Fatty Acid Synthase and Hormone-sensitive Lipase Expression in Liver Are Involved in Zinc-α2-glycoprotein-induced Body Fat Loss in Obese Mice

Objective To explore the effects of zinc-0t2-glycoprotein （ZAG） on body weight and body fat in high-fat-diet （HF1））-induced obesity in mice and the possible mechanism. Methods Thirty-six male mice were fed with standard food （SF） （n=9） and HFD （n=27）, respectively. Five weeks later, 9 mice fed with HFD were subjected to ZAG expression plasmid DNA transfection by liposome transfection method, and another 9 mice to negative control plasmid transfection. Two weeks later, serum ZAG level in the mice was assayed by Western blot, and the effects of ZAG over-expression on body weight, body fat, serum biochemical indexes, and adipose tissue of obese mice were evaluated. The mRNA expressions of fatty acid synthase （FAS） and hormone sensitive lipase （HSL） in liver tissue were deterlnined by reverse transcription-polymerase chain reaction. Results Serum ZAG level significantly lowered in simple HFD-fed mice in comparison to SF-fed mice （0.51±0.10 AU vs. 0.75±0.07 AU, P〈0.01）. Further statistical analysis demonstrated that ZAG level was negatively correlated with body weight （r =-0.56, P〈0.001）, epididymal fat mass （r=-0.67, P〈O. 001）, percentage of epididymal fat （r= 0.65, P〈0.001）, and increased weight （r= 0.57, P〈0.001） in simple SF- and HFD fed mice. ZAG over-expression in obese mice reduced body weight and the percentage of epididyreal fat. Furthermore, FAS mRNA expression decreased （P〈0.01） and HSL mRNA expression increased （P〈0.001） in the liver in ZAG over-expressing mice. Conclusions ZAG is closely related to obesity. Serum ZAG level is inversely correlated with body weight and percentage of body fat. The action of ZAG is associated with reduced FAS expression and increased HSL expression in the liver of obese mice.

Zinc-α2-glycoprotein 1 attenuates non-alcoholic fatty liver disease by negatively regulating tumour necrosis factor-α

BACKGROUND Zinc-α2-glycoprotein 1 (AZGP1) plays important roles in metabolism-related diseases. The underlying molecular mechanisms and therapeutic effects of AZGP1 remain unknown in non-alcoholic fatty liver disease (NAFLD). AIM To explore the effects and potential mechanism of AZGP1 on NAFLD in vivo and in vitro. METHODS The expression of AZGP1 and its effects on hepatocytes were examined in NAFLD patients, CCl4-treated mice fed a high fat diet (HFD), and human LO2 cells. RESULTS AZGP1 levels were significantly decreased in liver tissues of NAFLD patients and mice. AZGP1 knockdown was found to activate inflammation;enhance steatogenesis, including promoting lipogenesis [sterol regulatory elementbinding protein (SREBP)-1c, liver X receptor (LXR), fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC), and stearoyl CoA desaturase 1 (SCD)-1], increasing lipid transport and accumulation [fatty acid transport protein (FATP), carnitine palmitoyl transferase (CPT)-1A, and adiponectin], and reducing fatty acid β-oxidation [farnesoid X receptor (FXR) and peroxisome proliferator-activated receptor (PPAR)-α];accelerate proliferation;and reverse apoptosis in LO2 cells. AZGP1 overexpression (OV-AZGP1) had the opposite effects. Furthermore, AZGP1 alleviated NAFLD by blocking TNF-α-mediated inflammation and intracellular lipid deposition, promoting proliferation, and inhibiting apoptosis in LO2 cells. Finally, treatment with OV-AZGP1 plasmid dramatically improved liver injury and eliminated liver fat in NAFLD mice. CONCLUSION AZGP1 attenuates NAFLD with regard to ameliorating inflammation, accelerating lipolysis, promoting proliferation, and reducing apoptosis by negatively regulating TNF-α. AZGP1 is suggested to be a novel promising therapeutic target for NAFLD.

Thrombophilia Caused by Beta2-Glycoprotein I Deficiency： In Vitro Study of a Rare Mutation in APOH Gene

This study aimed to explore the mechanism of a novel mutation （p.Lys38Glu） in apolipoprotein H （APOH） gene causing hereditary beta2-glycoprotein I （β2GPI） deficiency and thrombosis in a proband with thrombophilia. The plasma level of β2GPI was measured by ELISA and Western blotting, and anti-β2GPI antibody by ELISA. Lupus anticoagulant （LA） was assayed using the dilute Russell viper venom time. Deficiency of the major natural anticoagulants including protein C （PC）, protein S （PS）, antithrombin （AT） and thrombomodulin （TM） was excluded from the proband. A mutation analysis was performed by amplification and sequencing of the APOH gene. Wild type and mutant （c.112A〉G） APOH expression plasmids were constructed and transfected into HEK293T cells. The results showed that the thrornbin generation capacity of the proband was higher than that of the other family members. Missense mutation p.Lys38Glu in APOH gene and LA coexisted in the proband. The mutation led to β2GPI deficiency and thrombosis by impairing the protein production and inhibiting the platelet aggregation. It was concluded that the recurrent thrombosis of the proband is associated with the coexistence ofp.Lys38Glu mutation in APOH gene and LA in plasma.

Relationship and significance between anti-b2-glycoproteinⅠantibodies and platelet activation state in patients with ulcerative colitis

AIM： To study the relationship between anti-β2- glycoprotein Ⅰ （aβ2GPⅠ） antibodies and platelet activation state in patients with ulcerative colitis （UC） and its significance. METHODS： Peripheral blood samples were collected from 56 UC patients （34 males and 22 females, aged 43.5 years, range 21-66 years）, including 36 at active stage and 20 at remission stage, and 25 sex-and age-matched controls. The level of aβ2GP Ⅰ was measured by ELISA. The platelet activation markers, platelet activation complex- Ⅰ （PAC- Ⅰ ） and P-selectin （CD62P） were detected by flow cytometry. RESULTS： The A value for IgG aβ2GP Ⅰ in the active UC group was 0.61 ± 0.13, significantly higher than that in the remittent UC and control groups （0.50 ± 0.13 and 0.22 ± 0.14, P 〈 0.01）. There was a significant difference between the two groups （P 〈 0.01）. The A value for IgM aβ2GP Ⅰ in the active and remittent UC groups was 0.43 ± 0.13 and 0.38 ± 0.12, significantly higher than that in the control group （0.20 ± 0.12, P 〈 0.01）. However, there was no significant difference between the two groups （P 〉 0.05）. The PAC- Ⅰ positive rate for the active and remittent UC groups was 30.6% ± 7.6% and 19.6% ± 7.8% respectively, significantly higher than that for the control group （6.3% ± 1.7%,P 〈 0.01）. There was a significant difference between the two groups （P 〈 0.01）. The CD62P positive rate for the active and remittent UC groups was 45.0% ± 8.8% and 31.9% ± 7.8% respectively, significantly higher than that for the control group （9.2% ± 2.7%, P 〈 0.01）. There was a significant difference between the two groups （P 〈 0.01）. In the active UC group, the more severe the state of illness was, the higher the A value for IgG aβ2GP Ⅰ was, and the positive rate for PAC-Ⅰ and CD62P was positively correlated with the state of illness （Faβ2GP Ⅰ = 3.679, P 〈 0.05; FPAC-Ⅰ （%） = 5.346, P 〈 0.01; and FCD62P （%） = 5. 418, P 〈 0.01）. Meanwhile, in the same state of illness, the A value for IgG aβ2GP Ⅰ was positively correlated to the positive rates for PAC-Ⅰ and CD62P. CONCLUSION： aβ2GP Ⅰ level, platelet activation state and their relationship of them are closely correlated with the pathogenesis and development of UC.

High levels of Zinc-α-2-Glycoprotein among Omani AIDS patients on combined antiretroviral therapy

Objective:To investigate the levels of zinc-α-2-glycoprotein(ZAG) among Omani AIDS patients receiving combined antiretroviral therapy(cART).Methods:A total of 80 Omani AIDS patients(45 males and 33 females),average age of 36 vears.who were receiving cART at the Saltan Qaboos University Hospital(SQUH).Muscat,Oman,were tested for the levels of ZAG.In addition,SO healthy blood donors(46 males and 34 females),average age of 26 years,attending the SOUH Blood Bank,were tested in parallel as a control group.Measurement of the ZAG levels was performed using a competitive enzyme—linked immunosorbent assay and in accordance with the manufacturer's instructions.Results:The ZAG levels were found to he significantly higher among AIDS patients compared to the healthy individuals(P=0.033).A total of 56(70%) of the AIDS patients were found to have higher levels of ZAG and 16(20%) AIDS patients were found to have high ZAG levels,which are significantly(P>0.031) associated with weight loss.Conclusions:ZAG levels are high among Omani AIDS patients on cART and this necessitales the measurement of ZAG on routine basis,as it is associated with weight loss.

利拉鲁肽联合锌alpha2糖蛋白对棕榈酸诱导的HepG2细胞凋亡、脂质代谢和炎症因子表达的影响

目的探讨利拉鲁肽联合锌alpha2糖蛋白(ZAG)对棕榈酸诱导的HepG2细胞凋亡、脂质代谢和炎症因子表达的影响。方法实时定量PCR(RT-qPCR)和蛋白质印迹法分析ZAG在非酒精性脂肪性肝病(NALFD)患者肝组织以及棕榈酸诱导的HepG2细胞中的表达水平。将HepG2细胞分为对照组、棕榈酸组、棕榈酸+利拉鲁肽组、棕榈酸+pcDNA-ZAG组、棕榈酸+利拉鲁肽+ZAG shRNA组、棕榈酸+利拉鲁肽+pcDNA-ZAG组。流式细胞术分析细胞凋亡情况。试剂盒测定细胞中总胆固醇(TC)、甘油三酯(TG)水平,同时测定细胞培养液上清中白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)水平。结果NALFD患者肝组织中ZAG mRNA和ZAG蛋白表达水平显著降低(P <0.05)。棕榈酸处理显著下调ZAG mRNA和ZAG蛋白表达,增加TC、TG、IL-6和TNF-α水平,增加HepG2细胞凋亡率(P <0.05)。利拉鲁肽处理或过表达pcDNA-ZAG显著抑制棕榈酸诱导的HepG2细胞凋亡,并降低TC、TG、IL-6和TNF-α水平(P <0.05)。沉默ZAG显著减弱利拉鲁肽对棕榈酸诱导的HepG2细胞凋亡、TC、TG、IL-6和TNF-α水平的影响(P <0.05)。过表达ZAG显著增强利拉鲁肽对棕榈酸诱导的HepG2细胞凋亡、TC、TG、IL-6和TNF-α水平的影响(P <0.05)。结论利拉鲁肽联合ZAG可抑制棕榈酸诱导的HepG2细胞凋亡、脂质沉积和炎症反应。

锌-α2-糖蛋白水平变化与维持性血液透析患者营养不良的关系

目的探讨锌-α2-糖蛋白（ZAG）水平变化与维持性血液透析患者营养不良的关系。方法选取50例维持性血液透析患者为血液透析（MHD）组，30例对照组选自健康体检者（肾小球滤过率〉60ml／min）。应用酶联免疫吸附试验（ELISA）法检测受试者血清ZAG水平、相关生化及人体测量学指标。采用改良定量主观整体评估法（MQSGA）将MHD组患者分为营养不良组与营养正常组，比较两组患者间血清ZAG水平的差异。结果血清ZAG水平MHD组患者（90．10±17．70）mg／L较对照组（50．10±14．80）mg／L明显升高（t=10．86，P〈0．01）。对照组血清ZAG水平与瘦素（LP）呈负相关（r=-0．492，P〈0．01），与甘油三酯（TG）呈正相关（r=0．445，P〈0．05），MHD患者组血清ZAG与体质量指数（BMI）、白蛋白（Alb）、LP均呈负相关（r=-0．386，-0．362，-0．461，P均〈0．01），与MQSGA评分呈正相关（r=0．276，P〈0．05）。MHD患者中营养不良组血清ZAG（98．00±20．60）mg／L比营养正常组血清ZAG（82．90±10．50）mg／L明显升高（t=3．22，P〈0．01）。结论MHD患者组血清ZAG水平明显高于对照组，血清ZAG水平的升高在营养不良患者中表现更为明显，与MHD患者营养不良的发生密切相关，可作为MHD患者营养风险的独立预测指标。

锌alpha2糖蛋白在小鼠NAFLD发展中的动态变化

目的建立高脂饲料喂养的非酒精性脂肪性肝病(NAFLD)小鼠模型,动态观察锌alpha2糖蛋白(ZAG)的表达变化,初步探讨ZAG与NAFLD的关系。方法将48只8周龄C57BL/6小鼠随机分成高脂饮食组(HFD组)和标准饮食组(SD组),分别给予高脂饲料和标准饲料饲养4周、8周、12周和16周。HE染色分析肝脏组织病理学改变。酶联免疫吸附法(ELISA)检测肝脏甘油三酯(TG)含量、血清ZAG以及炎症因子水平。荧光实时定量聚合酶链反应(PCR)检测肝脏ZAG的mRNA表达。结果 4周开始,HFD组小鼠血清肿瘤坏死因子α(TNF-α)较SD组小鼠明显升高(P<0.05),且随非酒精性脂肪性肝病的发展而持续增加。8周起HFD组小鼠白细胞介素6(IL-6)、IL-8以及IL-1β水平逐渐升高(P<0.05)。4周起,HFD组小鼠肝脏ZAG mRNA表达逐渐下降(P<0.05),而血清ZAG浓度从8周开始明显降低(P<0.05)。结论肝脏ZAG表达水平降低可能参与NAFLD的发生。

锌α2糖蛋白与脂代谢和胰岛素抵抗

锌α2糖蛋白存在于人血浆和多种体液中，最近发现也可由脂肪细胞分泌，是一种新的脂肪因子。锌α2糖蛋白对于脂代谢有重要的影响，能促进脂肪分解和利用；同时也与其他细胞因子相互作用。脂代谢障碍和脂肪因子所导致的胰岛素抵抗在肥胖病、代谢综合征、糖尿病等代谢性疾病中有重要作用。因此，锌α2糖蛋白的研究有助于为这些疾病诊疗提供新思路。

锌alpha2糖蛋白与非酒精性脂肪性肝病

随着肥胖与代谢综合征在全球的流行,非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)已成为严重威胁人类健康的公共卫生问题。脂肪在肝脏过度蓄积是NAFLD的主要特点。了解调节肝脏三酰甘油(triglyceride,TG)合成所涉及的过程是预防和治疗NAFLD新视角。锌alpha2糖蛋白(Zinc alpha2 glycoprotein,ZAG)是一种能强烈促进脂肪分解、减少脂肪蓄积的新型脂肪细胞因子。ZAG可能在维持肝脏脂质代谢平衡中发挥重要作用。

The receptor for β2GPⅠon membrane of hepatocellular carcinoma cell line SMMC-7721 is annexin Ⅱ

AIM: To evaluate the receptor protein which can specifically bind to β2GPⅠon the membrane of hepatocellular carcinoma (HCC) cell line SMMC-7721, and to study the biological function of the receptor.METHODS: Through β2GPⅠ-affinity chromatography column, the peptid-polysome-mRNA complex, which can specially bind to β2GPⅠ, stayed with the column and was separated from the whole polysome of liver cells, and then eluted and collected. Using cDNA synthesis kit and cDNA PCR kit, the corresponding cDNA was obtained and sequenced. RT-PCR was used to amplify annexinⅡ, and flow cytometry was used to study the competitive binding of annexinⅡ with β2GPⅠto SMMC-7721.RESULTS: A total of 1.1 kb of the cDNA fragment of the specific binding protein of β2GPⅠon liver cell membrane was obtained. The sequence of cDNA shared high homology with human annexinⅡ (98%). AnnexinⅡ was expressed on the membrane of SMMC-7721, and could compete with β2GPⅠfor combining with SMMC-7721.CONCLUSION: The receptor for β2GPⅠon membrane of SMMC-7721 cells is annexinⅡ, which might bridge HBV to infect hepatocytes.

ZAG对结肠腺癌癌症恶病质小鼠脂质代谢的影响

目的:通过小鼠结肠腺癌癌症恶病质(CC)模型探索ZAG对CC机体脂质代谢的影响。方法:BALB/c小鼠皮下接种小鼠结肠癌colon-26细胞,构建结肠腺癌模型,BALB/c小鼠根据造模情况分为健康对照组(HC组)、肿瘤组(TC组)和肿瘤恶病质组(CC组),每组6只。取TC组和CC组的癌旁组织和肿瘤组织检测ZAG mRNA和蛋白表达水平;3组小鼠皮下脂肪组织检测ZAG蛋白浓度和m RNA表达水平,并分析TC组和CC组ZAG mRNA表达水平与体质量减轻的相关性;取3组小鼠血清,检测ZAG和脂联素的mRNA水平,并分析TC组和CC组两者的相关性。构建小鼠结肠腺癌CC模型,分别注射pcDNA、pcDNA-ZAG,观察并记录小鼠体质量变化情况。结果:CC组肿瘤组织和脂肪组织中ZAG m RNA和蛋白水平显著高于TC组(P<0.05),且脂肪组织中ZAG mRNA水平与体质量减轻值之间呈正相关;CC组血清中ZAG和脂联素的m RNA水平显著高于TC组(P<0.05),且两者呈正相关;注射pc DNA-ZAG的结肠腺癌CC小鼠体质量明显降低(P<0.05)。结论:ZAG能显著促进脂肪分解,影响结肠腺癌CC小鼠体质量。

Expression and Function of Zinc-α2-Glycoprotein

Zinc-α2-glycoprotein(ZAG), encoded by the AZGP1 gene, is a major histocompatibility complex I molecule and a lipid-mobilizing factor. ZAG has been demonstrated to promote lipid metabolism and glucose utilization, and to regulate insulin sensitivity. Apart from adipose tissue, skeletal muscle, liver, and kidney, ZAG also occurs in brain tissue, but its distribution in brain is debatable. Only a few studies have investigated ZAG in the brain. It has been found in the brains of patients with Krabbe disease and epilepsy, and in the cerebrospinal fluid of patients with Alzheimer disease, frontotemporal lobe dementia, and amyotrophic lateral sclerosis. Both ZAG protein and AZGP1 m RNA are decreased in epilepsy patients and animal models, while overexpression of ZAG suppresses seizure and epileptic discharges in animal models of epilepsy, but knowledge of the specific mechanism of ZAG in epilepsy is limited. In this review, we summarize the known roles and molecular mechanisms of ZAG in lipid metabolism and glucose metabolism, and in the regulation of insulin sensitivity, and discuss the possible mechanisms by which it suppresses epilepsy.

肥胖儿童青少年血清指标表达的影响因素分析

目的探讨肥胖儿童青少年血清锌-α2-糖蛋白(ZAG)、肥胖抑制素(obestatin)及血浆摄食抑制因子(NUCB2/Nesfatin-1)表达水平的影响因素。方法选取2015年2月至2017年1月在上海市奉贤区奉城医院门诊就诊及奉城镇中小学生单纯性肥胖症共125例作为肥胖组,选择同时期在该院体检的正常儿童青少年共90例作为对照组。比较两组的身体质量指数(BMI)、腰臀比(WHR)、体内脂肪百分比(Fat%)、空腹血糖(FBG)、糖化血红蛋白(HbAlc)、空腹胰岛素(FINS)、胰岛素抵抗指数(HOMA-IR)、高密度脂蛋白(HDL-C)、低密度脂蛋白(LDL-C)、总胆固醇(TC)、甘油三酯(TG)、Obestatin、ZAG、NUCB2/Nesfatin-1,Obestatin、ZAG、NUCB2/Nesfatin-1,经Spearman相关性分析后进一步做logistic回归分析,分别得出其表达水平的影响因素。结果肥胖组BMI、Fat%、WHR、FBG、HbAlc、HDL-C、LDL-C、TC、TG明显高于对照组(χ~2/t值分别为5.37、7.11、9.10、7.02、5.00、4.60、4.82、3.99、8.21,均P<0.05),但肥胖组ZAG、NUCB2/Nesfatin-1、Obestatin明显低于对照组(t值分别为8.00、6.68、7.19),差异均有统计学意义(均P<0.05)。经Logistic回归分析得到:BMI、WHR是影响ZAG表达水平的独立危险因素,BMI、HDL-C是影响Obestatin表达水平的独立危险因素,BMI、HOMA-IR是影响NUCB2/Nesfatin-1表达水平的独立危险因素。结论肥胖儿童青少年患者血清存在ZAG、Obestatin及NUCB2/Nesfatin-1表达水平的变化,ZAG、Obestatin及NUCB2/Nesfatin-1可能参与了儿童青少年肥胖的发生发展。

Proteomic analysis of human serum from diabetic retinopathy

AIM: To establish and compare serum proteomic of diabetic retinopathy (DR) patients in various phases and discuss pathogenesis of DR so as to find out possible serum specific molecular markers for early diagnosis of DR. METHODS: Thirty-two subjects were divided into four groups: one group of eight type 2 diabetes mellitus (T2DM) patients without apparent DR (No-DR, NDR), one group of eight T2DM patients with non-proliferative diabetic retinopathy (NPDR), one group of eight T2DM patients with proliferative diabetic retinopathy(PDR) and one group of eight healthy volunteer participants. Two dimensional fluorescence difference gel electrophoresis (2D-DIGE) was applied to establish differential protein expression profiles in four groups. Matrix-assisted laser desorption/ionization time of flight tandem mass spectrometry (MALDI-TOF-TOF MS) was applied to identify mass spectrometry of differential proteins and analyze follow-up bioinformatics. RESULTS: 2D-DIGE maps of serum protein were satisfactory obtained from NDR, NPDR, PDR and normal control groups. Twenty-six different proteins spots were screened(the volume ratio was > 1.5 based on DeCyder software analysis). Twenty-four of them were verified and two of them were not. Fifteen proteins were verified. Most of them were high-abundant proteins in serum. The four relatively low-abundant ones were beta 2-glycoprotein I (beta (2)-GPI), alpha2-HS-glycoprotein (AHSG), alpha1-acid glycoprotein (alpha(1)-AGP) and apolipoprotein A-1 (apo A-1). beta (2)-GPI expression was gradually increased in the development of DR but unrelated to the severity of DR. The volume ratio of beta (2)-GPI is 1.54, 2.43, and 2.84 in NDR, NPDR and PDR group respectively compared with normal control group. CONCLUSION: Serum proteomic analysis of 2D-DIGE combined with MALDI-TOF-TOF MS is feasible to be applied in the study of DR. 13 2-GPI probably takes part in the process of DR occurrence and development and it could be a candidate biomarker on DR diagnosis in early phase.

T cells from induced and spontaneous models of SLE recognize a common T cell epitope on β2-glycoprotein Ⅰ

Systemic lupus erythematosus is a prototypic model for B-cell epitope spread in autoimmunity.Autoantibodies to numerous molecularly distinct self-antigens emerge in a sequential manner over several years,leading to disease manifestations.Among the earliest autoantibodies to appear are those targeting phospholipid-binding proteins,particularlyβ2-glycoprotein Ⅰ.Notably,mice immunized with β2-glycoprotein Ⅰ and lipopolysaccharide develop a strong T cell response to β2-glycoprotein Ⅰ that is associated with autoantibody production and renal disease,similar to that seen in human SLE.Here we hypothesized that mice with murine systemic lupus erythematosus,whether induced or spontaneous,should have T cells that recognizeβ2-glycoprotein I.We evaluated the response of splenic T cells from mice with induced(C57BL/6 and C3H/HeN)and spontaneous(MRL/lpr)systemic lupus erythematosus to peptides spanning the entire sequence of humanβ2GPI.We found that mice with induced and spontaneous systemic lupus erythematosus recognize a common T cell epitope(peptide 31;LYRDTAVFECLPQHAMFG)in domain Ⅲ ofβ2-glycoprotein I.β2GPI-reactive CD4^(+)T cells from the two models differed primarily in cytokine production:T cells from mice with induced SLE expressed IFN-γ,while T cells from MRL/lpr mice expressed both IL-17 and IFN-γ,indicating that IL-17-expressing T cells are not necessary for generating aβ2GPI-reactive T cell response.These data suggest that the generation of aβ2-glycoprotein Ireactive T cell response is shared by both induced and spontaneous models of systemic lupus erythematosus and that this T cell response may mediate epitope spread to autoantibodies in both models.

Role of β2-glycoprotein I in the pathogenesis of the antiphospholipid syndrome

Antiphospholipid syndrome(APS)is typically characterized by increased levels of three classes of antiphospholipid antibodies,namely lupus anticoagulant,anti-β2-glycoprotein I(anti-β2GPI),and anticardiolipin antibodies.β2-Glycoprotein(β2GPI)is a phospholipid-binding protein composed of five domains(DI-V)and a major antigen in APS.β2GPI is expressed on the surfaces of several cell types,including endothelial cells,monocytes,trophoblast cells,and platelets.Its binding to the anti-β2GPI antibody triggers downstream signaling events and ultimately exerts a variety of cellular effects.β2GPI modulates hemostasis and the complement system,as well as playing an important role in APS-associated vascular injury.Therefore,studying β2GPI will help elucidate the pathogenesis of APS and improve the treatment of patients with this condition.This review will mainly focus on the structure and function of β2GPI,as well as its implication in the pathogenesis of APS.

LRG1 promotes epithelial-mesenchymal transition of retinal pigment epithelium cells by activating NOX4

AIM:To investigate the effect of leucine-rich-alpha-2-glycoprotein 1(LRG1)on epithelial-mesenchymal transition(EMT)in retinal pigment epithelium(RPE)cells,and to explore the role of NADPH oxidase 4(NOX4).METHODS:RPE cells(ARPE-19 cell line)were treated with transforming growth factor-β1(TGF-β1)to induce EMT.Changes of the m RNA and protein expression levels of LRG1 were tested in the TGF-β1 treated cells.The recombinant human LRG1 protein(r LRG1)and si RNA of LRG1 were used to establish accumulation of exogenous LRG1 model and the down-regulation of LRG1 model in ARPE-19 cells respectively,and to detect EMT-related markers including fibronectin,α-smooth muscle actin(α-SMA)and zonula occludens-1(ZO-1).The m RNA and protein expression level of NOX4 were measured according to the above treatments.VAS2870 was used as a NOX4 inhibitor in r LRG1-treated cells.EMT-related markers were detected to verify the effect of NOX4 in the process of EMT.RESULTS:TGF-β1 promoted the expression of LRG1 at both the m RNA and protein levels during the process of EMT which showed the up-regulation of fibronectin andα-SMA,as well as the down-regulation of ZO-1.Furthermore,the r LRG1 promoted EMT of ARPE-19 cells,which manifested high levels of fibronectin andα-SMA and low level of ZO-1,whereas knockdown of LRG1 prevented EMT by decreasing the expressions of fibronectin andα-SMA and increasing the expression of ZO-1 in ARPE-19 cells.Besides,the r LRG1 activated and LRG1 si RNA suppressed NOX4 expression.EMT was inhibited when VAS2870 was used in the r LRG1-treated cells.CONCLUSION:These results for the first time demonstrate that LRG1 promotes EMT of RPE cells by activating NOX4,which may provide a novel direction to explore the mechanisms of subretinal fibrosis.

2型糖尿病患者血清锌α2糖蛋白水平的变化及其相关因素分析

目的探讨T2DM患者血清锌α2糖蛋白(ZAG)的水平与胰岛素抵抗(IR)的相关性,探索其潜在的作为肥胖或代谢综合征血清指标的可能性。方法 166名受试者分为T2DM组和正常糖耐量(NGT)组,根据BMI又分为正常体重组(NW)和超重肥胖组(OW-OB)两个亚组,均测定空腹血清ZAG水平和相关临床指标。结果 T2DM组比NGT组空腹血清ZAG水平明显升高(t=2.228,P=0.028)。T2DM组患者空腹血清ZAG水平分别与HOMA-IR,WC,HbA1c,TG正相关(r分别为0.543,0.442,0.385,0.437,P<0.05),与HDL-C,TNF-a负相关(r分别为-0.447,-0.501,P<0.05)。多元逐步线性回归显示在T2DM组,WC和HDL-C是其的独立影响因素。结论 T2DM患者血清ZAG显著高于NGT患者,超重肥胖和正常体重亚组间差异无统计学意义。T2DM组中ZAG与HOMA-IR,WC,HbA1c,TG正相关,与HDL-C,TNF-α负相关;其中WC和HDL-C是血清ZAG的独立影响因素。

美洲大蠊提取物对前列腺癌细胞的抑制作用及相关机制研究

目的:研究美洲大蠊提取物(PA-CII-3)抗前列腺癌的作用及相关机制。方法:将对数生长期的PC3细胞分为对照组及实验组,在实验组细胞中依次加入1 000、1 200、1 400、1 600、1 800、2 000μg/mL的PA-CII-3溶液进行干预,采用CCK-8法筛取合适的药物浓度和作用时间。然后以1 400μg/mL的PA-CII-3溶液对PC3细胞持续干预72 h,采用CCK-8法和平板集落形成实验检测细胞增殖能力,采用细胞划痕实验检测细胞迁移能力,分别采用Real-time qPCR法和Western blot法检测锌α2糖蛋白(ZAG)的mRNA和蛋白表达情况。结果:PA-CII-3干预后,PC3细胞的增殖和迁移能力均受到显著抑制(P<0.05,P<0.01),并促进了PC3细胞中ZAG的mRNA和蛋白表达(P<0.05)。结论:PA-CII-3可能通过抑制PC3细胞的增殖和迁移及上调ZAG发挥抗前列腺癌作用。