Thrombomodulin gene polymorphism and thrombomodulin expression in essential hypertension

Patients with hypertension have the characteristics of abnormalities of vessel wall,blood constituents and blood flow. These abnormalities may confer a prothrombotic or hypercoagulable state and are related to the damage of target organs and long-term prognosis. Soluble thrombomodulin （sTM） as abnormalities of levels of specific plasma markers of endothelial damage or dysfunction may relate with the complications of hypertension and the determination of blood pressure itself. TM plays a critical role as a co-factor in the protein C pathway, 1 which is important in regulating coagulation as well as inflammation. Thus we hypothesized that the -33G〉A polymorphism alter thrombomodulin expression and/or impair anticoagulant function, which can predispose to the damage of the target organs during the progress of hypertension. Then, we investigated a possible association of sTM, TM on monocytes and the -33G〉A polymorphism with essential hypertension and cardiovascular disease （CVD） in the Chinese Han ethnic population.     

Altered expression of adipose differentiation-related protein gene in placental tissue of pre-eclampsia

Objective: To investigate the altered expression of lipid metabolism-related gene adipose differentiation-related protein (ADRP) in pre-eclampsia. Methods: Semi-quantitative RT-PCR and Western blotting were used to validate the altered expression of ADRP gene between pre-eclamptic placentas (pre-eclampsia group) and normotensive placentas (control group) respectively. In situ hybridization (ISH) was used to localize ADRP mRNA in pre-eclamptic placentas. Results: There was a significant difference in the levels of placental ADRP mRNA between pre-eclampsia group and control group (1. 98±0. 50 vs 1. 09±0. 20, P<0. 01). Western blotting showed that placentas both in pre-eclampsia group and control group expressed the special ADRP band at 48. 1 kD. The relative levels of ADRP protein in pre-eclampsia group were significantly higher than those of control group (0. 40 ±0. 19 vs 0. 19 ±0. 09, P<0. 01 ). ADRP mRNA was diffusely distributed in pre-eclamptic placentas. Their positive staining existed in cytoplasm of trophoblast. Conclusion: Abnormal expression of ADRP gene in pre-eclamptic placenta may be associated with the pathogenesis of pre-eclampsia.     

Serum resistin and adiponectin concentrations in patients with overweight and obesity

Objective： To explore the serum levels of resistin and adiponectin in patients with overweight and obesity. Methods： Fifty-eight cases with normal weight and 24 patients with overweight and obesity have taken fasting blood samples for measurements of plasma glucose, plasma lipids, insulin, C-peptide, thyroid hormones, C-response protein, interleukin-6, TNF-α, leptin, adiponectin and resistin. Results： The concentrations of resistin in cases with overweight and obesity were significant higher than those in the normal weight cases （16. 01 ± 8. 60 vs 11. 63 ± 9. 05 ng/ml, P = 0. 047）. Pearson relation analysis showed that serum resistin concentrations were positively correlated with age （r= 0. 476, P = 0.019）, but negatively correlated with C-peptide （r=-0.45, P=0. 024）, and adiponectin concentrations were positively correlated with HDL-c （r=0. 463, P=0. 023） and systolic blood pressure （r=0. 409, P=0. 047） in overweight and obesity cases. Conclusion： The concentrations of resistin in cases with over-weight and obesity are higher, and there is no correlation between resistin and blood glucose, blood lipids and insulin, while the serum adiponectin concentrations positively correlated with HDL-c and systolic blood pressure.     

Expression of Resistin Protein in Normal Human Subcutaneous Adipose Tissue and Pregnant Women Subcutaneous Adipose Tissue and Placenta

Resistinisanovelpolypeptidehormoneintheadiposetissue,whichcharacterizedbyacysteine richmotif(CX12CX8CXCX3CX10CXCXCX9CC).Resistinismainlyexpressedinthewhiteadiposetissueandweakinthebrownadiposetissue.Re sistinlinksobesitytodiabetes[1,2].Serumresistinlevelsismarkedlyincreasedinob/obanddb/dbmouse[1].Thiazolidinedionescanobviouslyreduceresistinexpressioninmouse.Duringthepregnan cy,pregnantwomenhavephysiologicalinsulinre sistance,decreaseglucosetoleranceandinsulinsensitivityandareeasytoget…     

Expression of Resistin mRNA in Adipose Tissue of Rat Model with Polycystic Ovarian Syndrome and Its Implication

Polycysticovarysyndrome(PCOS),oneofthemostcommondisordersinpremenopausalwomen,involvesmenstrualdisturbance,chronicanovulation,andhyperandrogenism.WomenwithPCOSarefrequentlyinsulinresistant(IR),andhavehyperinsulinemia.Althoughithasbeendifficulttoevaluateth…     

Interaction of Dietary Composition and PYY Gene Expression in Diet-induced Obesity in Rats

Dietary obesity ,induced by exposure to highfat food,is the rodent model that most closely re-sembles human obesity . Comparing rats that areresistant or prone to diet-induced obesity serves thedual purpose of illustrating the molecular adapta-tions that prevent obesity and identifying genesthat become dysregulated by high fat diets .Neuropeptide Y ( NPY) and peptide YY(PYY) are closely related polypeptides which arecomposed of 36 amino acides and share considera-ble homology . While NPYa…     

Altered expression of renal bumetanide-sensitive sodium-pota-ssium-2 chloride cotransporter and Cl- channel -K2 gene in angiotensin Ⅱ-infused hypertensive rats

Background Little information is available regarding the effect of angiotensin Ⅱ (Ang Ⅱ) on the bumetanide-sensitive sodium-potassium-2 chloride cotransporter (NKCC2), the thiazide-sensitive sodium-chloride cotransporter (NCC), and the Cl- channel (CLC)-K2 at both mRNA and protein expression level in Ang Ⅱ-induced hypertensive rats. This study was conducted to investigate the influence of Ang Ⅱ with chronic subpressor infusion on nephron-specific gene expression of NKCC2, NCC and CLC-K2.Methods Sprague Dawleys rats were treated subcutaneously with either Ang Ⅱ (100 ng·kg-1·min-1 ) or vehicle for 14 days. Expression of NKCC2, NCC and CLC-K2 mRNA in kidneys was determined by real time polymerase chain reaction (PCR). Western blotting analysis was used to measure NKCC2 and NCC protein expression. Results Ang Ⅱ significantly increased blood pressure and up-regulated NKCC2 mRNA and protein expression in the kidney. Expression of CLC-K2 mRNA in the kidney increased 1.6 fold (P&lt;0.05).There were no changes in NCC mRNA or protein expression in AngII-treated rats versus control.Conclusions Chronic subpressor Ang Ⅱ infusion can significantly alter NKCC2 and CLC-K2 mRNA expression in the kidney, and protein abundance of NKCC2 in kidney is positively regulated by Ang Ⅱ. These effects may contribute to enhanced renal Na+ and Cl- reabsorption in response to Ang Ⅱ. Chin Med J 2005; 118(23):1945-1951     

Small interfering RNA inhibits SARS-CoV nucleocapsid gene expression in cultured cells and mouse muscles

SARS-CoV isa newly identified coronavirus that causes severe acute respiratory syndrome （SARS）. Currently, there is no effective method available for prophylaxis and treatment of SARS-CoVinfections. In the present study, the influence of small interfering RNA （siRNA） on SARS-CoV nucleocapsid （N） protein expression was detected in cultured cells and mouse muscles. Four siRNA expression cassettes driven by mouse U6 promoter targeting SARS-CoV N gene were prepared, and their inhibitory effects on expression of N and enhanced green fluorescence protein （EGFP） fusion protein were observed.     

Effects of fatty acid regulation on visfatin gene expression in adipocytes

Background The levels of long-term elevated serum or intracellular free fatty acid （FFA） induce insulin resistance associated with central obesity. The insulin-mimetic protein visfatin is preferentially produced by visceral adipose tissues and has been implicated in obesity and insulin resistance. To identify that FFA is capable of inducing insulin resistance and to clarify the role of FFA on visfatin, we examined the effect of monounsaturated FFA oleate （C 18： 1） and saturated FFA palmitate （C 16：0） on glucose transport and visfatin gene expression in cultured 3T3-L1 adipocytes or preadipocytes. Methods FFA-free DMEM/F12, 0.125 mmol/L, 0.5 mmol/1 and 1.0 mmol/L oleate or palmitate was added to cultured 3T3-L1 adipocytes or preadipocytes and incubated overnight. Glucose transport was assessed as 3H- 2-deoxy-glucose uptake. Total RNA was extracted and subjected to RT-PCR for the measurement of visfatin mRNA levels. Statistical comparisons between control group and other groups were performed with the two-tailed paired t test, and one-way ANOVA was used to compare the mean values among the groups. Results Insulin increased specific membrane glucose transport in 3T3-L1 preadipocytes. Upregulation was evident from 15 minutes to 1 hour exposure to insulin. However, after 6-hour exposure to insulin, there was a downregulation in the response to insulin. Dose response studies demonstrated that 2-deoxy glucose transport was increased by 336% at 50 nmol/L insulin （P〈0.01）, and reached a maximal effect at 100 nmol/L insulin （P〈0.01）. Oleate and palmitate treatment did not influence basal glucose transport （without insulin stimulation）, whereas insulin-stimulated glucose transport was inhibited after overnight oleate and palmitate treatment in preadipocytes and adipocytes. In 3T3-L1 preadipocytes, insulin resistance could be achieved at 0.125 mmol/L oleate or palmitate （P〈0.05, respectively）, and the inhibition was dose dependent. In adipocytes, the inhibition was noted at 0.5 mmol/L oleate or 1.0 mmol/L palmitate. Visfatin mRNA expression increased during differentiation more than 1.5-fold. Bovine serum albumin （BSA） did not influence visfatin mRNA expression compared with the control group. Dose-response studies demonstrated that addition of 0.125 mmol/L oleate and palmitate to 3T3-L1 adipocytes decreased visfatin mRNA expression significantly （78%, 77%, respectively, relative to untreated control, P〈0.05）, and further to 65% （relative to untreated control, P〈0.05） and 55% （relative to untreated control, P〈0.01） at 1.0 mmol/L FFA. Furthermore, the suppression on preadipocytes was similar to that of adipocytes, which reached a maximal reduction of 44% （oleate, P〈0.05） and 47% （palmitate, P〈0.05） at 1.0 mmol/L FFA. Conclusions Oleic acid and palmitic acid may induce insulin resistance in 3T3-L1 adipocytes and preadipocytes. Downregulation of visfatin mRNA may contribute to impair insulin sensitivity caused by oleate and palmitate.     

PLASMA RESISTIN LEVELS AND SINGLE-NUCLEOTIDE POLYMORPHISMS IN RESISTIN GENE 5＇ FLANKING REGION IN PATIENTS WITH STROKE

OBJECTIVE: To analyze the role of resistin in insulin resistance (IR) through investigating the variation of plasma resistin levels and single-nucleotide polymorphisms (SNPs) in resistin gene 5' flanking region in stroke patients. METHODS: In 103 atherothrombotic cerebral infarction (ACI) patients, 85 lacunar infarction (LI) patients, 70 intracerebral hemorrhage (ICH) patients, and 86 healthy controls, plasma resistin and insulin levels were measured by ELISA, SNPs in resistin gene 5' flanking region were detected by PCR and direct DNA sequencing. The subjects' body height and weight, the body mass index, quantitative insulin sensitivity check index (QUICKI), blood pressure, and the concentration of fasting plasma glucose, triglyceride, total cholesterol, creatinine, low-density lipoprotein, and high-density lipoprotein were also determined. RESULTS: QUICKI was significantly lower in the ACI and ICH patients (0.316 +/- 0.037 and 0.309 +/- 0.032, respectively) than that in the controls (0.342 +/- 0.043, P < 0.001), while plasma resistin level was significantly higher in the ACI and ICH patients (6.36 +/- 3.79 and 7.15 +/- 4.27 ng/mL, respectively) than that in the controls (5.28 +/- 2.56 ng/mL, P < 0.05), but such difference was not observed in the LI patients compared with controls. There was a statistically negative correlation between plasma resistin level with QUICKI (r = -0.228, P < 0.001). The distributions of allele and genotype frequencies of resistin gene - 420C > G and - 537A > C SNPs were not significantly different among the different groups, and those SNPs were not correlated with other clinical and biochemical parameters. CONCLUSIONS: Plasma resistin is associated with stroke by participating in the development of IR. The SNPs in resistin gene 5' flanking region has no impact on the plasma resistin level.     

2型糖尿病患者大网膜与皮下脂肪组织抵抗素蛋白表达的研究

目的了解2型糖尿病(type 2 diabetes mellitus,T2DM)患者大网膜与皮下脂肪组织抵抗素蛋白表达的情况,探讨肥胖和糖尿病间的关系.方法Western bolt法检测大网膜及皮下脂肪组织抵抗素表达情况;磁性分离酶联免疫法测定空腹血胰岛素水平,葡萄糖氧化酶法测定血糖浓度,并计算胰岛素敏感指数;测量身高、体重、臀围、收缩压(SBP)、舒张压(DBP),计算腰臀比(WHR)、体重指数(BMI)、体内脂肪百分比(BF%).结果以OD值表示,正常对照组(57411±1225.59)大网膜与皮下脂肪组织抵抗素蛋白表达明显较单纯肥胖组(41671±884.38)(P＜0.01)、非肥胖T2DM组(42132±1286.15)(P＜0.01)\肥胖T2DM组(42677±1751.10)(P＜0.05)表达增高,后三组抵抗素蛋白表达差异无显著性.四组自身的大网膜与皮下脂肪组织抵抗素蛋白表达无显著差异.逐步回归分析示WHR、BMI、FBG对抵抗素表达的作用显著,方程为v=116338-38562×WHR-1154.99×BMI-1243.63×FBG.结论正常对照组大网膜与皮下脂肪组织抵抗素蛋白表达明显较单纯肥胖组、非肥胖T2DM组、肥胖T2DM组表达增高,抵抗素与腰臀比、体重指数、空腹血糖呈显著负相关提示人体抵抗素不是肥胖和糖尿病的联系所在.     

不同体重指数不同糖耐量状态下人的血清抵抗素水平

目的：探讨不同体重指数（BMI）、不同糖耐量（GT）状态下人血清抵抗素水平与胰岛素抵抗（IR）的关系。方法：2型糖尿病（T2DM）患者、正常糖耐量者（NGT）及糖调节受损者（IGR）分别按BMI≥25kg／m。分为肥胖与非肥胖，采用酶联免疫分析法检测空腹血清抵抗素水平，并以稳态模型（HOMA-model）计算胰岛素抵抗指数（HOMA-IR）。结果：T2DM组和NGT组中的肥胖者血清抵抗素水平均高于非肥胖者，T2DM组有统计学意义（P〈0.05）；IGR组血清抵抗素水平高于T2DM组及NGT组。空腹血清抵抗素与BMI、HOMA-IR呈正相关，与甘油三酯、空腹胰岛素及餐后2h胰岛素呈正相关；与空腹血糖、腰臀比无明显相关性。结论：抵抗素可能在肥胖发展为糖尿病的过程中，成为预测糖尿病发病的一个指标。     

高脂诱导胰岛素抵抗大鼠糖代谢及血抵抗素、脂联素水平的变化

目的 探讨高脂饮食诱导的胰岛素抵抗大鼠糖代谢及抵抗素、脂联素水平的变化。方法 采用大鼠扩展胰岛素钳夹和3-^3H标记葡萄糖示踪技术，估计高脂诱导胰岛素抵抗（IR）大鼠肝糖及外周组织糖代谢的变化，同时采用酶免法检测血浆抵抗素及脂联素水平。结果 高脂喂养组（HF组）大鼠基础血浆游离脂肪酸（FFA）水平明显增高。钳夹稳态时，HF组血浆胰岛素水平明显高于N组，差异有统计学意义（P〈0．01），FFA也明显高于N组，差异有统计学意义（P〈0．01）。HF组葡萄糖输注率（GIR）较N组减少38％，胰岛素介导的葡萄糖利用率（GRd）较N组亦明显减少21％。N组大鼠肝糖输出（HGP）明显抑制达71％，而在HF组胰岛素介导的HGP的抑制作用受到明显障碍（仅抑制30％）。钳夹结束时，N组血浆抵抗素水平与基础值相比明显降低，差异有统计学意义（P〈0．05），而HF组明显高于N组（P〈0.05）。两组血浆脂联素水平在钳夹后均明显下降差异有统计学意义（P〈0.05）。结论 高脂饮食诱导大鼠产生了外周和肝脏的IR。IR的形成可能与血浆FFA升高和在高胰岛素状态下较高的血浆抵抗素水平有关。     

Resistin protein expression in abdominal subcutaneous adipose tissue and placenta in pregnant women

JNANJINGMEDUNIV2006;20(2):113115INTRODUCTION Resistinisanewlydiscoveredpolypeptidehormoneintheadiposetissue,whichischaracterizedbya cysteine richmotif(CX12CX8CXCX3CX10CXCXCX9CC),andmainlyexpressedinthewhiteadiposetissueand weakinthebrownadiposetissue.Resistinlinksobesity todiabetes[12].Duringpregnancy,pregnantwomenwith physiologicalinsulinresistance(IR),impairedglucose tolerance,decreasedinsulinsensitivityareeasytoget gestationaldiabetesmellitus(GDM)[3].Inthepresent study,t…     

代谢综合征患者空腹血清抵抗素水平与胰岛素抵抗的关系

目的 探讨代谢综合征患者空腹血清抵抗素水平与胰岛素抵抗的相关性.方法 用ELISA方法 测定40例代谢综合征患者和10名正常健康人空腹血清抵抗素浓度,同时分别测定FBS、2h-BS、FINS、Homa-IR、血压、血脂等参数.结果 代谢综合征组BMI、WHR、SBP、DBP、FBS、2h-BS、FINS、TC、TG与正常对照组比较,明显高于正常对照组,且差异有显著性(P＜0.01),两组LDL-C比较代谢综合征组也高于正常对照组,差异也有显著性(P＜0.05);代谢综合征组空腹血清抵抗素水平高于正常对照组,差异具有显著性(P＜0.05).Homa-IR也高于正常对照组.差异也具有显著性(P＜0.01):抵抗素浓度与Homa-IR、FINS呈显著正相关(P＜0.01),经多因素逐步回归分析发现,Homa-IR仍与抵抗素浓度呈显著正相关.结论 代谢综合征患者血清抵抗素水平明显升高,它可能是引起机体产生胰岛素抵抗的重要因素.     

血清抵抗素水平与2型糖尿病周围神经病变相关关系的研究

目的研究血清抵抗素与2型糖尿病合并周围神经病变的相关关系及影响因素。方法按腓神经传导速度(SCV)将2型糖尿病患者分为伴有或不伴有周围神经病变组，对照组为健康体检人群。3组研究对象均采用ELISA法测定血清抵抗素水平，同时测定血糖、血脂、血清胰岛素、糖化血红蛋白(HbA1c)等生化指标，并计算胰岛素敏感指数(HOMA—IR)，作为评价胰岛素抵抗程度的指标。结果2型糖尿病患者血清抵抗素水平明显高于对照组，合并周围神经病变组高于无周围神经病变组；血清抵抗素水平与HOMA—IR、HbA1c、胰岛素、甘油三酯水平呈正相关。与血糖及其他脂代谢指标无明显相关性。结论血清抵抗素与2型糖尿病周围神经病变相关，参与周围神经病变形成的因素可能也影响了血清抵抗素的水平，同时抵抗素也影响着周围神经病变的发生和发展。     

妊娠糖尿病患者抵抗素水平的改变及临床意义

[目的]探讨妊娠糖尿病患者抵抗素水平的改变及与胰岛素抵抗（IR）的关系。[方法]对42例妊娠糖尿病患者（gestational diabetes mellitus,GDM）、50例糖耐量正常的孕妇,进行葡萄糖耐量试验（OGTT）、胰岛素释放试验（IRT）,并测定血清抵抗素水平,同时稳态模型（HOMA）计算胰岛素抵抗指数（HOMA-IR）,分析血清抵抗素与GDM患者IR的关系。[结果]（1）GDM组抵抗素水平为（13.7±3.4）μg/L,对照组为（10.2±2.6）μg/L,两组比较差异有统计学意义（P〈0.05）。（2）GDM组HOMA-IR为（3.0＋1.1）,对照组为（2.6＋0.7）,两组比较差异有统计学意义（P〈0.05）。（3）GDM组Homa-IR与孕前体重指数（BMI）、空腹胰岛素、抵抗素呈正相关（P〈0.05或P〈0.01）。[结论]GDM患者血清抵抗素水平明显升高,且与GDM患者IR密切相关。     

吡格列酮和非诺贝特对高脂饲养大鼠脂联素与胰岛素抵抗的影响

目的探讨吡格列酮、非诺贝特及两者联合应用对高脂饲养大鼠脂联素表达与胰岛素抵抗(IR)的影响。方法将75只SD大鼠随机分为普通饮食组(NC组)15只,高脂饮食组(高脂组)60只。高脂组随机分为非诺贝特组(FF组)、吡格列酮组(PIO组)、联合组(FF-PIO组)和高脂对照组(HF组),各15只。分别测定体重(BW)、血清游离脂肪酸(FFA)、甘油三酯(TG)、总胆固醇(TCH)、空腹血糖(FPG)、空腹胰岛素(FINS)等指标。采用Westernblot方法对各组大鼠腹部皮下脂肪中脂联素蛋白水平进行检测。结果HF组的体重、FFA、TG、FPG、FINS显著高于NC组,皮下脂肪中脂联素蛋白表达明显低于NC组(P<0.01)。与HF组比较,FF组的体重减轻、FFA、TG、TCH、FPG、FINS降低(P<0.01),皮下脂肪中脂联素增加,但差异无显著性(P>0.05);PIO组FFA、FPG、FINS降低(P<0.01),皮下脂肪中脂联素增加(P<0.05);FF-PIO组的体重、FFA、TG、TCH、FPG、FINS降低(P<0.05或P<0.01),皮下脂肪中脂联素增加(P<0.01)。结论吡格列酮、非诺贝特及联合应用可以一定程度地减轻肥胖对大鼠皮下脂肪中脂联素蛋白表达的抑制作用,改善肥胖引起的IR。