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与非糖尿病患者相比,砷暴露对2型糖尿病患者血清中AS3MT蛋白水平的影响。

Effect of Arsenic Exposure on AS3MT Protein Levels in Serum of Type 2 Diabetic Patients Compared to Non-diabetics.

作者信息

Shokat Saima, Iqbal Riffat, Ali Muhammad Ahmed, Yaqub Atif, Aslam Shaista, Riaz Samreen

机构信息

Government College University, Lahore, Pakistan.

National University of Medical Sciences, Rawalpindi, Pakistan.

出版信息

Biol Trace Elem Res. 2024 Dec 19. doi: 10.1007/s12011-024-04448-2.

Abstract

This research explores the impact of arsenic exposure on serum protein profiles in type 2 diabetes patients, with an emphasis on the AS3MT protein as a biomarker. Utilizing Bradford protein assay, SDS-PAGE, HPLC, and mass spectrometry, we quantified and analyzed variations in serum protein levels, focusing on differences between control groups (82.94 ± 8.03 µg/mL) and diabetic patients (96.95 ± 5.02 µg/mL) of high arsenic exposed in areas Kasur and Lahore, Punjab, Pakistan. The study revealed a significant increase in total serum proteins and specifically identified elevated levels of AS3MT in the diabetic group compared to controls. By using 15% gel, proteins were separated, and bands were visible at 42KD. Further investigations using HPLC provided a detailed chromatographic profile of AS3MT, isolating this protein effectively and displaying its heightened abundance through a marked peak within the sample chromatograms. Additionally, intact mass and tryptic digestion profiles analyzed by mass spectrometry (molecular weight of 41,747.79 D) further corroborated the identity and modification of AS3MT in the context of arsenic exposure. ELISA was used for the quantification of AS3MT protein concentration, and a 260% increase was confirmed in the diabetic group exposed to arsenic. These findings suggested that arsenic exposure significantly alters AS3MT protein and serum protein levels in diabetic patients, supporting the hypothesis that AS3MT can serve as a biomarker for arsenic-induced diabetic conditions.

摘要

本研究探讨了砷暴露对2型糖尿病患者血清蛋白质谱的影响,重点关注AS3MT蛋白作为生物标志物的情况。利用考马斯亮蓝蛋白测定法、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)、高效液相色谱(HPLC)和质谱分析法,我们对血清蛋白水平的变化进行了量化和分析,重点关注巴基斯坦旁遮普省卡苏尔和拉合尔高砷暴露地区的对照组(82.94±8.03μg/mL)和糖尿病患者(96.95±5.02μg/mL)之间的差异。研究发现,糖尿病组的总血清蛋白显著增加,并且与对照组相比,特别鉴定出AS3MT水平升高。通过使用15%的凝胶分离蛋白质,在42KD处可见条带。使用HPLC进行的进一步研究提供了AS3MT的详细色谱图,有效地分离了该蛋白,并通过样品色谱图中的明显峰显示其丰度增加。此外,通过质谱分析的完整质量和胰蛋白酶消化谱(分子量为41,747.79 D)进一步证实了在砷暴露情况下AS3MT的身份和修饰。酶联免疫吸附测定(ELISA)用于定量AS3MT蛋白浓度,并且证实暴露于砷的糖尿病组增加了260%。这些发现表明,砷暴露显著改变了糖尿病患者的AS3MT蛋白和血清蛋白水平,支持了AS3MT可作为砷诱导糖尿病状况生物标志物的假设。

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