Serum malondialdehyde levels,myeloperoxidase and catalase activities in patients with nephrotic syndrome

Abstract

Objectives

Some studies have indicated the pathophysiological importance of reactive oxygen species (ROS) in patients with nephrotic syndrome. Myeloperoxidase (MPO) is a leukocyte-derived enzyme-generating ROS that has been proposed to exert a wide array of pro-atherogenic effects throughout all stages of the atherosclerotic process. The aim of this study was to investigate the serum malondialdehyde (MDA) levels, MPO and catalase activities in patients with adult nephrotic syndrome.

Patients and Methods

Twenty-four patients with nephrotic syndrome and 24 healthy controls were enrolled. Serum MPO activity, catalase activity, and MDA levels were assessed.

Results

Serum MPO activity and MDA levels were signi?cantly higher in patients with nephrotic syndrome than controls (both, P < 0.001), while catalase activity was signi?cantly lower (P < 0.001). Serum catalase activity was found to be significantly correlated with MPO activity (r = ?0.417, P = 0.003) and MDA levels (r = ?0.532, P = 0.007). The serum MDA levels were also found to be significantly correlated with MPO activity (r = 0.419, P = 0.003).

Conclusions

We concluded that serum MPO activity and oxidative stress were increased and that serum catalase activity was decreased in patients with adult nephrotic syndrome. In addition, these results indicate that increased MPO activity is associated with an oxidant–antioxidant imbalance that may contribute to atherosclerosis in patients with adult nephrotic syndrome.     

抗动脉粥样硬化核酸疫苗安全性的初步研究

目的：研究抗动脉粥样硬化核酸疫苗的安全性。方法：观察该质粒在小鼠组织中的分布和急性毒性,井在正常给药的情况下对新西兰大白兔进行6个月毒性试验。采用肌肉注射20μg/只和100μg/只,用P（温法检测外源基因在小鼠组织的分布和存留时间;同样采用肌肉注射,对最高剂量达到到50kg/kg体重的小鼠进行急性毒性试验。结果：实验证明该核酸疫苗在注射部位可存留时间为8周,其他组织如心、肝、脾、肺、肾、脑和生殖器官也有低水平分布,急毒试验显示,在剂量达到有效荆量的60倍的情况下,无可观察到的毒性反应发生,小鼠血液学和生化指标亦无异常;新西兰大白兔毒性观察,体内生化指标以及解剖学研究结果表明,该药无可观察到的毒性反应发生。结论：该核酸疫苗无可观察到的明显毒性反应,安全性良好。     

巨噬细胞的自噬和极化抑制其泡沫化进程

目的:巨噬细胞是动脉粥样硬化斑块中最丰富的免疫细胞,巨噬细胞泡沫化加速动脉粥样硬化,本研究探讨巨噬细胞自噬与极化对泡沫化的影响。方法:分离培养小鼠腹腔巨噬细胞,免疫荧光检测巨噬细胞的标记物F4/80。不同浓度雷帕霉素处理巨噬细胞,western blot检测自噬标记物LC3II,经典激活的巨噬细胞(Classically activated macrophages, M1)标记物白细胞介素6(interleukin 6, IL-6)和替代激活的巨噬细胞(Alternatively activated macrophages, M2)标记物转化生长因子β(transform growth factor,TGF-β)的表达。用ox-LDL诱导巨噬细胞泡沫化,油红O染色鉴定泡沫细胞形成及巨噬细胞泡沫化情况。结果:免疫荧光结果显示,小鼠腹腔巨噬细胞F4/80阳性率达87.6%;雷帕霉素处理巨噬细胞24 h,western blot结果显示LC3II表达增加,M1标记物IL-6表达增加,而M2标记物TGF-β表达减少,对其条带进行统计分析结果显示都具有显著性差异(P0.05);油红O染色结果显示雷帕霉素明显减少巨噬细胞泡沫化形成。结论:雷帕霉素能诱导巨噬细胞自噬,促进其向M1型极化,从而抑制巨噬细胞泡沫化。     

miR-19靶向PTEN并介导HMGB1影响小鼠动脉粥样硬化进程的机制研究

摘要 目的：探究miR-19靶向PTEN并介导HMGB1影响小鼠动脉粥样硬化进程的机制研究。方法：SPF级C57BL/6J ApoE^-/-雄性小鼠根据研究目的将实验小鼠分为对照组、AS模型组和miR-19抑制剂组。通过RT-PCR分析小鼠主动脉组织中miR-19的mRNA表达。通过蛋白印迹分析小鼠主动脉PTEN、HMGB1和AKT的蛋白表达。通过荧光素酶活性检测miR-19a与PTEN的靶向关系。通过组织学和红油O染色分析小鼠胸腹主动脉和主动脉窦中的AS斑块面积。通过RT-PCR分析小鼠主动脉主动脉弓内膜中促炎细胞因子和趋化因子的mRNA表达。通过蛋白印迹分析主动脉弓内膜中ICAM-1和VCAM-1的蛋白表达。结果：AS模型组miR-19mRNA表达较对照组升高（P<0.05）,miR-19抑制剂组miR-19mRNA表达较AS模型组降低（P<0.05）。AS模型组PTEN蛋白表达较对照组降低,HMGB1和AKT蛋白表达较对照组升高（P<0.05）,miR-19抑制剂组PTEN蛋白表达较AS模型组升高,miR-19抑制剂组HMGB1和AKT蛋白表达较AS模型组降低（P<0.05）。AS模型组主动脉和主动脉窦的斑块面积较对照组增加（P<0.05）,miR-19抑制剂组主动脉和主动脉窦的斑块面积较AS模型组减少（P<0.05）。AS模型组TNF-α、IL-β、IL-6和CXCL2的mRNA表达较对照组升高（P<0.05）,miR-19抑制剂组TNF-α、IL-6、IL-β和CXCL2的mRNA表达较AS模型降低（P<0.05）。AS模型组ICAM-1和VCAM-1的蛋白表达较对照组升高（P<0.05）,miR-19抑制剂组ICAM-1和VCAM-1的蛋白表达较AS模型组降低（P<0.05）。结论：miR-19通过靶向调控PTEN表达激活HMGB1/PI3K/Akt信号通路,这可能会促进VSMCs的异常增殖、迁移和炎症反应,有助于AS的进展。     

冠心病合并糖尿病患者外周血巨噬细胞中脂质稳态与动脉粥样硬化指数的关联研究

摘要 目的：探究冠心病合并糖尿病患者外周血巨噬细胞中脂质稳态与动脉粥样硬化指数的关联机制。方法：2015年1月至2020年9月,筛选78名2型冠心病合并糖尿病患者,根据血糖变异度分为3组：A组、B组及C组。检测总胆固醇（TC）,甘油三酯（TG）,低密度脂蛋白胆固醇（LDL-C）、高密度脂蛋白胆固醇（HDL-C）、空腹胰岛素（FINS）和超敏C反应蛋白（hs-CRP）。评估动脉粥样硬化指数（AI）和冠状动脉狭窄程度,并分析其相关性及独立危险因素。结果：B组较A组TC、LDL-C和AI升高,HDL-C降低,B组较C组TC、LDL-C和AI降低,HDL-C升高（P<0.05）。B组较A组hs-CRP、FINS、TG值升高,B组较C组hs-CRP、FINS、TG值降低（P<0.05）。B组较A组HOMA-IR值升高,B组较C组HOMA-IR值降低（P<0.05）。B组较A组AI和冠脉Gensini评分值升高,B组较C组AI和冠脉Gensini评分降低（P<0.05）。Person线性相关分析显示：hs-CRP、TG、Gensini评分、HOMA-IR值与AI正相关（P<0.05）。在逻辑回归模型中,结果显示：hs-CRP、TG、Gensini评分、HOMA-IR值是AI的独立危险因素。结论：Gensini评分与AI呈正相关,且hs-CRP、TG、Gensini评分、HOMA-IR值AI的独立危险因素。     

A Solute Carrier Family 22 Member 3 Variant rs3088442 G→A Associated with Coronary Heart Disease Inhibits Lipopolysaccharide-induced Inflammatory Response

Recent genome-wide association studies have identified single-nucleotide polymorphism (SNPs) within the SLC22A3 (solute carrier family 22 member 3) gene associated with coronary heart disease (CHD) in the Caucasian population. We performed molecular analysis to investigate the potential role of SLC22A3 variants in CHD. Our study showed that the common polymorphism rs3088442 G→A, which is localized in the 3′ UTR of the SLC22A3 gene, was associated with a decreased risk of CHD in the Chinese population by a case control study. In silico analysis indicated that G→A substitution of SNP rs3088442 created a putative binding site for miR-147 in the SLC22A3 mRNA. By overexpressing miR-147 or inhibiting endogenous miR-147, we demonstrated that SNP rs3088442 G→A recruited miR-147 to inhibit SLC22A3 expression. Moreover, SLC22A3 deficiency significantly decreased LPS-induced monocytic inflammatory response by interrupting NF-κB and MAPK signaling cascades in a histamine-dependent manner. Notably, the expression of SLC22A3^A was also suppressed by LPS stimulus. Our findings might indicate a negative feedback mechanism against inflammatory response by which SLC22A3 polymorphisms decreased the risk of CHD.     

Understanding the fluid mechanics behind transverse wall shear stress

The patchy distribution of atherosclerosis within arteries is widely attributed to local variation in haemodynamic wall shear stress (WSS). A recently-introduced metric, the transverse wall shear stress (transWSS), which is the average over the cardiac cycle of WSS components perpendicular to the temporal mean WSS vector, correlates particularly well with the pattern of lesions around aortic branch ostia. Here we use numerical methods to investigate the nature of the arterial flows captured by transWSS and the sensitivity of transWSS to inflow waveform and aortic geometry. TransWSS developed chiefly in the acceleration, peak systolic and deceleration phases of the cardiac cycle; the reverse flow phase was too short, and WSS in diastole was too low, for these periods to have a significant influence. Most of the spatial variation in transWSS arose from variation in the angle by which instantaneous WSS vectors deviated from the mean WSS vector rather than from variation in the magnitude of the vectors. The pattern of transWSS was insensitive to inflow waveform; only unphysiologically high Womersley numbers produced substantial changes. However, transWSS was sensitive to changes in geometry. The curvature of the arch and proximal descending aorta were responsible for the principal features, the non-planar nature of the aorta produced asymmetries in the location and position of streaks of high transWSS, and taper determined the persistence of the streaks down the aorta. These results reflect the importance of the fluctuating strength of Dean vortices in generating transWSS.     

Contrasting effects of stanniocalcin-related polypeptides on macrophage foam cell formation and vascular smooth muscle cell migration

Stanniocalcin (STC) is a calcium- and phosphate-regulating hormone secreted by the corpuscles of Stannius, an endocrine gland of bony fish. Its human homologues, STC1 and STC2 showing 34% amino acid identity each other, are expressed in a variety of human tissues. To clarify their roles in atherosclerosis, we investigated the effects of their full-length proteins, STC1(18–247) and STC2(25–302), and STC2-derived fragment peptides, STC2(80–100) and STC2(85–99), on inflammatory responses in human umbilical vein endothelial cells (HUVECs), human macrophage foam cell formation, the migration and proliferation of human aortic smooth muscle cells (HASMCs) and the extracellular matrix expression. All these polypeptides suppressed lipopolysaccharide-induced expressions of interleukin-6, monocyte chemotactic protein-1, and intercellular adhesion molecule-1 in HUVECs. Oxidized low-density lipoprotein-induced foam cell formation was significantly decreased by STC1(18–247) and increased by STC2(80–100) and STC2(85–99), but not STC2(25–302), in human macrophages. Expression of acyl-CoA:cholesterol acyltransferase-1 (ACAT1) was significantly suppressed by STC1(18–247) but stimulated by STC2(80–100) and STC2(85–99). Expression of ATP-binding cassette transporter A1 was significantly stimulated by STC1(18–247). Neither STC1(18–247) nor STC2-derived peptides significantly affected CD36 expression in human macrophages or HASMC proliferation. STC2(80–100) and STC2(85–99) significantly increased HASMC migration, whereas STC1(18–247) significantly suppressed the angiotensin II-induced HASMC migration. Expressions of collagen-1, fibronectin, matrix metalloproteinase-2, and elastin were mostly unchanged with the exception of fibronectin up-regulation by STC2(80–100). Our results demonstrated the contrasting effects of STC1 and STC2-derived peptides on human macrophage foam cell formation associated with ACAT1 expression and on HASMC migration. Thus, STC-related polypeptides could serve as a novel therapeutic target for atherosclerosis.     

编者按: 脂代谢与心血管疾病

心血管病是我国成年人致死的最主要的疾病,而脂代谢异常是心血管疾病的独立危险因素。因此,阐述研究揭示脂代谢异常在心血管疾病发生发展中的作用及其机制,具有重要的理论意义和临床实用价值。本期专题主要一方面综合评述了脂蛋白组分与脂质代谢,以及甘氨酸的心血管疾病保护作用等相关领域的研究新进展,同时并分别展示了国内学者有关于PCSK9/LDLR通路、模拟人apoE结构域的小分子多肽EpK、多不饱和脂肪酸、脂联素、LXRα- ABCA1途径和普罗布考等在脂代谢中的作用及相应分子机制等方面的研究成果,以期让更多的人进一步深入了解脂质代谢以及,了解心血管病发病的复杂机制和及研究现状。     

Oxidized LDL induces phosphorylation of non-muscle myosin IIA heavy chain in macrophages

Oxidized LDL (oxLDL) performs critical roles in atherosclerosis by inducing macrophage foam cell formation and promoting inflammation. There have been reports showing that oxLDL modulates macrophage cytoskeletal functions for oxLDL uptake and trapping, however, the precise mechanism has not been clearly elucidated. Our study examined the effect of oxLDL on non-muscle myosin heavy chain IIA (MHC-IIA) in macrophages. We demonstrated that oxLDL induces phosphorylation of MHC-IIA (Ser1917) in peritoneal macrophages from wild-type mice and THP-1, a human monocytic cell line, but not in macrophages deficient for CD36, a scavenger receptor for oxLDL. Protein kinase C (PKC) inhibitor-treated macrophages did not undergo the oxLDL-induced MHC-IIA phosphorylation. Our immunoprecipitation revealed that oxLDL increased physical association between PKC and MHC-IIA, supporting the role of PKC in this process. We conclude that oxLDL via CD36 induces PKC-mediated MHC-IIA (Ser1917) phosphorylation and this may affect oxLDL-induced functions of macrophages involved in atherosclerosis. [BMB Reports 2015; 48(1): 48-53]