Aromatase deficiency caused by mutation of CYP19A1 gene: A case report / 中南大学学报(医学版)

Aromatase deficiency (AD) is a rare autosomal recessive genetic disease caused by loss-of-function mutations in aromatase gene (CYP19A1), leading to congenital estrogen deficiency syndrome. Both mothers of AD patients during pregnancy and female AD fetus show virilization, while male patients are usually diagnosed in adulthood due to continued height increase and metabolic abnormalities. In 2019, a patient with AD was admitted in the Second Xiangya Hospital. The patient was a 37-year-old adult male who continued to grow linearly after adulthood. His estradiol was below the measurable line, the follicle-stimulating hormone (FSH) increased, bone age delayed, epiphysis unfused, and the bone mass reduced. CYP19A1 gene detection showed that c.1093C>T, p.R365W was homozygous mutation. This disease is rare in clinic. Clinicians need to raise awareness of the disease for early diagnosis and treatment to improve the long-term prognosis of patients.

Expression of sclerostin in alveolar bone remodeling of ovariectomized rats / 口腔疾病防治

Objective@# To investigate the expression and distribution of sclerostin in the alveolar bone of rat in the absence of estrogen, and to provide evidence for the analysis of the histological correlation between sclerostin and alveolar bone remodeling in rats. @*Methods @#The experimental subjects of this study were 32 8-week-old female Wistar rats. Among them, 16 rats were ovariectomized (OVX), and 16 rats were subjected to a sham operation (Sham). These rats were sacrificed 1, 2, 3, and 4 weeks after the operation, and the mandibles were removed and embedded. The mesial and distal sections of the rat,s mandibular first molars were selected and stained with anti-tartrate phosphatase (TRAP), sclerostin immunostaining, multiple immunostainings, RANKL and TRAP double staining, and silver-plated multiple staining. @*Results @#As the postoperative time in rats increased, the TRAP-positive osteoclasts counts in the OVX group in the interalveolar septum of mandibular first molar increased significantly, and statistical difference was noted between the groups (P ＜ 0.05). The OVX 2w, 3w, and 4w groups exhibited more TRAP-positive osteoclasts compared with the Sham group at the corresponding time point, and the results were statistically different (P ＜ 0.05). Sclerostin immunostaining revealed that the proportion of positive bone cells in the mesial side of the periodontal ligament area of mandibular first molar in the OVX group gradually decreased. Statistical differences were noted between the OVX 3w group and the OVX 4w group as well as the OVX 1w group and, the OVX 2w group (P ＜ 0.05). In the comparison between the area near the periodontal ligament and the central area of the alveolar bone septum of the mandibular first molar in the same group, the positive expression ratio of sclerostin in the OVX 3w and OVX 4w groups in the area near the periodontal ligament was reduced compared with that in the central area of the alveolar bone septum. The results were statistically significant (P ＜ 0.05). A larger number of osteoblasts was noted around the osteoclasts in the OVX 4w group compared with the Sham 4w group based on ALP/ TRAP /sclerostin multiple staining, whereas less sclerostin-positive osteoblasts were noted in the OVX 4w group. Sclerostin/TRAP/silver plating staining showed that the bone tubules around the sclerostin positive bone cells mostly exhibited a parallel and neat arrangement, and the bone tubules around sclerostin negative bone cells were more irregular and disorderly arranged in the OVX 4w group@*Conclusion@#Sclerostin protein is involved in alveolar bone remodeling in estrogen-deficient rats.

Lovastatin combined with insulin effects on fracture healing in rat models of bilateral ovariectomized type 2 diabetic mellitus / 中国组织工程研究

BACKGROUND: Osteoporotic fracture combined with type 2 diabetic mellitus in female patients is often accompanied by dyslipidemia and hyperglycemia. In addition to insulin treatment, statins are often prescribed for combination therapy, but the combined effect of these two drugs on fracture healing has not been reported in such patients. OBJECTIVE; To investigate the effect of lovastatin combined with insulin on the fracture healing of bilateral ovariectomized rats suffering from type 2 diabetic mellitus. METHODS: The study was approved by the Ethical Committee of North China University of Science and Technology. Thirty-two female Sprague-Dawley rats were divided into control, diabetic ovariectomized, insulin and combined groups. A model of type 2 diabetes and osteoporosis fracture was established in all rats except for the control group. At 7 days after the type 2 diabetic model was successfully established by injection of streptozotocin, the rats in the insulin and combined groups received the subcutaneous injection of insulin (2-4 U in the morning, and 4-6 U in the evening) until the end of the experiment. The rats in the combined group were given 20 mg/kg lovastatin via gavage daily, and those in the other two groups were not treated. All rats were sacrificed at 3 weeks after fracture. Radiographic, clinical and histomorphometric detections of the callus were performed. The expression levels of bone morphogenetic protein 2, vascular endothelial growth factor and collagen type II were detected. All above results were used to analyze the fracture healing in each group. RESULTS AND CONCLUSION: (1)The radiographic score, micro-CT index, histologic score and the expression levels of vascular endothelial growth factor and collagen type II in the diabetic ovariectomized group were significantly poorer than those in the control group (P < 0.05). (2) All above indexes in the insulin group were significantly improved compared with the diabetic ovariectomized group (P < 0.05), which promoted the fracture healing of model rats. (3) After combined with lovastatin, although the expression levels of vascular endothelial growth factor and bone morphogenetic protein 2 in the callus were significantly increased (P < 0.05), there was no significant improvement in the radiographic appearance and microstructure of the callus.

Effects of ovariectomy on the secretory apparatus in the right atrial cardiomyocytes of middle-aged mice

OBJECTIVE : The aim of the present study was to evaluate the effects of ovariectomy on the secretory apparatus of natriuretic peptides in right atrial cardiomyocytes. METHODS: Nine-month-old mice underwent bilateral ovariectomy or sham surgery. The blood exam of the ovariectomized mice showed results consistent with castrated females. Systolic blood pressure was measured after ovariectomy (9 mo of age) and at the moment of sacrifice (12 mo of age). Fragments of the right atrium were collected and prepared for electron microscopy examination. The following variables were quantified: the quantitative density and area of the natriuretic peptide granules, the relative volume of euchromatin in the nucleus, the number of pores per 10 μm of the nuclear membrane and the relative volumes of the mitochondria and Golgi complex. RESULTS: The cardiomyocytes obtained from ovariectomized mice indicated that the quantitative density and the area of secretory granules of natriuretic peptides were significantly lower compared with the sham-operated mice. Furthermore, there was a decrease in the relative volume of euchromatin, a lower density of nuclear pores, and lower relative volumes of the mitochondria and Golgi complex in the ovariectomized mice compared with the sham-operated mice. These findings suggest a pool with a low turnover rate, i.e., low synthesis and elimination of natriuretic peptides. CONCLUSION: A lack of estrogen caused hypotrophy of the secretory apparatus in right atrial cardiomyocytes that could explain the weak synthesis of natriuretic peptides in mice. Furthermore, one of the mechanisms of blood pressure control was lost, which may explain, in part, the elevated blood pressure in ovariectomized mice. .

Experimental study of correlation between T-cell paradigm and bone loss induced by estrogen deficiency / 中国免疫学杂志

Objective:To demonstrate the relationship between the Th 1/Th2/Th17/Treg paradigm and the bone loss induced by estrogen deficiency and looking for potential target for clinical treatment.Methods:30 BALB/c mice were divided randomly into the normal control group , the sham operation group , and the ovariectomy group.The serum estradiol ( E2 ) was assessed by ELISA.Bone mineral density (BMD) of thigh bone was measured with dual energy X ray absorptiometry.Meanwhile,the T-cell subsets (Th1:CD4+TNFα+, Th2: CD4+IL-4+, Th17: CD4+IL-17 A+, Treg: CD4+CD25+Foxp3+) in spleen lymphocytes were analyzed by flow cytometry.Results:Compared with the normal group and the sham operation group , both E2 and BMD in the ovariectomy group decreased significantly ( P<0.05 ).The percentage of Th 1 and Th17 subset increased while the percentage of Th 2 and Treg decreased significantly in ovariectomy mice compared with sham operation mice.Correlation analysis showed that BMD was positively related to E 2 level and the percentage of Th 2 and Treg subset;however ,BMD was negatively related to the percentage of Th 1 and Th17 subset ( P<0.05 ).Conclusion: Conclusion: T-cell paradigm was involved in the bone loss induced by estrogen deficiency.Modifying T-cell paradigm may become a potential target for reducing bone loss induced by estrogen deficiency .

Tocotrienol supplementation in postmenopausal osteoporosis: evidence from a laboratory study

OBJECTIVE: Accelerated bone loss that occurs in postmenopausal women has been linked to oxidative stress and increased free radicals. We propose the use of antioxidants to prevent and reverse postmenopausal osteoporosis. This study aimed to examine the effects of tocotrienol, a vitamin E analog, on bone loss due to estrogen deficiency. Our previous study showed that tocotrienol increased the trabecular bone volume and trabecular number in ovariectomized rats. In the current study, we investigated the effects of tocotrienol supplementation on various biochemical parameters in a postmenopausal osteoporosis rat model. MATERIALS AND METHODS: A total of 32 female Wistar rats were randomly divided into four groups. The baseline group was sacrificed at the start of the study, and another group was sham operated. The remaining rats were ovariectomized and either given olive oil as a vehicle or treated with tocotrienol at a dose of 60 mg/kg body weight. After four weeks of treatment, blood was withdrawn for the measurement of interleukin-1 (IL1) and interleukin-6 (IL6) (bone resorbing cytokines), serum osteocalcin (a bone formation marker) and pyridinoline (a bone resorption marker). RESULTS: Tocotrienol supplementation in ovariectomized rats significantly reduced the levels of osteocalcin, IL1 and IL6. However, it did not alter the serum pyridinoline level. CONCLUSION: Tocotrienol prevented osteoporotic bone loss by reducing the high bone turnover rate associated with estrogen deficiency. Therefore, tocotrienol has the potential to be used as an anti-osteoporotic agent in postmenopausal women. .

Effect of Trifolium pratense isoflavones on osteoporosis rats induced by estrogen deficiency / 中国药学杂志

OBJECTIVE: To examine the effects of Trifolium pratense isoflavones (TPIF), the main active component of Trifolium pratense L., on ovariectomy-induced osteoporosis in rats. METHODS: A total of 60 six-month-old female rats were randomly assigned to a sham-operated group and five ovariectomized(OVX) groups, ie, OVX with vehicle(10 mL · kg-1 · d-1, ig), OVX with graded doses of TPIF(30, 60, and 90 mg · kg-1 · d-1, ig), and OVX with nilestriol(2.5 mg · kg-1 · week-1, ig). The animals were sacrificed after 12-week treatment. RESULTS: Compared to the OVX group, TPIF significantly inhibited the increase of body weight. In addition, TPIF administration significantly decreased serum levels of alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP), and increased serum level of estrogen. CONCLUSION: TPIF may be a potential alternative medicine for the prevention and treatment of postmenopausal osteoporosis. Its mechanism may be related to increasing estrogen level, inhibiting bone resorption, and promoting bone formation.

Fresh soy oil protects against vascular changes in an estrogen-deficient rat model: an electron microscopy study

OBJECTIVE: To observe the effects of consuming repeatedly heated soy oil on the aortic tissues of estrogen-deficient rats. METHODS: Thirty female Sprague Dawley rats (200- 250 g) were divided equally into five groups. One group served as the normal control (NC) group. The four treated groups were ovariectomized and were fed as follows: 2 percent cholesterol diet (OVXC); 2 percent cholesterol diet + fresh soy oil (FSO); 2 percent cholesterol diet + once-heated soy oil (1HSO); and 2 percent cholesterol diet + five-times-heated soy oil (5HSO). After four months, the rats were sacrificed, and the aortic tissues were obtained for histological studies. RESULTS: After four months of feeding, the NC, FSO and 1HSO groups had a lower body weight gain compared to the OVXC and 5HSO groups. The tunica intima/media ratio in the 5HSO group was significantly thicker (p < 0.05) compared to the NC, OVXC and FSO groups. Electron microscopy showed that endothelial cells were normally shaped in the FSO and NC groups but irregular in the 1HSO and 5HSO groups. A greater number of collagen fibers and vacuoles were observed in the 5HSO group compared to the other treatment groups. CONCLUSIONS: Fresh soy oil offered protection in the estrogen-deficient state, as these rats had similar features to those of the NC group. The damage to the tunica intima and the increase in the ratio of tunica intima/media thickness showed the deleterious effect of consuming repeatedly heated soy oil in castrated female rats.

The effect of estrogen deficiency on rat pulpodentinal complex / 대한치과보존학회지

The purpose of this study was to investigate the effects of estrogen deficiency on pulpodentinal complex of tooth in ovariectomized rats. Thirty female Sprague-Dawley rats, 10 weeks old, were used. Rats were grouped into two groups. One group (n = 15) was subjected to sham surgery (SHAM) and the other group (n = 15) was ovariectomized bilaterally (OVX). Animals were sacrificed 12 weeks later, and their mandibular molars and associated periodontal supporting tissues were dissected out, and fixed in 10% buffered formalin. For comparison of groups, immunostained for osteonectin. Histomorphometrical measurement of change of teeth was performed using an image analysis system and paired t-test was used and the level of significance for overall differences was set at p < 0.05. In immunostaining of osteonectin, they were significantly different from each other. The predentin thickness in OVX rats was wider than in SHAM rats. And in SHAM rats, osteonectin was more specifically stained in predentin areas than in OVX rats. These results indicate that estrogen deficiency increased the unmineralized predentin areas and decreased osteonectin content in pulpal tissues in rats. If our result is applicable to human studies, odotoblast is affected by estrogen deficiency.

Crucial Role of Tissue-specific Apoptosis on the Development of Primary Sjögren's Syndrome / Oral Science International

Primary Sjögren's syndrome is an autoimmune disorder characterized by lymphocytic infiltrates and destruction of the salivary and lacrimal glands, and systemic production of autoantibodies to the ribonucleoprotein (RNP) particles SS-A/Ro and SS-B/La, leading to clinical symptoms of dryness of the mouth and eyes (sicca syndrome). Autoreactive T cells bearing the CD4 molecule may recognize an unknown self antigen, triggering autoimmunity in the salivary and lacrimal glands. Although several candidate autoantigens including α-fodrin have been reported in Sjögren's syndrome, the pathogenic roles of the autoantigens in initiation and progression of SS are still unclear. It is possible that individual T cells activated by an appropriate self antigen can proliferate and form a restricted clone. Recent evidence suggests that the apoptotic pathway plays a central role in making T cells tolerant to tissue-specific self antigen, and may drive the autoimmune phenomenon. We recently reported that tissue-specific apoptosis in estrogen-deficient mice may contribute to autoantigen cleavage, leading to the development of autoimmune exocrinopathy. The studies reviewed imply that tissue-specific apoptosis and caspase-mediated α-fodrin proteolysis are involved in the progression of autoimmune lesions in Sjögren's syndrome. Moreover, Fas ligand (FasL) and its receptor Fas are essential in the homeostasis of the peripheral immune system. It is considered that a defect in activation-induced cell death (AICD) of effector T cells may result in the development of autoimmune exocrinopathy in Sjögren's syndrome.

Menopause and hormone replacement therapy.

The global population in the 21st century has reached 6.2 billion people, by the year 2025 it is to be around 8.3-8.5 billion, and will increase further. Elderly people are expected to grow rapidly than other groups. The fastest increase in the elderly population will take place in Asia. Life expectancy is increasing steadily throughout developed and developing countries. For many menopausal women, increased life expectancy will accompanied by many health problems. The consequences of estrogen deficiency are the menopausal symptoms. The treatment of menopause related complaints and diseases became an important socioeconomic and medical issue. Long term symptoms, such as the increase in osteoporosis fractures, cardio and cerebrovascular disesses and dementia, created a large financial burden on individuals and society. All these health problems can be lreated or prevented by hormone replacement therapy (HRT). Natural HRT is usually prefened. Synthetic estrogen in oral contraceptives (oc) are not recommended for HRT. Many contra-indications for oc, but now it is widely usedfor HRT. The main reasons for discontinuing HRT are unwanted bleeding, fear of cancer, and negative side effects. Until now there are sill debates about the rebrtonship between HRT and the incidence of breast cancer. Many data showed that there were no clear relationship between the use of HRT and breast cancer. ThereÎore, nwny experts advocate the use of HRTfrom the first sign of climacteric complaints until death.

Relaxation Response of the Rabbit Clitoral Corpus Cavernosal Smooth Muscle in Chronic Estrogen-Deficiency State / 대한비뇨기과학회지

PURPOSE: To investigate involvement of chronic estrogen deficiency in impairment of relaxation response of the clitoral cavernous smooth muscle. MATERIALS AND METHODS: New Zealand white female rabbits were randomly divided control (sham operation, n=6) and experimental groups; 1) bilateral oophorectomy (n=8), 2) oophorectomy with estradiol replacement (n=7), 3) oophorectomy with flutamide treatment (n=6). All the rabbits were sacrificed 12 weeks after the operations. Blood levels of estradiol and lipid fractions were measured just before the operations and sacrifice. Clitoral cavernous strips were obtained to observe relaxation responses to endothelium-dependent and -independent vasodilators, and electrical field stimulation (ESF). RESULTS: The uterus weight and serum estradiol level decreased significantly (p<0.01) in the oophorectomy group compared with the baseline data. The serum levels of total cholesterol, triglyceride, and low density lipoprotein increased significantly (p<0.01) in the oophorectomy group. The relaxations of the clitoral strips to acetylcholine were significantly attenuated in the oophorectomy group compared with those of the control and estrogen replacement groups. However, sodium nitroprusside- and nonadrenergic noncholinergic-induced relaxations were not significantly different among the 4 groups. No significant histologic changes were noted in the clitoral tissues of the oophorectomy group. CONCLUSIONS: Chronic estrogen deficiency in the rabbits may cause impairment of endothelium-dependent relaxation of clitoral corpus cavernosal smooth muscles. Further studies are needed to determine whether the hypoesterogenemia itself and/or secondary hypercholesterolemia due to the chronic estrogen deficiency may result in the impairment.