Ovariectomy Group Research Articles

The Differences Between the Effect of Oral and Intraperitoneal Induction of Aluminum Chloride (AlCl3) on the Memory Function of White Rats (Rattus norvegicus strain wistar) Menopause Model

The decline in estrogen at menopause leads to a loss of neuroprotective function. One of the effects is Alzheimer’s disease (AD), with clinical manifestations of dementia, decreased memory function, language, thinking, and learning. The pathology of AD is widely studied but not completely understood. Various animal models have been used but have not been able to represent the pathology of AD fully. This study aimed to determine the effect of oral and intraperitoneal induction of aluminium chloride (AlCl3) on the memory function of menopausal model white rats (Rattus norvegicus strain wistar). This study used 16 female rats, divided into four groups, namely the negative control group (K1), the ovariectomy group (OVX, K2), and the OVX + AlCl.6H20 group 100 mg/kg BW orally (K3), and the OVX + AlCl3.6H20 group. 70 mg/kg body weight intraperitoneally. AlCl3.6H20 induction was given for six weeks. In the last five days, the Morris water maze test was carried out. From the MANOVA test, the p-value of F values in Pillai’s trace, Wilks‘ Lambda, Hotelling’a Trace, and Roy’s Largest Root were < 0.05. Based on the post hoc Bonferroni test on the fifth day of the trial, there was a significant difference between the standard and OVX groups with the AlCl3 induction group, and there was no significant difference between the oral and intraperitoneal induction groups. Conclusion: There is no significant effect of oral and intraperitoneal induction of Aluminum Chloride (AlCl3) on the memory function of white rats (Rattus norvegicus strain wistar) menopausal model.
 Keywords: menopause, Alzheimer’s disease, memory test

MicroRNA-29b-3p/sirtuin-1/peroxisome proliferator-activated receptor γ suppress osteogenic differentiation.

Osteoporosis is described as an age-associated impairment of bone formation. microRNA (miR)-29b-3p was thought to be linked to osteoblast differentiation; however, the underlying molecular pathways are yet unknown. The study's goal was to look into the involvement of miR-29b-3p in osteoporosis and the pathophysiological mechanisms. A murine model of estrogen deficiency-induced bone loss was established to simulate postmenopausal osteoporosis. Reverse transcription-quantitative PCR (RT-qPCR) was performed to assess the level of miR-29b-3p of bone tissue. Additionally, miR-29b-3p/sirtuin-1 (SIRT1)/peroxisome proliferator-activated receptor γ (PPARγ) axis in the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) was examined. Osteogenesis-related markers, including alkaline phosphatase (ALP), osteocalcin (OCN), and runt-related transcription factor 2 (RUNX2), were assessed at protein and molecular levels. ALP staining and Alizarin Red staining were used to detect ALP activity and calcium deposition. The ovariectomy group was shown to express miR-29b-3p at higher levels in vitro, and miR-29b-3p mimics suppressed osteogenic differentiation and protein/mRNA expression levels of osteogenesis-related markers in vivo. SIRT1 was identified as a target of miR-29b-3p using luciferase reporter assays. Overexpression of SIRT1 reduced the inhibition of osteogenic differentiation by miR-29b-3p. Rosiglitazone, an activator of PPARγ signaling, was able to reverse the downregulation of the osteogenic differentiation of BMSCs and the protein expression of PPARγ caused by miR-29b-3p inhibitors. The results revealed that osteogenesis was suppressed by miR-29b-3p, which blocks the SIRT1/PPARγ axis. These results suggested that postmenopausal osteoporosis could be treated by targeting miR-29b-3p SIRT1/PPARγ.

Effects of retinoid-loaded hyaluronic acid nanomicelles on vaginal epithelium in a murine menopause model.

This study aimed to develop hyaluronic acid (HA)-based, retinoic acid (RA)-containing nanomicelles and to investigate the effects of these newly developed nanomicelles on regeneration of the vaginal epithelium and aquaporin 3 (AQP3) expression in a murine menopause model. The HA-based, RA-loaded nanomicelles were developed, and the RA-loading rate, encapsulation efficiency, and hydrodynamic diameter were measured. Female BALB/c mice (8 weeks; n=30) were divided into control and experimental groups. Menopause was established in the experimental group by removing both ovaries. The experimental group was further divided into an ovariectomy group, an HA-C18 vehicle group, and an HA-C18-RA group (2.5 µg per mouse); vaginal administration of HA-C18 or HA-C18-RA was performed once daily. After 4 weeks of treatment, murine vaginal tissue was removed, and histological analysis was performed. Three drug-loaded nanomicelles were synthesized: the RA content in HA-C18-RA-10, HA-C18-RA-20, and HA-C18-RA-30 was 3.13%, 2.52%, and 16.67%, respectively, and the RA encapsulation efficiency was 95.57%, 83.92%, and 93.24%, respectively. In the experimental versus control group, serum estrogen levels were significantly reduced, and the vaginal mucosal epithelial layer was significantly thinner. After 4 weeks of treatment, the thickness of the vaginal mucosal epithelial layer and AQP3 expression was increased in the HA-C18-RA group compared with the HA-C18 vehicle group. The newly developed HA-based nanomicelles containing RA resulted in vaginal epithelial recovery and increased AQP3 expression. The results may contribute to the development of functional vaginal lubricants or moisturizers for the treatment of vaginal dryness.

Effects of whole-body vibration at different periods on lumbar vertebrae in female rats.

The current study aimed to investigate the effects of whole-body vibration (WBV) before and after ovariectomy on lumbar vertebrae, and to observe whether the positive effects of WBV before and after ovariectomy on lumbar vertebrae in rats could be maintained after vibration stopped. Three-month-old female rats were divided into four groups (n=45/group): control (CON), ovariectomy (OVA), WBV before ovariectomy (WBV-BO), and WBV after ovariectomy (WBV-AO) groups. For 1-8 weeks, WBV-BO group was subjected to vertical WBV. At the 9th week, the rats in WBV-BO, WBV-AO, and OVA groups were ovariectomized. During 11-18 weeks, WBV-AO group was subjected to vibration. For 19-26 weeks, no intervention was done for rats. The lumbar vertebrae were examined by Micro-CT, compressive test, creep test, and microindentation test. At the 8th week, the displacement of the L1-L2 annulus fibrosus in WBV-BO group was 18% smaller compared with CON group (p<0.05). At the 18th week, the elastic modulus of the L5 vertebral body in WBV-BO and WBV-AO groups was 53% and 57% higher than that in CON group, respectively (p<0.05); the displacement of the L1-L2 annulus fibrosus in WBV-BO group was 25% smaller than those in the other groups (p<0.05). At the 26th week, there was no significant difference in the displacement of the L1-L2 annulus fibrosus between WBV-BO group and other groups (p>0.05); the elastic modulus of the L5 vertebral body had no significant difference between WBV-AO group and CON group (p>0.05). Our results demonstrated that WBV before ovariectomy effectively prevented disc degeneration with significant effects up to 8 weeks after ovariectomy. The vertebral mechanical properties could be significantly improved by WBV after ovariectomy, but the residual effect did not maintain after WBV stopped.

4D label-free quantitative proteomics analysis to screen potential drug targets of Jiangu Granules treatment for postmenopausal osteoporotic rats.

Background: Postmenopausal osteoporosis (PMOP) is a disease with a high prevalence in postmenopausal women and is characterized by an imbalance in bone metabolism, reduced bone mass, and increased risk of fracture due to estrogen deficiency. Jiangu granules (JG) is a compound prescription used in traditional Chinese medicine to treat PMOP. However, its definitive mechanism in PMOP is unclear. This study used a 4D label-free quantitative proteomics method to explore the potential therapeutic mechanism of JG in an ovariectomy (OVX) rats' model. Materials and methods: A rat model of PMOP was established by removing the ovaries bilaterally. Nine 3-month-old specific-pathogen-free female SD rats. The nine rats were randomly divided into 3 groups (n = 3 in each group): the sham-operated group (J), the ovariectomy group (NC), and the JG treatment (ZY) group. Proteins extracted from the bone tissue of the lumbar spine (L3, L4) of three groups of rats were analyzed by 4D label-free quantitative proteomics, and proteins differentially expressed after JG treatment and proteins differentially expressed after de-ovulation were intersected to identify proteins associated with the mechanism of PMOP by JG treatment. Result: There were 104 up-regulated and 153 down-regulated differentially expressed proteins (DEPs) in the J group vs. NC group, 107 up-regulated and 113 down-regulated DEPs in the J group vs. ZY group, and 15 up-regulated and 32 down-regulated DEPs in the NC group vs. ZY group. Six potential target proteins for JG regulation of osteoblast differentiation in OVX rats were identified by taking intersections of differential proteins in the J group vs. NC group and NC group vs. ZY group. Conclusion: JG may exert therapeutic effects by modulating the expression levels of target proteins associated with osteoblast differentiation to enhance osteoblast differentiation in OVX rats. These results further uncovered the target proteins and specific mechanisms of JG in treating PMOP, providing an experimental basis for the clinical application of JG in treating PMOP.

Aloe polysaccharide promotes osteogenesis potential of adipose-derived stromal cells via BMP-2/Smads and prevents ovariectomized-induced osteoporosis

BackgroundAloe polysaccharide (AP) is a type of an active macromolecule of Aloe vera, which contributes to its function. However, whether AP possesses anti-osteoporosis properties is unknown.MethodsAdipose-derived stromal cells were treated with different concentrations of AP. Early and late osteogenesis were, respectively, evaluated by ALP and Alizarin Red S staining. The effect of AP on the processes of adipogenesis inhibition in ADSCs was analyzed by oil red O staining. Western blot was used to assess the expression of osteogenic and adipogenic related factors. Then, Noggin was administered to further confirm the mechanism by which AP promotes the osteogenesis of ADSCs. Finally, 40 female SD rats were classified into a bilateral laparotomy group (Sham group) and three bilateral ovariectomy groups: OVX group, OVX + AP group, and OVX + AP + Noggin group. The bilateral rat femurs were collected to perform micro-CT scanning, HE, Masson trichrome, and Oil red O staining.ResultsThe results indicated that AP could increase ALP expression and calcium deposition. Through molecular mechanisms, AP promotes the protein expression of COL1A1, OPN, and ALP in ADSCs, but downregulates the expression of PPARγ. Also, AP directs ADSCs’ fate by stimulating the BMP2/Smads signaling pathway. In vivo, the rat AP-treated had more trabecular bone than the OVX rat, indicating partial protection from cancellous bone loss after treatment with AP.ConclusionOur results show that AP may promote osteogenesis of ADSCs through BMP-2/Smads signaling pathway and inhibits lipogenic differentiation. Thus, AP might be a promising alternative medicine to treat postmenopausal osteoporosis.

Effect of Lactobacillus plantarum on Osteoporosis in the Ovariectomized Rat

Osteoporosis is a growing global health concern primarily associated with decreased estrogen in postmenopausal women. Recently, some strains of probiotics were examined for potential anti-osteoporotic effects. This study intended to evaluate the impacts of Lactobacillus plantarum CJNU 3002 strain (LP) in ovariectomized rats. For this purpose, twelve weeks old female Wistar rats (n=21; 250–300 g) were divided into 3 groups; ovariectomy (OVX) group, OVX/LP group and Sham group (control). In these groups; two went through respective OVX and one had daily LP administration through oral gavage. Prior to 16 weeks after OVX, we collected blood samples and extracted the tibiae. We scanned the extracted tibiae by ex-vivo micro-computed tomography (micro-CT) and evaluated pathology by hematoxylin and eosin (H&amp;E) and Masson’s trichrome staining. The serum levels of C-telopeptide of type I collagen (CTX) and osteocalcin (OC) were examined. The OVX/LP group showed increases in bone mineral density, trabecular bone volume, trabecular number, and trabecular thickness (Tb.Th), and a decrease in trabecular spacing than the OVX group. However, OVX/LP group and control group were measurably comparable in Tb.Th. Micro-CT, H&amp;E, and Masson’s trichrome findings exhibited increased preservation and maintenance of trabecular bone structure in the OVX/LP group in comparison to the OVX group. In serum, the levels of CTX and OC were significantly different among the OVX and OVX/LP groups (p&lt;0.05). Taken together, L. plantarum CJNU 3002 could be helpful for the treatment of osteoporosis.

Global phosphoproteomic profiling of skeletal muscle in ovarian hormone-deficient mice.

Protein phosphorylation is important in skeletal muscle development, growth, regeneration, and contractile function. Alterations in the skeletal muscle phosphoproteome due to aging have been reported in males; however, studies in females are lacking. We have demonstrated that estrogen deficiency decreases muscle force, which correlates with decreased myosin regulatory light chain phosphorylation. Thus, we questioned whether the decline of estrogen in females that occurs with aging might alter the skeletal muscle phosphoproteome. C57BL/6J female mice (6 mo) were randomly assigned to a sham-operated (Sham) or ovariectomy (Ovx) group to investigate the effects of estrogen deficiency on skeletal muscle protein phosphorylation in a resting, noncontracting condition. After 16 wk of estrogen deficiency, the tibialis anterior muscle was dissected and prepped for label-free nano-liquid chromatography-tandem mass spectrometry phosphoproteomic analysis. We identified 4,780 phosphopeptides in tibialis anterior muscles of ovariectomized (Ovx) and Sham-operated (Sham) control mice. Further analysis revealed 647 differentially regulated phosphopeptides (Benjamini-Hochberg adjusted P value < 0.05 and 1.5-fold change ratio) that corresponded to 130 proteins with 22 proteins differentially phosphorylated (3 unique to Ovx, 2 unique to Sham, 6 upregulated, and 11 downregulated). Differentially phosphorylated proteins associated with the sarcomere, cytoplasm, and metabolic and calcium signaling pathways were identified. Our work provides the first global phosphoproteomic analysis in females and how estrogen deficiency impacts the skeletal muscle phosphoproteome.

Laparoscopic versus Open Ovariectomy in Bitches: Changes in Cardiorespiratory Values, Blood Parameters, and Sevoflurane Requirements Associated with the Surgical Technique.

Simple SummaryLaparoscopic neutering techniques are widely used in veterinary medicine due to their numerous advantages compared to open ones. However, laparoscopy requires the creation of pneumoperitoneum to establish a working space, and it has been related to the occurrence of pathophysiological alterations. The objective of this study was to evaluate the potential global systemic changes associated to the establishment of the pneumoperitoneum in laparoscopic ovariectomy and to compare them with those determined in conventional open surgery in dogs. Additionally, we evaluated the gas anesthetic requirements based on the surgical technique. The results of our study suggested that the cardiorespiratory alterations related to pneumoperitoneum seemed to be well-tolerated and reversible after CO2 deflation. Regarding the anesthetic gas evaluation, we determined higher requirements during ovarian resection in the open ovariectomy group. In conclusion, laparoscopic neutering procedures should be currently recommended by veterinarians due to their lower invasiveness and resulting complications. Nevertheless, a thorough understanding of the changes associated with pneumoperitoneum is essential to plan and obtain an adequate perioperative management.The aim of this study was to examine the cardiorespiratory and blood changes associated with pneumoperitoneum (PNP) in laparoscopic ovariectomy (LAP Ove), as well as sevoflurane requirements, comparing them to those determined in open surgery (LPT Ove). The study was performed in 16 bitches submitted to LAP or LPT Ove. The cardiorespiratory and end-tidal sevoflurane concentration values were recorded as follows: at the beginning of surgery (T1), after the right ovary resection (T2), after the left ovary resection (T3), and after surgical closure (T4). Blood samples were taken before and after PNP. Among the cardiorespiratory parameters, no differences were observed in the values of end-tidal CO2, minute volume, and heart rate. In the LAP Ove group, a significant increase in inspiratory pressures and a decreased compliance were identified at T2 and T3. Significant higher arterial pressure values were observed in both groups at T2 and T3, with this increase especially marked at T2 in the LPT Ove group. Sevoflurane requirements were significantly higher in the LPT group during ovarian resection. Finally, in terms of the hematochemical parameters, statistical differences were recorded between pre- and post-operative assessments, but not between both surgical groups. The pathophysiological effects associated with PNP seemed to be transient and well-tolerated by healthy dogs.

Influence of ovarian-sparing surgery and ovariectomy on prognosis in early cervical adenocarcinoma: a systematic review and meta-analysis.

The effect of ovarian-sparing surgery versus ovariectomy on prognosis in early cervical adenocarcinoma is controversial. The aim of this study was to compare the effect of ovary preservation versus ovariectomy on the prognosis of patients with cervical adenocarcinoma. A literature search was conducted of the PubMed, Excerpta Medica Database, Medline, Central, China National Knowledge Infrastructure databases, and China Science Periodical Database. The subjects of the literature study were patients with cervical adenocarcinoma. The literature compared the prognostic impact of ovary-sparing versus ovariectomy surgery. The Newcastle-Ottawa Scale was used to evaluate the quality of the articles. The Chi-square test was used to test the heterogeneity of the articles, and the random-effects model was used if the results indicated heterogeneity. A subgroup analysis and sensitivity analysis were used to examine the source of heterogeneity. If there was no heterogeneity among the articles, a fixed-effects model was used. Publication bias was evaluated using funnel plots and Egger test. A total of 3,467 patients with stage IA-IB cervical adenocarcinoma from 5 articles were included in the meta-analysis, of whom 995 had ovarian preservation and 1,895 had ovariectomy. There was no statistically significant difference in the 5-year overall survival (OS) between the stage IA-IIB cervical adenocarcinoma patients in the ovariectomy group and the ovarian preservation group (P=0.14). Additionally, there was no heterogeneity among these articles, and no publication bias (P>0.05). There was no significant difference in the 5-year progression free survival (PFS) between the stage IA-IIB cervical adenocarcinoma patients in the ovariectomy group and the ovarian preservation group (P=0.11). Additionally, there was no heterogeneity among these articles, and no publication bias (P>0.05). There was no significant difference in the 5-year disease specific survival (DSS) between the stage IA-IIB cervical adenocarcinoma patients in the ovariectomy group and the ovarian preservation group (P=0.48). Additionally, there was no heterogeneity among these articles, and no publication bias (P>0.05). There was no statistically significant difference in 5-year OS, PFS and DSS between ovarian-sparing surgery and oophorectomy for early-stage cervical adenocarcinoma. High-quality randomized controlled trials are still needed to verify this conclusion.

Evaluation of early bilateral ovariectomy in mice as a model of left heart disease.

Postmenopausal women tend to have worse cardiovascular outcomes in a manner that is associated with osteoporosis severity. In this study, we performed the first evaluation of the left ventricle and aortic valve phenotype of ovariectomized mice aged on Western diet to 1 yr. Disease was monitored in vivo using echocardiography and dual X-ray absorptiometry imaging and ex vivo using quantitative histological and immunostaining analysis. Mice had decreased bone mineral density in response to ovariectomy and increased fat mass in response to Western diet. Ovariectomized mice had a significantly increased left ventricle mass compared with control animals, absent of fibrosis. There was a slight increase in aortic valve peak velocity but no change in mean pressure gradient across the valve in the ovariectomy group. There was no evidence of leaflet hypertrophy, fibrosis, or calcification. This model of ovariectomy may present a novel method of studying left ventricle hypertrophy in female populations but does not have a phenotype for the study of aortic stenosis. This is particularly useful as it does not require genetic manipulation or drug treatment and more faithfully mimics aging, high-cholesterol diet, and postmenopausal osteoporosis that many female patients experience potentially resulting in a more translatable disease model.NEW & NOTEWORTHY This article uses in vivo and ex vivo analysis to track the development of osteoporosis and left heart cardiovascular disease in an aged, high-cholesterol diet, mouse ovariectomy model. Mice develop early left ventricle hypertrophy without concurrent fibrosis or aortic valve stenosis. These findings allow for a new model of the study of left ventricle hypertrophy in postmenopausal osteoporosis that more closely mimics the natural progression of disease in female patients.

Early effects of ovariectomy on bone microstructure, bone turnover markers and mechanical properties in rats

BackgroundFragility fracture is one of the most serious consequences of female aging, which can increase the risk of death. Therefore, paying attention to the pathogenesis of postmenopausal osteoporosis (PMOP) is very important for elderly women.Methods and materialsForty 12-week-old female rats were divided into two groups including the ovariectomy (OVX) group and the control group. Four rats in each group were selected at 1, 4, 8, 12 and 16 weeks after operation. Vertebral bones and femurs were dissected completely for micro-Computed Tomography (micro-CT) scanning, biological modulus detection and histomorphological observation.ResultsIn OVX group, bone volume/total volume (BV/TV), bone trabecular connection density (Conn.D) and trabecular bone number (Tb.N) decreased significantly with time (P < 0.05). The elastic modulus of femur in OVX group was lower than that in control group, but there was no significant difference between them (P > 0.05). Over time, the tartrate resistant acid phosphatase (TRAP), osteocalcin (BGP), type I procollagen amino terminal propeptide (PINP) and type I collagen carboxy terminal peptide (CTX-I) in OVX group increased significantly (P < 0.05). The micrographs of the OVX group showed sparse loss of the trabecular interconnectivity and widening intertrabecular spaces with time.ConclusionThe bone loss patterns of vertebral body and femur were different in the early stage of estrogen deficiency. The bone turnover rate of OVX rats increased, however the changes of biomechanical properties weren’t obvious.

Effect of oligonol, a lychee-derived polyphenol, on skeletal muscle in ovariectomized rats by regulating body composition, protein turnover, and mitochondrial quality signaling.

Oligonol is a low‐molecular‐weight polyphenol product derived from lychee (Litchi chinensis Sonn.) fruits. This study was focused on the effects of oligonol on the skeletal muscle of ovariectomized rats. We randomly divided female Sprague–Dawley rats into three groups: a sham surgery control group (Sham), an ovariectomy (OVX) group, and an OVX group treated with oligonol (OVX + Oligonol). Oligonol was intraperitoneally administrated at 30 mg/kg daily for 6 weeks. Oligonol treatment after OVX decreased body weight and fat mass, regulated lipid metabolism in skeletal muscle, without loss of lean mass and bone. Bone turnover was not affected by oligonol. In protein synthesis and degradation, oligonol increased the levels of the mammalian target of rapamycin and its downstream targets, eukaryotic initiation factor 4E‐binding protein 1 and 70‐kDa ribosomal protein S6 kinase, and it stimulated the expression of ubiquitin‐proteasome pathway proteins, the forkhead box transcription factors of the class O and the muscle ring‐finger protein‐1. Moreover, oligonol treatment enhanced mitochondrial biogenesis and dynamics. Thus, our results indicated that oligonol treatment had beneficial effects on the skeletal muscle in an estrogen‐deficiency rat model.

Genistein Daidzein in Kudzu Root (Pueraria lobata) Extract Decreased Malondialdehyde Plasma Levels in Hypoestrogenic Rats

Introduction: Menopause can reduce women's quality of life and cause health issues. The decline in estrogen during menopause is thought to trigger oxidative stress. When the creation of antioxidants does not match the synthesis of reactive oxygen species (ROS) as an antidote, oxidative stress ensues, characterized by malondialdehyde. The aim of the research is to evaluate the effect of the dose of isoflavone genistein daidzein powder in Kudzu root (Pueraria lobata) extract on the decrease in plasma malondialdehyde (MDA) plasma levels in hypoestrogenic rats. Methods: 30 rats Wistar rats were separated into five groups, namely: the control group, the ovariectomy (OVX) group, waited two weeks to produce hypoestrogenic rats. Of the four groups, the positive group was not given a Pueraria lobata powder containing the isoflavones genistein and daidzein (IGD), and the other group was given 15 mg, 30 mg, 60 mg per kg body weight per day, respectively. Twenty-one days later, the mice were killed, and plasma was taken to be checked for MDA levels using spectrophotometry. Results: Genistein Daidzein in kudzu root extract was effective in reducing the oxidative stress. All IGD doses significantly reduced plasma MDA levels. At a dose of 60mg/kg BW, the plasma MDA levels were comparable to those in the control group, it could prevent a significant increase in plasma MDA levels in hypoestrogenic rats. Conclusion: Isoflavone Genistein and Daidzein of kudzu root (Pueraria lobata) extract reduced MDA plasma levels in hypoestrogenic rats.

Experimental study of the effects of hypoxia simulator on osteointegration of titanium prosthesis in osteoporotic rats

BackgroundPoor osseointegration is the key reason for implant failure after arthroplasty,whether under osteoporotic or normal bone conditions. To date, osseointegration remains a major challenge. Recent studies have shown that deferoxamine (DFO) can accelerate osteogenesis by activating the hypoxia signaling pathway. The purpose of this study was to test the following hypothesis: after knee replacement, intra-articular injection of DFO will promote osteogenesis and osseointegration with a 3D printed titanium prosthesis in the bones of osteoporotic rats.Materials and methodsNinety female Sprague–Dawley rats were used for the experiment. Ten rats were used to confirm the successful establishment of the osteoporosis model: five rats in the sham operation group and five rats in the ovariectomy group. After ovariectomy and knee arthroplasty were performed, the remaining 80 rats were randomly divided into DFO and control groups (n = 40 per group). The two groups were treated by intraarticular injection of DFO and saline respectively. After 2 weeks, polymerase chain reaction (PCR) and immunohistochemistry were used to evaluate the levels of HIF-1a, VEGF, and CD31. HIF-1a and VEGF have been shown to promote angiogenesis and bone regeneration, and CD31 is an important marker of angiogenesis. After 12 weeks, the specimens were examined by micro-computed tomography (micro-CT), biomechanics, and histopathology to evaluate osteogenesis and osseointegration.ResultsThe results of PCR showed that the mRNA levels of VEGF and CD31 in the DFO group were significantly higher than those in the control group. The immunohistochemistry results indicated that positive cell expression of HIF-1a, VEGF, and CD31 in the DFO group was also higher. Compared with the control group, the micro-CT parameters of BMD, BV/TV, TB. N, and TB. Th were significantly higher. The maximal pull-out force and the bone-to-implant contact value were also higher.ConclusionsThe local administration of DFO, which is used to activate the HIF-1a signaling pathway, can promote osteogenesis and osseointegration with a prosthesis in osteoporotic bone.

A high dose of estrogen can improve renal ischemia-reperfusion-induced pulmonary injury in ovariectomized female rats.

Renal ischemia-reperfusion injury (RIRI) as a pathological process induces remote organ injury such as lung complications and it is regulated in a hormone-dependent manner. This study investigates the effect of estrogen on RIR-induced pulmonary injury in ovariectomized (OV) rats. A total of 60 female Wistar rats were divided into six groups: (i) intact sham, (ii) OV sham, (iii) OV sham + estradiol valerate (E), (iv) intact ischemia, (v) OV ischemia, and (vi) OV ischemia + E. Bilateral ischemia was performed for 45min in all groups except sham. Before the ischemia, OV groups received an intramuscular (i.m.) injection of E. After reperfusion, blood samples were collected for serum analysis and kidney and lung tissue were separated for pathological experiment and malondialdehyde (MDA) and nitrite measurement. The left lung was weighed to measure pulmonary edema. Estrogen deficiency caused a greater increase in blood urea nitrogen and creatinine levels during IRI. Ischemia reduced nitrite of serum and lung tissue. The increased level of MDA during ischemia, returned to normal levels via estrogen injection. The severity of renal and lung damage in ischemic groups increased significantly, and estrogen improved this injury. Estrogen as an antioxidant agent can reduce oxidative stress and may improve renal function and ameliorating lung damage caused by RIR.

Transient Receptor Potential Melastatin 2 Thermosensitive Neurons in the Preoptic Area Involved in Menopausal Hot Flashes in Ovariectomized Mice

Introduction: Menopausal hot flashes are related to hypothalamic preoptic area (POA) dysfunction. Thermosensitive transient receptor potential channels (ThermoTRPs) are involved in temperature sensing and regulation of thermosensitive neurons (TSNs) in the POA. Whether ThermoTRP-TSNs in the POA, particularly the non-noxious thermoreceptor, transient receptor potential melastatin 2 (TRPM2), are involved in the occurrence of hot flashes is still unclear. Methods: Twenty wild-type and 50 Trpm2-Cre adult female mice were randomly divided into sham (SHAM) and ovariectomy (OVX) groups. In the POA, ERα, ERβ, GPR30, TRPA1, TRPM8, TRPM2, and TRPV1 expression was detected by Western blot or/and quantitative real-time polymerase chain reaction and the number of TSNs expressing TRPM2 (TRPM2-TSNs) by immunofluorescence. Before and after TRPM2-TSN activation/inhibition, back (BST) and tail skin temperature (TST) and the proportion of glutamatergic and GABAergic neurons among TRPM2-TSNs were recorded. Results: Compared with SHAM, the expression of ERα, ERβ, TRPM2, and TRPM8 in the POA of the OVX group decreased, with a significantly larger change range for TRPM2 than TRPM8. In addition, the number of TRPM2-TSNs showing TRPA1, TRPM8, and TRPV1 expression in the OVX group decreased, and the proportion of glutamatergic and GABAergic neurons in TRPM2-TSNs decreased and increased, respectively. Meanwhile, BST and TST increased. After activating or inhibiting TRPM2-TSNs, the proportions of glutamatergic and GABAergic neurons in TRPM2-TSNs changed, along with the BST and TST. Conclusion: In menopause, the abnormal quantity and function of TRPM2-TSNs in the POA is key for the development of hot flashes, characterized by an imbalance in heat dissipation and production due to the corresponding imbalance in glutamatergic and GABAergic neurons.

Phytoestrogen in cowpea (Vigna unguiculata L. Walp) (Fabaceae) extract reduces vaginal oxidative stress and increases proliferation of fibroblast in ovariectomized rats

Purpose: To evaluate the effects of a phytoestrogen extract of cowpea (Vigna unguiculata) on malondialdehyde (MDA) and superoxide dismutase (SOD) levels, mitogen-activated protein kinase (MAPK) activity, as well as proliferation of fibroblasts in ovariectomized rat vagina. This is with a view to identify the optimal dose of a phytoestrogen supplement for use during the menopausal period of women.&#x0D; Methods: Wistar rats (Rattus norvegicus) were divided into five groups: negative control, ovariectomy, and ovariectomy groups treated with 1.25, 2.5, or 5 mg/kg of cowpea extract, respectively. The three doses were used to identify the optimal dose required to reduce vaginal oxidative stress and increase fibroblast proliferation in ovariectomized rats. After treatment, the ovariectomized rat vagina was assessed for SOD and MDA levels, MAPK activity, and fibroblast proliferation.&#x0D; Results: Cowpea extract at a dose of 2.5 and 5 mg/kg caused significant reduction in SOD levels in the vaginal tissue of rats compared with control (p &lt; 0.05). In contrast, cowpea extract at 1.25 mg/kg resulted in MDA levels similar to that of control rats (p &lt; 0.05). Cowpea extract treatment had positive effects on SOD and MDA levels and on fibroblast proliferation. Increase in SOD levels in rat vagina was induced by increased MDA levels, which is characteristic of oxidative stress, while ovariectomy resulted in decreased MAPK activity.&#x0D; Conclusion: Overall, the cowpea extract has no significant effect on MAPK activity or fibroblast proliferation. However, it has a significant effect on SOD and MDA levels and on fibroblast proliferation at a dose of 2.5 mg/kg.

Ginkgetin aglycone exerts anti-osteoporotic effect via regulation of NOX4/Akt/PI3K pathway

Purpose: To investigate the protective effect of Ginkgetin aglycone (GA) on ovariectomy-induced osteoporosis in rats, as well as the mechanism of action involved.&#x0D; Methods: Adult female Wistar rats (n = 40) were separated into four group: normal control, ovariectomy (OVR), 100 mg GA/kg dose, and 200 mg GA/kg dose. The rats were ovariectomized using standard procedures, except for those in normal control group. Rats in the two treatment groups received 100 or 200 mg GA/kg orally for a period of 12 weeks. Biochemical assays were performed on the urine and blood. Markers of bone formation and mediators of inflammation were assessed. Bone microarchitectural changes were examined using micro-CT scanner, while Western blotting was used to determine the expressions of NOX4, NF-κB p65, PI3K, Akt and JNK proteins in rat femurs.&#x0D; Results: Phosphorus and calcium levels in the serum varied among different groups. Levels of calcium, phosphorus and creatinine decreased (p &lt; 0.01) significantly to a greater extent in the urine of GA group than in that of OVR group (p &lt; 0.05). Interleukin-1β (IL-1β), tumor necrosis factor α (TNF-α) and osteocalcin (OC) levels and the activity of alkaline phosphatase (ALP) decreased more in GA group than in OVR group. In GA-treated group, bone mineral density (BMD) was enhanced in a dose dependent manner than OVR group (p &lt; 0.05). Treatment with GA ameliorated altered bone microarchitecture in OVR rats. Treatment of osteoporotic rats with GA led to significant and dosedependent decrease in the expressions of JNK, NOX4, NF-κB p65 and PI3K, and (p &lt; 0.05) increase in the expression of Akt in femur tissue.&#x0D; Conclusion: In conclusion, result of study proves the anti-osteoporotic activity of GA is exerted via regulation of NOX4/PI3K/Akt pathway.

The relative contribution of bone microarchitecture and matrix composition to implant fixation strength in rats.

Bone microarchitectural parameters significantly contribute to implant fixation strength but the role of bone matrix composition is not well understood. To determine the relative contribution of microarchitecture and bone matrix composition to implant fixation strength, we placed titanium implants in 12-week-old intact Sprague-Dawley rats, ovariectomized-Sprague-Dawley rats, and Zucker diabetic fatty rats. We assessed bone microarchitecture by microcomputed tomography, bone matrix composition by Raman spectroscopy, and implant fixation strength at 2, 6, and 10 weeks postimplantation. A stepwise linear regression model accounted for 83.3% of the variance in implant fixation strength with osteointegration volume/total volume (50.4%), peri-implant trabecular bone volume fraction (14.2%), cortical thickness (9.3%), peri-implant trabecular crystallinity (6.7%), and cortical area (2.8%) as the independent variables. Group comparisons indicated that osseointegration volume/total volume was significantly reduced in the ovariectomy group at Week 2 (~28%) and Week 10 (~21%) as well as in the diabetic group at Week 10 (~34%) as compared with the age matched Sprague-Dawley group. The crystallinity of the trabecular bone was significantly elevated in the ovariectomy group at Week 2 (~4%) but decreased in the diabetic group at Week 10 (~3%) with respect to the Sprague-Dawley group. Our study is the first to show that bone microarchitecture explains most of the variance in implant fixation strength, but that matrix composition is also a contributing factor. Therefore, treatment strategies aimed at improving bone-implant contact and peri-implant bone volume without compromising matrix quality should be prioritized.