Impact of 1 25(OH) 2 D 3 on TG content in liver of rats with type 2 diabetes

Abstract Purpose: To evaluate the effects of 1,25 dihydroxy vitamin D3 (1,25(OH)2D3) on the content of triglyceride (TG), as well as on the gene and protein expressions of adiponectin receptor 2 (AdipoR2), p38 mitogen-activated protein kinase (P38MAPK), and lipoprotein lipase (LPL) in the liver of rats with type 2 diabetes mellitus (T2DM) so as to provide theoretical basis for exploring the mechanism by which 1,25(OH)2D3 regulates TG. Methods: Wistar rats were divided into four groups (n=25), with different treatments and detected the gene and protein expressions of AdipoR2, p38MAPK, and LPL in the liver tissue by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting. Meanwhile, the content of TG in the liver tissue was detected by the Enzyme-linked immunosorbent assay. Results: The expression of AdipoR2, p38MAPK, LPL gene and protein in the liver of VitD intervention group was significantly higher than that in T2DM group (P <0.05), while the TG content was significantly lower than that in T2DM group (P <0.05). Conclusion: 1,25(OH)2D3 can decrease the content of TG in the liver, and its mechanism may be achieved by upregulating the expressions of AdipoR2, p38MAPK, and LPL in the liver.

The effects of excessive fluoride on expression of osteocalcin in osteoblasts and glucolipid metabolism in mice / 中华地方病学杂志

Objective To explore excessive fluoride on expression of osteocalcin (OC) in osteoblast and glucolipid metabolism in mice.Methods 3T3-L1 preadipocyte and MC3T3-E1 osteoblast were conventionally cultured,MC3T3-E1 cells were stimulated with O,2,10 and 50 mg/L sodium fluoride (NaF) to establish fluorine cell model,and levels of OC were tested.The corresponding NaF concentration resulted in the highest OC level was used as the optimal fluorine concentration.Cell experiments were divided into four groups:3T3-L1,3T3-L1 + NaF,3T3-L1 + MC3T3-E1 and 3T3-L1 + MC3T3-E1 + NaF.The treatment groups were respectively or jointly treated with the optimal concentration of NaF (50 mg/L) or MC3T3-E1 osteoblast,enzyme linked immunosorbent assay (ELISA) was used to detect OC and adiponectin (APN) levels.At the same time,40 C57BIL/6 mice were numbered by weight,randomly divided into control and fluoride groups,20 per group,control group drunk pure water,fluoride group drunk 100 mg/L NaF solution,changes of teeth and body weight [M (P25,P75)] of the mice were observed.Serum OC and APN levels were tested by ELISA at 12 weeks after modeling.The fasting plasma glucose (FPG) and the fasting plasma insulin (FINS) were detected by glucose oxidase and chemiluminescence methods,and insulin resistance (HOMA-IR) was evaluated.Results The APN levels of 3T3-L1,3T3-L1 + NaF,3T3-L1 + MC3T3-E1 and 3T3-L1 + MC3T3-E1 + NaF groups were (0.94 ± 0.18),(1.07 ± 0.21),(1.76 ± 0.20),and (2.49 ± 0.43) μg/L,MC3T3-E1 osteoblast with NaF had promote collaborative interaction effect on APN levels (F =14.519,P ＜ 0.01).The body weight of mice in fluoride group [27.5 (25.8,28.3) g] was significantly lower than that of the control group [31.4 (30.3,32.6) g,Z =-4.695,P ＜ 0.01].The levels of FPG [(7.78 ± 1.86) mmol/L],FINS [(3.22 ± 0.75) mU/L],OC [(6.11 ± 1.49) μμg/L],APN [(8.65 ± 1.78) μg/L] and HOMA-IR (1.15 ± 0.49) were higher than those of control groups [(5.40 ± 1.51) mmol/L,(2.45 ± 0.64) mU/L,(3.14 ± 0.92) μg/L,(4.03 ± 1.45) μg/L,0.62 ± 0.31],the differences were statistically significant (t =-4.466,-3.518,-7.560,-9.002,-4.182,P ＜ 0.01).OC levels in mice were positively correlated with FPG,FINS,APN and H-OMA-IR (r =0.868,0.707,0.911,0.818,P ＜ 0.01).Conclusion The OC of osteoblast in mice exposed to fluoride is increased significantly,OC levels in mice are closely related to blood glucose and APN,it is one of the key molecules in lipid metabolism.

The effects of excessive fluoride on expression of osteocalcin in osteoblasts and glucolipid metabolism in mice / 中华地方病学杂志

Objective To explore excessive fluoride on expression of osteocalcin (OC) in osteoblast and glucolipid metabolism in mice.Methods 3T3-L1 preadipocyte and MC3T3-E1 osteoblast were conventionally cultured,MC3T3-E1 cells were stimulated with O,2,10 and 50 mg/L sodium fluoride (NaF) to establish fluorine cell model,and levels of OC were tested.The corresponding NaF concentration resulted in the highest OC level was used as the optimal fluorine concentration.Cell experiments were divided into four groups:3T3-L1,3T3-L1 + NaF,3T3-L1 + MC3T3-E1 and 3T3-L1 + MC3T3-E1 + NaF.The treatment groups were respectively or jointly treated with the optimal concentration of NaF (50 mg/L) or MC3T3-E1 osteoblast,enzyme linked immunosorbent assay (ELISA) was used to detect OC and adiponectin (APN) levels.At the same time,40 C57BIL/6 mice were numbered by weight,randomly divided into control and fluoride groups,20 per group,control group drunk pure water,fluoride group drunk 100 mg/L NaF solution,changes of teeth and body weight [M (P25,P75)] of the mice were observed.Serum OC and APN levels were tested by ELISA at 12 weeks after modeling.The fasting plasma glucose (FPG) and the fasting plasma insulin (FINS) were detected by glucose oxidase and chemiluminescence methods,and insulin resistance (HOMA-IR) was evaluated.Results The APN levels of 3T3-L1,3T3-L1 + NaF,3T3-L1 + MC3T3-E1 and 3T3-L1 + MC3T3-E1 + NaF groups were (0.94 ± 0.18),(1.07 ± 0.21),(1.76 ± 0.20),and (2.49 ± 0.43) μg/L,MC3T3-E1 osteoblast with NaF had promote collaborative interaction effect on APN levels (F =14.519,P ＜ 0.01).The body weight of mice in fluoride group [27.5 (25.8,28.3) g] was significantly lower than that of the control group [31.4 (30.3,32.6) g,Z =-4.695,P ＜ 0.01].The levels of FPG [(7.78 ± 1.86) mmol/L],FINS [(3.22 ± 0.75) mU/L],OC [(6.11 ± 1.49) μμg/L],APN [(8.65 ± 1.78) μg/L] and HOMA-IR (1.15 ± 0.49) were higher than those of control groups [(5.40 ± 1.51) mmol/L,(2.45 ± 0.64) mU/L,(3.14 ± 0.92) μg/L,(4.03 ± 1.45) μg/L,0.62 ± 0.31],the differences were statistically significant (t =-4.466,-3.518,-7.560,-9.002,-4.182,P ＜ 0.01).OC levels in mice were positively correlated with FPG,FINS,APN and H-OMA-IR (r =0.868,0.707,0.911,0.818,P ＜ 0.01).Conclusion The OC of osteoblast in mice exposed to fluoride is increased significantly,OC levels in mice are closely related to blood glucose and APN,it is one of the key molecules in lipid metabolism.

Global adiponectin suppress the high expression of monocyte chemotactic protein-1 induced by high glucose in NRK52E cells / 中华肾脏病杂志

Objective To investigate the effect of globular adiponectin on the high expression of monocyte chemotactic protein-1 (MCP-1) induced by high glucose in rat renal tubular epithelial cells (NRK52E),and its relationship with adiponectin receptors and p38MAPK.Methods NRK52E cells were cultured in vitro and divided into six groups:normal glucose group (NG,5.6 mmol/L glucose),high glucose group(HG,25 mmol/L glucose),gAd groupl (HG+gAd 2 mg/L),gAd group2 (HG+gAd 5 mg/L),gAd group3 (HG+gAd 10 mg/L),p38MAPK antagonist group:(SB,HG+SB203580 10 μmol/L).The protein expression of phosphorylated p38MAPK (p-p38MAPK),total p38MAPK (t-p38MAPK),MCP-1 and AdipoR1/AdipoR2 were examined by western blotting.The mRNA expression of MCP-1 and AdipoR1/AdipoR2 were detected by RT-PCR and real-time PCR respectively.Results Compared with NG group,the mRNA and protein expression of MCP-1 increased significantly in HG group (all P＜ 0.05).The phosphorylation of p38MAPK increased (P＜ 0.05) with no change in t-p38MAPK protein.The addition of gAd or SB203580 inhibited the unregulation of MCP-1 and p-p38MAPK induced by HG.Two kinds of adipoR,adipoR1 and adipoR2,were all detectable in NG group,and mRNA and protein expression of adipoR1 was higher than that of adipoR2 (P＜ 0.01).Compared with NG group,the expression of adipoR decreased in HG group,but the difference had no statistical significance(P ＞ 0.05).Compared to HG group,the mRNA and protein expression of adipoR1 increased in gAd groups (all P ＜ 0.01).Conclusion The gAd can dose-dependently attenuate the overexpression of MCP-1 induced by high glucose,and this protective effect may be mediated by adipoR1 and p38MAPK.

Relationship between aoiponectin receptor 1 gene + 10225C/G variant and type 2 diabetes / 中国基层医药

Objective To investigate the relationship between single nucleotide polymorphism (SNP) at position + 10225 in adiponectin receptor 1 gene and type 2 diabetes.Methods The genotypes of + 10225C/G of adiponectin receptor 1 were detected by polymerase chain reaction with sequence specific primers (PCR-SSP) in 200 type 2 diabetes and 100 health controls.Fasting blood samples of all cases were obtained to extract DNA and detect genotype.Statistical software spss 13.0 was used to analyzed.Results Frequency of G-type allele in type 2 diabetes was 40.5％,that in normal controls was 23.5％,there was significant diffierences in the genotype frequencies of SNP+10225 between type 2 diabetes and health controls (x2 =128.0,P＜0.01).Conclusion AdipoR1 + 10225C/G polymorphism was probably associated with type 2 diabetes,G-type allele might be a genetic risk factor of type 2 diabetes.

Preliminary study of adiponectin receptor Ⅰ gene polymorphisms and polycystic ovary syndrome / 中国医师杂志

Objective To study the correlation of the adiponectin receptor 1 gene rs1139646C/G and rs10920531C/A single nucleotide polymorphisms with Chinese Han women with polycystic ovary syndrome (PCOS).Methods For PCOS group (n =80) and control group (n =80),polymerase chain reaction (PCR) was used to amplify the upstream and downstream 250 bases of rsl139646C/G and rs10920531C/A,respectively.PCR products were directly sequenced.Results For rs1139646C/G loci sites between PCOS and control groups,all were the homozygous genotype CC,and no statistical significance was found for genotype frequency and allele frequency between two groups.For rs10920531C/A loci sites between PCOS and control groups,no statistical significance was found for genotype frequency and allele frequency between two groups (P =0.683,0.580),and no statistical significance was found for two loci with genetic linkage disequilibrium analysis (x2 =0.307,Fisher 's P =0.579610,Pearson' s P =0.579608).Conclusions Adiponectin receptor 1 gene rs1139646C/G and rs10920531C/A single nucleotide polymorphisms have nothing to do with Chinese Han PCOS women.Genetic linkage disequilibrium analysis has nothing to do with PCOS.The rs1139646C/G in Chinese Han population is relatively stable site,and no locus mutation is found.

Expression of adiponectin and adiponectin receptor 2 in the livers of patients with hepatitis B virus-related liver failure / 中华传染病杂志

Objective To explore the expressions of adiponectin and adiponectin receptor 2 (AdipoR2) in the liver of patients with hepatitis B virus (HBV)-related liver failure and their clinical implications in the pathogenesis of liver failure.Methods The healthy controls (HC),patients with chronic hepatitis B (CHB) and patients with HBV-related liver failure (HBV-LF) were enrolled in the study.Each group had 20 participants.The liver tissues from 11 CHB patients who were diagnosed by liver biopsy,6 patients with HBV-LF and 6 liver donors during liver transplantation were collected.Histological specimens were observed by hematoxylin-eosin staining and Masson trichrome staining under microscope.The mRNA and protein expressions of adiponectin and AdipoR2 in the liver were measured by semi-quantitative reverse transcription and polymerase chain reaction (SqRT-PCR) and immunohistochemical staining,respectively.The level of serum adiponectin was detected by enzymelinked immunsorbent assay.Serum biochemical parameters and HBV DNA levels were also measured.The pairwise comparison between groups was done by Student-Newman-Keuls and mann whitney U test.The relationship between two variables was analyzed using Spearman correlation.Results The level of serum adiponectin in HBV-LF group [(0.86 ± 0.15) ng/mL] was higher than those in HC [(0.59±0.15) ng/mL] and CHB groups [(0.62±0.13) ng/mL,Z=3.963,Z=3.774,both P＜0.01)],but showing no difference between CHB and HC groups (P＞0.05).The expressions of adiponectin and AdipoR2 mRNA in the liver were significantly higher in HBV-LF group (0.251 ±0.028 and 0.223 ± 0.021,respectively) than those in HC (0.091 ± 0.018 and 0.072 ± 0.020,respectively) and CHB (0.121±0.019 and 0.097±0.017,respectively) groups (q=18.428,17.069,19.807,18.673,respectively; all P＜0.01).Also,the expressions of adiponectin and AdipoR2 mRNA in CHB group were significantly higher than those in HC group (q=3.895,3.860,both P＜0.05).The expressions of adiponectin and AdipoR2 proteins in the liver were significantly higher in HBV-LF group (8.482±0.772 and 7.654±0.272,respectively) than those in HC (4.045± 0.815 and 2.804±0.623,respectively) and CHB (5.545±0.613 and 4.775±0.458,respectively) groups (q=15.327,11.542; Z=2.802,3.266; respectively; all P＜0.01).Also,the expressions of adiponectin,AdipoR2 proteins in CHB group were significantly higher than those in HC group (q=5.894,Z=3.266,both P＜0.01).In HBV-LF group,serum adiponectin levels as well as the expressions of adiponectin mRNA and protein in the liver were negatively correlated with serum albumin (r=-0.815,-0.886,-0.943; P＜0.01 or P＜0.05),but positively correlated with serum alanine aminotransferase (r=0.701,0.886,0.943; P＜0.01 or P＜0.05).The expression of AdipoR2 mRNA in the liver was negatively correlated with serum albumin (r=-0.943,P＜0.01),but positively correlated with serum aspartate aminotransferase (r=0.829,P＜0.05).Conclusions The expressions of adiponectin and AdipoR2 are up-regulated during HBV infection,especially in patients with HBV-LF,which might reflect the degree of necroinflammation in the liver.

Correlation between adiponectin and breast cancer / 国际肿瘤学杂志

Adiponectin is a kind of adipocyte-specific protein which is only inversely related with obesity by now. It is involved in enhancing insulin sensitivity,anti-atherogenic and antiinflammatory activities.Recent studies have reported that adiponectin is closely correlated with the genesis and development of a variety of obesity related malignant tumors,especially postmenopausal breast cancer.Obesity is an independent risk factor for the development of breast cancer.Recently researches about the correlation between adiponectin and breast cancer relations and the action mechanisms,signaling pathways have made a progress.

Myocardial protection of adiponectin /adiponectin receptor 1 signal pathway in rats with limb ischemic preconditioning / 中华老年医学杂志

Objective To investigate the myocardial protection of adiponectin (ADP) /adiponectin receptor 1 (ADPR1) related-signal pathway in rats with limb ischemic preconditioning.Methods Thirty SD male rats were randomly divided into sham-operation group,myocardial ischemia reperfusion injury (MIRI) group,limb ischemic preconditioning (LIPC) group,LY294002 (the PI3-specific inhibitor) pretreatment group and LY294002+LIPC group (n=10 each).The mRNA level of myocardial ADP and ADPR1,the protein expressions of phosphatidylinositol 3-kinase (PI3k)phosphorylated Akt (p-Akt) were determined by RT-PCR and Western blot,respectively. Results As compared with sham-operation group,the mRNA levels of ADP and ADPR1 in MIRI group were significantly decreased (0.53 ± 0.07 vs.0.74 ± 0.08 and 0.52 ± 0.02 vs.0.72 ± 0.04,P＜0.05).Compared with MIRI group,the mRNA levels of ADP (0.72±0.21) and ADPRI (0.80±0.023) in LIPC group were increased,ADP(0.49±0.07) and ADPR1 (0.52± 0.02) mRNA were decreased in LY294002 group (both P＜0.05),but there were no difference in ADP(0.70±0.16) and ADPR1(0.78±0.05) mRNA between LY294002+LIPC group and MIRI group.The protein levels of Pl3k and p-Akt were lower in MIRI group than in sham-operation group (3.85±0.23 vs.2.83±0.22and 3.77±0.32 vs.2.66±0.29,P＜0.05).In contrast to MIRI group,the yield of PI3k (2.65±0.32)and p-Akt(2.26±0.27) protein (P＜0.05) were increased in LIPC group,but there were unproductive protein of PI3k (3.75 ± 0.65) and p-Akt (4.01 ± 0.71) in LY294002 group with no differences versus the levels of PI3k (3.23 ± 0.48) and p-Akt (3.17 ± 0.54) in LY294002 + LIPC group. Conclusions Limb ischemic preconditioning may protect myocardium by promoting serum adiponectin levels,improving myocardial mRNA expressions of ADP and ADPR1,activating the ADP/PI3k/Akt signaling pathway in reperfusion injury.

Adiponectin and ischemic stroke / 国际脑血管病杂志

Adiponectin (APN) is an insulin sensitizing protein with protective effect secreted by adipose tissue.It has the effects of regulating glucose and lipid metabolism,protecting vascular endothelial function,promoting angiogenesis,and anti-inflammation,etc.It plays an important protective effect in cerebrovascular diseases.This article reviews APN and its effect in ischemic stroke.

Globular adiponectin protects human umbilical endothelial cells against apoptosis induced by advanced glycation end products through increasing adiponectin receptor 1 / 中国病理生理杂志

AIM: To investigate the effects of globular adiponectin (gAd) on the expression of adiponectin receptor 1 (AdipoR1) in human umbilical vein endothelial cells (HUVECs), and on the apoptosis induced by advanced glycation end products (AGEs). METHODS: HUVECs were treated with the indicated concentrations of AGEs for 24 h or 48 h in vitro. The cells in gAd treatment group were pretreated with gAd for 24 h, and then were treated with AGEs for another 24 h or 48 h. Cell variability was quantified by MTT assay. Apoptotic cells were detected by flow cytometry with Annexin-FITC/PI double staining. AdipoR1 mRNA in the cells was determined by quantitative real time reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: The viability of HUVECs treated with AGEs decreased significantly as compared to the cells treated with HSA (control, P<0.05). Under the same condition of AGEs exposure, the viability of the cells treated with gAd was greatly higher than that of the cells without gAd treatment (P<0.01). The apoptotic rate of HUVECs was significantly elevated by AGEs treatment vs HSA treatment observed by Annexin V-FITC/PI double staining analysis with flow cytometry (P<0.05). Under the condition of AGEs stimulation, the apoptosis of HUVECs was decreased by pretreatment with gAd as compared to that of the cells without gAd treatment (P<0.01). Measured by quantitative real time PCR, AGEs decreased the expression level of AdipoR1 mRNA and gAd increased the expression of AdipoR1 mRNA contrarily (P<0.05). CONCLUSION: AGEs increase the apoptotic rate of HUVECs in a concentration dependent manner and gAd promotes the AdipoR1 mRNA expression.

Effects of high levels of glucose on the expression of adiponectin receptors in human kidney proximal tubular cells / 中国医师杂志

Objective To investigate the effect of in vitro high glucose stimulation on the expression of adiponectin receptor (adipoR) in human kidney proximal tubular cells.Methods The HK-2 cells were cultured in the low glucose DMEM culture medium containing 10% fetal bovine serum until the cells were adherent and 80% confluence. After cultured in the serum-free DMEM for 24 hours, these cells were stimulated with glucose-containing 1mg/ml, 2mg/ml, 4mg / ml, 6mg/ml, 8mg/ml serum-free DMEM for 48 hours. Then RT-PCR and western blot were used to analyze adipoR ( R1, R2) expression levels. The HK-2 cells were cultured respectively in high glucose (4mg/ml) , low glucose (1mg/ml) DMEM culture medium containing 10% fetal bovine serum to cultivate 0h, 12h, 24h, 48h, 72h, 96h, then RT - PCR was applied to analyze adipoR (R1, R2) mRNA expression levels semi-quantitatively. Results Two kinds of adiponectin receptor gene were both expressed in HK-2 cells, and the quantity of gene expression of adipoR1 (0. 63 ±0. 12) was 3. 9 times to adipoR2 (0. 16 ±0.03) , the difference was statistically significant ( P<0. 01). The different concentrations of glucose and different time of high glucose on HK-2 cells had no significant effect ( P>0. 05 ) on adipoR gene expression. Expression of adipoR 1 protein in HK-2 cells was detected by western blot, and it was not affected by glucose concentration ( P>0. 05).Conclusion adi-poR1 and adipoR2 gene were both expressed in HK-2 cells, and the adipoR1 was the major one, which suggested that adipoR1 played a more significant role in kidney disease. The expression of adipoRl/R2 of HK-2 cells was not affected by high glucose concentration.

Distribution and expression of adiponectin receptors in different tissues of normal Wistar rats / 解放军医学杂志

Objective To investigate the distribution and expression of adiponectin receptors in different tissues of normal Wistar rats. Methods The rats were euthanized and the total RNA was extracted from equal weight tissue samples of cardiac muscle, skeletal muscle, liver, kidney, testicle and fat. RT-PCR was employed to amplify the isolated cDNA, and the expression of adiponectin receptors in different tissues was semi-quantitatively analyzed. Results AdipoR1 was observed in rat cardiac muscle, skeletal muscle, liver, kidney, testicle and fat, with the highest expression in testicle and fat (P