Nallagangula Krishna Sumanth, Shashidhar K N, Lakshmaiah V, Muninarayana C
Department of Biochemistry, Sri Devaraj Urs Medical College, SDUAHER, Tamaka, Kolar, Karnataka, India.
Pract Lab Med. 2017 Oct 7;9:53-57. doi: 10.1016/j.plabm.2017.10.002. eCollection 2017 Dec.
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.
肝硬化是一种病理状态，其中肝脏功能因慢性肝脏损伤而受损。由于合成能力下降，血浆蛋白在血流中的表达往往会降低。丝氨酸蛋白酶抑制剂（Serpins）是一类由肝脏表达的血浆蛋白，具有结构相似性和多种功能。SERPINA4（激肽释放酶抑制蛋白）是一种由肝脏表达的多功能丝氨酸蛋白酶抑制剂A类蛋白，血清中的浓度反映了肝功能障碍的程度。
确定肝硬化患者和健康受试者中其他丝氨酸蛋白酶抑制剂与SERPINA4的免疫交叉反应对其的干扰。
从印度卡纳塔克邦科拉尔的R.L.贾拉帕医院和研究中心的20名受试者（10名肝硬化患者，10名健康人）采集血样。通过SDS-PAGE进行蛋白质分离。使用蛋白质印迹法分析交叉反应性研究。
通过SDS-PAGE分离了肝硬化患者和健康受试者血清中的蛋白质。在（肝硬化患者和健康人）聚偏二氟乙烯（PVDF）膜上均未检测到条带。然而，重组激肽释放酶抑制蛋白出现了一条明显的条带。
由于肝硬化患者和健康受试者中存在不同的表位，结构相似但氨基酸序列相似度较小的丝氨酸蛋白酶抑制剂与SERPINA4未显示任何免疫交叉反应。本研究表明，在激肽释放酶抑制蛋白的定量估计中，丝氨酸蛋白酶抑制剂对免疫反应没有干扰，这需要进一步验证。
