Yi-Qun Peng

Epidemiology and management of osteoporosis in the People’s Republic of China: current perspectives

With the progressive aging of the population, osteoporosis has gradually grown into a global health problem for men and women aged 50 years and older because of its consequences in terms of disabilities and fragility fractures. This is especially true in the People’s Republic of China, which has the largest population and an increasing proportion of elderly people, as osteoporosis has become a serious challenge to the Chinese government, society, and family. Apart from the fact that all osteoporotic fractures can increase the patient’s morbidity, they can also result in fractures of the hip and vertebrae, which are associated with a significantly higher mortality. The cost of osteoporotic fractures, moreover, is a heavy burden on families, society, and even the country, which is likely to increase in the future due, in part, to the improvement in average life expectancy. Therefore, understanding the epidemiology of osteoporosis is essential and is significant for developing strategies to help reduce this problem. In this review, we will summarize the epidemiology of osteoporosis in the People’s Republic of China, including the epidemiology of osteoporotic fractures, focusing on preventive methods and the management of osteoporosis, which consist of basic measures and pharmacological treatments.

Cellular and molecular responses in progressive pseudorheumatoid dysplasia articular cartilage associated with compound heterozygous WISP3 gene mutation.

Progressive pseudorheumatoid dysplasia (PPD) is characterized by continuous degeneration and loss of articular cartilage, which has been attributed to mutations in the gene encoding WISP3. We collected a PPD family and analyzed their WISP3 genes mutation. Articular chondrocytes (ACs) were purified from the femurs of a PPD patient after hip replacement surgery. Cell growth, proliferation, and viability were examined. Gene expression profiling and analyses of matrix metalloproteinases (MMP)-1, -3, and -13 proteins were carried out using cDNA differential microarrays, real-time reverse transcriptase-polymerase chain reaction (RT-PCR), immunohistochemistry, and Western blot analysis. We found that two probands carried a deletion (840delT) mutation in maternal allele, which leads to truncated WISP3 protein missing 43 residues in C terminus; and a 1000T>C substitution in paternal allele, which was also passed on to four other members in the PPD kindred. PPD ACs were heterogeneous in size with an enhanced rate of cell proliferation and viability compared with the normal ACs. MMP-1, -3, and -13 mRNA expressions were dereased in PPD ACs. MMP-1, -3, and -13 protein levels, however, were increased in cell lysates from PPD ACs, but markedly decreased in the supernatants from cultured ACs. WISP3 mRNA expression in PPD ACs was also decreased. Our results show, for the first time, a compound heterozygous mutation of WISP3 and a series of cellular and molecular changes disturbing the endochondral ossification in this PPD patient.

Stimulation of RANKL and inhibition of membrane-type matrix metalloproteinase-1 expression by parathyroid hormone in normal human osteoblasts.

Receptor activator of NF‐κB (RANK) ligand (RANKL), expressed by cells of the osteoblast lineage binds to RANK, induces signaling and a gene expression cascade that leads to osteoclast differentiation and activation. Recently, osteoblast‐derived membrane‐type matrix metalloproteinases‐1 (MT1‐MMP) have been implicated in the process of bone resorption by degrading bone matrix. In the present study, we investigated the effects of parathyroid hormone [PTH (1–34)] on RANKL and MT1‐MMP production in cultured normal human osteoblast‐like cells (hOB). In reverse transcription polymerase chain reaction studies, we observed that PTH (1–34) induced RANKL messenger ribonucleic acid (mRNA) expression. Activity assays demonstrated that PTH (1–34) simultaneously inhibited MT1‐MMP protein expression in a dose‐ and time‐dependent manner. The effect of PTH (1–34) on MT1‐MMP production was parallel to that on RANKL expression, suggesting a tight inverse relationship between MT1‐MMP and RANKL expression. Our findings indicated that the decreased MT1‐MMP expression by PTH may be involved in RANKL signaling in osteoblasts and activation of osteoclasts.

Taurine restores Axl/Gas6 expression in vascular smooth muscle cell calcification model

Our previous studies demonstrated that taurine inhibits osteoblastic differentiation of vascular smooth muscular cells (VSMCs) via the mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) signaling pathway, but the underlying mechanism is not elucidated. The tyrosine kinase receptor Axl and its ligand growth arrest-specific protein 6 (Gas6) are expressed in VSMCs. Axl/Gas6 signaling system is known to inhibit VSMCs calcification. We herein showed that taurine partially restored Axl and Gas6 expression in β-glycerophosphate (β-GP)-induced VSMC calcification model. Taurine also induced activation of ERK, but not other two MAPKs including c-jun N-terminal Kinase (JNK) and p38 in VSMCs. Either knockdown of the taurine transporter (TAUT) or treatment with the ERK-specific inhibitor PD98059 blocked the activation of ERK by taurine and abolished taurine-induced Axl/Gas6 expression and calcium deposition reduction in β-GP-induced VSMC calcification model. These results demonstrate for the first time that taurine stimulates expression of Axl and Gas6 via TAUT/ERK signaling pathway in β-GP-induced VSMC calcification model.

Connective tissue growth factor induces osteogenic differentiation of vascular smooth muscle cells through ERK signaling

Connective tissue growth factor (CTGF) plays an important role in the pathogenesis of atherosclerosis by promoting vascular smooth muscle cell (VSMC) growth, migration, apoptosis, adhesion and the secretion of matrix components. The osteogenic differentiation of VSMCs is essential in the development of vascular calcification. However, the role of CTGF in the transdifferentiation and calcification of VSMCs is unclear. In the present study, we examined whether CTGF stimulates VSMC transdifferentiation. Primary VSMCs were obtained from mouse thoracic aortas by enzymatic digestion and identified by immunostaining for smooth muscle specific α-actin antibody (α-SMA). VSMC calcification was induced by the addition of CTGF to the osteogenic mediaum containing 5-10% FBS in the presence of 0.25 mM ascorbic acid and 10 mM β-glycerophosphate for 14 days. Calcified cells were determined by Alizarin Red S staining. Our results revealed that CTGF induced the expression of several bone markers, including alkaline phosphatase (ALP), osteocalcin (OC), osteoprotegerin (OPG) and core-binding factor subunit α1 (Cbfα1)/runt-related transcription factor 2 (Runx2), as well as calcification. However, the inhibition of extracellular signal-regulated kinase (ERK) activity using the ERK-specific inhibitor, PD98059, blocked the induction of these proteins and VSMC calcification. Based on these data, we conclude that CTGF stimulates the transdifferentiation of VSMCs into osteoblasts and that the ERK signaling pathway appears to play a critical role in this process.

L-carnitine and taurine synergistically inhibit the proliferation and osteoblastic differentiation of vascular smooth muscle cells

AbstractAim:To investigate the synergistic action of L-carnitine (LC) and taurine (TAU) on the proliferation and osteoblastic differentiation of vascular smooth muscle cells (VSMCs).Methods:DNA and protein synthesis of VSMCs were assessed using scintillation counting. Alkaline phosphatase (ALP) activity and calcium content were determined to investigate the effects of LC and TAU on the osteoblastic differentiation and mineralization of VSMCs. TAU uptake by VSMCs was assayed. RNA interference was used to down-regulate the expression of the TAU transporter (TAUT) in rat VSMCs.Results:LC and TAU synergistically inhibited the proliferation and β-glycerophosphate (β-GP)-induced osteoblastic differentiation of VSMCs as evidenced by the decreased [3H]thymidine incorporation, ALP activity and calcium deposition. Furthermore, LC stimulated the TAU uptake and TAUT expression in VSMCs. Suppression of TAUT with short hairpin RNA (shRNA) abolished the synergistic action of LC and TAU in VSMCs.Conclusion:The synergistic inhibitory action of LC and TAU on the proliferation and osteoblastic differentiation of VSMCs is attributable to the up-regulation of TAUT expression and TAU uptake by LC.

Relationship between age-related serum concentrations of TGF-β1 and TGF-β2 and those of osteoprotegerin and leptin in native Chinese women

BACKGROUND Transforming growth factor-beta 1 (TGF-beta1), TGF-beta2, osteoprotegerin (OPG), and leptin are important cytokines in the regulation of bone remodeling. We investigated the relationship of TGF-beta1 and TGF-beta2 concentrations with those of OPG and leptin in Chinese females. METHODS The serum concentrations of TGF-beta1, TGF-beta2, OPG, and leptin were measured by ELISA in 459 healthy Chinese females aged 25-80 y. RESULTS The mean values (+/-SD) of the serum concentrations of TGF-beta1, TGF-beta2, OPG, and leptin in Chinese females were 29.7+/-1.69 microg/l, 13.7+/-3.86 microg/l, 3.81+/-1.96 pmol/l, and 10.5+/-2.01 microg/l, respectively. Further, the serum TGF-beta1 concentrations of postmenopausal women were significantly lower than those of perimenopausal and premenopausal women (24.3+/-1.59 vs 33.4+/-1.69 and 37.6+/-1.64, respectively), while the TGF-beta2 concentrations of postmenopausal women were significantly higher than those of perimenopausal and premenopausal women (14.6+/-3.91 vs 13.5+/-3.93 and 11.7+/-2.68, respectively). The serum TGF-beta1 concentration was found to be significantly negatively correlated with age (r=-0.335, P=0.000) and the TGF-beta2 concentration, to be significantly positively correlated with age (r=0.230, P=0.000). The TGF-beta1 concentration was found to be significantly negatively correlated with both TGF-beta2 (r=-0.261, P=0.000) and OPG (r=-0.313, P=0.000) concentrations; a significantly positive correlation was found between the TGF-beta1 and leptin concentrations (r=0.164, P=0.000) and between TGF-beta2 and OPG concentrations (r=0.432, P=0.000). CONCLUSION These results provide age-related reference values of TGF-beta1 and TGF-beta2 in Chinese adult women, and reveal the relationships between these cytokines.

WISP3 suppresses insulin-like growth factor signaling in human chondrocytes

WISP3 is essential for maintaining cartilage integrity mainly by regulating the expression of collagen II, and mutations of WISP3 linked to spondyloepiphyseal dysplasia tarda with progressive arthropathy (SEDT-PA) can compromise this function and lead to cartilage loss. The aim of this study was to evaluate the effect of WISP3 on insulin-like growth factor (IGF) signaling in human chondrocytes, investigate whether WISP3 up-regulates collagen II through the IGF signaling pathway, and compare IGF signaling between wild-type and mutant WISP3. Experimental results suggest that WISP3 up-regulates collagen II expression and inhibits the activation of IGF-IR, IRS-1, and ERK kinase in human chondrocytes, and mutation of WISP3 augments IGF signaling in human chondrocytes. In addition to the IGF signaling pathway, WISP3 might up-regulate collagen II expression through an IGF-independent signaling cascade.

The relationship between the levels of gonadotropic hormones and OPG, leptin, TGF-β1 and TGF-β2 in Chinese adult women

BACKGROUND The relationship between the levels of gonadotropic hormones and bone metabolism-related cytokines in Chinese women is unclear. We investigated the relationship between FSH and LH and OPG, leptin, TGF-beta1, and TGF-beta2 in Chinese women. METHODS A cross-sectional study of 694 Chinese women, aged 20 to 82y was conducted. Levels of serum FSH, LH, OPG, leptin, TGF-beta1, and TGF-beta2 were determined. RESULTS In premenopausal females, serum follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels seemly showed no correlation with the cytokine levels. In perimenopausal females, serum FSH and LH levels showed significant positive correlation with osteoprotegerin (OPG) and transforming growth factor-beta2 (TGF-beta2) levels (r=0.286 to 0.405, all P=0.000), whereas they showed negative correlation with TGF-beta1 levels (r=-0.413 and -0.354, all P=0.000). In postmenopausal females, FSH and LH levels showed positive correlation with OPG levels (r=0.247 and 0.241, all P=0.000), negative correlation with leptin and TGF-beta1 levels (r=-0.234 to -0.319, all P=0.000), and no correlation with TGF-beta2 levels. Multiple linear regression stepwise analysis revealed the following results. In premenopausal females, 2.0% and 1.5% of the changes in LH could be explained by OPG and leptin, respectively, while 1.9% of the changes in OPG could be explained by LH. In perimenopausal females, the determinants of OPG and TGF-beta1 on FSH were 10.9% and 17.0%, respectively, and the determinants of OPG, TGF-beta1 and TGF-beta2 on LH were 4.5%, 4.9% and 16.4%, respectively. The determinants of FSH and LH on OPG were 14.5% and 2.5%, respectively. The determinant of FSH on TGF-beta1 was 4.5%, while the determinant of LH on TGF-beta2 was 16.4%. In postmenopausal females, the determinants of leptin and OPG on FSH were 10.2% and 2.8%, respectively, and the determinants of OPG and TGF-beta1 on LH were 5.8% and 2.3%, respectively. The determinant of FSH on OPG, leptin and TGF-beta1 were 6.1%, 3.4% and 9.2%. CONCLUSIONS These results indicate that age-related gonadotropic hormone levels are associated with changes in OPG, TGF-beta1, TGF-beta2 and leptin, and change with menopausal status.

Oestrogen Inhibits Arterial Calcification by Promoting Autophagy

Arterial calcification is a major complication of cardiovascular disease. Oestrogen replacement therapy in postmenopausal women is associated with lower levels of coronary artery calcification, but its mechanism of action remains unclear. Here, we show that oestrogen inhibits the osteoblastic differentiation of vascular smooth muscle cells (VSMCs) in vitro and arterial calcification in vivo by promoting autophagy. Through electron microscopy, GFP–LC3 redistribution, and immunofluorescence analyses as well as measurement of the expression of the autophagosome marker light-chain I/II (LC3I/II) and autophagy protein 5 (Atg5), we show that autophagy is increased in VSMCs by oestrogen in vitro and in vivo. The inhibitory effect of oestrogen on arterial calcification was counteracted by 3-methyladenine (3MA) or knockdown of Atg5 and was increased by rapamycin. Furthermore, the inhibitory effect of oestrogen on arterial calcification and the degree of autophagy induced by oestrogen were blocked by a nonselective oestrogen receptor (ER) antagonist (ICI 182780), a selective oestrogen receptor alpha (ERα) antagonist (MPP), and ERα-specific siRNA. Our data indicate that oestrogen inhibits the osteoblastic differentiation of VSMCs by promoting autophagy through the ERα signalling pathway in vitro and arterial calcification in vivo by increasing autophagy. Our findings provide new insights into the mechanism by which oestrogen contributes to vascular calcification in vitro and in vivo.