Interaction proteomics of long noncoding rna uc431+ gene in postmenopausal osteoporosis / 中国组织工程研究

BACKGROUND: Overexpression of LncRNA TUG1 promotes osteoclast proliferation and inhibits apoptosis. Knockdown with siRNA has achieved the opposite result, indicating that knockdown of LncRNA TUG1 may become a potential target for osteoporosis treatment. OBJECTIVE: To study the proteins interacting with the non-coding RNA uc431+ associated with postmenopausal osteoporosis and carry out bioinformatics analysis. METHODS: Human monocytic leukemia cell line (THP-1) cells crosslinked with formaldehyde were broken by ultrasound. The experimental group was hybridized with magnetic beads combined with biotin labeled specific probes. The control group was hybridized with the magnetic beads of non-specific probes. The obtained peptides were identified using mass spectrometry. The data were searched and quantified by MaxQuant. The quantitative results of the two sets of samples were statistically analyzed, and the corresponding enrichment proteins were obtained. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, protein-protein interaction analysis and display were performed, and the protein-protein interaction network was constructed. RESULTS AND CONCLUSION: The total number of peptides obtained was 918, with 271 proteins in total, and the total number of proteins after filtration was 241. Compared with the control group, there were 10 differential proteins in the experimental group, including DDOST, DMBT1, HPD, IGLL5, IGK, LTF, LYZ, MUC5AC, PIGR, and RPL23. GO enrichment analysis showed that they were involved in biological processes such as defense reaction, leukocyte activation, ribosomal rRNA binding, lysozyme activity and other molecular functions. KEGG pathway analysis predicted that they were involved in ubiquinone and other terpenoid-quinone biosynthesis, phenylalanine metabolism, ribosome and salivary secretion. Combined with the analysis of proteomics and bioinformatics, it is predicted that uc431+, a gene related to postmenopausal osteoporosis, may be involved in immune regulation and bone metabolism by interacting proteins.

The effects of Xuling-Jiangu formula on the mRNA expression of osteoprotegerin and receptor activator of nuclear factor κB ligand in osteoporosis model rats / 国际中医中药杂志

Objective To study the effects and mechanism of Xuling-Jiangu formula on bone mineral density in the osteoporosis model rats.Methods According to the random number table method,50 SD female rats were randomly divided into the sham group,the model group,the low dose group of Xuling-Jiangu formula group,the medium dose group and the high dose of group.In addition to the sham group,the other groups were osteoporosis model.After 30 days,low dose group received intragastric Xuling-Jiangu formula solution 7.5 g/kg;medium dose group and high dose group received 15 and 30 g/kg,respectively.And the sham group and the model group received normal saline 10 ml/kg.After 12 weeks treatment,the bone mineral density of the left tibia was measured by double energy X ray.Estrone,osteocalcin in serum had been detected by Enzyme linked immunosorbent assay (ELISA).The mRNA expression of osteoprotegerin (OPG),receptor activator of nuclear factor κB ligand (RANKL) in lumbar were measured by quantitative real-time PCR.Results The bone mineral density (0.215 ± 0.010 g/cm2,0.222 ± 0.013 g/cm2 vs.0.196 ± 0.016 g/cm2) of the Xuling-Jiangu formula group were significantly higher than that of the model group (P＜0.01 or P＜0.05).The estrone in low,middle and high dose groups showed an upward trend,but there was no significant difference compared with the model group.The OC (87.0 ± 8.9 ng/L vs.100.5 ± 16.8 ng/L) of high dose group was significantly lower than that of the model group (P＜0.05).Compare with the model group,OPG mRNA level (0.97 ± 0.23 vs.0.78 ± 0.17) in high dose group increased (P＜0.05),the RANKL mRNA level decreased significantly (1.12 ± 0.17,0.97 ± 0.38,1.04 ± 0.29 vs.1.31 ± 0.18) in three Xuling-Jiangu formula groups (P＜0.05).Conclusions The bone mineral density of rats with osteoporosis can be improved by treating Xuling-Jiangu formula.It may be related to the increase of mRNA expression of OPG,and the reduction of RANKL.

Effects of liver-soothing therapy on the luteotropic hormone receptor and follicle stimulating hormone receptor of perimenopausal syndrome rats with liver qi stagnation / 国际中医中药杂志

Objective To explore the mechanism of Liver-soothing therapy on the Luteotropic hormone receptor (LHR) and Follicle stimulating hormone receptor (FSHR) in Perimenopausal Syndrome (PMS). Methods A total of 30 nature aging rats (13-month-old) were randomly assigned into three groups;PMS Liver-qi stagnation model (n=8), PMS Liver-qi stagnation model (n=8) treated with Chaihu-Shugan powder (4.0 g/kg?d) and PMS Liver-qi stagnation model (n=8) treated with Danzhi-Xiaoyao powder (4.0 g/kg?d). The PMS Liver-qi stagnation syndrome rat model were established by immobilizing the nature aging rats. Twelve-week-old female rats (n=8) were used as normal controls. Water decoctions of Chaihu-Shugan powder or Danzhi-Xiaoyao powder were administered respectively for 3 weeks while the rat models established. The serum E2, FSH, LH level were measured by radioimmunoassay. The LHR, FSHR in ovary were measured by quantitative real-time PCR. Results Compared with Liver-qi stagnation syndrome rat model, FSH in the Chaihu-Shugan powder group (4.32 ± 0.33 mIU/ml vs. 5.24 ± 0.45 mIU/ml) decreased (P<0.01), and LH in the Danzhi-Xiaoyao powder group (6.76 ± 0.52 mIU/ml vs. 8.08 ± 0.59 mIU/ml) decreased (P<0.01). Compared with normal controls, LHR mRNA, FSHR mRNA level increased in PMS Liver-qi stagnation model. Compared with Liver-qi stagnation syndrome rat model, LHR mRNA, FSHR mRNA level decreased (7.42 ± 2.54,4.91 ± 1.76 vs. 3.80 ± 1.36) in the ovary of Danzhi-Xiaoyao powder group (P<0.01), but there was no remarkable FHR, LHR expression changes in Chaihu-Shugan powder group. Conclusions The mechanism of Liver-soothing therapy may be related to the regulation of endocrine and decrease of LHR, FSHR.

The effects ofXuling-Jianguformula on bone mineral density and the bone biomechanic in osteoporosis model rats / 国际中医中药杂志

Objective To study the effects ofXuling-Jiangu formula on bone mineral density and the bone biomechanic in the osteoporosis model rats.Methods According to the random number table method, 40 SD female rats were randomly divided into the caltrate D group, theXuling-Jianguformula group, the model group and the sham group,10 rats each group. In addition to the sham group, the other groups were osteoporosis model. After 30 days, the Caltrate D group received intragastric caltrate D mixed suspension 0.126 g/kg; the Xuling-Jiangu formula group receivedXuling-Jianguformula solution 15 g/kg, and the sham group and the model group received normal saline 10 ml/kg. After 12 weeks treatment, detection of left tibia bone mineral density andthree-point bending method was used for biomechanical testing.Results The mineral density of the Xuling-Jiangu formula group (0.244 ± 0.022 g/cm2,0.195 ± 0.017 g/cm2vs. 0.223 ± 0.013 g/cm2) were significantly higher than the model group and caltrate D group (P<0.01); compared with the model group, the bone biomechanictests in theXuling-Jiangu formula group (0.072 ± 0.036 kN vs.0.041 ± 0.015 kN; 843.754 ± 428.722 N/mm2vs. 482.084 ± 176.646 N/mm2) were significantly higher (P<0.05).ConclusionXuling-Jiangu formula canimprove the bone mineral density and the bone lbiomechanic of osteoporosis rats.

Effects of liver-soothing therapy on the expression of estrogen receptors mRNA in perimenopausal syndrome rats with liver qi stagnation / 国际中医中药杂志

Objective To explore the effect of liver-soothing therapy on the expression of estrogen receptors mRNA in perimenopausal syndrome (PMS) rats with liver qi stagnation. Methods A total of 30 nature aging rats are assigned into control groups (n=8), model groups (n=8),Chaihu-Shugan San (CHSGS group,n=8) andDanzhi-Xiaoyao San (DZXYS group, n=8), according to the random number table. The PMS liver-Qi stagnation syndrome rat models were established by the methods of isolation raised and chronic bondage in all the groups except the control group. CHSGS group were administered 4.0 g/kg water decoctions ofChaihu ShuganSan, and DZXYS group 4.9 g/kg water decoctions ofDanzhi XiaoyaoSan respectively for 3 weeks after the rat models established. The model group and control group were administered with equal volume of normal saline. The open field test was used for the behavior test. The serum E2, FSH, LH level were measured by radioimmunoassay. The ERα, ERβ in ovary were measured by quantitative real-time PCR. Results Compared with model group on the 21st days, the CHSGS and DZXYS groups showed a significantly increase in crossings (49.6 ± 6.0, 51.6 ± 5.8vs. 40.0 ± 4.6,P0.05) increaed in the DZXYS group, but there was no remarkable difference. Conclusion The Liver-soothing therapy can improve the behavior of PMS rats with liver-Qi stagnation, and the mechanism may be related to the regulation of endocrine and ovarian estrogen receptors.

Effect of Chaihushugan San on expression of the Raf/mitogen-activated protein kinase/extracellular signal-regulated kinase pathway in the hippocampi of perimenopausal rats induced by immobilization stress.

OBJECTIVE: We wished to study the impact of Chaihushugan San (CSS) on the behavior of perimenopausal rats with liver-Qi stagnation (LQS) and to investigate the effect of CSS on signal transduction of the Raf/mitogen-activated protein kinase (MEK)/ extracellular signal-regulated kinase (ERK) cascade in the hippocampi of rats induced by immobilization. METHODS: Twenty 52-week-old female rats were divided into two groups by the random number table method: model control group (MCG) and CSS group (CSSG), with 10 rats in each group. Ten-week-old female rats were used as the normal control group (NCG). CSS effects were assessed using rats exposed to immobilization stress by measuring body weight and sucrose consumption, serum hormone levels, and observing performance in the open field test (OFT). Molecular mechanisms were examined by measuring the effect of CSS on expression of Raf1, MEK1/2 and ERK1/2 mRNA in hippocampi using quantitative real-time polymerase chain reaction and by measuring levels of these proteins and related phospho-proteins using Western blotting. RESULTS: Perimenopausal rats with LQS had decreased locomotor activity; reduced sucrose consumption; and increased serum levels of corticotropin releasing hormone (CRH) and corticosterone (CORT). Activation of hippocampal Raf/MEK/ERK cascade was suppressed significantly in the MCG, and activation was increased after 21 days of CSS treatment. CONCLUSION: CSS has significant effects upon relief of the symptoms of LQS in immobilization-induced rats. The mechanism underlying this action might (at least in part) be mediated by reversal of disruption of the Raf/MEK/ERK pathway.

The association between polymorphism of parathyroid hormone gene and bone mineral density in Fuzhou postmenopausal women / 中国组织工程研究

BACKGROUND:Studies have shown the bone mineral density of postmenopausal women is closely related to parathyroid hormone. But there are differences in different areas. OBJECTIVE:To investigate the association between BstBⅠ polymorphism of parathyroid hormone gene with bone mineral density in postmenopausal women from Fuzhou area. METHODS:The bone mineral densities of the lumbar spine, femoral neck, trochanter and Ward’s triangle were measured in 150 postmenopausal women by dual energy X-ray absorptiometry. The genotype of parathyroid hormone gene was determined by polymerase chain reaction-restriction fragment length polymorphism. RESULTS AND CONCLUSION:(1) The distribution of parathyroid hormone genotypes were BB genotype 68.8%, Bb 24.1%, and bb 7.1%. The B al elic gene frequencies reached 81%, while b was 19%. The distribution fol owed the Hardy-Weinberg equilibrium. (2) Analysis of the relationship between the genotypes and bone mineral density:There was no significant difference in the bone mineral densities of the lumbar spine, femur, neck, trochanter and Ward’s triangle among the three genotypes (P>0.05). BstBⅠ gene polymorphism of parathyroid hormone gene is not correlated to bone mineral density, and there is no enough evidence to support genotype of parathyroid hormone gene as a genetic marker in predicting the risk of developing osteoperosis in Fuzhou postmenopausal women.

The effect of the PXh haplotype combining estrogen receptor gene Xba Ⅰ, Pvu Ⅱ polymorphisms and osteocaicin gene Hind Ⅲ polymorphism on bone mineral density in postmenopausal women / 中华老年医学杂志

Objective To investigate the correlation between bone mineral density(BMD)and the PXh haplotype combining estrogen receptor (ER) gene Xba Ⅰ , Pvu Ⅱ polymorphisms and osteocalcin gene Hind Ⅲ polymorphism in postmenopausal women.Methods In 307 subjects,the BMD of lumbar vertebrae and proximal femur were measured by dual energy X-ray absorptiometry and the Xba Ⅰ and Pvu Ⅱ polymorphisms of ER gene and the Hind Ⅲ potymorphism of osteocalcin gene were detected by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP).Results (1)The BMD of greater trochanter was significantly lower in XX genotype group than in xx genotype group ( P<0.05).The BMD of femoral neck, greater trochanter and Ward's triangle were lower in Xx genotype group[(0.695±0.087)g/cm2 , (0.592±0.106)g/cm2, (0.500±0.115) g/cm2] and X allele group[(0.697±0.088)g/cm2 , (0.594±0.105)g/cm2, (0.505±0.123)g/cm2] than in xx genotype group[(0.737±0.108) g/cm2,(0.653±0.119)g/cm2 ,(0.554±0.130)g/cM2] and non-X allele group[(0.737 ± 0.108) g/CM2, (0.653 ± 0.119) g/cm2 , (0.554 ± 0.130) g/cm2] ,respectively (all P<0.05 ).(2)The BMD of Ward's triangle was lower in PP genotype group and P allele group than in pp genotype group and non-P allele group (P<0.05).(3)The BMD of femoral neck, greater trochanter and Ward's triangle were lower in hh genotype group and h allele group than in I-IH genotype group, and were lower in non-h allele group than in HH genotype group(all P<0.05).(4)Women carrying PX, PXh haplotypes combining ER gene and osteocalcin gene had lower BMD at femoral neck than those not carrying PX,not carrying PXh haplotypes, respectively (all P<0.05).ConclusionsER gene(Xba Ⅰ) polymorphism and osteocalein gene(Hind Ⅲ) polymorphism are associated with BMD in postmenopausal women.The presence of X allele or h allele　shows negative influence on the BMD of postmenopausal women.The PXh haplotype is a suitable　genetic marker of postmenopausal women osteoporosis in Fuzhou area.

Association of genetic polymorphisms in several vitamin D receptor gene sites with bone mineral density and biochemical markers of bone turnover in postmenopausal women / 中国组织工程研究

OBJECTIVE: To study the association of the vitamin D receptor gene BSM Ⅰ, TAQ Ⅰ and APA Ⅰ genetic polymorphisms with bone mineral density and biochemical markers of bone turnover in postmenopausal women.METHODS: ①total of 576 postmenopausal Han ethnic women of 48-84 (62.17±6.37) years old in Fuzhou city were investigated, on the basis of their informed consent, through random sampling method from January 2007 to December 2008. ②The subjects were recorded regarding to their age, menopause duration, body mineral index and postmenopausal fracture incidence. ③Dual energy X-ray absorptiometry was used for measuring the bone mineral density of vertebrae L<,2-4>, left femoral neck, trochanter and Ward's triangle. ④The genetic polymorphisms of vitamin D receptor gene BSM Ⅰ, TAQ Ⅰ and APA Ⅰ were detected using polymerase chain reaction-rastriction and fragment length polymorphism (PCR-RFLP) technique. ⑤The biochemical markers of bone turnover (serum bone gla protein, serum bone alkaline phosphatase, urinary pyddinoline and urinary deoxypyridinoline) were detected with enzyme linked immunosorbent assay.RESULTS: A total of 561 subjects up to standard were involved in the result analysis. ①There was no significant difference in bone mineral density among genotypes of vitamin D receptor gene BSM Ⅰ, TAQ Ⅰ and APA Ⅰ polymorphisms (P > 0.05). ②There was no significant difference in the biochemical markers of bone tumover among genotypes of BSM Ⅰ, TAQ Ⅰ and APA Ⅰ polymorphisms (P > 0.05). ③There was no significant difference in the incidence of osteoporosis among genotypes of BSM Ⅰ, TAQ Ⅰ and APA Ⅰ polymorphisms (P > 0.05). ④There was no significant difference in the incidence of postmenopausal fracture among genotypes of BSM Ⅰ, TAQ Ⅰ and APA Ⅰ polymorphisms (P > 0,05).CONCLUSION: BSM Ⅰ, TAQ Ⅰ and APA Ⅰ polymorphisms of the vitamin D receptor gene are not obviously associated with osteoporesis in postmenopausal women, and accordingly can not be taken as genetic markers of osteoporosis in postmenopausal women in Fuzhou.