group was 0.98 +/- 0.19 cm, significantly different to that in the control group (0.85 +/- 0.21 cm, P < 0.01). Twenty-five patients in the observed group (41.7%), and fourteen patients in the control group (24.1%), respectively got pregnancy, the pregnant rate in the former was obviously higher than that in the latter (P < 0.05).

CONCLUSIONYHCD can ameliorate hypoestrinemia induced by clomiphene and increase the pregnant rate in patients.

Adult ; Clomiphene ; adverse effects ; therapeutic use ; Drugs, Chinese Herbal ; therapeutic use ; Estrogens ; blood ; deficiency ; Female ; Humans ; Infertility, Female ; drug therapy ; Ovulation ; drug effects ; Ovulation Induction ; Phytotherapy

OBJECTIVETo explore the effects of environmental estrogens (n-4-noniphenol, NP; bisphenol, BisA; and dibutylphthalate, DBP) on apoptosis induced by estrogen depletion in breast cancer T47D cells.

METHODSHuman T47D breast cancer cells were grown in DMEM medium containing 10% bovine serum. Four days before adding the test compounds, the cells were washed in phosphate-buffered saline, and the medium was substituted with a phenol red-free DMEM medium containing 5% dextral charcoal-stripped FBS. Respective test compound was added in fresh medium and the control cell received only the vehicle (ethanol). Apoptotic features in T47D cell were analyzed by light microscope that was commonly used to define apoptosis. DNA integrity of T47D cells was examined by agarose gel electrophoresis. Hypodiploid population was detected by flow cytometry.

RESULTSThe typical characters of apoptosis in T47D cells were observed after estrogen deletion and then disappeared following exposure to T47D cells at 32 x 10(-7) mol/L Np and 32 x 10(-7) mol/L BisA respectively. Inhibition of apoptosis at 32 x 10(-6) mol/L DBP was not shown in our study.

CONCLUSIONN-4-noniphenol and Bisphenol A could inhibit apoptosis induced by estrogen deletion in breast cancer T47D cells. This result suggests that these environmental estrogens might involve in signal transduction connected with apoptosis.

Apoptosis ; drug effects ; Benzhydryl Compounds ; Cell Line, Tumor ; drug effects ; metabolism ; Dibutyl Phthalate ; pharmacology ; Estrogens ; deficiency ; Estrogens, Non-Steroidal ; pharmacology ; Female ; Flow Cytometry ; Humans ; Phenols ; pharmacology

OBJECTIVES: There are many interesting reports suggesting that magnesium(Mg) deficiency is deleterious in patients with congestive heart failure (CHF). It is paradoxical that the most important cause of Mg deficiency in these persons is maybe use of therapeutics including diuretics. Authors investigated the trend of serum and 24 hour urine Mg with other relating electrolytes in Mg homeostasis prospectively, in the management of CHF. And we assessd the effects of medications and many variables in .CHF on serum Mg, and the usefulness of serum Mg representing the body content. METHODS: Fifty three patients who were diagnosed as CHF by clinical finding and echocardiogaphy were prescribed conventional doses of diuretics as furosemide 40mg and spironolactone 50mg daily, with or without angiotensin converting enzyme(ACE) inhibitor and digitalis. And then, serial serum and 24 hour urine Mg, sodium, potassium and calcium were obtained at admission, 2nd day, 5th day, and discharge. RESULTS: The patients group with chronic CHF, which was defined as long-term use of diuretics over 6 months, showed higher prevalence of low level of serum Mg concentration than the group with acute one(11 of 28, 39% vs. 2 of 25. 8%, P< 0.01). Of those two groups, the latter showed upward trend of serum Mg from admission to discharge, but the former showed no change. In 24 hour urine Mg excretion, the amount of the patients with CHF was larger than that of control group. In the chronic CHF group, the effect of digitalis on decreasing serum Mg was evident. Serum Mg of acute CHF group correlated with serum BUN(r=0.5609). Whereas, that of chronic group with ejection fraction(r=-0.4742) and plasma renin activity(r=-0.3791), with serum potassium(r=0.4673) and creatinine(0.5846). Serum Mg may be useful indicator of Mg homeostasis, especially in chronic CHF patients. CONCLUSION: Because patients with chronic CHF were prone to deficiency of Mg in the management, maintaining the adequate serum Mg through long- term replacement seems very important in decreasing the morbidity and mortality of these persons.
Angiotensins ; Calcium ; Digitalis ; Diuretics ; Electrolytes ; Estrogens, Conjugated (USP)* ; Furosemide ; Heart Failure* ; Homeostasis ; Humans ; Magnesium Deficiency ; Magnesium* ; Mortality ; Plasma ; Potassium ; Prevalence ; Prospective Studies ; Renin ; Sodium ; Spironolactone