Cirrhosis of liver: Interference of serpins in quantification of SERPINA4 - A preliminary study.
Cirrhosis of liver: Interference of serpins in quantification of SERPINA4 - A preliminary study.
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DOI：
10.1016/j.plabm.2017.10.002
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发表时间：
2017-12
影响因子：
1.9
通讯作者：
Muninarayana C
中科院分区：
文献类型：
--
作者：
Nallagangula KS;Shashidhar KN;Lakshmaiah V;Muninarayana C
肝硬变是一种慢性肝衰竭导致肝功能受损的病理状态。由于合成能力的下降，血液中血浆蛋白的表达往往会减少。丝氨酸蛋白酶抑制因子(Serpins，Serpins)是一类在肝脏中表达的血浆蛋白，结构相似，功能多样。SERPINA4(Kalistatin)是一种由肝脏表达的多功能丝氨酸A蛋白，血清中SERPINA4的浓度是反映肝功能障碍程度的指标。通过与SERPINA4的免疫交叉反应，确定其他蛇毒对肝硬变和健康人的干扰。血液样本来自印度卡纳塔克邦科拉尔的R.L.Jalappa医院和研究中心的20名受试者(10名肝硬变，10名健康人)。用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法分离蛋白质。用免疫印迹法分析交叉反应性。用十二烷基硫酸钠-琼脂糖凝胶电泳法分离肝硬变患者和健康人血清中存在的蛋白质。在肝硬变和健康的PVDF(聚偏二氟乙烯)膜上均未检测到条带。然而，重组kallistatin出现了一条明显的条带。由于肝硬变患者和健康人的表位不同，结构相似、氨基酸序列相似性较小的蛇毒与SERPINA4没有表现出任何免疫交叉反应。目前的研究表明，蛇毒对免疫反应的干扰不存在于kalistatin的定量估计中，这一点有待进一步验证。通过免疫学上的相互作用排除结构相似的Serpins对SERPINA4定量估计的干扰。肝硬变生物标志物卡力斯汀评价方法的初步研究病例对照研究采用SDS-PAGE和Western印迹分析方法。
Cirrhosis of liver is a pathological condition, wherein functions of liver are impaired by chronic liver exploitations. Due to decrease in synthetic capacity, expressions of plasma proteins tend to decrease in blood stream. Serpins (Serine protease inhibitors) are class of plasma proteins expressed from liver with structural similarities and diverse functions. SERPINA4 (Kallistatin) is a multifunctional serpin clade A protein expressed from liver and concentration in serum is the reflection of extent of liver dysfunction. To identify interference of other serpins by immunological cross reactivity with SERPINA4 in cirrhotic liver and healthy subjects. Blood samples were collected from 20 subjects (10 cirrhotic liver, 10 healthy) from R.L. Jalappa Hospital and Research Centre, Kolar, Karnataka, India. Separation of proteins was carried out by SDS-PAGE. Cross reactivity study was analyzed using western blot. Proteins present in cirrhotic liver and healthy subject's serum were separated by SDS PAGE. There was no band detection on both (cirrhotic liver and healthy) PVDF (polyvinylidene diflouride) membranes. However, a significant band was observed with recombinant kallistatin. Structurally similar serpins with minor amino acid sequence similarities did not show any immunological cross reactivity with SERPINA4 due to non identical epitope in cirrhotic liver and healthy subjects. Present study revealed that there is no interference of serpins for immunological reactions in quantitative estimation of kallistatin which needs further validation. To rule out structurally similar Serpins interference in SERPINA4 quantitative estimations by immunological interactions. A preliminary study for the evaluation of method in evolution of biomarker Kallistatin for Cirrhosis of Liver. Case control study using analytical methods SDS-PAGE and western blot.