Calcium-fortified fresh milk ameliorates postmenopausal osteoporosis via regulation of bone metabolism and gut microbiota in ovariectomized rats

The aging of the global population has made postmenopausal osteoporosis prevention essential;however,pharmacological treatments are limited.Herein,we evaluate the effect of calcium-fortified fresh milk(FM)in ameliorating postmenopausal osteoporosis in a rat model established using bilateral ovariectomy.After 3 months of FM(containing vitamin D,and casein phosphopeptides,1000 mg Ca/100 g)or control milk(110 mg Ca/100 g milk)supplementation,bone changes were assessed using dual-energy X-ray absorptiometry,microcomputed tomography,and bone biomechanical testing.The results revealed that FM can regulate bone metabolism and gut microbiota composition,which act on bone metabolism through pathways associated with steroid hormone biosynthesis,relaxin signaling,serotonergic synapse,and unsaturated fatty acid biosynthesis.Furthermore,FM administration significantly increased bone mineral content and density in the lumbar spine and femur,as well as femoral compressive strength,while improving femoral trabecular bone parameters and microarchitecture.Mechanistically,we found that the effects may be due to increased levels of estrogen,bone formation marker osteocalcin,and procollagen typeⅠN-propeptide,and decreased expression of the bone resorption marker C-telopiptide and tartrate-resistant acid phosphatase 5b.Overall,the findings suggest that FM is a potential alternative therapeutic option for ameliorating postmenopausal osteoporosis.

Erxian decoction potentially prevents postmenopausal osteoporosis by modulating miR-335: a study based on bioinformatics analysis and preliminary clinical case validation

Background:miRNAs are closely related to bone metabolism.Studies have shown that Erxian decoction can improve bone metabolism,possibly achieving this regulatory effect through miRNA targets.Netinfer was used to predict the miRNA targets of Erxian decoction for the treatment of postmenopausal osteoporosis,and the results were validated by clinical trials.Methods:In this study,we identified possible targets of Erxian decoction in osteoporosis by means of network pharmacological analysis and bioinformatic prediction.Fifteen cases of postmenopausal osteoporosis with kidney Yin and Yang deficiency(In traditional Chinese medicine,kidney Yin nourishes and moistens the tissues of the internal organs of the body,while kidney Yang promotes and warms the tissues of the internal organs of the body.)were treated with Erxian decoction for four weeks,and serum bone metabolism indices(P1NP,osteocalcin,andβ-CTX)and miRNA-335-5p expression were measured before and after treatment.Results:The constructed miRNA postmenopausal osteoporosis related gene network of the effective compound of the Erxian decoction has 296 points and 981 edges.The 39 postmenopausal osteoporosis related genes regulated by miRNA-335-5p were enriched in ossification,while the signaling pathways were enriched in rheumatoid arthritis,the Toll signaling pathway,the HIF-1 signaling pathway,and the MAPK signaling pathway.After taking Erxian decoction,the expression of the serum bone formation index(P1NP,osteocalcin)and miRNA-335-5p gene expression levels increased significantly.The alterations in P1NP and osteocalcin were correlated with the changes in miRNA-335-5p.Conclusion:Circulating miRNA-335-5p may serve as an important target of Erxian decoction in the treatment of postmenopausal women.The effect of Erxian decoction on bone formation is significant,but the underlying mechanism requires further investigation.

Postmenopausal Osteoporosis and Osteopenia Management with a Combination of Once-Monthly Oral Ibandronate and Cholecalciferol—A Systematic Review

Postmenopausal osteoporosis and osteopenia are chronic and uncurable conditions that invariably lead to an increased risk of vertebral, hip, and femoral neck fracture if left untreated. Clinical guidelines establish, in general, pharmacological combinations allied to lifestyle changes as the mainstay of their management, and also increasing bone marrow density, lowering fracture risk, and improving quality of life are their main therapeutic goals. The objective of this systematic review was to analyze the available data in the scientific medical literature regarding the role of the ibandronate and cholecalciferol combination in postmenopausal osteoporosis and osteopenia management. Based on our results, we concluded that the above combination is safe and feasible for the clinical control of both conditions.

A Clinical Study of Yigu Capsule (益骨胶囊) in Treating Postmenopausal Osteoporosis

Objective: To observe the efficacy and safety of Yigu capsule (益骨胶囊, YGC), a Chinese herbal compound preparation, in treating postmenopausal osteoporosis (PMO) and to explore its possible mechanism. Methods: The clinical study was conducted in a prospective, randomized, double blinded method lasting for 6 months with placebo and positive control. Two hundred and ten PMO patients with confirmed diagnosis were assigned into the YGC group, the calciferol group and the placebo group. Besides being administered element calcium, they were treated with YGC, calciferol capsule and placebo capsule respectively. And such symptoms as newly found fracture and ostealgia, bone mineral density (BMD) of the 2nd-4th lumbar vertebrae (L_ 2-4 ) and upper femur, blood and urinary indexes for bone metabolism, sex hormone level and adverse reaction that occurred in patients were observed.Results: In the YGC group, the total effective rate was 95.50%, with no new occurrence of fractures, which was significantly better than that in the other two groups (P<0.05). Moreover, in the YGC group,the increase rate of BMD was 9.83% in L_ 2-4 , 4.09% in femoral neck, 4.60% in Wards triangle, 3.00% in greater trochanter, which was also better than that in the placebo group (P<0.05, P<0.01). As compared with the placebo group, levels in the YGC group of urinary oxyproline hydroxyproline/creatinine, urinary calcium/creatinine were significantly lower, serum and bone alkaline phosphatase, osteocalcin, estradiol and estradiol/testosterone were significantly higher, but no difference was shown in the comparison of testosterone level. In the observation period, no abnormality in blood or urine routine, liver or renal function was found. Only mild, transient gastro-intestinal response occurred in individual patients, but it did not affect the treatment. Conclusion: YGC could treat PMO effectively, as it could obviously increase the BMD of lumbar vertebrae and coxafemoral bone, elevate the alleviating rate of ostealgia and incessant motion time, yet causing no newly found compressive fracture of vertebrae, or and any related adverse reaction. YGC could not only promote the formation, but also inhibit the absorption of bone as well as increase the sex hormone level. Therefore, it is a pure Chinese herbal compound preparation worthy of further research and development.

Efficacy and Safety of Risedronate Sodium in Treatment of Postmenopausal Osteoporosis

Summary： To evaluate the efficacy and safety of risedronate sodium in treatment of postmenopausal osteoporosis, one-year randomized, double blind clinical trial was performed among 54 women with postmenopausal osteoporosis. The changes were compared in bone mineral density （BMD）, bone metabolism markers and adverse events after 12 months oral administration of risedronate sodium. BMD was measured by dual energy X-ray absorptionmetry （DEXA） and bone turnover marker was detected. The results showed that there was a significant increase in BMD of the lumbar spine （3.29 %±41.18 %, 4.51%±1.64 % respectively） after 6 and 12 months in the risedronate treatment group versus placebo control group （-0.62 %±0.24 %, 0.48 %±0. 18 % respectively）. Bone turnover was decreased to a stable nadir over 6 and 12 months for resorption markers [N-Telopeptide （NTx）, P〈0.05] and over 12 months for formation marker （ALP, P〈0.05; BGP, P〈0. 05）. The safety profile of risedronate sodium was similar to that of placebo. There were no trends toward increased frequency of any adverse experience except for gastrointestinal symptoms （7.1%）, rash （7.1%） and hematuria （3.6 %）, which were usually mild, transient, and resolved with continued treatment. It was concluded that risedronate was an efficacious and safe drug in treatment of postmenopausal osteoporosis.

Network pharmacological study of Zuogui pill and Yougui pill for the treatment of postmenopausal osteoporosis

Background:Postmenopausal osteoporosis(PMO)is the most common primary osteoporosis in older women.This condition imposes a huge economic and medical burden on society.Aside from western medicine,traditional Chinese medicine is also widely used for the treatment of PMO,especially in Asian countries.Zuogui pill(ZGP)and Yougui pill(YGP)are classical formulas for the treatment of PMO with potent clinical effects.This study aimed to explore the potential active compounds,potential targets,and potential mechanisms of ZGP and YGP for the treatment of PMO.Methods:Compounds from ZGP and YGP were collected from three online databases,and these compounds were screened by oral bioavailability and drug-likeness.Their corresponding targets were determined from the Medicine Systems Pharmacology Database and Analysis platform.The corresponding targets for PMO were obtained from two disease databases.The target protein-protein interaction was identified through the STRING platforms and the core targets were ascertained by analyzing the protein-protein interaction network by using Cytoscape software.PMO-related genes were identified via Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses to identify specific biological processes,cellular components,molecular functions and key signalling pathways of ZGP and YGP for PMO treatment.Results:A total of 68 compounds were screened from ZGP with 168 putative potential target genes,whereas 99 compounds were screened from YGP with 214 potential target genes associated with PMO.Most of the core components included quercetin and kaempferol,and most of the core targets were TP53,AKT1,MAPK1,JUN,TNF,HSP90AA1,APP,IL6,VEGFA,RELA,MAPK8,NR3C1,EGFR,CXCL8,ESR1,FOS and MAPK14.Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analyses revealed that ZGP and YGP treat PMO primarily via regulation of bone formation and resorption,anti-inflammatory immunity and hormonal regulation.Conclusion:Our data provided insights into the mechanisms,by which ZGP and YGP treat PMO.This study points out a direction for further research into ZGP and YGP monomers and pathways and offers a new clinical treatment option for non-traditional Chinese medicine clinicians in the treatment of PMO.

Association of Serum Dkk-1 Levels with β-catenin in Patients with Postmenopausal Osteoporosis

Wnt signaling plays an important role in the bone development and remodeling. The Wnt antagonist Dkk-1 is a potent inhibitor of bone formation. The aims of this study were firstly to compare the serum Dkk-1 levels in postmenopausal osteoporosis patients with age-matched healthy controls, and secondly, to assess the possible relationship between Dkk-1 and β-catenin, sclerostin, or bone turnover markers [CTX, PINP, N-MID-OT and 25（OH）D] in the setting of postmenopausal osteoporosis. A total of 350 patients with postmenopausal osteoporosis and 150 age-matched healthy controls were enrolled, and the serum levels of Dkk-1, β-catenin, sclerostin, OPG, and RANKL were detected by ELISA, and bone turnover markers [CTX, PINP, N-MID-OT and 25（OH）D] were measured by Roche electrochemiluminescence system in two groups. Serum Dkk-1 levels were significantly higher in postmenopausal osteoporosis group than in control group（P〈0.001）. Univariate analyses revealed that serum Dkk-1 levels were weakly negatively correlated to β-catenin（r=–0.161, P=0.003） and OPG（r=–0.106, P=0.047）, while multiple regression analysis showed a negative correlation between serum Dkk-1 levels with β-catenin（β=–0.165, P=0.009） and BMD（β=–0.139, P=0.027）, and a positive correlation between serum Dkk-1 levels and CTX（β=0.122, P=0.040） in postmenopausal osteoporosis group. No similar correlations ware observed in control group. The results provided evidence for the role of Dkk-1 in bone metabolism and demonstrated the link of Dkk-1 and Wnt/β-catenin in some ways.

Repurpose dasatinib and quercetin:Targeting senescent cells ameliorates postmenopausal osteoporosis and rejuvenates bone regeneration

Clinical therapies developed for estrogen-deficiency-driven postmenopausal osteoporosis(PMO)and related diseases,such as bone degeneration,show multiple adverse effects nowadays.Targeting senescent cells(SnCs)and the consequent senescence-associated secretory phenotype(SASP)with a combination of dasatinib and quercetin(DQ)is a recently developed novel therapy for multiple age-related diseases.Herein,we found that estrogen deficiency induced-bone loss was attributed to a pro-inflammatory microenvironment with SASP secretions and accelerated SnC accumulation,especially senescent mesenchymal stem cells(MSCs)characterized by exhaustion and dysfunction in middle aged rats.Systematically targeting SnCs with DQ strikingly ameliorated PMO and restored MSC function.Local administration of DQ and bone morphogenetic protein 2(BMP2)in combination promoted osteogenic differentiation of MSCs and rejuvenated osteoporotic bone regeneration.Our results repurposed DQ as an attractive therapy for treating PMO and related diseases.

Liuwei Dihuang Pill(六味地黄丸)Treats Postmenopausal Osteoporosis with Shen(Kidney) Yin Deficiency via Janus Kinase/Signal Transducer and Activator of Transcription Signal Pathway by Up-regulating Cardiotrophin-Like Cytokine Factor 1 Expression

Objectives： To investigate the mechanism of Liuwei Dihuang Pill （六味地黄丸, LDP） in treating postmenopausal osteoporosis （PMOP） with Shen （Kidney） yin deficiency. Methods： In this study, 205 cases of PMOP were divided into the PMOP Shen-yin deficiency group （Group A）, PMOP Shen-yang deficiency group （Group B）, PMOP without Shen deficiency group （Group C）, and control group （Group N）. Real-time polymerase chain reaction （RT-PCR） and Western blot techniques were used to observe the effects of LDP treatment on the cardiotrophin-like cytokine factor 1 （CLCF1）, ankyrin repeat and SOCS box containing 1 （ASB1）, and proldneticin 2 （PROK2） genes and the Janus kinase/signal transducer and activator of transcription （JAK/STAT） signaling pathway. Results： The mRNA （P〈0.05） and protein （P〈0.01） expression levels of the CLCF1 gone in Group A were significantly lower than the corresponding levels in Group N. After LDP treatment for 3 months, the mRNA expression levels of the CLCF1 gone were obviously up-regulated （P〈0.01）. After 6-month treatment, the expression levels of CLCF1 mRNA and protein were significantly up-regulated （both P〈0.01）, and the average bone density of the top femur had significantly increased （P〈0.05）. In vitro, CLCF1 overexpression resulted in a significant increase in the total protein and phosphorylated protein levels of JAK2 and STAT3. Conclusions： The CLCF1 gone is an important gone associated with PMOP Shen-yin deficiency and the therapeutic effects of LDP may be mediated by up-regulation of CLCF1 gone expression and activation of the JAK/STAT signaling pathway.

Serum sclerostin levels associated with lumbar spine bone mineral density and bone turnover markers in patients with postmenopausal osteoporosis

Background Sclerostin, expressed exclusively by osteocytes, is a negative regulator of bone formation. To gain insights into the action of sclerostin in postmenopausal osteoporosis, we evaluated serum sclerostin levels in postmenopausal women and investigated its possible associations with bone turnover markers in patients with postmenopausal osteoporosis. Methods We detected serum sclerostin, and measured lumbar spine bone mineral density in 650 Chinese postmenopausal women. We also assessed serum levels of 13-isomerized C-terminal crosslinking of type I collagen, intact N-terminal propeptide of type I collagen, N-mid fragment of osteocalcin, 25-hydroxyvitamin D, and estradiol. Results Serum sclerostin levels were lower in postmenopausal osteoporotic women compared with non-osteoporotic postmenopausal women （（38.79＋7.43） vs. （52.86＋6.69） pmol/L, P 〈0.001）. Serum sclerostin was positively correlated with lumbar spine bone mineral density （r=0.391, P 〈0.001） and weakly negatively correlated with [3-isomerized C-terminal crosslinking of type I collagen, intact N-terminal propeptide of type I collagen, N-mid fragment of osteocalcin （t= -0.225, P 〈0.001; r= -0.091, P=0.046; r= -0.108, P=0.018; respectively） in postmenopausal osteoporosis. There was no significant association of serum sclerostin with age, body mass index, 25-hydroxyvitamin D, and estradiol （r= -0.004, P=0.926; r=0.067, P=0.143; r=0.063, P=0.165; r= -0.045, P=0.324; respectively）.Conclusion Sclerostin may be involved in the pathogenesis of postmenopausal osteoporosis and may play a role in bone turnover.

Clinical Biochemical Observation on the Treatment ofPostmenopausal Osteoporosis with Gusong (骨松)Ⅱ

Objective: To assess the effect of Gusong II in treating postmenopausal osteoporosis and try to reveal its mechanism. Methods: Thirty-five women, 49 to 62 years old, with 5. 6 years menopause, were treated as the subjects and followed-up for 6 months, 12 months or 24 months. The course of treatment ranged from6 months to 25 months with an average of 14 months. Some related biochemical parameters, such as serumestradiol (E2 ), testosterone (T), the ratio of fasting urinary calcium to creatinine (Ca/Cr), the ratio of 24hurinary calcium to creatinine (24h Ca/Cr), 24h urinary E2, and 24h urinary hydroxyproline (HOP) were measured before and after treatment. Results: After 6～9 months or 1～2 years of treatment, the levels of serum E2and 24h urinary E2 were raised significantly, while the levels of serum T and urinary Ca/Cr fell evidently afterone or two year's treatment. The levels of serum E2 and 24h urinary E2 were significantly correlated with thelevels of fasting urinary Ca/Cr, 24h urinary Ca/Cr, serum T, and 24h urinary HOP respectively. Conclusions:Gusong-II can elevate the serum E2 levels of postmenopausal women, reduce the discharge of urinary calcium andHOP, and inhibit bone resorption. The rise of the serum EZ level might be correlated with the conversion of T toE2.

Observation on Therapeutic Effect of Erxian Decoction on Relieving Low Back Pain after PVP of PMOP-derived Vertebral Fracture

[Objectives]To explore the clinical effect of Erxian decoction on relieving low back pain after percutaneous vertebroplasty(PVP)of vertebral compression fracture caused by postmenopausal osteoporosis(PMOP).[Methods]Ninety patients who were treated in Suzhou TCM Hospital from September 2021 to January 2023 were randomly divided into three groups:traditional Chinese medicine group(n=30),western medicine group(n=30)and blank group(n=30).The patients in all the three groups were treated with basic anti-osteoporosis drugs.The patients in the traditional Chinese medicine group were treated with Erxian decoction after PVP,and those in the western medicine group were treated with celecoxib to relieve pain after operation.The visual analogue(VAS)score,Oswestry dysfunction index(ODI)score,TCM syndrome score and serum indexes such as interleukin-6(IL-6)and estrogen E2 were recorded before treatment and 2 weeks,1 month and 3 months after treatment.[Results](i)In terms of pain relief,the VAS score of the western medicine group was lower than that of the traditional Chinese medicine group after 2 weeks of treatment,but there was no significant difference in VAS score between the two groups after 1 month and 3 months,and the pain improvement of the two groups was better than that of the blank group.(ii)After 3 months of treatment,the ODI score in the traditional Chinese medicine group was lower than that in the western medicine group,and the improvement of TCM syndrome in the traditional Chinese medicine group was better than that in the other two groups 1 and 3 months after treatment(P<0.05).(iii)The level of IL-6 in the western medicine group was lower than that in the other groups after 2 weeks,and there was no significant difference between the two groups after 3 months of treatment.After 3 months of treatment,the level of E2 in the traditional Chinese medicine group was higher than that before treatment and higher than that in the western medicine group and the blank group,but there was no significant difference between the two groups(P>0.05).[Conclusions]Both Erxian decoction and non-steroidal anti-inflammatory drugs can relieve residual low back pain after PVP,and their long-term effects are similar,but Erxian decoction has more advantages in alleviating pain,muscle and joint pain and sensory abnormalities in postmenopausal women.Moreover,it is safe and reliable,and is worthy of clinical application.

Effect of Gengnianchun Recipe (更年春方) on Bone Mineral Density, Bone Biomechanical Parameters and Serum Lipid Level in Ovariectomized Rats

Objective： To observe the effect of Gengnianchun Recipe （更年春方, GNC） on bone mineral density （BMD）, bone biomechanical parameters and serum lipid level in the bilaterally ovariectomized （OVX） rats and to explore the prophylactic and therapeutic action of GNC on ovariectomy induced osteoporosis and hyperlipidemia. Methods： OVX SD rats, 10- 12 months old, were divided into different groups and fed with GNC 2 g/d, GNC 1 g/d and Nilestriol 0. 125 mg/week, respectively for 4 months to observe the change of BMD and bone biomechanical parameters of the lumbar vertebrae, and the serum levels of total cholesterol （TO）, triglyceride（TG）, high-density lipoprotein cholesterol （HDL-C） and low-density lipoprotein cholesterol （LDL-C）, and to compare the effect of the two drugs on the morphology of the uterus. Results： There was marked reduction in BMD and biomechanical parameters in lumbar vertebrae （ P〈0. 01 ） and increase of serum TO and LDL-C levels （ P〈0. 01 ） in rats after OVX. GNC or Nilestriol significantly improved the decreased BMD and biomechanical parameters of the lumbar vertebrae （ P〈0.05 or P〈0. 01 ）, and reduced the serum TO and LDL-C levels （ P〈0. 01 ）. In the Nilestriol group, the wet weight of uterus got increased obviously （ P〈0.01 ）, the number of uterine glands increased, uterine columnar epithelium thickened, and the mitotic figures in the epithelial stroma and myointimal cells augmented. But no such effect in wet weight and morphology of uterus was found in the GNC group. Conclusion： GNC could increase the BMD and biomechanical parameters of the lumbar vertebrae, reduce the serum TO and LDL-C levels, yet produce no adverse reaction in stimulating proliferation and hypertrophy of uterus.

Iron accumulation and its impact on osteoporotic fractures in postmenopausal women

Postmenopausal osteoporosis is a kind of degenerative disease,also described as“invisible killer.”Estrogen is generally considered as the key hormone for women to maintain bone mineral content during their lives.Iron accumulation refers to a state of human serum ferritin that is higher than the normal value but less than 1000μg/L.It has been found that iron accumulation and osteoporosis could occur simultaneously with the decrease in estrogen level after menopause.In recent years,many studies indicated that iron accumulation plays a vital role in postmenopausal osteoporosis,and a significant correlation has been found between iron accumulation and fragility fractures.In this review,we summarize and analyze the relevant literature including randomized controlled trials,systematic reviews,and meta-analyses between January 1996 and July 2022.We investigate the mechanism of the effect of iron accumulation on bone metabolism and discuss the relationship of iron accumulation,osteoporosis,and postmenopausal fragility fractures,as well as the main clinical treatment strategies.We conclude that it is necessary to pay attention to the phenomenon of iron accumulation in postmenopausal women with osteoporosis and explore the in-depth mechanism of abnormal bone metabolism caused by iron accumulation,in order to facilitate the discovery of effective therapeutic targets for postmenopausal osteoporosis.

Estrogen receptor gene polymorphisms and bone mineral density in Chinese postmenopausal women

Objective To investigate the relationships between the polymorphisms of estrogen receptor (ER) gene, bone mineral density (BMD) and bone biochemical markers in Chinese postmenopausal women. Methods BMD of lumbar spine and femoral neck were measured using dual-energy X-ray absorptiometry (DEXA)in 186 Chinese postmenopausal women. The PvuⅡ and XbaⅠ polymorphisms of the ER gene were detected using polymerase chain reaction (PCR). Bone biochemical markers, serum alkaline phosphatase, osteocalcin and pyridinoline were measured by ELISA. Results The femoral neck(FN) BMD (Z score) was higher in pp compared to Pp (-0.01±0.12 vs. -0.35±0.09, P<0.05) while lumbar spine BMD (Z score) was higher in XX type compared to Xx and xx genotypes (0.01±0.45 vs -1.53±0.17, -1.29±0.10, P<0.001 and 0.001, respectively). Women without Px haplotype (n=79) had a higher BMD Z-score for the lumbar spine (-1.03±0.14 vs -1.45±0.11, P<0.05) and femoral neck (-0.01±0.11 vs -0.31±0.09, P<0.05) than those who had it (n=107). Conclusions The present study suggested that the pp and XX genotypes of ER gene might play a certain role in maintaining FN and lumbar spine BMD. ER genotypes without Px haplotype might be favorable to bone mass, while those with it might exert some harmful effect on bone mineral density.

Benefit period using alendronate to increase bone mineral density in women with osteoporosis?

Background Alendronate, a nitrogen-containing bisphosphonate is a specific inhibitor of bone resorption and now in the forefront of treatment of osteoporosis. In this study, we reported a significant increase in bone mineral density (BMD) of the spine and the hip in postmenopausal women taking alendronate at 10 mg/d for 1, 2 and 3 years. Methods Participants had received daily, oral, 10 mg dose of alendronate for one to three years and placed into one of three groups according to alendronate treatment duration: 41 women received alendronate for 1 year (group Ⅰ), 46 received alendronate for 2 years (group Ⅱ), and 30 received alendronate for 3 years (group Ⅲ). Measurements of bone density had been made by dual energy X-ray absorbtiometry once each year. Results The differences in L2-L4, L2, L4, femoral neck and trochanter BMD values before and after treatment for first group were significantly different. In second group, significant differences between initial and after treatment were found at the other sites except at the Ward’s triangle. In the third group, only a significant increase in the L2-L4, L2, L3, L4, trochanter BMD values between before treatment and at the end of third year was found. Comparisons between groups were performed with Student’s t test. ANOVA was used to test the age, menopause age, menopause duration and initial BMD values between the three groups. Calculated P values of less than 0.05 were considered statistically significant. Conclusions Alendronate had increased BMD significantly at the spine and hip in postmenopausal women over three years. Increases of BMD in third group were significant during the first and second years. However, continued therapy with alendronate had been required to maintain the gain in BMD over the third year.

Zuogui Wan(左归丸) improves trabecular bone microarchitecture in ovariectomy-induced osteoporosis rats by regulating orexin-A and orexin receptors

OBJECTIVE:To investigate the protective effects of Zuogui Wan(左归丸,ZGW)on bone loss induced by ovariectomy(OVX)and its mechanism via orexin-A and orexin receptors in the osteoporosis rat model.METHODS:Fifty Sprague-Dawley female rats were randomly divided into sham-operated(sham)group and four OVX subgroups.Rats subjected to sham and OVX were treated with the vehicle(OVX,1 mL/100 g weight,n=10),17β-estradiol(E2,50μg·kg-1·d-1),and ZGW at the doses of 2.3(ZGW-L)and 4.6(ZGW-H)g/kg/day lyophilized powder daily for 3 months,respectively.The serum biochemical parameters of 17β-estrogen(17β-E2),tartrate-resistant acid phosphatase(TRACP-5 b)and bone alkaline phosphatase(BALP)were measured by enzyme-linked immunosorbent assay.Hematoxylin-eosin staining was used to detect the changes in the morphological structure in bones.Microcomputed tomography was used to evaluate the bone mineral density and microarchitecture of the distal femur.The gene or protein expression of orexin-A,orexin receptor 1(OX1R),orexin receptor 2(OX2R),osteoprotegerin(OPG)and receptor activator of nuclear factor-κB ligand(RANKL)were assayed by either quantitative polymerase chain reaction or Western blot analysis.RESULTS:Compared with the OVX group,ZGW could reduce the serum level of TRACP-5b and increased the serum levels of BALP and 17β-E2(P<0.01).Meanwhile,ZGW could prevent bone loss and improved bone trabecular microarchitecture by increasing the trabeculae structure thickness and trabecular number,and arranging the trabeculae structure properly.Compared with the OVX group,it was upregulated for the orexin-A and OX2R mRNA or protein expression from the hypothalamus and tibiae,and OPG in the tibiae of ZGW groups(P<0.01,<0.05),while downregulated for the OX1R mRNA and protein expression in the tibiae and hypothalamus and RANKL from the tibiae(P<0.01).CONCLUSION:ZGW exhibited a protective effect for PMOP that may be mediated via orexin-A and orexin receptors regulation.