OVX Control Group Research Articles

Freeze-Dried Watermelon Supplementation Has Modest Effects on Bone and Lipid Parameters of Ovariectomized Mice.

This study investigated the effects of two doses of freeze-dried watermelon (WM) on bone and lipid parameters in ovariectomized (OVX) mice, a model of post-menopausal osteoporosis. Three-month-old C57BL/6 female mice (n=46) were sham-operated (SHAM) or OVX and randomly assigned to the control or WM diets for 12 weeks: SHAM-control, OVX-control, OVX+1%, or 10% (wt/wt) freeze-dried WM. All diets were isocaloric and isonitrogenous, and had the same calcium and phosphorus concentrations. Freeze-dried WM supplementation was not able to prevent the decrease in whole body, tibial, and lumbar bone mineral density due to estrogen deficiency. Micro-computed tomography analyses showed that WM was also not able to modulate changes in tibial trabecular and cortical bone microarchitecture due to ovariectomy. However, the lumbar trabecular micro-architecture analyses revealed that the WM-10% group had a similar connectivity density, trabecular number, trabecular separation, and structure model index as the SHAM group. Supplementation with 10% WM reduced plasma cholesterol and total liver lipids to the level of the SHAM group but was still similar to that of the OVX-control group. Supplementation with 10% WM increased liver catalase (CAT) mRNA levels but had no effects on mRNA levels of glutathione peroxidase (GPX) and the pro-inflammatory cytokine interleukin-6. There were no differences in plasma activity of the antioxidant enzymes GPX and CAT between all treatment groups. Our findings demonstrate some positive effects of watermelon for modulating lipids and attenuating lumbar vertebral bone loss arising from ovarian hormone deficiency.

Qu Feng Zhi Tong capsule increases mechanical properties of cortical bone in ovariectomised rats

Abstract Backgroud/objective Qu Feng Zhi Tong capsule (QF, Buchang Pharma, Xi'an, Shanxi, China) is officially approved Fufang formula in China for the treatment of rheumatoid arthritis (RA). As a Traditional Chinese Medicine formula, it consists of seven herbs, among which Erodium stephanianum, Dipsacus asper, and Viscum Coloratum were reported to be able to prevent bone loss and promote bone healing. Moreover, osthole from Angelica pubescens exerted anabolic effect on cortical bone by activating β-catenin-bone morphogenetic protein (BMP) signalling. So, we hypothesise that the formula QF has beneficial potential on preventing osteoporosis. Method In this study, high-performance liquid chromatography (HPLC) was used to identify the main constituents in QF, and SD female rats were ovariectomised (OVX) to examine antiosteoporotic efficacy. Thirty OVX rats were orally administrated with QF (162 and 648 ​mg/kg/day, L-QF, and H-QF) or estradiol (1 ​mg/kg/day, E2) for 12 weeks. Sham and OVX control group were treated with vehicle. Results Eleven compounds in QF extract were identified using HPLC, and the sum of their peak areas was covered 44.2% of total peak area of QF extract. As to in vivo results, compared to OVX group, H-QF had significantly higher ultimate load (P ​ Conclusion QF promoted cortical mechanical properties because of its positive effects on cortical thickness, porosity, and skeletal muscle. H-QF increased cortical thickness by promoting bone formation of periosteum and inhibiting bone absorption of endosteum in OVX rats. The translational potential of this article This study provided theoretical basis for QF as a conventional drug in osteoporosis to enhance cortical bone mechanical and avoid long bone fracture. As cortical diaphyseal bone is an important target of RA, this study also provides a possibility that QF is beneficial to RA by its positive effects on cortical bone to prevent bone erosions in RA patients.

Enhanced Bone Formation in Osteoporotic Mice by a Novel Transplant Combined with Adipose-derived Stem Cells and Platelet-rich Fibrin Releasates

Osteoporotic fracture is the main complication of osteoporosis (OP) and accounts for millions of injuries annually. Local intervention by intra-marrow injection has been a good option for preventing osteoporotic bone loss when the osteoporotic femoral fracture has been treated. In this study, tail vein transplantations were examined to evaluate the cell-based therapeutic approach for treating OP with adipose-derived stem cells (ADSCs) and platelet-rich fibrin releasates (PRFr) in an ovariectomized (OVX) mice model. Thirty-six 12-wk-old female ICR mice were randomly divided into six groups: untreated control; sham-operated; OVX-control; OVX-ADSCs; OVX-PRFr; and OVX-ADSCs+PRFr. Starting 8 wk after ovariectomy, the OVX mice received tail vein injections once each week for four consecutive weeks, then were evaluated radiographically and histopathologically 8 wk after the first injection. We also assessed changes to bone trabeculae in the proximal tibial growth plate. In OVX mice treated with ADSCs or PRFr alone, or with a combination of ADSCs and PRFr, the trabecular bone mineral density (BMD), bone volume ratios (BV/TV), and numbers (Tb.N) in the proximal tibia areas were significantly higher than that in the OVX-control group. Significant differences between OVX-treated mice and OVX controls were found for trabecular separation, but not for trabecular thickness. These results indicate that ADSCs or PRFr treatment enhances bone microarchitecture in OP. The treatment of bone loss of OVX mice with ADSCs+PRFr induced greater bone consolidation with bone tissue production (P < 0.01) when compared to the others. Thus, we conclude that the transplantation of ADSCs combined with PRFr might provide an alternative strategy for the treatment of various bone disorders in OP with an unlimited source of cells and releasates.

A stereological study of the effects of antidepressants on postmenopausal rat kidney

ABSTRACTMany factors can cause depression including genes (DNA), brain chemistry or stress. Antidepressant drugs affect the brain, heart, liver and kidney. We investigated the effects of the antidepressant drugs, amitriptyline (AMI) and paroxetine (PARO) on kidney. We used 24 adult female rats that were ovariectomized bilaterally 7 days before the experiment. The ovariectomized (OVX) animals and healthy control rats were divided into four equal groups for 4 weeks: control group, OVX control group (sham), AMI group and PARO group. Following the experimental period, the Cavalieri method was applied to sections of the kidney. PARO produced adverse effects on distal and proximal tubule volume, but AMI had no effect on the volume of distal and proximal tubules. Both PARO and AMI decreased the volume of Bowman spaces. PARO also damaged the kidney tubules and cells.

Therapeutic Effects of Low-Intensity Pulsed Ultrasound on Osteoporosis in Ovariectomized Rats: Intensity-Dependent Study

This study investigated the effects of low-intensity pulsed ultrasound (LIPUS) of different spatial-average-temporal-average intensity (ISATA) ranging from 15–150 mW/cm2 on the treatment of osteoporosis in ovariectomized rats. Healthy 3-mo-old female Sprague–Dawley rats were randomly divided into nine groups (n = 12 per group): sham-ovariectomy (OVX) control group, OVX control group and OVX groups treated with LIPUS at seven different intensities (ISATA: 15, 30, 50, 75, 100, 125 and 150 mW/cm2, respectively). LIPUS was applied to bilateral femurs 12 wk post-OVX for 20 min/d for 6 wk. Micro-computed tomography, biomechanical tests, serum biochemical analysis and grip strength tests were performed to evaluate the therapeutic effects of LIPUS at different intensities. Results revealed that LIPUS intensity yielded strong correlations with bone mineral density and bone microstructure (R2 = 0.57–0.83) and bone mechanical strength (R2 = 0.80–0.97), and that the intensity of 150 mW/cm2, instead of the 30 mW/cm2 widely used in bone fracture healing, was most effective in maintaining bone mass among all the LIPUS signals between 15 and 150 mW/cm2. This suggests that higher ultrasound intensity (i.e., 150 mW/cm2) may be more effective than lower intensity in mitigation of osteopenia and osteoporosis.

Ojayeonjonghwan, an oriental medicine composed of five seeds, protects against vasomotor and neurological disorders in estrogen-deficient rats.

Menopausal symptoms impair the quality of life of many women, and although conventional treatments are often effective, their use is limited by adverse effects. Ojayeonjonghwan, OJa, is a traditional Oriental medicine that is used for both male and female reproductive health and has a long history of safe use. We evaluated the effectiveness of two variations of OJa (OJa1 and OJa2) for treating menopausal symptoms in ovariectomized (OVX) rats. Both OJa preparations were effective for relieving indicators of hot flashes and depression, and for preventing loss of bone mineral density and lean body mass. Only OJa 2 prevented memory dysfunction. These results show that the traditional Oriental medicine, Ojayeonjonghwan, has the potential to relieve menopausal symptoms in women and should be further evaluated in human clinical trials as an alternative to convention therapies in women for whom conventional therapies are not indicated or found to be ineffective.

Effect of Long-Term Green Tea Polyphenol Supplementation on Bone Architecture, Turnover, and Mechanical Properties in Middle-Aged Ovariectomized Rats.

We investigated the effects of 6-month green tea polyphenols (GTP) supplementation on bone architecture, turnover, and mechanical properties in middle-aged ovariectomized (OVX) rats. Female rats were sham-operated (n = 39, 13/group) or OVX (n = 143, 13/group). Sham-control and OVX-control rats (n = 39) receiving no GTP were assigned for sample collection at baseline, 3, or 6months. The remaining OVX rats (n = 104) were randomized to 0.15%, 0.5%, 1%, and 1.5% (g/dL) GTP for 3 or 6months. Blood and bone samples were collected. Relative to the OVX-control group, GTP (1% and 1.5%) lowered serum procollagen type 1 N-terminal propeptide at 3 and 6months, C-terminal telopeptides of type I collagen at 3months, and insulin-like growth factor-I at 6months. GTP did not affect bone mineral content and density. At 6months, no dose of GTP positively affected trabecular bone volume based on microCT, but a higher cortical thickness and improved biomechanical properties of the femur mid-diaphysis was observed in the 1.5% GTP-treated group. At 3 and 6months, GTP (0.5%, 1%, and 1.5%) had lower rates of trabecular bone formation and resorption than the OVX-control group, but the inhibitory effects of GTP on periosteal and endocortical bone mineralization and formation at the tibial midshaft were only evident at 3months. GTP at higher doses suppressed bone turnover in the trabecular and cortical bone of OVX rats and resulted in improved cortical bone structural and biomechanical properties, although it was not effective in preventing the ovariectomy-induced dramatic cancellous bone loss.

“Osteo-protective effect of Curcumin; Fenugreek and their combination on ovariectomized female rats”.

THE MOST common type of osteoporosis is the bone loss associated with ovarian hormonal deficiency at menopause. It is characterized by an imbalance between bone resorption and formation, resulting in a decrease of bone mass and a deterioration of bone microarchitecture. Purified curcumin, Fenugreek extract and their combination were evaluated for improvement of bone loss on ovariectomized rats as a model of postmenopausal osteoporosis. 60, female Wistar rats weighing 130150-g were ovariectomized (OVX) except sham-operated (SH) group. Rats fed a soya bean deficient diet and randomly divided into 5 groups. After 3 months of ovariectomy, SH group and OVX control group was received 1 % DMS0 (1 ml/kg P.O.), while, other OVX groups were received curcumin (100mg/kg p.o.), alcoholic fenugreek extract (200mg/kg p.o.) and their combination (Received the same doses of drugs). Fumer, tibia and total bone mineral density showed significant decrease in OVX control vs. sham control, while the combination of curcumin and fenugreek showed significant increase compared with OVX control group. Data obtained showed that curcumin, significantly, decreased the bone loss. In addition, curcumin combined with alcoholic fenugreek extract showed a synergistic effect that consequently improved both bone turnover and bone strength.

LncRNA LINC00311 Promotes the Proliferation and Differentiation of Osteoclasts in Osteoporotic Rats Through the Notch Signaling Pathway by Targeting DLL3

Osteoporosis is a commonly occurring condition marked by a loss of bone density. Previous evidence has highlighted the roles played by microRNAs as potential treatment tools for the disease. At present, the influence of long non-coding RNAs (lncRNAs) on the progression of osteoporosis remains largely unclear. Thus, an investigation was conducted into the target relationship between LINC00311, which has been reported to be highly expressed in osteoporosis, and delta-like 3 (DLL3), which is involved in the Notch signaling pathway, in connection with a series of bioinformatic methods. An osteoporotic rat model was established by means of ovariectomy (OVX) to evaluate the influence exerted by DLL3-binding LINC00311 on osteoclasts through the Notch signaling pathway. Osteoclasts were extracted from osteoporotic rats and transfected with the LINC00311-vector, shRNA-LINC00311, Notch activator, or a combination of the Notch activator and LINC00311-vector. Western blotting and RT-qPCR techniques were applied to determine the expression levels of LINC00311, DLL3, Notch1, Notch2, Jagged1, Hes-1 and TRAP in tissues and cells, while cell activity was detected by MTT assay. The cell cycle as well as the rate of apoptosis was detected by flow cytometry. The successfully established osteoporotic rats were designated into the OVX-siRNA, OVX-LINC00311 and OVX-control groups to observe the effects of LINC00311 on the proliferation and differentiation of osteoclasts. Cells transfected with the LINC00311-vector exhibited increased expression levels of Notch2 and TRPA as well as increased cell activity, while decreased expression levels of DLL3, Notch1, Jagged1 and Hes-1, along with a decreased cell apoptosis rate, were observed. The opposite tendencies of these parameters were observed in the cells treated with shRNA-LINC00311. A key observation was made when the Notch signaling pathway was activated, in that the cell activity was decreased while the rate of apoptosis increased. In comparison with the OVX-control group, the expression levels of LINC00311, Notch2 and TRAP as well as the positive expression rate of TRAP all exhibited reductions, while those of DLL3, Jagged1 and Notch1 were elevated in the OVX-siRNA group. Compared with those in the sham group, in the OVX-control and OVX-LINC00311 groups, LINC00311 and the expression levels of Notch2 and TRAP were increased; however, decreased levels of DLL3, Jagged1 and Notch1 were noted. Taken together, the key findings of the present study suggest that LINC00311 induces proliferation and inhibits apoptosis of osteoclasts via the regulation of the Notch signaling pathway by inhibiting DLL3 expression, ultimately demonstrating that LINC00311 and its target gene DLL3 may serve as independent factors in cases of osteoporosis.

Synergistic Effects of Genistein and Zinc on Bone Metabolism and the Femoral Metaphyseal Histomorphology in the Ovariectomized Rats.

In this study, we evaluated the synergistic effects of genistein and zinc on bone metabolism, bone mineral density, and the femoral metaphyseal histomorphology in the ovariectomized rats. Sixty female Sprague-Dawley rats, aged 13weeks, were divided into sham-operated group (SHAM), ovariectomized group (OVX), genistein-treated group (OVX-Gen), zinc-treated group (OVX-Zn), genistein combined zinc-treated group (OVX-Gen-Zn), and 17β-estradiol-treated group (OVX-E) (n=10). Genistein, zinc, and 17β-estradiol injections were started immediately after OVX and continued for 10weeks. The OVX-Gen group was subcutaneous injections of genistein (5mg/kg once every day). The OVX-Zn group was given intragastric ZnSO4 (0.25mg/kg once every day) after bilateral ovariectomy. The OVX-Gen-Zn group was subcutaneous injections of genistein (5mg/kg), at the same time intragastric ZnSO4 (0.25mg/kg once every day). OVX-E group were intraperitoneally injected with 17β-estradiol (10μg/kg) once every other day. The results showed that genistein and zinc did not alter body weight in OVX rats. The soluble ZnSO4 had no effect to uterus weight loss. Genistein and zinc supplementation significantly increased the BMD of the femur and lumbar spine in OVX rats (P<0.05, vs. OVX control group). The BMD of the lumbar spine and femur in the OVX-Gen-Zn group is higher than that in the OVX-Gen and OVX-Zn groups (P<0.05), and there were no differences of BMD among the OVX-Gen-Zn, OVX-E, and SHAM groups (P>0.05). The femoral metaphyseal morphology and bone histomorphometric parameters revealed that the trabecular volume and thickness in the OVX-Gen-Zn and OVX-E groups were similar to that of SHAM group, and the OVX-Gen-Zn group had more trabecular volume than the OVX-Gen and OVX-Zn groups. Serum zinc was found to be significantly reduced in the OVX group when compared with SHAM. Serum zinc levels were enhanced in the OVX-Zn, OVX-Gen-Zn, and OVX-E groups and restored to normal levels. Serum calcium and serum phosphorus in the OVX-Gen-Zn group was greater than that in the OVX-Gen and OVX-Zn groups (P<0.05); the serum calcium, phosphorus, and ALP in the OVX-Gen-Zn group were similar to that of SHAM and OVX-E groups (P>0.05). The OVX group had higher osteocalcin, CTX, and RANKL than the SHAM group (P<0.01). They were lower in the OVX-Gen and OVX-Zn groups (P<0.05), and they were significantly lower in the OVX-Gen-Zn and OVX-E groups, than in the OVX group. There were no significant differences of osteocalcin, CTX, and RANKL among the OVX-Gen-Zn, OVX-E, and SHAM groups (P>0.05). OPG and OPG/RANKL ratios were significantly higher in the OVX-Gen and OVX-Zn groups than that in the OVX group (P<0.05). The OPG and OPG/RANKL ratio in the OVX-Gen-Zn group is higher than that in the OVX-Gen and OVX-Zn groups (P<0.05, vs. OVX-Gen group; P<0.01, vs. OVX-Zn group). The differences of the OPG and OPG/RANKL ratios between the OVX-Gen-Zn, OVX-E, and the SHAM groups were not significant (P>0.05). The results above indicate that the genistein and zinc have synergistic effects on both stimulation of bone formation and suppression of bone resorption in ovariectomized rats, thereby increasing bone mass.

Effect of in vivo low-level laser therapy on bone marrow-derived mesenchymal stem cells in ovariectomy-induced osteoporosis of rats

Postmenopausal osteoporosis (PMOP) is considered by decreased bone strength that escalates the threat of fractures. Positive effects of photobiomodulation (PBM) with pulse wave have been demonstrated in cell culture and animal models. The aim of this study was to assess the in vivo effects of PBM on viability and calcium ion release of ovariectomy induced osteoporosis (OVX) - bone marrow derived mesenchymal stem cells (BMMSCs). Material and Methods18 female rats were distributed into the following groups: 1) control healthy, 2) LASER-healthy (890nm, 80Hz, 1.5J/cm2, three days weekly, 60days), 3) control OVX, 4) LASER-OVX, 5) Alendronate (Alen.)-OVX [0.5mg/kg, 5days per week, 60days], and 6) Alen.+LASER-OVX. Ovariectomy was done on rats of groups 3, 4, 5 and 6. After that all rats were euthanized and their MSC harvested and cultured in complete osteogenic medium. In all groups, BMMSC viability, and calcium colorimetric assay were performed. ResultsWe observed a significant increase in optical density (OD) of BMMSCs viability in LASER healthy group compared to control-OVX, Alen.-OVX, LASER-OVX, LASER+Alen.-OVX, groups. LASER+Alen.-OVX group displayed a significant escalation in OD of BMMSCs viability compared to LASER-OVX, Alen.-OVX, and control-OVX groups. There were a significant increase in calcium ion release of LASER-healthy group compared to control healthy, control-OVX, Alen.-OVX, LASER-OVX, and LASER+Alen.-OVX groups. LASER+Alen.-OVX group displayed a significant escalation in calcium ion release compared to LASER-OVX, Alen.-OVX, and control-OVX groups. ConclusionPulse wave (PW) PBM significantly stimulated viability and cell proliferation of healthy BMMSCs compared to those of control-OVX, OVX-alendronate, OVX-LASER, and LASER+alendronate-OVX. In addition stimulatory effect of LASER+alendronate on viability and cell proliferation of OVX-BMMSCs compared to those of control-OVX, alendronate-OVX, and LASER-OVX groups were found.

Antiosteoporotic activity of a syringic acid diet in ovariectomized mice.

In recent years, the number of patients with osteoporosis has risen with the increase in average longevity. Therefore, the chemoprevention of osteoporosis using food materials or food components has become an increasingly important target. Syringic acid (SA) is a phenolic compound present in the fruit of the açaí palm Euterpe oleracea and the mycelium of the shiitake mushroom Lentinula edodes. This compound has no affinity for estrogen receptors and is potentially useful for disease prevention. However, little is known about the effects of a SA diet on bone metabolism, particularly bone resorption in vivo. Here, we demonstrated the effects of a SA diet on bone loss and uterine weight loss in ovariectomized (OVX) mice. Ten-week-old OVX mice were fed SA-containing diets (100mg/kg body weight/day) for 10weeks. After 10weeks of dietary SA, the body weight, food intake, and uterine weight of the OVX mice were unaffected; however, femoral bone mineral density (cortical bone density, cancellous bone density, and total bone density) was higher in the SA-fed groups than in the OVX-control group. Furthermore, histomorphometric analysis revealed that the number of osteoclasts and osteoblasts was decreased and increased, respectively, in the SA-fed groups. These results suggest that a SA diet suppresses bone loss by downregulating bone resorption and upregulating bone formation without affecting the uterus in OVX mice. Although further studies are needed, SA may be a compound that can be used to prevent or retard osteoporosis.

Antiosteoporotic effect of Hemidesmus indicus Linn. on ovariectomised rats

Pharmacological relevanceRoot of Hemidesmus indicus (L). R. Br., an herbal drug of India is traditionally used to prevent or treat female hormonal disturbance related disorders and assuaging symptoms of menopausal conditions. Aim of the studyLack of scientific data for potential use in bone loss, the present study was undertaken to investigate the effects of ethanol extract of Hemidesmus indicus root for osteoporosis in ovariectomised rats. Materials and methodsThirty wistar female rats were randomly divided into five groups. One group was sham operated and other four groups were undergone dorsal ovariectomy (OVX). After four weeks of post-operative recovery, three OVX groups, viz. Group1, Group2 and Group3 were treated for ninety days with standard raloxifene (RLX) 1mg/kg/day, Hemidesmus indicus root extract (HIRE) 100mg/kg/day and HIRE 200mg/kg/day respectively. Sham and OVX control groups were fed with equal 1% vehicle volume. Serum and urine biochemistry (alkaline phosphatase, tartrate resistant acid phosphatase, triglyceride, total cholesterol, calcium, phosphorous and hydroxyproline), weight of body, uterus and vagina, femur parameters, three point bending of tibia and fourth lumbar vertebra compression were examined. Furthermore, bone architecture and uterine hypertrophy were examined by histopathological studies. ResultsApplication of HIRE has increased bone strength and restored bone turnover markers such as alkaline phosphatase, tartrate resistant acid phosphatase and hydroxyproline moreover, improved femur parameters indicated mineralized bones, similar to the results observed with raloxifene treatment. Histopathology studies showed fibrocartilaginous proliferation of trabecular bone and absence of uterine hypertrophy. ConclusionThe results strongly suggest that HIRE prevents bone loss in OVX induced osteoporosis without estrogen like side effects. It might be a potential remedy like raloxifene for postmenopausal or estrogen deficiency caused osteoporosis.

Effects of exercise on mature or precursor brain‑derived neurotrophic factor pathways in ovariectomized rats.

Ovariectomy (OVX) is a method used to block estrogen in female rats that induces hippocampal dysfunction and affects brain‑derived neurotrophic factor (BDNF) pathways. The majority of previous studies investigating OVX focused on BDNF expression in the hippocampus and cognitive function. The present study focused on the pathways of each BDNF type, precursor (proBDNF) and mature (mBDNF), and the effects of regular exercise in the hippocampus of ovariectomized rats. Female Sprague‑Dawely rats were used and OVX surgery was performed. After 1week of recovery from surgery, two groups of rats that received OVX surgery were subjected to regular treadmill exercise for 8weeks. The results of protein levels by western blotting indicated that the expression of proBDNF, p75 neurotrophin receptor (p75NTR) and c‑Jun N‑terminal protein kinase (JNK) was increased, and mBDNF, tropomyosin‑related kinase B (TrkB) and nuclear factor‑κB expression was significantly reduced in the OVX control group compared with the sham control group SC (P<0.05). Thus, the survival pathway by mBDNF was impaired and the pro‑apoptotic response was activated by increased JNK expression due to proBDNF‑p75NTR binding in the hippocampus of ovariectomized rats. By contrast, exercise reduced activation of the pro‑apoptotic response and increased mBDNF‑TrkB expression in the hippocampus of ovariectomized rats. Thus, regular exercise may increase the activation of survival pathways via mBDNF and reducing the activation of the pro‑apoptotic pathway of proBDNF in the hippocampus of ovariectomized rats.

Preparation and in vivo evaluation of an orally available enteric-microencapsulated parathyroid hormone (1-34)-deoxycholic acid nanocomplex.

The N-terminal 34-amino-acid peptide fragment of human parathyroid hormone PTH (1-34), is used clinically to treat osteoporosis; however, it is currently administered by a once-daily subcutaneous injection, resulting in poor patient compliance. We have developed enteric microcapsules containing an ionic nanocomplex between PTH (1-34) and lysine-linked deoxycholic acid (LysDOCA) for the oral delivery of PTH (1-34). We measured the particle size of the PTH/LysDOCA complex and assessed its biological activity by determining the cAMP content in MC3T3-E1 cells. We also assessed its permeability across a Caco-2 cell monolayer and the bioavailability of the intrajejunally administered PTH/LysDOCA complex compared with PTH (1-34) in rats. In addition, the antiosteoporotic activity of the PTH/LysDOCA complex, encapsulated in an enteric carrier by coaxial ultrasonic atomization, was evaluated after it was orally administered to ovariectomized (OVX) rats. The formation of an ionic complex between PTH (1-34) and LysDOCA produced nanoparticles of diameter 33.0±3.36 nm, and the bioactivity of the complex was comparable with that of PTH (1-34). The Caco-2 cell permeability and AUClast value of the PTH/LysDOCA (1:10) nanocomplex increased by 2.87- and 16.3-fold, respectively, compared with PTH (1-34) alone. Furthermore, the OVX rats treated with oral PTH/LysDOCA-loaded enteric microcapsules showed an increase in bone mineral density (159%), bone volume fraction (175%), and trabecular number (174%) compared with those in the OVX control group. Therefore, the PTH/LysDOCA nanocomplex oral delivery system is a promising treatment modality for osteoporosis because it improves osteogenesis and trabecular connectivity.

Hawthorn Fruit Extract Elevates Expression of Nrf2/HO-1 and Improves Lipid Profiles in Ovariectomized Rats.

The purpose of this study was to investigate the effects of hawthorn (Crataegus pinnatifida Bunge) extract on the lipid profiles and antioxidant properties in ovariectomized (OVX) rats. After ovariectomy, the rats were randomly divided into four groups: the non-OVX control (Sham), the OVX-control (OVX), the OVX + 100 mg/kg b.w. of hawthorn extract (OL), and the OVX + 200 mg/kg b.w. of hawthorn extract (OH). The final body weights of the OVX group were significantly increased, but the increment was significantly decreased in hawthorn groups (p < 0.05). The serum total and low-density lipoprotein (LDL) cholesterol levels were significantly elevated in the OVX group, whereas the hawthorn groups showed a significant decrease in these levels (p < 0.05). The hepatic triglyceride (TG) and malondialdehyde (MDA) levels were significantly reduced in the hawthorn groups compared with the OVX group (p < 0.05). The mRNA expression of nuclear factor erythroid 2–related factor (Nrf2), heme oxygenase-1 (HO-1), and glutathione peroxidase (GPx) were significantly decreased in the OVX group, whereas the hawthorn groups exhibited a significant increase in expression (p < 0.05). The protein expressions of Nrf2, HO-1, and GPx were lower in the OVX group than the Sham group (p < 0.05). The oral administration of hawthorn extract reversed the suppression of protein levels. These results suggest that hawthorn extract could have protective effects in OVX rats by improving lipid profiles, decreasing oxidative stress, and improving the antioxidant defense system.

Hypolipidemic and antioxidant effects of Hawthorn extract in pre‐ and postmenopausal rats

Even though many previous studies have reported the antioxidative, hypolipidemic and cardiovascular protective effects of hawthorn, none of them were tested using ovariectomized model. The aim of the present study was to investigate lipid profiles and antioxidant properties of hawthorn extract in pre‐ and postmenopausal models of rat. Forty‐two female Sprague‐Dawley rats were randomly divided into six groups; non‐OVX control(Sham), Sham+100mg/kg b.w of hawthorn extract(SL) and Sham+200mg/kg b.w of hawthorn extract(SH), OVX‐control(OVX), OVX+100mg/kg b.w of hawthorn extract(OL) and OVX+200mg/kg b.w of hawthorn extract(OH). Treatment groups were administered with hawthorn extract every day for 8 weeks. Final body weight of OVX group were significantly higher than Sham group(p&lt;0.05), however the increment was significantly lower in both OL and OH group when compared with OVX group(p&lt;0.05). Compared to Sham group, uterine weight and 17β‐estradiol level were significantly decreased in all OVX groups(p&gt;0.05). These results show that ovariectomy is induced properly as intended. Serum total cholesterol(TC) level was significantly elevated in OVX group compared with Sham group(p&lt;0.05), whereas OL and OH groups showed significant decrease(p&lt;0.05). Hepatic triglyceride(TG) level was significantly reduced in both SL and SH group compared with Sham group(p&lt;0.05). In addition, OH group showed the lowest hepatic TG level and highest HDL‐cholesterol level compared with OVX group(p&lt;0.05). The OL and OH groups had significantly lower non‐HDL cholesterol level than OVX group(p&lt;0.05). Serum glutathione peroxidase(GPx) activity of OH group was significantly higher than OVX group(p&lt;0.05). The mRNA expression of GPx was highest in OL group(p&lt;0.05) and the mRNA expression of catalase(CAT) was higher in both SL and SH group than Sham group(p&lt;0.05). These results suggest that hawthorn extract could have protective effects on pre‐ and postmenopausal rats by improving lipid profiles and increasing antioxidant enzyme activity.

Effects of 18-month treatment with bazedoxifene on enzymatic immature and mature cross-links and non-enzymatic advanced glycation end products, mineralization, and trabecular microarchitecture of vertebra in ovariectomized monkeys

Bazedoxifene (BZA) is used for the treatment of post-menopausal osteoporosis. To elucidate changes in collagen, mineralization, and structural properties and their relationship to bone strength after treatment with BZA in ovariectomized (OVX) monkeys, the levels of collagen and enzymatic immature, mature, and non-enzymatic cross-links were simultaneously examined, as well as trabecular architecture and mineralization of vertebrae. Adult female cynomolgus monkeys were divided into 4 groups (n=18 each) as follows: Sham group, OVX group, and OVX monkeys given either 0.2 or 0.5mg/kg BZA for 18months. Collagen concentration, enzymatic and non-enzymatic pentosidine cross-links, whole fluorescent advanced glycation end products (AGEs), trabecular architecture, mineralization, and cancellous bone strength of vertebrae were analyzed. The levels of enzymatic immature and mature cross-links, bone volume (BV/TV), and trabecular thickness (Tb.Th) in BZA-treated groups were significantly higher than those in the OVX control group. In contrast, the trabecular bone pattern factor (TBPf), the structure model index (SMI), the enzymatic cross-link ratio, and the levels of pentosidine and whole AGEs in BZA-treated groups were significantly lower than those in the OVX control group. Stepwise logistic regression analysis revealed that BV/TV, Tb.Th, TbPf, and pentosidine or whole AGEs independently affected ultimate load (model R2=0.748, p<0.001) and breaking energy (model R2=0.702). Stiffness was affected by Tb.Th, enzymatic immature cross-link levels and their ratio (model R2=0.400). Treatment with BZA prevented OVX-induced deterioration in the total levels of immature enzymatic cross-links and AGEs accumulation and structural properties such as BV/TV, Tb.Th, and TbPf, which contribute significantly to vertebral cancellous bone strength.

Oxidant/antioxidant status, paraoxonase activity, and lipid profile in plasma of ovariectomized rats under the influence of estrogen, estrogen combined with progesterone, and genistein.

IntroductionThe aim of this study was to investigate whether estradiol (E2), E2 combined with progesterone (Prog) (E2/Prog), and genistein (Gen) treatment had antioxidative and anti-hyperlipidemic effects in the plasma of ovariectomized (OVX) rats.Materials and methodsAdult female Sprague–Dawley rats were divided into five groups. Rats in all groups, except for those in a sham group, underwent bilateral ovariectomy under general anesthesia. The groups were as follows: sham group; control OVX group; group treated with estrogen (0.014 mg/kg 17-β E2); group treated with a combination of E2 and Prog (0.014 mg/kg 17-β E2 plus 0.028 mg/kg drospirenone), and group treated with Gen (10 mg/kg/day). Plasma of rats of each treatment group was analyzed to determine the total antioxidant status, total oxidant status, paraoxonase activity, lipid profile, high-density lipoprotein (HDL-chol), low-density lipoprotein (LDL-chol), total cholesterol (Total-C), triacylglycerols, lipoprotein (a), and oxidative stress index.ResultsPlasma Total-C levels and body weight increased in all the OVX groups compared with the sham group (P<0.005). The group treated with E2 had significantly elevated total oxidant status, oxidative stress index, LDL-chol, and Total-C compared with the control group (P<0.005). Gen treatment might lead to lower LDL-chol and Total-C levels compared with E2 treatment.ConclusionsGen treatment might be preferred to E2 treatment for treatment of menopausal symptoms in patients at risk for cardiovascular diseases. However, considering the small sample size of this study, larger studies are needed in this area.

The effect of Lycii Radicis Cortex extract on bone formation in vitro and in vivo.

Osteoporosis is a common skeletal disease caused by decreased bone mass; it enhances the risk of bone fracture. This study aimed to discover novel herbal extract(s) for the treatment of osteoporosis. We screened 64 ethanol extracts of edible plants native to Korea for their ability to increase the cellular proliferation and differentiation of two osteoblastic cell lines: C3H10T1/2 and MC3T3-E1. We selected a Lycii Radicis Cortex (LRC), Lycium Chinese root bark as the primary candidate. Treatment with LRC extract showed enhanced alkaline phosphatase activity and increased expression of bone metabolic markers Alpl, Runx2, and Bglap genes in both osteoblastic cell lines. There was no effect on the osteoclastic differentiation of primary-cultured monocytes from the mouse bone marrows. Furthermore, the study examined the effect of LRC extract in vivo in ovariectomizd (OVX) mice for 8 weeks and 16 weeks, respectively. Bone mineral density (BMD) was significantly higher in LRC extract-administered group than in the non-LRC-administered OVX control group. The results indicated that LRC extract prevented the OVX-induced BMD loss in mice via promoting the differentiation of osteoblast linage cells. These results suggest that LRC extract may be a good natural herbal medicine candidate for the treatment of osteoporosis.