S100A8/A9 aggravates post-ischemic heart failure through activation of RAGE-dependent NF-κB signaling

The extracellular heterodimeric protein S100A8/A9 activates the innate immune system through activation of the receptor of advanced glycation end products (RAGE) and Toll-like receptors. As activation of RAGE has recently been associated with sustained myocardial inflammation and heart failure (HF) we studied the role of S100A8/A9 in the development of post-ischemic HF. Hypoxia led to sustained induction of S100A8/A9 accompanied by increased nuclear factor (NF-)κB binding activity and increased expression of pro-inflammatory cytokines in cardiac fibroblasts and macrophages. Knockdown of either S100A8/A9 or RAGE rescued the induction of pro-inflammatory cytokines and NF-κB activation after hypoxia. In a murine model of post-ischemic HF both cardiac RNA and protein levels of S100A8/A9 were elevated as soon as 30 min after hypoxia with sustained activation up to 28 days after ischemic injury. Treatment with recombinant S100A8/A9 resulted in reduced cardiac performance following ischemia/reperfusion. Chimera experiments after bone marrow transplantation demonstrated the importance of RAGE expression on immune cells for their recruitment to the injured myocardium aggravating post-ischemic heart failure. Signaling studies in isolated ventricles indicated that MAP kinases JNK, ERK1/2 as well as NF-κB mediate signals downstream of S100A8/A9-RAGE in post-ischemic heart failure. Interestingly, cardiac performance was not affected by administration of S100A8/A9 in RAGE^?/?-mice, which demonstrated significantly improved cardiac recovery compared to WT-mice. Our study provides evidence that sustained activation of S100A8/A9 critically contributes to the development of post-ischemic HF driving the progressive course of HF through activation of RAGE.     

核因子κB通路参与缺氧诱导心肌细胞分泌肿瘤坏死因子α

目的:探讨缺氧刺激能否诱导体外培养心肌细胞分泌肿瘤坏死因子α(TNF-α),核因子κB(NF-κB)通路是否参与其中,并发挥调节作用。方法:1%O2物理缺氧刺激原代SD大鼠乳鼠心肌细胞,在不同时间点Real-time PCR检测TNF-αmRNA水平;ELISA检测培养上清中TNF-α蛋白水平;Western blot检测胞质NF-κB相关通路蛋白IKKα,IKKβ,IKKBα蛋白及磷酸化蛋白水平以及胞核NF-κB p65蛋白表达;电泳迁移率实验检测NF-κB DNA结合能力。缺氧同时给予NF-κB抑制剂四氢化吡咯二硫代氨基甲酸酯(PDTC)刺激心肌细胞,检测细胞核p65蛋白表达,TNF-αmRNA及蛋白水平。结果:缺氧诱导原代心肌细胞表达TNF-α,TNF-αmRNA水平从缺氧1h开始升高,12h及24h达峰值;其蛋白分泌从缺氧6h开始升高,12h和24h达峰值。缺氧激活NF-κB通路:缺氧5min胞质pIKKα/IKKβ表达开始上调,30min达峰值,持续6h;IKKBα表达缺氧5min开始降低,而pIKKBα缺氧30min开始增加;胞核p65蛋白水平缺氧5min上调,1h达峰值,持续6h;NF-κB DNA结合能力缺氧5min后开始增强。PDTC抑制NF-κB活性,有效抑制TNF-αmRNA及其蛋白表达。结论:缺氧通过激活NF-κB通路,诱导心肌细胞自分泌TNF-α,NF-κB在缺氧诱导心肌细胞自分泌TNF-α过程中起正向调节作用。     

The effects of cardiac myocytes on interstitial fibroblasts in toxic iron overload

Iron deposits preferentially in myocytes in mixed cultures of cardiac myocytes and nonmyocytic fibroblasts. In vivo, iron overload is associated with cardiac fibrosis. Therefore, we examined whether iron loading of cardiac myocytes in culture could trigger a response in nonmyocytes characteristic of a fibrogenic phenotype. We found that the nonmyocytes adopted a myofibroblast phenotype in culture. The rate of DNA synthesis (measured by [³H]thymidine incorporation) by the nonmyocytes was decreased by the myocyte-conditioned medium, compared to that of the unconditioned medium, and this activity was retained in >10-kDa fractions. The rate was partially restored when the medium was obtained from iron-loaded myocytes, and in this medium, the >10-kDa fraction was even more effective in reversing the suppression of proliferation. This suppression suggests a decreased secretion of a growth-inhibitory substance in the iron-loaded myocytes, and this effect was partially reversed when the iron-loaded cells were treated with the iron chelator, deferoxamine. This indicates that cardiac myocytes may play a paracrine role in suppressing the proliferation of myofibroblasts that is partially overcome when the myocytes are iron overloaded. The myocyte-conditioned medium also affects the myofibroblast phenntype, increasing the cells' fibronectin mRNA content and decreasing α-smooth-muscle actin mRNA. The myocyte-conditioned medium increases transforming growth factor-β (TGF-β) secretion by myofibroblasts, but the TGF-β content of the conditioned medium was found to play, at most, a minor role in determining the response of the myofibroblast.     

HDL和阿托伐他汀对oxLDL刺激下脂肪细胞炎症反应的影响

目的 观察高密度脂蛋白（HDL）及阿托伐他汀对氧化型低密度脂蛋白（oxLDL）刺激下3T3-L1脂肪细胞肿瘤坏死因子-α（TNFα）分泌及mRNA表达的影响,并探讨其可能的作用机制.方法 3T3-L1脂肪细胞促分化成熟后,oxLDL刺激脂肪细胞,给予不同浓度的HDL（10～100 μg/mL）和阿托伐他汀（0.1～10 μM）,及H-89（10 μM）＋HDL（100 μg/mL）干预,收集细胞,测定脂肪细胞TNFα水平、TNFα mRNA表达水平、核因子-κB（NF-κB）活性及NF-κB抑制单位（IκB）蛋白浓度.结果 oxLDL刺激使3T3-L1脂肪细胞TNFα分泌、mRNA表达水平及NF-κB活性明显增强.阿托伐他汀浓度依赖性降低TNFα 分泌及mRNA表达,抑制NF-κB活化.10 μM阿托伐他汀使oxLDL诱导的脂肪细胞TNFα mRNA表达降低56.5%,NF-κB活性减少41.2%.HDL也呈浓度依赖性抑制TNFα分泌及mRNA表达,降低NF-κB活性,减少IκB降解.与oxLDL刺激组比较,100 μg/ml HDL使TNFα mRNA表达降低64.5%,NF-κB活性减少49%,并明显增加IκB蛋白水平.HDL的这些抗炎效应能被蛋白激酶A（PKA）抑制剂（应放在H89第一次出现之处）H-89部分抑制.结论 HDL能抑制oxLDL诱导的3T3-L1脂肪细胞TNFα分泌和mRNA表达,PKA-IκB-NF-κB信号通路可能是其中作用途径之一,该效应不需要HDL与oxLDL的直接接触作用.阿托伐他汀亦通过NF-κB途径抑制oxLDL诱导的3T3-L1脂肪细胞TNFα分泌和mRNA表达.HDL的抗炎作用强度与阿托伐他汀相似.     

The TIR/BB-loop mimetic AS-1 prevents cardiac hypertrophy by inhibiting IL-1R-mediated MyD88-dependent signaling

Activation of NF-κB contributes to cardiac hypertrophy and the interleukin-1 receptor (IL-1R)-mediated MyD88-dependent signaling pathway predominately activates NF-κB. Recent studies have shown that the TIR/BB-Loop mimetic (AS-1) disrupted the interaction of MyD88 with the IL-1R, resulting in blunting of NF-κB activation. We have examined the effects of AS-1 on the IL-1β-induced hypertrophic response using cultured neonatal cardiac myocytes in vitro and transverse aortic constriction (TAC) pressure overload-induced cardiac hypertrophy in vivo. Neonatal cardiac myocytes were treated with AS-1 15?min prior to IL-1β stimulation for 24?h. AS-1 treatment significantly attenuated IL-1β-induced hypertrophic responses of cardiac myocytes. In vivo experiments showed that AS-1 administration prevented cardiac hypertrophy and dysfunction induced by pressure overload. AS-1 administration disrupted the interaction of IL-1R with MyD88 in the pressure overloaded hearts and prevented activation of NF-κB. In addition, AS-1 prevented increases in activation of the MAPK pathway (p38 and p-ERK) in TAC-induced hypertrophic hearts. Our data suggest that the IL-1R-mediated MyD88-dependent signaling pathway plays a role in the development of cardiac hypertrophy and AS-1 attenuation of cardiac hypertrophy is mediated by blocking the interaction between IL-1R and MyD88, resulting in decreased NF-κB binding activity and decreased MAPK activation.     

Ras同源基因家族成员Ⅰ基因在胃癌细胞中下调核因子-κB信号转导通路转录活性

目的 研究Ras同源基因家族成员Ⅰ(ARHⅠ)基因与NF-κB之间的相互关系以及在胃癌发生发展中的可能机制.方法 利用前期构建的pIRES2-EGFP-ARH Ⅰ重组质粒和实验室保存的NF-κB报告基因质粒共转人胚胎肾293T细胞,根据荧光素酶反应测定NF-κB转录激活能力.通过siRNA方法干扰胃癌MKN-28细胞株ARH Ⅰ的表达,Western印迹法检测其NF-κB磷酸化水平以及细胞核内NF-κB的变化情况.结果 使293T细胞中ARH Ⅰ过表达可以明显下调NF-κB的转录激活能力;在MKN-28细胞株中抑制ARHⅠ表达,可使NF-κB磷酸化水平及细胞核内NF-κB蛋白水平均明显上调.结论 ARH Ⅰ可以下调NF-κB的转录激活能力,且能使胃癌细胞株中NF-κB的磷酸化降低、减少入核从而发挥抑癌作用.     

醛固酮在乳鼠心肌细胞表达肿瘤坏死因子-α中的作用

目的:观察醛固酮对培养心肌细胞表达肿瘤坏死因子(TNF-α)的影响。方法:在原代培养的新生大鼠心肌细胞,采用免疫组化和RT-PCR,评价10-6M、10-5M醛固酮及10-5M醛固酮拮抗剂螺内酯对心肌细胞表达TNF-α的影响。用凝胶滞留实验(EMSA)验证NF-κB是否参与TNF-α的转录调控,筛选TNF-α可能启动子的位点。结果:TNF-α的免疫组化染色在对照组心肌细胞呈阴性,10-5和10-6M醛固酮作用心肌细胞48h,心肌细胞胞浆中出现棕色颗粒,10-5M醛固酮与螺内酯共同孵育的心肌细胞内TNF-α染色呈阴性。与单纯培养的对照组心肌细胞相比,10-5M,10-6醛固酮刺激心肌细胞24h致心肌细胞TNF-α表达的mRNA显著增高,与螺内酯共同培养的心肌细胞中TNF-αmRNA水平与对照组相比没有明显差异。醛固酮组心肌细胞的核蛋白与TNF-α上游含有潜在NF-κB结合位点的2段寡核苷酸(-619~-591 and-508~-481)的结合强于对照组,螺内酯能阻断这一效应。结论:正常情况下培养的新生鼠心肌细胞内无TNF-α合成,醛固酮通过醛固酮受体促使培养的心肌细胞表达TNF-α,NF-κB核转移是醛固酮激活心肌细胞表达TNF-α的一条通路。     

The effect of enantiomers of beta-agonists on myofibroblast-derived vascular endothelial growth factor and other matrix components in the presence of dust-mite extract.

Beta(2)-adrenergic receptor agonists have been shown to modulate airway epithelial cell and smooth muscle release of cytokines and growth factors transforming growth factor (TGF) beta, associated with remodeling, is known to up-regulate the synthesis of vascular endothelial growth factor (VEGF) and stimulate differentiation of fibroblasts to the myofibroblast phenotype. VEGF and fibronectin can promote angiogenesis and (S)-albuterol can induce VEGF secretion from normal human lung fibroblasts (NHLF). We hypothesize that (S)-albuterol could stimulate myofibroblast secretion and expression of VEGF and fibronectin in the presence of Dermatophagoides pteronyssinus extract. Cultured NHLFs were stimulated with IL-1beta, TGF-beta, D. pteronyssinus, and treated with (R)- and (S)-enantiomers of albuterol. VEGF and fibronectin and basic fibroblast growth factor (bFGF) were measured by ELISA and mRNA. VEGF secretion by fibroblasts was twofold higher with 10(-7) M of (R) relative to (S) (p < 0.05). Myofibroblast secretion of VEGF was increased twofold over fibroblasts, but there was no difference between enantiomers. (S)-albuterol at 10(-8)-10(-4) M caused an increase in VEGF mRNA that paralleled VEGF secretion relative to 10(-8)-10(-4) M. Fibronection secretion by myofibroblasts but not fibroblasts was increased by 10(-5) M of (S) relative to (R) in the presence of recombinant interleukin 1 (rhIL-1)beta and D. pteronyssinus (S)-albuterol at 10(-6) M increased bFGF. The 10(-6) M of (S)-albuterol, but not (R)-albuterol, may promote angiogenesis. Increased fibronectin or bFGF by (S)-albuterol could enhance matrix deposition and remodeling in a subset of asthmatic patients.     

塞来昔布对大鼠胰腺腺泡细胞炎症损伤的作用

背景：急性胰腺炎（AP）的发病始于胰腺腺泡细胞内胰酶的激活,造成腺泡细胞损伤。环氧合酶-2（COX-2）和核因子-κB（NF．KB）在AP的炎症反应中起重要作用。目的：观察雨蛙肽和选择性COX-2抑制剂塞来昔布对离体大鼠胰腺腺泡细胞COX-2和NF—κB表达的影响．探讨塞来昔布对腺泡细胞炎症损伤的作用。方法：分离大鼠胰腺腺泡细胞,分为对照组、雨蛙肽组（1×10^-7mol／L）和塞来昔布干预组（100μmol／L,15min后加入雨蛙肽）,分别培养1、3、6、12h。测定腺泡细胞活力、淀粉酶分泌率和乳酸脱氢酶（LDH）漏出率,逆转录聚合酶链反应（RT-PCR）和免疫细胞化学染色检测COX-2、NF—κBmRNA和蛋白表达。结果：与对照组相比,雨蛙肽组各时间点腺泡细胞活力均显著降低,淀粉酶分泌率和LDH漏出率显著增高,COX-2和NF—κBmRNA表达量显著增高,蛋白表达阳性率亦增加（P〈0．05）。塞来昔布干预组各时间点腺泡细胞活力、淀粉酶分泌率和LDH漏出率均较雨蛙肽组显著改善（心O．05）,COX-2mRNA和蛋白表达显著降低（P〈0．05）,NF—κBmRNA和蛋白表达与雨蛙肽组无明显差异。结论：塞来昔布可抑制大鼠胰腺腺泡细胞中雨蛙肽刺激的COX-2活性,从而减轻细胞炎症损伤。     

高糖通过激活人脐静脉内皮细胞ⅠκB-α/NF-κB途径诱导MCP-1的表达

目的观察高糖诱导人脐静脉内皮细胞NF-κB的活性和单核细胞趋化蛋白1（MCP-1）的表达,探讨糖尿病并发动脉粥样硬化的发病机制。方法在培养的人脐静脉内皮细胞中加入不同浓度葡萄糖,检测内皮细胞中MCP-1mRNA、MCP-1蛋白的表达,NF-κB的活性及IκB-α的磷酸化水平。结果高糖明显地诱导了血管内皮细胞NF-κB的活性、MCP-1的表达及IκB-α的磷酸化;NF-κB活性抑制剂明显地降低了高糖所诱导的MCP-1的表达。结论高糖通过诱导血管内皮细胞IκB-α的磷酸化激活NF-κB,从而诱导了MCP-1的表达。提示在糖尿病并发动脉粥样硬化的发病过程中,高血糖通过激活血管内皮细胞IκB-α/NF-κB途径诱导MCP-1的表达,进而发挥了重要的作用。     

Tumor necrosis factor alpha induces human atrial myofibroblast proliferation, invasion and MMP-9 secretion: inhibition by simvastatin

OBJECTIVE: Tumor necrosis factor alpha (TNFalpha) is implicated in myocardial remodeling, a process in which activated cardiac fibroblasts (myofibroblasts) secrete matrix-degrading metalloproteinases (MMPs) and undergo increased proliferation and invasion. Statins are cholesterol-lowering drugs that also have direct cellular effects, which may underlie their ability to reduce myocardial remodeling. This study investigated the effect of TNFalpha on human cardiac myofibroblast proliferation, invasion and MMP-9 secretion, and determined whether these properties were modulated by simvastatin. METHODS: Human cardiac myofibroblasts were cultured from right atrial appendage. TNF receptor expression was quantified by immunoblotting. Cell proliferation, invasion, MMP-9 secretion and MMP-9 mRNA expression were determined by cell counting, Matrigel-coated modified Boyden chamber assays, gelatin zymography and RT-PCR, respectively. RESULTS: Human atrial myofibroblasts expressed the TNF-RI and TNF-RII receptor subtypes. TNFalpha (1 ng/ml) induced a 23.1+/-3.9% increase in cell number after 4 days (P<0.001). Additionally, TNFalpha (1-10 ng/ml) significantly (P<0.01) increased myofibroblast invasion, with a concomitant increase in MMP-9 secretion, that was due to increased MMP-9 mRNA levels. Using TNF-R-specific neutralizing antibodies, we determined that these cellular effects of TNFalpha were predominantly TNF-RI-mediated. Simvastatin (0.1-10 mumol/l) concentration dependently inhibited TNFalpha-induced myofibroblast proliferation, invasion and MMP-9 secretion. CONCLUSIONS: TNFalpha, acting predominantly via the TNF-R1 receptor, increased human atrial myofibroblast proliferation, invasion and MMP-9 secretion, all of which were inhibited by simvastatin. Inhibition of cytokine-induced cardiac myofibroblast activation by statins provides a rationale for their use in patients with cardiac pathologies characterized by adverse myocardial remodeling.     

Norepinephrine Upregulates Vascular Endothelial Growth Factor in Rat Cardiac Myocytes by a Paracrine Mechanism

Norepinephrine has growth-promoting effects in cardiac myocytes. The present study in cultured neonatal rat cardiac myocytes tested the hypothesis that norepinephrine also stimulates expression of vascular endothelial growth factor (VEGF), an important angiogenic factor. As assessed by polymerase chain reaction cardiac myocytes and non-myocytes expressed all three isoforms of rat VEGF, with the short isoform (VEGF 121 ) preferentially expressed in non-myocytes. When cardiac myocytes were stimulated with 1 micro M norepinephrine for 24 h in the presence or absence of the specific alpha - and beta -adrenoceptor antagonists prazosin and propranolol, respectively, VEGF mRNA levels and splice variant pattern did not change, whereas atrial natriuretic peptide mRNA levels increased 3 to 4-fold. CoCl(2) increased VEGF mRNA levels in cardiac myocytes five-fold. When cardiac myocytes were cultured with conditioned medium from non-myocytes that had been stimulated with norepinephrine for 24 h VEGF mRNA increased 2-fold. The increase was blocked by antibodies neutralizing TGF beta. These data suggest that norepinephrine stimulates myocardial angiogenesis by a paracrine mechanism that involves cardiac non-myocytes and TGF beta.     

Toll样受体4和核因子-κB信号通路在溶血磷脂酸致动脉粥样硬化中的作用

目的探讨Toll样受体4(TLR4)/NF-κB信号通路在溶血磷脂酸(LPA)致动脉粥样硬化中的作用。方法以不同浓度LPA(0¹0μmol/L)刺激人单核细胞株THP-1细胞4h,以及LPA 1μmol/L处理THP-1细胞不同时间(010μmol/L)刺激人单核细胞株THP-1细胞4h,以及LPA 1μmol/L处理THP-1细胞不同时间(0⁸h),荧光定量RT-PCR法测定TLR4mRNA表达,Western blot检测TLR4蛋白、细胞核NF-κB p65表达变化,ELISA法测定细胞因子TNF-α,随后在LPA 1μmol/L条件下,TLR4单抗干预THP-1细胞,观察其对LPA诱导的细胞核NF-κB p65表达及TNF-α分泌水平的影响。结果当LPA 1μmol/L时,TLR4mRNA和蛋白及细胞核NF-κB p65表达较0μmol/L、0.1μmol/L、0.5μmol/L、5μmol/L、10μmol/L LPA明显增高,差异有统计学意义(P<0.01)。LPA 1μmol/L处理THP-1细胞4h时,THP-1细胞TLR4mRNA和蛋白及细胞核NF-κB p65表达水平明显高于0、1、2、8h(P<0.01)。与TLR4单抗干预前比较,TLR4干预后LPA诱导的THP-1细胞NF-κB p65表达及TNF-α分泌水平明显升高,差异有统计学意义(P<0.01)。结论 LPA可显著上调THP-1细胞TLR4表达及促进NF-κB的活化,LPA致动脉粥样硬化作用可能部分是由TLR4/NF-κB信号途径介导的。     

Human foetal intestinal fibroblasts are hyper-responsive to lipopolysaccharide stimulation

Background

Intestinal myofibroblasts contribute to immune regulation in adults with inflammatory bowel disease but have not been characterised in neonatal intestinal inflammatory diseases.

Aims

To compare lipopolysaccharide (LPS)-stimulated interleukin-8 (IL-8) production between human foetal and mature intestinal myofibroblasts in vitro.

Methods

Foetal, neonatal and adult cells were stimulated with increasing concentrations of E. coli LPS. In LPS stimulated foetal myofibroblasts, Toll-like receptor 4 mRNA expression was assessed by real-time PCR whilst Toll-like receptor 4 receptor activity was determined using anti-Toll-like receptor 4 antibody. Mitogen activated protein kinase pathway activity was assessed using chemical inhibitors and Western blotting. IL-8 production was measured by quantitative ELISA.

Results

IL-8 production by LPS stimulated foetal myofibroblasts occurred in a dose dependent manner. Toll-like receptor 4 expression was constitutive and Toll-like receptor 4 receptor blockade reduced IL-8 production by 42% (P = 0.0262). C-Jun N-terminal kinase, p38 and NF-κB inhibitors significantly attenuated LPS stimulated IL-8 production by 42%, 33% and 2%, respectively. Mitogen activated protein kinase activity was confirmed by the presence of phosphorylated proteins on Western blots.

Conclusion

These data demonstrate increased IL-8 production by foetal myofibroblasts that is partially mediated by Toll-like receptor 4, mitogen activated protein kinase and NF-κB cell signalling pathways. Intestinal myofibroblasts cells may contribute to the dysregulated inflammatory response in the immature intestine and may form targets that lead to new therapies to prevent neonatal intestinal inflammatory bowel diseases.     

内毒素对肝星状细胞醛固酮分泌及核因子-κBP65 mRNA表达的影响

目的 探讨内毒素对大鼠肝星状细胞(HSC)醛固酮的分泌及核因子-κB P65(NF-κBP65)mRNA表达的影响.方法 传代培养大鼠HSC-T6,不同浓度内毒素处理细胞后分组.放射免疫法检测HSC分泌醛固酮的情况,半定量RT-PCR方法检测HSC表达NF-κB P65 mRNA的水平.应用方差分析、t检验和Pearson直线相关分析法进行统计学分析.结果 浓度为0.01、0.1、1.0和10.0 mg/L内毒素组醛固酮的分泌分别为(4.95±0.35)、(5.52±0.32)、(6.04±0.60)和(5.16±0.46)μg/L,显著高于对照组的(3.65±0.51)μg/L(t=2.9745,5.8725,6.8465,3.2065;均P＜0.05).浓度为0.01、0.1、1.0和10.0 mg/L内毒素组NF-κB P65 mRNA的表达分别为0.825±0.06、1.07±0.07、1.23±0.06和0.96±0.05,也明显高于对照组的0.43±0.04(t=5.4776,6.8084,7.9382,7.5136;均P＜0.01).10.0 mg/L内毒素组醛固酮的分泌和NF-κB P65 mRNA的表达均较1.0 mg/L内毒素组有明显降低(t=4.3865,3.7246;均P＜0.05).HSC中醛固酮的分泌与NF-κB P65 mRNA的表达存在正相关(r=0.886,P＜0.01).结论 一定浓度范围的内毒素可以上调大鼠HSC醛固酮的分泌和NF-κB P65 mRNA的表达,两者存在正相关和一定的剂量效应关系,这可能是引起肝纤维化的重要因素之一.     

脂联素与2型糖尿病乳腺癌发生的相关性及其机制研究

目的:探讨脂联素与2型糖尿病（T2DM）患者乳腺癌发生的相关性及其可能机制。方法:选取2016年1至6月于安徽医科大学第一附属医院甲状腺与乳腺外科住院的乳腺癌患者50例作为研究对象。按照是否合并T2DM将纳入患者分为合并T2DM的乳腺癌组（20例）以及单纯乳腺癌组（30例）。检测空腹血糖和胰岛素、血脂、总脂联素、高分子...