OVX Control Group Research Articles

Maltobionic acid protects against ovariectomy‐induced osteoporosis by suppressing bone resorption

AbstractBackgroundOsteoporosis is a debilitating disease characterized by decreased bone density. In this study, we evaluated the anti‐osteoporotic effect of maltobionic acid (MB), one of the components of honey, and calcium maltobionate (MBCa) on bone density and metabolism using a mouse model of osteoporosis. The underlying mechanisms of MB and MBCa action were also investigated.ResultsOvariectomized (OVX) mice were fed diet containing MB or MBCa for 80 days, and femoral bone mineral content and mineral density (BMD) were evaluated. As expected, OVX reduced BMD; however, the administration of MB and MBCa significantly prevented this decrease. Furthermore, MB and MBCa treatment significantly reduced the serum levels of tartrate‐resistant acid phosphatase 5b (TRACP‐5b), a bone resorption marker, and significantly increased the levels of serum calcitonin, compared to those in the OVX control group. In vitro, the relatively high‐dose levels of MB and MBCa inhibited osteoclast differentiation by decreasing the protein expression of nuclear factor of activated T‐cell cytoplasmic 1 (NFATc1), the master transcription factor of osteoclastogenesis.ConclusionOur results showed that osteoporotic mice treated with MB and MBCa had improved bone density and bone metabolism compared to the OVX controls. Moreover, it was hypothesized herein that the suppressive effect on osteoclast differentiation slightly contributed to these results concomitantly. These findings suggest that the intake of MB or MBCa may contribute to the maintenance of bone health, including the prevention of primary osteoporosis.

Phaleria macrocarpa Fruit Flavonoid Extract on Estrogen Receptor Alpha Expression in Ovariectomized Mice

Menopause signifies the cessation of ovarian function, presenting significant health challenges for women globally. Hormonal shifts during menopause impact various systems, including cardiovascular and reproductive health. Changes in reproductive organs increase susceptibility to health issues, necessitating effective management strategies. This study used a true experimental method with a randomized post-test-only control group design to investigate the potential of Phaleria macrocarpa fruit extract, which contains flavonoids, to modulate Estrogen Receptor Alpha (ERa) expression in menopausal mice. Following ovariectomy (OVX), mice treated with flavonoid showed increased ERa expression, with the highest expression observed at a dose of P2 (7.5 mg/mice/day), approaching those in non-OVX control (K-) with a percentage difference of 29.82%. Compared to the OVX control group (K+), the percentage increase was 2387.74%. Therefore, the P2 dose is considered the optimum dose. Regression analysis revealed a significant correlation of 43.3% between flavonoid dosage and ERa expression (p < 0.05). These findings suggest flavonoids as promising natural Selective Estrogen Receptor Modulators (SERMs) for managing menopausal symptoms and reducing endometrial disease risk with ERa modulation. Further research is needed to elucidate the mechanisms of flavonoids, including molecular pathways such as ligand binding and transcription, interactions with other hormones like progesterone, long-term effects, and potential clinical applications. To initiate clinical trials, the author established the P1 dose (3.75 mg/mice/day), which demonstrated expression levels approaching the effects produced by P2. Overall, flavonoids offer potential therapeutic benefits in addressing menopausal health concerns, with the identified optimal dosage providing valuable guidance for future studies and clinical interventions.

Silkworm (Bombyx mori L.) Has Beneficial Effects on Menopausal Symptoms by Enhancing Estrogen Receptor Signaling in Ovariectomized Mice.

Existing hormone replacement therapy for menopause has drawbacks, necessitating new treatment agents. Silkworms have demonstrated estrogenic properties, offering promising alternatives. We assessed the therapeutic effects of freeze-dried silkworm powder (SWP) on menopausal symptoms using an ovariectomized (OVX) mouse model. The experimental design comprised a sham surgery group (Sham), an OVX control group, a low-dose SWP group post-OVX (80 mg/kg, OVX-SWP-L), a high-dose SWP group post-OVX (160 mg/kg, OVX-SWP-H), and an estradiol treatment group post-OVX (OVX-E2). Treatments were administered orally thrice weekly over eight weeks; body weight was monitored weekly. The SWP-treated groups (SWP-L and SWP-H) exhibited less weight gain and increased uterine thickness than the OVX control. Molecular analyses demonstrated that SWP significantly enhanced the phosphorylation of estrogen receptor alpha (ERα), ERK, and AKT. Furthermore, biochemical assays revealed reduced serum neutral lipids across all SWP treatment groups. Notably, HDL-cholesterol levels were significantly increased in the SWP-L group compared to the OVX group. Serum estradiol concentrations were elevated in all the SWP groups, with significant increases in the high-dose group. These findings indicate that SWP may promote the activation of estrogen receptor signaling and improve symptoms associated with estrogen deficiency during menopause.

Virgin Coconut Oil and Palm Tocotrienol Supplementation: Effects on Lipid Parameters of Experimental Rats

Virgin coconut oil (VCO) and palm tocotrienol (TT) possess cholesterol-regulating properties, but their combined effects remained unexplored. Thus, this study aims to determine their individual and combination effects on lipid parameters in ovariectomised (OVX) rats on hypercholesterolemic diet. Female Sprague-Dawley rats were ovariectomised except for the sham group, assigned into (i) sham-operated fed with basal rat diet, (ii) OVX control, (iii) V1 (OVX + VCO 1.43 mL/kg), (iv) V2 (OVX + VCO 4.29 mL/kg), (v) TT (OVX + TT 30 mg/kg), (vi) V1+TT (OVX + VCO 1.43 mL/kg + TT 30 mg/kg) and (vii) V2+TT (OVX + VCO 4.29 mL/kg + TT 30 mg/kg). Groups (ii) to (vii) were fed with 2% cholesterol mixed with five-time heated palm oil. VCO and TT alone or in combination reduced food intake, visceral fat weight, thiobarbituric acid reactive substances and HMG-CoA reductase activity significantly (p<0.05 vs the OVX control). HDL was reduced significantly in V1, V2, and V1+TT compared to the TT and V2+TT (p<0.05). The increase in LDL was the lowest in V1 compared to other groups (p<0.05). V2 and TT significantly reduced total cholesterol compared to other supplementations (p<0.05). All supplementations were found to reduce triglyceride compared to the OVX control group (p<0.05). The increase in apolipoprotein A was higher in V2+TT than other groups (p<0.05). The reduction in apolipoprotein B was higher in V1+TT and V2+TT than the V1, V2 and TT. VCO and TT exerted beneficial effects on lipid parameters, but the efficacy was not better than individual agents.

Bone strength is reduced in a neonatal androgenized rat model

BackgroundWhether polycystic ovary syndrome (PCOS) affects bone health during a woman's lifespan remains controversial. An androgenized rodent model replicated many metabolic and reproductive features of women with PCOS, and we aimed to use it to investigate the impact of androgens on microarchitecture (by micro-CT), bone mechanical strength, bone formation and resorption markers in rats with intact ovaries (SHAM) who underwent oophorectomy. MethodsWistar rats (Rattus norvegicus albinus) were employed for the experiments in this study. The protocol of androgenization consisted of the application of 1.25 mg s.c. testosterone propionate beteween days 2–5 of life, while the controls received the same amount of corn oil s.c. as previously established. Androgenized SHAM rats exhibited chronic anovulation identified by vaginal cytology and a reduction in the proportion of corpus luteum in the ovary in comparison to control SHAM rats. The realization of the ovariectomy or SHAM procedure occurred on Day 100 of life. All groups (n = 8) were followed-up for 180 days to address the study endpoints. ResultsMicro-CT from androgenized female rats (SHAM) showed a divergence between the trabecular and cortical bone profiles. Compared to SHAM controls, these rats had an increase in trabecular bone mass with a diminution in bone resorption C-terminal telopeptide of type 1 collagen (CTX) (p < 0.05), a concomitant decrease in cortical area and thickness in the femur, and a reduction in the strength of the femur on the mechanical test (p < 0.01). ConclusionsOur results suggest that a reduction in the cortical thickness and cortical area observed in PCOS model rats was associated with a reduced strength of the femur, despite increased trabecular formation. Ovariectomy in the androgenized OVX group limited the progression rate of cortical bone loss, resulting in bone resistance and cortical thickness comparable to those observed in the control OVX group.

The bone-protective benefits of amino-conjugated calcium in an ovariectomized (OVX) rat model

Low bone density, fragility, and microarchitectural disintegration are the symptoms of osteoporosis. An imbalance between bone growth and resorption can lead to osteoporosis. This study evaluated the effects of amino-calcium (AC) on bone protection in ovariectomized control group (NC) rats. Amino-calcium (AC) was characterized using Fourier-transform infrared spectroscopy (FT-IR), energy-dispersive X-ray spectroscopy (EDS), and nuclear magnetic resonance spectroscopy analyses (NMR). After determining the biocompatibility of amino-calcium (AC) with MC3T3-E1 cells, alkaline phosphatase staining revealed significant changes on day 7. Three of the four groups underwent ovariectomy, whereas one group received a placebo. On micro-computed tomography, in vivo, data showed increased bone volume fraction in the femoral head and shaft areas in the amino-calcium (AC) group. Hematoxylin and eosin staining showed a bone mass and architectural protection in the amino-calcium (AC) group compared with the calcium carbonate and OVX control group. RNA sequencing analysis revealed high expression of osteogenesis-related genes in MC3T3-E1 cells. RNA sequencing revealed a significant fold change in the expression of integrin-binding sialoprotein (IBSP), bone gamma-carboxyglutamate proteins 1 and 2(BGLAP1 and BGLAP2), and periostin (POSTN). The study concluded that supplementing the OVX rats with calcium enhanced bone protection.

Effects of the Germinated Glycine max and Angelica gigas Nakai Mixture on Hepatic Lipid Metabolism and Bone Turnover Balance in Ovariectomized Rats

Women going through menopause experience changes in their hormone levels, such as decreased estrogen secretion. Those changes can lead to weight gain and steatosis (fatty liver) due to abnormal lipid metabolism and bone turnover. Natural herbal medicines have been developed to treat and prevent menopausal symptoms. The aim of this study was to evaluate how a mixture of germinated Glycine max (GG) and Angelica gigas Nakai (AG) affected serum lipid profile levels, hepatic damage, inflammation, and bone turnover in ovariectomized (OVX) rats. The animals were randomly allocated into six groups: Sham control group (Sham), OVX control group (OVX), OVX + 50 mg/kg b.w. of GG (GG50) group, and OVX + 25, 50, 100 mg/kg b.w. of GG and AG mixture (GAM; GAM25, GAM50, GAM100) groups. After four weeks of treatment, the GAM groups exhibited decreases in serum lipid profile levels (TC, TG, and Low Density Lipoprotein (LDL)) and increases in High Density Lipoprotein (HDL). Proinflammatory mediators (IL-1β, IL-6, TNF-α, and iNOS) were reduced after the administration of GAM, and the degree of liver damage (ALT, AST) also decreased. Bone resorption (CTX1, NTX1, osteoclasts in H&amp;E staining) decreased in the GAM-treated groups, and bone morphometric markers (BMD, BV/TV, Tb.N) improved compared with the OVX group. Additionally, increased bone formation (ALP, mineralization) and decreased bone resorption (TRACP) were confirmed in in vitro experiments. These results suggest that GAM has anti-obesity and anti-inflammatory effects by preventing dyslipidemia and that it alleviates bone mass loss in OVX rats by inhibiting osteoclastogenesis.

The probiotic Lactiplantibacillus plantarum attenuates ovariectomy-induced osteoporosis through osteoimmunological signaling.

Osteoporosis is characterized by low bone mass and bone tissue microarchitectural deterioration with increased fracture risk in numerous populations. Probiotics are reported to be a potential biotherapeutic for the prevention and treatment of osteoporosis. In this study, the IL-10 secretion properties of probiotics were simulated in vitro and the potential applications of the novel strain Lactiplantibacillus plantarum 622 were investigated in an in vivo osteoporosis model. Female Sprague-Dawley rats were ovariectomized (OVX) and orally administered Lp. plantarum GMNL-662 or alendronate for 14 weeks. The Lp. plantarum treatment group exhibited an increase in the level of fecal Lp. plantarum, Lactobacillus, and Lachnospiraceae. Bone marker analysis indicated improvements in the levels of osteocalcin and N-terminal telopeptides in the Lp. plantarum treatment group. Compared with the OVX control group, the Lp. plantarum treatment group exhibited marked improvements in femur bone mineral density, trabecular bone volume, trabecular number, and lumbar vertebrae. Moreover, biomechanical three-point bending testing indicated considerably higher improvements in femur maximum load, stiffness, and energy to maximum load in the Lp. plantarum treatment group than in the OVX control group. Quantitative polymerase chain reaction analysis indicated reduced expression levels of OVX-induced IL-1, IL-6, TNFα, and RANKL and increased expression levels of IL-10, TGF-β, and osteoprotegerin in the Lp. Plantarum treatment group. In summary, Lp. plantarum GMNL-662 exhibits high probiotic potential and potentially influences osteoimmunity through the modulation of proinflammatory cytokines and bone metabolism-related markers.

Impact of ovariectomy and estradiol-17β (E2) replacement on the brain steroid levels of the Indian stinging catfish Heteropneustes fossilis

In the present study, effects of ovariectomy (Ovx) and estradiol-17β (E2) replacement on the brain steroid levels were investigated in the resting and prespawning phases of the female catfish Heteropneustes fossilis. The study showed that Ovx resulted in significant decreases in plasma levels of estradiol-17β (E2) and its precursors, and the effect varied with the reproductive stage and duration. Our study showed that E2 supplementation reversed the effect of Ovx in 3-week Ovx fish group, either restored or increased according to the dose. In contrast, E2 levels increased in the brain after Ovx in the resting stage and produced a biphasic effect of significant decreases on week 1 and 2, and significant increases on week 3, 4 and 5 in the pre-spawning stage. The brain E2 levels significantly inhibited in E2 supplementation groups. The testosterone (T) levels showed biphasic effects, an initial decrease (up to 3 weeks Ovx) and an increase later (week 4, 5) in the resting stage. In the pre-spawning stage, the T levels increased significantly after Ovx. The E2 supplementation significantly inhibited the T levels, more severely in the low dose group in the resting and pre-spawning stages. Brain 17-hydroxyprogesterone(17-OHP4) levels increased (week 1, 2, 3) but decreased on week 4 and 5 of Ovx in the resting stage and increased throughout Ovx in the pre-spawning stage except in the 5th week (though compared to other steroids, level was very low; almost negligible). The E2 replacement significantly decreased the steroid levels further down compared to the Ovx control group. Brain progesterone (P4) and pregnenolone (P5) levels decreased initially (week 1 and 2 Ovx) and recovered later. The E2 replacement significantly decreased P4 levels in the Ovx fish in the pre-spawning stage and in the lower dose group in the resting stage. The P5 levels remained inhibited after the E2 supplementation in the resting stage but unchanged or increased it in the pre-spawning stage. Brain cortisol (F) levels were inhibited initially (1 and 2 weeks and later increased after OVX in the resting stage. In the pre-spawning stage, the F levels increased in the week 1, 2 and 3 of Ovx and decreased in the 4th and 5th weeks. The E2 supplementation inhibited the cortisol (F) levels in both phases. The results show that ovarian steroids influence neurosteroidogenesis in a reproductive stage-dependent manner.

Estradiol Supplement or Induced Hypertension May Attenuate the Angiotensin II Type 1 Receptor Antagonist-Promoted Renal Blood Flow Response to Graded Angiotensin II Administration in Ovariectomized Rats.

Backgrounds Estrogen replacement therapy (ERT) and hypertension may influence females' renin-angiotensin system (RAS) and its components. The angiotensin II (Ang II) type 1 receptor (AT1R) antagonist (losartan) may promote renal blood flow (RBF), and it is widely used in the clinic to control hypertension. The main objective of this study was the effects of estradiol or induced hypertension on RBF response to Ang II in losartan-treated ovariectomized (OVX) rats. Methods Two groups of OVX rats were treated with placebo (group 1) and estradiol (group 2) for period of four weeks, and another group of OVX rats was subjected to induce hypertension by two-kidney one clip (2K1C) model (group 3). All the groups were subjected to the surgical procedure under anesthesia, and AT1R was blocked by losartan. RBF and renal vascular resistance (RVR) responses to Ang II administration were determined and compared. Results Mean arterial (MAP) and renal perfusion (RPP) pressures in group 3 and uterus weight (UT) in group 2 were significantly more than other groups (P < 0.05). Ang II infusion resulted in dose-related percentage change increase in RBF and decrease in RVR. However, these responses in the OVX-estradiol and OVX-hypertensive rats were significantly lower than in the OVX-control group (P < 0.05). For instance, at the dose of 1000 ng/kg/min of Ang II administration, the percentage change of RBF was 45.1 ± 10.4%, 17.9 ± 2.3%, and 16.7 ± 4.7% in the groups of 1 to 3, respectively. Conclusion Losartan prescription in some conditions such as hypertension or ERT could worsen RBF and RVR responses to Ang II.

Estrogenic Effect of Asparagus racemosus, Cissus quadrangularis, Punica granatum and Pueraria tuberosa in Post-menopausal Syndrome

Pharmacognosy Research,2021,13,4,238-245.DOI:10.5530/pres.13.4.17Published:October 2021Type:Original ArticleAuthors:Seema Singh, Santosh Kumar, Sunita Singh, Chetna Mishra, Dinesh Tripathi, and S K Verma Author(s) affiliations:Seema Singh1,*, Santosh Kumar1, Sunita Singh2, Chetna Mishra3, Dinesh Tripathi3, SK Verma1 1Department of Respiratory Medicine, King Georges Medical University, Lucknow, Uttar Pradesh, INDIA. 2Department of Microbiology, King Georges Medical University, Lucknow, Uttar Pradesh, INDIA. 3Department of Physiology, King Georges Medical University, Lucknow, Uttar Pradesh, INDIA. Abstract:Background: Asparagus racemosus, Cissus quadrangularis, Punica granatum and Pueraria tuberosa are employed for their supposed properties in treatment in alleviating symptoms of menopause traditionally from ages. The aim of the present study is to evaluate the estrogenic effect of Asparagus racemosus, Cissus quadrangularis, Punica granatum and Pueraria tuberosa in ovariectomized (OVX) rats. Materials and Methods: Female wistar rats were divided into sham control and ovariectomized (OVX), OVX rats receiving standard drug, raloxifene (5.4 mg/kg) and groups treated with 500 mg/kg and 700 mg/kg of test drug (containing equal quantity of Asparagus racemosus, Pueraria tuberose, Cissus quadrangularis and Punica granatum extracts) daily for 90 days. The vaginal cornification, uterine weight, bone loss, biomechanical, biochemical and histopathological observation were carried out to ascertain the effect of test drug in post-menopausal syndrome. Results: The experimental animals treated with mixture of extract of four drugs showed dose dependent activity. The significant increase in uterine weight, femur BMD, femur hardness, was observed. In addition, increased levels of calcium and phosphorus in serum and significant decreased in urine were observed as compared to control OVX group. The histopathological results also confirm the protective effect of extracts. Conclusion: The present findings strongly suggest that Asparagus racemosus, Cissus quadrangularis, Punica granatum and Pueraria tuberosa possess the potent estrogenic activity in ovariectomized rats and substantiate the ethnic use in treatment of postmenopausal osteoporosis. Keywords:Biochemical, Estrogenic, Histopathological, Menopause, Ovariectomized (OVX) rats, Post menopausal syndromeView:PDF (994.49 KB)

Dietary syringic acid reduces fat mass in an ovariectomy-induced mouse model of obesity

Postmenopausal women are at increased risk of metabolic diseases such as obesity and diabetes. Therefore, the chemoprevention of postmenopausal changes in health via dietary supplements is important. Syringic acid (SA) is a phenolic compound present in the fruit of the assai palm, Euterpe oleracea, and in the mycelium of the shiitake mushroom, Lentinula edodes. This compound shows no affinity for estrogen receptors and may exert disease-preventive effects. Reportedly, dietary SA ameliorates high-fat diet-induced obesity in mice; however, its effects on estrogen deficiency-induced obesity are still unclear. Therefore, in this study, we investigated whether and how dietary SA affects these factors in ovariectomized (OVX) mice. Ten-week-old OVX mice were fed SA-containing diets (100 mg/kg body weight/d) for 12 weeks. Their body weights, food intake, and uterus weights as well as other parameters were measured and comparisons were made with mice in the control group. Dietary SA did not affect the body weight, food intake, or uterus weight of OVX mice over the study period; however, the SA-fed group showed lower fat mass (ie, visceral, subcutaneous, and total fat) than the OVX-control group (11.1 ± 3.3 vs. 8.3 ± 2.4, P < 0.05; 7.9 ± 1.1 vs. 5.9 ± 1.6, P < 0.05; 19.0 ± 4.2 vs. 14.1 ± 3.8, P < 0.05, respectively). Furthermore, blood analysis revealed that SA-treatment resulted in a dose-dependent decrease and increase in serum triglyceride (59.2 ± 8.3 vs. 43.9 ± 12.2 mg/dL P < 0.05) and adiponectin (7.7 ± 0.3 vs. 9.5 ± 0.6 μg/mL, P < 0.05) levels, respectively. These results suggest that the SA diet improves lipid metabolism without affecting the uterus in OVX mice. Therefore, dietary SA has potential applicability for the prevention of postmenopausal obesity and type 2 diabetes.

Estrogenic and Antioxidant Activities of Pterocarpus soyauxii (Fabaceae) Heartwood Aqueous Extract in Bilateral Oophorectomized Wistar Rat.

Phytoestrogens are used to ease postmenopausal symptoms, a property probably due to estrogenic and antioxidant effects. Pterocarpus soyauxii (P. soyauxii) is empirically used in Cameroon to treat among others primary and secondary amenorrhea. The aim of this study is to evaluate estrogenic and antioxidant activities of P. soyauxii heartwood aqueous extract in bilateral oophorectomized Wistar rats. Firstly, a characterization of the extract was carried out. For that, flavonoids, phenols, and tannins levels in P. soyauxii extract were evaluated by colorimetric assays and UHPLC-MS analysis was realized. In vitro antioxidant analysis of P. soyauxii was conducted using DPPH, ABTS, and FRAP assays. Secondly, 2 sets of pharmacologic tests were carried out. The results revealed that P. soyauxii aqueous extract contains, respectively, 229.42 ± 3.62 mg EAG/g, 63.42 ± 2.16 mg EQ/g, and 27.88 ± 0.23 mg ETA/g of polyphenols, flavonoids, and tannins. UHPLC-MS enabled identifying seven components including mono(2-ethylhexyl) phthalate, cembrene, 3′,5′-dimethoxy-4-stilbenol, and linoleic acid. DPPH, ABTS, and FRAP assays revealed that P. soyauxii extract possessed a high antioxidant activity with IC50 value of 730.20 µg/mL, 892.90 µg/mL, and 765.75 mEAG/g of extract, respectively. In the uterotrophic assay, P. soyauxii extract induced significant increase of fresh uterine weight, uterine and vaginal epithelial size, and mammary glands differentiation compared to Ovx control. In the postmenopausal model, compared to the sham control, vagina and uterine dystrophies were observed in Ovx rats treated with distilled water. P. soyauxii aqueous extract expressed estrogenic-like effects on vagina and did not affect uterine epithelial height compared with vehicle groups. On the back of these vaginotrophic effects, the extract displayed antiatherogenic properties by reducing (p < 0.001) AI and LDL cholesterol level as compared to Ovx control group. The extract at 200 mg/kg significantly prevented the increase of MDA (p < 0.01) level and decreased nitrites (p < 0.001) and GSH (p < 0.01) levels compared to Ovx rats. These beneficial effects are related at least in part to the presence of compound such as mono(2-ethylhexyl) phthalate, 3′,5′-dimethoxy-4-stilbenol, and linoleic acid. Overall, P. soyauxii aqueous extract exhibits estrogenic and antioxidant effects which can inhibit postmenopausal symptoms by providing vaginal stratification, improving lipid profile and insulin sensitivity, and reducing oxidative stress without side effects on the endometrium and mammary gland in 84-day Ovx rats.

The effects of royal jelly protein on bone mineral density and strength in ovariectomized female rats.

[Purpose]Sex hormones deficiency leads to dramatically bone loss in particular postmenopausal women. Royal jelly has anti-osteoporosis effect due to maintain bone volume in that condition. We hypothesized that royal jelly protein (RJP, a latent residue after extracting royal jelly) also prevents bone deficient in ovariectomized (OVX) female rats, the animal model of postmenopausal women. [Methods]Female Sprague-Dawley rats (n = 30, 6 weeks age old) were sham operated (Sham; sham operated group, n = 7), OVX control group (OC, n = 7), OVX with low RJP intake group (ORL, n = 8), and OVX with high RJP intake group (ORH, n = 8) during 8 weeks experimental periods. In the end point of this experiment, the bone samples (lumbar spine, tibia, and femur) were surgically removed under anesthesia. These bone samples were evaluated bone mineral density (BMD) and bone strength.[Results]BMD of lumbar spine in RJP intake groups (ORL, ORH) were higher than that in OC group (p < 0.05 and p < 0.01) in RJP intake volume dependent manner. BMD of tibial proximal metaphysis and diaphysis in RJP intake groups were also higher than these in OC group (p < 0.01 and p < 0.01 / p < 0.05 and p < 0.001). In addition, breaking force of femur in RJP intake groups were significantly increase compared with that in OC group (p < 0.001 respectively). [Conclusion]These findings indicate that RJP contribute to prevent sex hormone related bone abnormality

Irisin Has a Protective Role against Osteoporosis in Ovariectomized Rats.

The reduction in estrogen levels results in a decrease in bone density at menopause. Irisin is a myokine that modulates the benefits of exercise, which may include bone health. This study was planned to examine irisin's impact in preventing osteoporosis after ovariectomy. 4 groups of female albino rats (10 rats/group): control, sham-operated, ovariectomized (OVX-control), and OVX-irisin-treated. Serum levels of bone markers [osteocalcin (OC), bone alkaline phosphatase (BALP), tartrate-resistant acid phosphatase (TRAP), calcium (Ca++), phosphorus (P)], glucose, and insulin were being measured. Body mass index, Homeostatic Model Assessment of Insulin Resistance (HOMA-IR), dry and ash femur weight, and bone contents of Ca++ and P were investigated. The femur was examined histopathologically. The OVX-control group showed an increase in serum levels of OC, BALP, TRAP, calcium, phosphorus, BMI, glucose, insulin, and HOMA-IR (P < 0.05) and a reduction in dry and ash weight of the femur, the concentration of calcium and phosphorus content in bone ash (P < 0.05). The OVX-irisin-treated group exhibited a decrease in serum levels of OC, BALP and TRAP, calcium, phosphorus, BMI, glucose, insulin, HOMA-IR (P < 0.05), and a rise in dry and ash weight of the femur, the concentration of calcium and phosphorus in bone ash (P < 0.05). Histological examination of the distal femur diaphysis of the OVX-irisin-treated group exhibited proper bone architecture and density compared with that of the OVX-control group. It is concluded that irisin treatment in the OVX rats safeguarded the regular bone architecture and normal levels of serum bone biomarkers. Irisin may be a possible novel target in the prohibition of postmenopausal osteoporosis.

Effect of combined treatment with pulsed electromagnetic field stimulation and sclerostin monoclonal antibody on changes in bone metabolism and pedicle screw augmentation in rabbits with ovariectomy-induced osteoporosis.

Both pulsed electromagnetic field (PEMF) stimulation and sclerostin monoclonal antibody are useful for treating osteoporosis, but whether the two therapies have synergistic effects on both screw fixation quality and bone metabolism of ovariectomy-induced osteoporosis has not been reported. We used ovariectomy to create a rabbit model of postmenopausal osteoporosis. Then, specimens were fixed with pedicle screws in the L4 vertebral body. Rabbits were randomly divided into an OVX control group, PEMF group, Scl-Ab group, and PEMF+Scl-Ab group. The PEMF group was given PEMF magnetic therapy, the Scl-Ab group was administered a subcutaneous Scl-Ab injection, and the PEMF+SclAb group received both therapies. The OVX group was injected subcutaneously with the same dose of saline instead. After eight weeks of treatment, the bone metabolism index, bone mineral density (BMD), and bone microstructural, biological, and biomechanical parameters were evaluated. BMD significantly decreased six months post-ovariectomy. Compared with that of the OVX group, the BMD of the PEMF, Scl-Ab, and PEMF+Scl-Ab groups increased by 20.3%, 19.9%, and 35.0%, respectively. The maximum pulling force of pedicle screws increased by 14.0%,15.0% and 19.1%, and the maximum failure power consumption of pedicle screws increased by 27.9%, 27.2% and 33.6%, respectively; these differences were statistically significant (P<0.05). The bone metabolism index and bone microstructure parameters of the PEMF+Scl-Ab group were more optimal than those in the single treatment group. Both Scl-Ab and PEMF therapy can enhance the BMD and the mechanical strength of pedicle screws in osteoporotic bones of rabbits with postmenopausal osteoporosis. However, combination of the two measures has achieved even better results, yielding potential clinical application value.

The effect of dehydroepiandrosterone administration on intestinal calcium absorption in ovariectomized female rats

[Purpose] Dehydroepiandrosterone (DHEA) administration reportedly recovers osteoporosis, a bone disorder associated with bone deficiency in postmenopausal women. However, the physiological mechanism of DHEA in osteoporosis remains elusive, especially in terms of intestinal calcium absorption. Therefore, we investigated the effect of DHEA administration on calcium absorption in ovariectomized (OVX) female rats using an estrogen receptor antagonist.[Methods] Female Sprague-Dawley rats (n=23, 6 weeks old) were randomized into three groups: OVX control group (OC, n=7), OVX with DHEA treatment group (OD, n=8), and OVX with DHEA inhibitor group (ODI, n=8) for 8 weeks.[Results] Intestinal calcium accumulation, as well as the rate of absorption, demonstrated no significant differences during the experimental period among investigated groups. The bone mineral density (BMD) of the tibia at the proximal metaphysis was higher in the OD group than that in the OC group (p<0.05); however, BMD of the ODI group showed no significant difference from investigated groups. Furthermore, the BMD of the tibia at the diaphysis did not significantly differ among these groups.[Conclusion] We revealed that DHEA administration does not involve intestinal Ca absorption, although this treatment improves BMD levels in OVX rats. These observations indicate that the effect of DHEA on the bone in postmenopausal women is solely due to its influence on bone metabolism and not intestinal calcium absorption.

The role of postnatal estrogen deficiency on cranium dimensions.

The aim of this study was investigate the cranium dimensions of adult female rats, who suffered estrogen deficiency during the prepubertal stage, to assess the impact of estrogen deficiency on craniofacial morphology. Twenty-two female Wistar rats were divided into ovariectomy (OVX) (n = 11) and sham-operated control (n = 11) groups. Bilateral ovariectomy were performed in both groups at 21 days old (prepubertal stage), and rats were euthanized at an age of 63 days (post-pubertal stage). Micro-CT scans were performed with rat skulls, and the cranium morphometric landmark measurements were taken in the dorsal, lateral, and ventral view positions. Differences in measurements between the OVX and sham control groups were assessed using t test with an established alpha error of 5%. The measures of the rats' skull showed that the inter-zygomatic arch width and anterior cranial base length were significantly larger in OVX group (p = 0.020 and p = 0.050, respectively), whereas the length of parietal bone was significantly higher in the sham group (p = 0.026). For the remaining measurements no significant differences between groups were detected (p > 0.05). This study provides evidence that ovariectomized rats had alterations in cranial bone dimensions, demonstrating that estrogens during puberty are important for skull morphology. To understand the role of estrogen on the postnatal cranium development will impact the clinical diagnose and therapy during childhood and adolescence.

Chitosan alleviated menopausal symptoms and modulated the gut microbiota in estrogen-deficient rats.

Menopause disturbs energy, glucose, and lipid metabolisms and changes the composition of the gut microbiota, but dietary fibers without phytoestrogens may ameliorate menopausal metabolic disorders. The objective of the present study was to assess whether consuming the prebiotics chitosan and citrus pectin can improve postmenopausal symptoms, possibly by modulating the gut microbiota in ovariectomized (OVX) rats, and the mechanism of action was examined. The OVX rats were given 4.5% cellulose (OVX-Control), chitosan (OVX-Chitosan), or citrus pectin (OVX-Pectin) in a 43% fat diet and the sham rats were given the same diet as the OVX-Control for 12weeks. Sham-operated rats had the same diet as OVX-Control (Normal-Control). Body-weight, visceral fat mass, tail skin temperature, serum 17β-estradiol, glucose intolerance, and insulin tolerance were determined. Gut microbiota in the fecal samples was measured by NGS and analyzed with PICRUSt2. Short-chain fatty acids (SCFA) and metabolomic characteristics of serum were also measured with UPLC-mass spectrometry. Chitosan and citrus pectin were selected because the incubation of rat feces with these two prebiotics in vitro had shown increased butyrate production. OVX-Chitosan reduced the weight, visceral fat content, and tail skin temperature, and OVX-Chitosan and OVX-Pectin improved glucose tolerance, compared to the OVX-Control. Both alleviated dyslipidemia, compared to the OVX-Control. OVX-Chitosan and OVX-Pectin elevated serum propionate and butyrate concentrations but only OVX-Chitosan lowered serum acetate concentrations. In PICRUSt2, chitosan upregulated the functional genes of gut microbiota involved in valine, leucine, and isoleucine biosynthesis, whereas the OVX-Control exhibited significantly upregulated lipopolysaccharide biosynthesis. OVX-Pectin exhibited increased α-diversity in the fecal bacteria. Metabolomic analysis revealed higher serum urate concentrations in the OVX-Control group than the other groups, and serum arginine and leucine concentrations were higher in the OVX-Chitosan group (P < 0.05). Chitosan and citrus pectin consumptions improved menopausal symptoms by improving the diversity and composition of the gut microbiota, and serum metabolites and SCFA originating from fecal bacteria. Chitosan was more effective for improving menopausal symptoms than citrus pectin.

Assessment of safety and efficacy of pine bark extract in normal and ovariectomized mice.

We evaluated the influence of pine bark extract (PBE) on organs, the cytochrome-P450 (CYP) activities in liver and estrogenic effects in normal and ovariectomized (OVX) female mice. The PBE did not affect organ weights and liver-function indexes (activities of alkaline phosphatase, aspartate amino transferase, and alanine amino transferase) at doses; 0.04%, 0.4%, and 2.0% PBE in the diet, in normal and OVX female mice. In the OVX mice, CYP1A1 activity was significantly higher in the 0.4% and 2.0% PBE groups than in the OVX control group, and in the 0.4% and 2.0% PBE groups were significantly higher than in the 0.04% PBE group. CYP1A2 and 3A4 activities were significantly higher in the 2.0% PBE group than in all other groups. The PBE did not affect uterine weight and femoral bone mineral density at all PBE doses. These results showed that the dose of PBE at the recommended human intake, had no toxic and estrogenic effects in normal female and OVX mice, however, it may need attention to use the excess intake of PBE with some drugs in postmenopausal women.