Postmenopausal Osteoporosis Group Research Articles

Gut microbiota and short-chain fatty acids signatures in postmenopausal osteoporosis patients: A retrospective study

Studies have shown that gut microbiota (GM) and its metabolites, short-chain fatty acids (SCFAs), are associated with the development of postmenopausal osteoporosis (PMO). This study explored the clinical and laboratory evidence of the relationship of GM and SCFAs to PMO and attempted to determine the potential mechanism of action. 18 patients (Collected from the First Affiliated Hospital of Guangdong Pharmaceutical University between January 2021 and August 2021) were included in this retrospective study, including 10 PMO women and 8 healthy young women as the healthy control (HC) group from Guangzhou, China. Bone mineral density (BMD) was determined by dual-energy X-ray absorptiometry. The composition of GM and its metabolites, SCFAs, in the fecal samples were measured by 16S rRNA gene sequencing and gas chromatography/mass spectrometry analysis, respectively. Compared with healthy control, PMO group had significantly decreased BMD in lumbar spines 1-4 (BMD_L) and femoral neck (BMD_F). 16S rRNA gene sequencing revealed that, compared with healthy control, PMO group had a markedly decreased abundance in Subdoligranulum, Norank_f_Muribaculaceae, and Alistipes at the genus level. Gas chromatography/mass spectrometry analysis indicated that the concentration of propanoic acid significantly dropped in PMO group. Additionally, we found that Subdoligranulum, Norank_f_Muribaculaceae, and Alistipes were positively correlated with BMD_L. Subdoligranulum and Norank_f_Muribaculaceae were also positively correlated BMD_F and propanoic acid, while Subdoligranulum is the only species that presented a strong correlation with the levels of acetic acid and butyric acid. Our findings indicated that, in postmenopausal women, there were evident changes in GM and SCFAs, and these changes were found correlated with patients’ BMD. These correlations provide novel insights into the underlying mechanism of PMO development, representative of early diagnostic markers and therapeutic targets that may improve the bone health in postmenopausal women.

Identification of biomarkers associated with oxidative stress-related genes in postmenopausal osteoporosis.

Osteoporosis (OP) is a prevalent metabolic disease, with aging and menopause being the major risk factors. Studies have shown that nearly one-third of postmenopausal women suffer from osteoporosis. However, there is a scarcity of research on antioxidant systems for the prevention and treatment of postmenopausal osteoporosis (PM-OP). To address this gap, we performed differential analysis using Limma to identify differentially expressed genes (DEGs) in PM-OP samples. We employed weighted correlation network analysis (WGCNA) to identify oxidative stress (OS)-related genes (OSRGs) highly correlated with PM-OP. The intersection of key modular genes and DEGs yielded differentially expressed OSRGs (DE-OSRGs) specific to PM-OP. We conducted GO and KEGG functional enrichment analyses on these genes. Additionally, we constructed a protein-protein interaction (PPI) network and utilized support vector machine recursive feature elimination (SVM-RFE) and random forest (RF) algorithms to identify signature genes. The diagnostic value of the signature genes was evaluated and validated using ROC curves. GSEA enrichment analysis was employed to explore the potential mechanisms associated with the signature genes. Finally, we constructed a regulatory network involving TF-miRNA-mRNA interactions for the signature genes and verified the biological roles of FOXO3 and DDIT3 in PM-OP and healthy groups using quantitative real-time PCR (qRT-PCR). Our analysis revealed 20 DE-OSRGs specific to PM-OP, obtained by intersecting modular and differential genes. The PPI network identified central genes (DDIT3, MAPK8, CDK2, SIRT1, and FOXO3) with more than 3 nodes. Through integration with machine learning algorithms, we identified DDIT3 and FOXO3 as signature genes. The ROC curve analysis indicated that the AUC value was greater than 0.7, suggesting the potential diagnostic value of these signature genes. Furthermore, GSEA results revealed their involvement in pathways related to the regulation of neutrophil activation, oxidative phosphorylation, MAPK signaling, mitochondrial matrix, and phagocytosis. Lastly, we constructed a regulatory network comprising 27 nodes (22 TFs, 3 miRNAs, and 2 mRNAs) and 28 edges. Additionally, qRT-PCR confirmed the significant up-regulation of FOXO3 and DDIT3 expressions in the PM-OP group compared to the healthy control group. In summary, this study employed bioinformatics analysis to identify OS-related biomarkers (DDIT3 and FOXO3) in PM-OP, providing new biological targets for clinical treatment.

Denosumab for osteoporosis in patients with primary hyperparathyroidism and mild-to-moderate renal insufficiency.

We aimed to assess the efficacy and safety of denosumab in postmenopausal women with primary hyperparathyroidism (PHPT)-related osteoporosis and chronic kidney disease (CKD). Women over 50 years of age with PHPT or postmenopausal osteoporosis (PMO) were retrospectively recruited into this longitudinal study. These PHPT and PMO groups were further categorized into subgroups based on the presence of CKD (Glomerular filtration rate (GFR) < 60 mL/min/1.73 m2). All patients were given denosumab over 24 months due to verified osteoporosis. The primary outcomes were changes in bone mineral density (BMD) and serum calcium levels. 145 postmenopausal women median age 69 [63;77] were recruited and assigned to one of the subgroups: PHPT patients with CKD (n = 22), PHPT patients without CKD (n = 38), PMO patients with CKD (n = 17) and PMO patients without CKD (n = 68). Denosumab treatment significantly increased BMD in patients with PHPT-related osteoporosis and CKD: median T-score L1-L4 from -2.0 to -1.35 (p < 0.001), femur neck from -2.4 to -2.1 (p = 0.012), radius 33% from -3.2 to -3 (p < 0.05)) at 24 months. Changes in BMD were similar in all four studied groups compared to baseline. A marked decline in calcium was noted in the primary study group of PHPT with CKD (median ΔCa = -0.24 mmol/L p < 0.001), compared to PHPT without CKD (median ΔCa = -0.08 mmol/L p < 0.001) and PMO with or without CKD. Denosumab treatment was well-tolerated with no serious adverse events. Denosumab treatment was similarly effective at increasing BMD in patients with PHPT and PMO with and without renal insufficiency. The calcium lowering effects of denosumab were most significant in patients with PHPT and CKD. The safety of denosumab did not differ among participants with and without CKD.

SCIMP: A Novel Targeted Gene for Postmenopausal Osteoporosis Progression.

The literature suggests that not all postmenopausal women suffer from osteoporosis, and the occurrence of postmenopausal osteoporosis is closely related to the genetic susceptibility of genes in the population and the cellular pathways of related genes. To systematically understand the functions of SCIMP gene for osteoporosis, both in vitro and in vivo experiments were analyzed in depth in this integrated study. The significantly differentially expressed genes of postmenopausal osteoporosis (PMOP) patients from GEO database were selected. Meanwhile, the primary target gene was also confirmed in clinically recruited individuals using ELISA method; 50 postmenopausal osteoporosis patients with a T-score of -2.5 were randomly enrolled; postmenopausal women with a T-score > -2.5 were included in the non-osteoporotic group (including osteopenia and normal bone mineral density). The associated processes and signaling pathways were deeply investigated with GO and KEGG enrichment analysis. The downstream signaling factors including Erk-1/2, Akt, and IkB-related signaling pathways for the potential gene were evaluated using MG-63 cell line; the MTT, CCK-8, and flow cytometry assays were performed to exam MG-63 cell viability, proliferation, as well as apoptosis, respectively, under different treatments. Based on the differentially expressed gene analysis for GEO database, PMOP patients displayed 845 differentially expressed genes, including 709 down-regulated and 136 up-regulated ones. Ten genes including SCIMP were significantly differentially expressed (at least three-fold difference). SCIMP was the most markedly decreased in PMOP patients' specimens. Using clinical recruited individuals, the concentration of SCIMP was 96.6 ± 20.8 ng/μL in the PMOP group compared with 168.8 ± 23.5 ng/μL in the control group (p < 0.05). At the same time, the osteoclast differentiation signaling pathway was significantly up-regulated while hedgehogs as well as other signaling pathways were down-regulated based on the KEGG analysis. The phosphorylation level of Akt was markedly blocked in si-SCIMP treatment. Up-regulation of SCIMP increased cell proliferation, inhibited cell apoptosis, and enhanced cell viability in MG-63 cells, which was markedly rescued by AKT phosphorylation inhibitor. Finally, in vivo experiments also confirmed that the upregulation of SCIMP enhanced the structural parameters of rat trabecular bone and the osteogenic activity of bone tissue. SCIMP plays a critical role in the pathogenesis of postmenopausal osteoporosis in women. SCIMP influences osteoclasts function through an akt-dependent molecular pathway, and subsequently influences the equilibrium process of bone metabolism. This provides a new insight into the pathogenesis of postmenopausal osteoporosis as well as the clinical treatment of osteoporosis.

Low Unsaturated Fatty Acids Level in the Vertebral Bone Marrow of Postmenopausal Osteoporosis: A Pilot 2D iDQC-MRS on 3.0 T Study.

Unsaturated fatty acids (UFAs) of bone marrow play a critical role in osteoporosis. However, it is difficult to resolve the UFA, especially in the presence of trabecular bone, using conventional magnetic resonance spectroscopy (MRS) methods. To preliminarily compare the bone marrow fatty acids (FAs) composition in the presence of trabecular bone of postmenopausal osteoporosis (PMOP) and healthy controls (HC). Prospective. Total thirty-six postmenopausal women were recruited with CT-confirmed PMOP (n=19) and HC (n=17). A 3 T scanner. Localized 2D intermolecular double-quantum coherence-based MRS (iDQC-MRS). In addition to the conventional water and fat peaks, another four crossing peaks of the FAs were well resolved from the L4 vertebral bone marrow using iDQC-MRS technique: allylic methylene (2.0 ppm), terminal methylene (2.2ppm), diallylic methylene (2.7ppm), and olefinic (5.3ppm). The monounsaturated fatty acids (MOFA) and polyunsaturated fatty acids (PUFAs) were then calculated. Differences between PMOP and HC were investigated using the analysis of a t-test, and the relationships were investigated using regression analysis. MOFAs and PUFAs fractions were significantly lower in the PMOP group compared to the HC group. In contrast, the saturated FAs fraction is significantly higher in the PMOP group. Additionally, decreased PUFAs, MOFAs were moderately negatively correlated with the volumetric bone mineral density (vBMD) in the PMOP group. Furthermore, increased SFAs in PMOP were strongly associated with vBMD. Using spectra resolution enhanced 2D iDQC-MRS technique, we observed low unsaturated FAs levels in the vertebral bone marrow of the PMOP patients. The reduced unsaturated FAs levels in PMOP may be associated with dysfunction of the balance between osteoblastogenesis and osteoclastogenesis. 1.

Zhuanggu Zhitong Capsule alleviates postmenopausal osteoporosis in ovariectomized rats by regulating autophagy through AMPK/mTOR signaling pathway.

BackgroundPostmenopausal osteoporosis (PMOP) is the most common primary osteoporosis, which is prone to fractures and affect the health and quality of life of the elderly and even shorten their lifetime. Traditional Chinese medicine can not only effectively improve osteoporosis and reduce fracture rate, but also have tonifying and analgesic effects. The purpose of this study was to investigate the effects of Zhuanggu Zhitong (ZGZT) Capsule on autophagy related genes and proteins in PMOP rats, so as to elucidate the molecular mechanism of tonifying deficiency and regulating stasis in the treatment of osteoporosis and analgesia.MethodsThe PMOP rat model was established by bilateral oophorectomy, and then the rats were randomly divided into control group, PMOP group, PMOP + ZGZT group and PMOP + E2 group. The changes of mechanical pain threshold of rats were detected by von Frey filaments, and the changes of mechanical pain threshold of rats in each group were compared. Computed tomography (CT) and dual-energy X-ray were used to measure the bone mineral density of lumbar bone tissue. Enzyme-linked immunosorbent assay (ELISA) and tartrate-resistant acid phosphatase (TRAP) staining were used to detect inflammatory factors and bone metabolism related indicators. Hematoxylin-eosin (HE) staining was used to observe the tissue morphology of lumbar vertebra tissue. Western blot (WB) and quantitative polymerase chain reaction (qPCR) were used to detect AMPK/mTOR pathway- and autophagy-related factor expression.ResultsZGZT can effectively restore the bone mineral density (BMD) of PMOP rats, improve the microstructure of lumbar vertebra of PMOP rats, restore the balance of bone metabolism, promote the expression of AMPK and autophagy related factors, inhibit the expression of mTOR and the release of inflammatory factors, and increase the mechanical pain threshold of PMOP rats, so as to effectively improve osteoporosis and relieving osteoporosis pain in PMOP rats.ConclusionsZGZT affects autophagy by regulating AMPK/mTOR pathway, restores the homeostasis of bone metabolism and inhibits the release of inflammatory factors. Moreover, the regulation of feedback pathways between bone metabolism and inflammatory factors finally plays the role of “bone strengthening” and “pain relieving”. ZGZT may be a new treatment for PMOP and relieving osteoporotic pain.

The Potential Role of Serum IGF-1 and Leptin as Biomarkers: Towards Screening for and Diagnosing Postmenopausal Osteoporosis.

PurposeTo investigate the differences of several serum markers among population with different bone mass and to explore the utility of new potential biomarker for the diagnosing and screening for postmenopausal osteoporosis (PMOP).Materials and MethodsA total of 1055 postmenopausal women were screened and gathered data on BMD screening, biological samples, and questionnaire information. A liquid chip assay was used to measure serum IL-6, IGF-1, BMP-2, VEGF, leptin and FGF23. The predictive value of the indicator panels was assessed using the area under the receiver-operator characteristic curve (AUC). Statistical analyses were conducted by using SAS 9.4 and R software 4.1.1. Figures were created in GraphPad Prism 8.0.ResultsWhen compared against the normal group, in addition to the vitamin D, the PMOP group showed a significant increase in median values for other indicators (P < 0.05), especially in P1NP and β-CTX. Among the six cytokines representing different osteoporosis mechanisms, currently, we found that only IGF-1 and leptin showed significant differences between the groups. Also, the liquid chip assay results showed that IGF-1 and leptin, as newer cytokines in osteoporosis, not only have significant differences between groups, but also have a strong correlation with each other (P < 0.05). Then, we reported the accuracy of different indicator combinations by using AUC and, moreover, we demonstrated that IGF-1 with leptin did significantly provide incremental value to the AUC of conventional indexes, it markedly improved diagnostic efficacy, displaying an IDI of 9.45% (P = 0.000).ConclusionIFG-1 and leptin seem to be key biomarker associated with PMOP. The high prevalence of PMOP makes these cytokines might bear the potential of becoming a very useful screening test also for clinical follow-up of patients.

The Effect of Postmenopausal Osteoporosis on Middle Ear Resonance Frequency.

Background: The effect of postmenopausal osteoporosis on the middle ear mechano-acoustic system is unknown. The aim of this study is to investigate whether or not middle ear resonance frequency is affected in females with postmenopausal osteoporosis.Methods:The study included postmenopausal women aged 45-60 years, separated into 2 groups as females with postmenopausal osteoporosis and healthy postmenopausal females (control group). A detailed anamnesis was taken from all subjects and then the ear, nose, and throat examinations were done followed by pure tone audiometry, tympanometry, and multifrequency tympanometry tests. The groups were compared in respect of pure tone average, bone conduction threshold, RF, static admittance, and tympanometric peak pressure values.Results: The mean age of the patients was 59.2 ± 4.53 years (range, 48-65 years) in the postmenopausal osteoporosis group and 57.11 ± 4.27 years (range, 48-65 years) in the control group (P > .05). The mean resonance frequency values for the postmenopausal osteoporosis and control group were 954.41 ± 127.47 and 935.29 ± 126.39 Hz (P > .05 ). The mean static admittance values for the postmenopausal osteoporosis and control group were 0.82 ± 0.33 and 0.85 ± 0.3 mmho, and mean tympanometric peak pressure values were −7.35 ± 18.52 and −6.94 ± 19.52 daPa (P > .05 for both static admittance and tympanometric peak pressure). The mean pure tone averagevalues for the postmenopausal osteoporosis and control group were 20.96 ± 6.82 and 15.60 ± 7.81 dB, and mean bone conduction threshold values were 17.57 ± 6.03 and 12.10 ± 6.52 dB (P < .05 for both pure tone average and bone conduction threshold).Conclusions: The results showed that the middle ear resonance frequency values were not affected in postmenopausal osteoporosis patients, but there was seen to be greater sensorineural hearing loss in females with postmenopausal osteoporosis compared to healthy postmenopausal females.

PTHR1 Genetic Polymorphisms Are Associated with Osteoporosis among Postmenopausal Arab Women.

The parathyroid hormone 1 receptor (PTHR1) plays a crucial role in calcium homeostasis and bone metabolism. However, its genetic role in regulating bone turnover markers (BTMs) in postmenopausal osteoporosis (PMO) remains unclear. Herein, we explored parathyroid hormone (PTH) and PTHR gene variant susceptibility to osteoporosis and their association with various circulating BTM and inflammatory markers in postmenopausal women of Arab ethnicity. In total, 600 postmenopausal Arab women (300-PMO and 300-control) were genotyped for selected SNPs in PTH (rs1459015, rs307253, rs6054, rs307247, rs10500783 and rs10500784), PTHR1 (rs6442037, rs1138518, and rs724449 SNPs) and PTHR2 (rs9288393, rs10497900, and rs897083). Anthropometrics, BTMs, and inflammatory markers were measured. Bone mineral density (BMD) was measured at the lumbar spine L1–L4 and the femoral neck using dual-energy X-ray absorptiometry (DXA). PTHR1 rs1138518 genotype C/T was found to be a significant risk factor for PMO (OR = 1.49, 95% CI 1.0-2.1, P = 0.03). The genotypes C/T and T/T of PTHR1 rs1138518 were associated with 25-hydroxy-vitamin D (25(OH)D) regulation. In the PMO group, carriers of the C/T genotype had significantly lower 25(OH)D levels than carriers of the same genotypes in the control group (59.9 (36.7-92.4) nmol/l and 66.4 (43.5-87.8) nmol/l, respectively; P = 0.048]. Our study concludes that the PTHR1 rs1138518 genotype could be a potential risk factor for osteoporosis and 25(OH)D regulation in Arab women with PMO.

Microarray analysis of circRNAs sequencing profile in exosomes derived from bone marrow mesenchymal stem cells in postmenopausal osteoporosis patients.

IntroductionBone marrow‐derived mesenchymal stem cells (BMSCs)‐derived exosomes are involved in the modulation of tissue repair and regeneration. CircRNAs play important roles in BMSCs exosomes. The current study sought to explore the role of circRNAs in exosomes derived from BMSCs of postmenopausal osteoporosis (PMOP) patients and the underlying mechanisms.MethodsRNA was extracted from BMSCs exosomes of PMOP and a control group. RNA microarray and bioinformatics analyses were used to explore the expression profile and functions circRNAs. Differentially expressed circRNAs from 20 PMOP and 20 controls were analyzed using RT‐qPCR.ResultsA total of 237 upregulated and 279 downregulated circRNAs were identified in the current study. The top‐10 most upregulated circRNAs in the PMOP group were hsa_circ_0069691, hsa_circ_0005678, hsa_circ_0006464, hsa_circ_0015813, hsa_circ_0000511, hsa_circ_0076527, hsa_circ_0009127, hsa_circ_0047285, hsa_circ_0027741, and hsa_circ_0090949. The top‐10 most downregulated circRNAs were hsa_circ_0048669, hsa_circ_0090247, hsa_circ_0070899, hsa_circ_0087557, hsa_circ_0045963, hsa_circ_0090180, hsa_circ_0058392, hsa_circ_0040751, hsa_circ_0067910, and hsa_circ_0049484. RT‐PCR verified dysregulation of 5 circRNAs including hsa_circ_0009127, hsa_circ_0090759, hsa_circ_0058392, hsa_circ_0090247, and hsa_circ_0049484. Moreover, a circRNA‐microRNA‐mRNA interaction network was developed based on differentially expressed circRNAs. Functional analysis showed that pathways involved in the regulation of autophagy, PI3K‐Akt signaling, FoxO signaling, and MAPK signaling were associated with the differentially expressed circRNAs in PMOP patients.ConclusionThe findings of this study show dysregulated circRNAs in BMSCs exosomes of PMOP patients, which may affect the progression of PMOP. These circRNAs can be used as predictive biomarkers and as therapeutic targets for the treatment of PMOP.

Overexpression of circ_0021739 in Peripheral Blood Mononuclear Cells in Women with Postmenopausal Osteoporosis Is Associated with Reduced Expression of microRNA-194-5p in Osteoclasts.

BackgroundPostmenopausal osteoporosis, a common disease among elderly women, is linked to estrogen deficiency, mechanical loading, and genotype. Circular RNAs (circRNAs) are formed through reverse splicing of the splice donor at the 3′ end and the splice accepter at the 5′ end in pre-mRNA and have been shown to be involved in the development of multiple diseases. Based on their high sequence conservation and stability, circRNAs may be useful biomarkers in different diseases. However, the roles of circRNAs in postmenopausal osteoporosis remain incompletely understood.Material/MethodsFifty-three postmenopausal women were assigned to either the postmenopausal osteoporosis group (n=28) or the control group (n=25). Reverse-transcriptase quantitative polymerase chain reaction (RT-qPCR) analysis was performed to determine the differential expression of circRNAs between the 2 groups. Receiver-operating characteristic (ROC) curve analysis was conducted to evaluate the clinical diagnostic value of circRNA. Prediction of the binding sites between circRNA and miRNAs was conducted using miRanda and RNAhybrid. The function of the circRNA in osteoclastogenesis was determined by circRNA overexpression followed by tartrate-resistant acid phosphatase staining and RT-qPCR analysis.ResultsAmong 4 circRNAs previously identified by RNA-sequencing analysis as differentially expressed in patients with postmenopausal osteoporosis, only hsa_circ_0021739 showed a significant difference in expression between the groups and was downregulated in patients with postmenopausal osteoporosis. The hsa_circ_0021739 expression level was determined to be correlated with the lumbar vertebra, femur, and forearm T-scores. Overexpression of hsa_circ_0021739 decreased the level of hsa-miR-502-5p and inhibited the differentiation of osteoclasts.ConclusionsThe circRNA hsa_circ_0021739 is a potential blood biomarker for postmenopausal osteoporosis. In addition, hsa-miR-502-5p is a likely target of hsa_circ_0021739, which acts to regulate the differentiation of osteoclasts.

Correlation of oxidative stress-related biomarkers with postmenopausal osteoporosis: a systematic review and meta-analysis.

Oxidative stress (OS) has been implicated in postmenopausal osteoporosis (PO). However, the relations between OS-related markers and PO are controversial. This study aimed to quantitatively and comprehensively assess the roles of OS-related biomarkers in PO. Relevant articles were retrieved from electronic databases. All OS-associated biomarkers with at least 2 independent study outcomes were meta-analyzed. The pooled standardized mean differences (SMD) with its 95% confidence intervals (CI) were presented. A total of 36 studies involving 16 OS-related biomarkers were investigated. The overall results showed that total oxidant status (TOS), superoxide dismutase (SOD), hydroperoxides (HY), paraoxonase (PON1), nitric oxide (NO), and homocysteine (Hcy) were not statistically different between the PO and control groups, whereas significantly increased levels of oxidative stress index (OSI), malondialdehyde (MDA), advanced oxidation protein products (AOPP), and vitamin B12, along with decreased total antioxidant status (TAS), total antioxidant power (TAP), catalase (CAT), glutathione peroxidase (GPx), uric acid (UA), and folate, were detected in the PO group. Subgroup analysis based on biological samples displayed significantly elevated NO in erythrocyte and Hcy in serum, along with decreased SOD in serum. Monitoring of certain OS-related biomarkers might be beneficial to the diagnosis and prognosis of PO.

Differentially expressed microRNA and target gene analysis in patients with postmenopausal osteoporosis

Objective To analyze the differentially expressed microRNA (miRNA) and their target genes in peripheral blood and bone tissue of postmenopausal osteoporosis (PMOP), and provide basis for the study of pathogenesis as well as biomarkers identification of PMOP. Methods Two miRNA datasets of PMOP from the public platform NCBI-GEO DataSets were obtained, including GSE64433 (the miRNA expression profile of peripheral blood samples, including 23 PMOP patients and 25 controls) and GSE74209 (the miRNA expression profile of the femoral neck bone tissue sample, including six PMOP patients and six controls). R/Bioconductor was performed for data analysis and differentially expressed miRNA screening, and miRNAs with fold change>2 & P<0.05 between osteoporosis and controls were selected as differentially expressed miRNA. The miRNA target gene prediction was performed by combining TargetScan, miRDB and miRTarBase databases. The miRNA-target gene regulatory network was constructed and analyzed by Cytoscape. Results There were 224 differentially expressed miRNAs (75 up-regulated miRNAs and 149 down-regulated miRNAs) in the peripheral blood samples of PMOP group (GSE64433 dataset) and 132 differentially expressed miRNAs (58 up-regulated miRNAs and 74 down-regulated miRNAs) in the femoral neck bone tissue of PMOP group (GSE74209 dataset). We combined the results from the two datasets and obtained 8 miRNAs down-regulated in both datasets, and the 8 miRNAs regulated a total of 327 target genes, and 10 of these target genes were co-regulated by two miRNAs. Conclusions The core miRNAs and the target genes regulated by multiple miRNAs in the regulatory network may play important roles in the pathogenesis of PMOP and have potential application values as molecular biomarkers of disease. These findings are meaningful for the studies of PMOP pathogenesis, biomarkers screening and prevention of osteoporotic fractures. Key words: Osteoporosis, postmenopausal; MicroRNAs; Regulatory network

The Neuropeptide Vasoactive Intestinal Peptide Levels in Serum are Inversely Related to Disease Severity of Postmenopausal Osteoporosis: A Cross-Sectional Study.

Background: The neuropeptide vasoactive intestinal peptide (VIP) has been identified as inhibiting osteoclastogenesis and suppressing inflammation. Objective: This study was conducted to examine serum VIP levels in postmenopausal osteoporosis (PMOP) patients and explore the correlation of serum VIP levels with disease severity of PMOP. Methods: A total of 106 postmenopausal women diagnosed as osteoporotic were enrolled in the study and 102 postmenopausal women with normal bone mineral density (BMD) were enrolled as controls. BMD at the femoral neck (FN), lumbar spine 1-4, and total hip were examined using dual-energy X-ray absorptiometry. Genant semiquantitative grading was used for vertebral morphometry and fracture. Serum VIP levels were tested using enzyme-linked immunosorbent assay. Serum inflammatory factor interleukin-1β (IL-1β), osteoclastic activity marker tartrate-resistant acid phosphatase 5b (TRACP-5b), and estrogen-2 (E2) were also examined. Receiver operating characteristic (ROC) analyses was performed to determine the diagnostic values of serum VIP, IL-1β, TRCAP-5, and E2 with regard to Genant grade. Results: Our findings demonstrated a reduction in the serum level of VIP expressed in PMOP patients compared with controls. In the PMOP group, patients with lumbar fracture had significantly lower serum VIP concentrations in comparison with healthy controls. Serum VIP concentrations were positively associated with BMD at the FN, lumbar spine 1-4, and total hip. We also observed that serum VIP levels were positively correlated with E2 levels but negatively correlated with IL-1β and TRCAP-5 levels. In addition, ROC analysis found that reduction of serum VIP in combination with elevation of TRACP-5b may serve as an indicator of a severe Genant grade. Conclusions: Attenuated serum VIP levels were linked to disease severity of PMOP and may act as a protective marker for PMOP.

Let-7a-5p inhibits BMSCs osteogenesis in postmenopausal osteoporosis mice

PurposeThe aim of this study was to investigate the mechanism of let-7a-5p in osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in postmenopausal osteoporosis (PMOP) mice. MethodsA mouse model of PMOP was established and osteoporosis model was identified by micro-CT scan. BMSCs in the sham group and PMOP group were cultured and osteogenic differentiation was induced. The expression of let-7a-5p in BMSCs was detected by qRT-PCR, and BMSCs was induced by osteogenic differentiation in sham and PMOP group. The BMSCs treated by let-7a-5p mimics, let-7a-5p inhibitor and negative control were named as let-7a-5p mimics group, mimics NC group, let-7a-5p inhibitor group and inhibitor NC group, respectively. ALP staining and alizarin red staining were used to detect osteogenic differentiation ability, qRT-PCR and western blot were used to detect the expression of Runt-related transcription factor 2 (Runx2) and Osterix. The targeting relationship between let-7a-5p and TGFBR1 were verificated by target scan and luciferase reporter gene assay. ResultsThe PMOP mouse model was successfully established. The expression of let-7a-5p in BMSCs of PMOP group was significantly higher than that in the sham group (P < 0.05). Let-7a-5p reduced the expression of ALP and the formation of calcified nodules, while also inhibited the expression of Runx2 and Osterix. TGFBR1 is the target gene of let-7a-5p. ConclusionLet-7a-5p might inhibit the osteogenic differentiation of BMSCs in PMOP mice by regulating TGFBR1.

Peculiar features of life quality in patients with type 2 diabetes mellitus and postmenopausal osteoporosis

Aim. To study the peculiarities of the quality of life in patients with type 2 diabetes mellitus (DM2) and postmenopausal osteoporosis (PO).&#x0D; Materials and methods. 104 postmenopausal women aged 49–67 were examined. They were divided into 3 groups: group 1 (38 persons) – patients with DM2 and PO, group 2 (24 persons) – patients with PO, group 3 (42 persons) – postmenopausal patients with DM2 without any osteoporosis signs. Their quality of life was assessed using SF-36 survey, sensations of pain – with four-component visual analogue scale.&#x0D; Results. Physical component of health (PH) was statistically significantly lower in DM2 and PO group. Most patients of this group had decreased psychical component of health. In DM2 and PO group as compared with PO group statistically significantly lower values according to physical status-conditioned role functioning scales (RP) (p = 0,018), emotional status-conditioned role functioning (p = 0,028), PH (p = 0,038) were detected. When compared with DM2 group, physical functioning (p = 0,04), RP (p = 0,014), pain intensity (p = 0,049), PH (p = 0,014) appeared to be statistically significantly lower. The factors, negatively influencing the quality of life in patients of DM2 and PO group were the following: remoteness of diabetes, comorbidity, level of pain sensations, presence of diabetic polyneuropathy, insulin therapy in treatment, unsatisfactory glycemic control.&#x0D; Conclusions. Osteoporosis in patients with type 2 diabetes mellitus worsens their quality of life, in particular at the expense of elevated intensity of pain sensations.

Effect of Primary Hypertension on Treatment Outcomes of Patients with Postmenopausal Osteoporosis: A 5 Year Follow Up Retrospective Study.

To investigate the efficacy of primary hypertension (HTN) on the treatment prognosis of patients with postmenopausal osteoporosis (PMOP). 45 patients who were diagnosed as PMOP with lumbar and/or femur neck bone mineral density screening (BMD) but have no history of PMOP treatment including calcium and vitamin D, have comorbid primary HTN and treated with a vasodilator antihypertensive drug at least a year were included to the study. Control group was constituted with 44 patients with PMOP at same age but have no comorbidity. Demographic features including age, height, weight, occupation, educational level menarche and menopause age, clothing style, daily intake of calcium, smoking and/or alcohol consumption, daily physical activity level, personal history of fragility fracture or in mother and duration of primary HTN diagnosis were recorded. Biochemical parameters were also recorded. Patients were treated with bisphosphonate, calcium and vitamin D and same parameters were evaluated at the end of first and fifth year. Demographic and disease characteristics were not different between groups before treatment (p > 0.05). In group analysis, there was significant improvement in lumbar and femur neck T scores of PMOP + HT and PMOP groups after 1 and 5years of treatment compared to baseline (p < 0.05) Lumbar and femur neck T score variations between the baseline, first and fifth years of treatment were not significantly different in PMOP + HT and PMOP groups (p < 0.05). Although the results vary between populations, primary HTN does not have an impact on the prognosis of PMOP treatment in Turkish population.

Effect of type 2 diabetes mellitus on treatment outcomes of patients with postmenopausal osteoporosis: a retrospective study.

The aim of this study is to investigate the effect of type 2 diabetes mellitus (T2DM) on the treatment outcomes of patients with postmenopausal osteoporosis (PMOP). Thirty-five patients who had been diagnosed as PMOP by lumbar and/or femoral neck bone mineral density screening (BMD) and who had comorbid T2DM were included in the study. Thirty-five patients who had been diagnosed as PMOP but who had no comorbidity including DM constituted the control group. Demographic features, biochemical parameters, femoral and lumbar T scores were all recorded. All patients were treated with bisphosphonate, calcium and vitamin D and the same parameters were evaluated at the end of the first and fifth year. Lumbar T scores and serum osteocalcin levels before treatment were significantly lower in the DM + PMOP group (p < 0,05). At the end of 5years, despite the lumbar T score having increased, the femoral T score was found to be significantly lower in the DM + PMOP group. In the PMOP group, there was significant improvement in the T scores and serum osteocalcin levels following a 5-year treatment period (p < 0,05). T2DM has unfavorable effects on treatment prognosis in patients with PMOP. Different risk factors of PMOP which differ in the general population maybe more important when evaluating fracture risk in patients wtih T2DM.

Association of Vitamin D Receptor Gene Variation With Osteoporosis Risk in Belarusian and Lithuanian Postmenopausal Women.

Vitamin D receptor (VDR) is one of the main mediators of vitamin D biological activity. VDR dysfunction might substantially contribute to development of postmenopausal osteoporosis (PMO). Numerous studies have revealed the effects of several VDR gene variants on osteoporosis risk, although significant variation in different ethnicities have been suggested. The main purpose of this work was to assess the frequency of distribution of VDR genetic variants with established effect and evaluate their haplotype association with the risk of PMO in a cohort of Belarusian and Lithuanian women. Case group included women with PMO (n = 149), the control group comprised women with normal bone mineral density (BMD) and without previous fragility fractures (n = 172). Both groups were matched for age, height, sex, and BMI—no statistically significant differences observed. VDR gene polymorphic variants (ApaI rs7975232, BsmI rs1544410, TaqI rs731236, and Cdx2 rs11568820) were determined using polymerase chain reaction and restriction fragment length polymorphism. The lumbar spine (L1-L4) and femoral neck BMD was measured using dual-energy X-ray absorptiometry. Association between each VDR variant and PMO risk was assessed using multiple logistic regression. The genotyping revealed statistically significant difference in the rs7975232 genotype frequencies between the patients and the controls (homozygous C/C genotype was overrepresented in patients, p = 0.008). Patients with osteoporosis were also three times more likely to carry the rs1544410 G/G genotype, when compared to controls. We found that rs7975232, rs1544410, and rs731236 variants were in a strong direct linkage disequilibrium (p < 0.0001), suggesting that risk alleles of these markers are preferably inherited jointly. For the bearers of C-G-C haplotype (consisting of rs7975232, rs1544410, and rs731236 unfavorable alleles), the risk of PMO was significantly higher (OR = 4.7, 95% CI 2.8–8.1, p < 0.0001) compared to controls. This haplotype was significantly over-represented in PMO group compared to all other haplotypes. Our findings highlight the importance of identified haplotypes of VDR gene variants. Complex screening of these genetic markers can be used to implement personalized clinical approach for prevention, treatment, and rehabilitation programs.

Postmenopausal osteoporosis is associated with elevated aldosterone/renin ratio.

Plasma aldosterone/renin ratio (ARR) is a useful method for primary aldosteronism (PA) screening. However some confounders, such as medications and dietary, affect plasma renin and aldosterone levels, resulting in false-negative or -positive plasma ARR. This study investigated the association between postmenopausal osteoporosis (PMO) and plasma ARR. Bone mineral density (BMD) was measured by dual-energy X-ray-absorptiometry (DXA) in 324 normotensive postmenopausal women. Based on clinical characteristics and BMD, 186 and 96 subjects were diagnosed as PMO and osteopenia respectively, and the remaining 42 subjects were grouped as normal BMD. Plasma aldosterone concentration (PAC), plasma renin concentration (PRC), parathyroid hormone (PTH), bone alkaline phosphatase (BALP) and 25-Hydroxyvitamin D(25-(OH)D) were determined. Subjects with PMO showed significantly higher levels of PAC (121.0 ± 78.8 vs. 81.8 ± 71.5 pg/ml, p < 0.01 and 121.0 ± 78.8 vs. 91.7 ± 56.2 pg/ml, p < 0.01) and ARR (32.0 ± 53.6 vs. 9.0 ± 9.3 pg/μU, p < 0.01 and 32.0 ± 53.6 vs. 16.3 ± 32.1 pg/μU, p < 0.01) compared to women with normal BMD and osteopenia, respectively. Using ARR ≥ 37.0 pg/μU as the cutoff for positive screening, more false-positive was found in the PMO group when compared to the normal BMD group (24 vs. 2%) and osteopenia group (24 vs. 7%), respectively. PAC was negatively associated with lumbar spine BMD T-score (r = -0.239, p < 0.001), femur neck BMD T-score (r = -0.234, p < 0.001) and total hip BMD T-score (r = -0.228, p < 0.001). PTH was positively associated with PAC (r = 0.119, p < 0.05) and ARR (r = 0.136, p < 0.05). PAC and ARR are elevated in women with PMO, which might increase the risk of false-positive for case detection of PA.