Sarnatskaya Veronika, Mikhailenko Victor, Prokopenko Igor, Gerashchenko Bogdan I, Shevchuk Oksana, Yushko Larysa, Glavin Alexei, Makovetska Lyudmila, Sakhno Larysa, Sydorenko Oleksii, Kozynchenko Oleksandr, Nikolaev Vladimir
R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology (IEPOR), NAS of Ukraine, Kyiv, Ukraine.
I. Horbachevsky Ternopil State Medical University, Ternopil, Ukraine.
Heliyon. 2020 Jan 6;6(1):e03126. doi: 10.1016/j.heliyon.2019.e03126. eCollection 2020 Jan.
The liver failure means inability to perform its normal synthetic, biotransformation and excretory functions. The disturbance of metabolic processes leads to the development of "metabolic endogenous intoxication" resulting in oxidative stress. Oxidative stress initiates the processes of oxidation of amino acid residues of blood plasma proteins causing the changes in their structure and functions. The effect of administration of highly activated porous carbonic enterosorbents on oxidative stress manifestations and molecular conformation of serum albumin in blood of experimental animals with acute liver failure induced by carbon tetrachloride (CCl) needs to be investigated. Two forms of activated carbonic enterosorbents such as AC1 (primary beads with the range of diameters of 125-250 μm) and AC2 (secondary granules prepared from micronized AC1 having the mean particle size of ~1 μm) derived from phenol-formaldehyde resin were used in rat model with CCl intoxication. The total level of reactive oxygen species (ROS) in blood plasma, the activity of catalase (CAT) in blood hemolysates; the content of reduced glutathione (GSH) in liver homogenates, and the level of oxidative modification of proteins (OMP) such as aldehyde-dinitrophenylhydrazone (A-DNPH) and ketone-dinitrophenylhydrazone (K-DNPH) derivatives in blood plasma and liver homogenates were determined. In addition, the level of pro/antioxidant ratio in blood hemolysates and the content of lipid peroxidation product - malondialdehyde (MDA), in blood plasma and liver were determined. Melting thermograms of blood plasma proteins (BPP) and molecular conformation changes of serum albumin were analyzed by biophysical methods (differential scanning microcalorimetry and spectrofluorimetry). The extent of CCl-induced oxidative damage in blood and liver of experimental animals was shown to be less expressed for AC1 in comparison with AC2 enterosorbent. However, AC2 used in the form of secondary granules positively influenced some biophysical properties of albumin molecule (temperature of melting, shape of melting endotherm and intrinsic fluorescence) after rats exposure to CCl. In general, administration of both AC1 and AC2 led to the reduction of oxidative stress manifestations and partial restoration of native molecular conformation of serum albumin. These observations are promising in terms of achieving recovery of detoxification potential of organism after severe liver injury.
肝衰竭意味着肝脏无法执行其正常的合成、生物转化和排泄功能。代谢过程的紊乱会导致“代谢性内源性中毒”的发生，进而引发氧化应激。氧化应激会启动血浆蛋白氨基酸残基的氧化过程，导致其结构和功能发生变化。需要研究给予高活性多孔碳质肠吸附剂对四氯化碳（CCl）诱导的急性肝衰竭实验动物血液中氧化应激表现及血清白蛋白分子构象的影响。两种由酚醛树脂制成的活性碳质肠吸附剂，如AC1（直径范围为125 - 250μm的初级珠粒）和AC2（由平均粒径约为1μm的微粉化AC1制备的次级颗粒）被用于CCl中毒的大鼠模型。测定了血浆中活性氧（ROS）的总水平、血液溶血产物中过氧化氢酶（CAT）的活性、肝脏匀浆中还原型谷胱甘肽（GSH）的含量，以及血浆和肝脏匀浆中蛋白质氧化修饰（OMP）如醛-二硝基苯腙（A-DNPH）和酮-二硝基苯腙（K-DNPH）衍生物的水平。此外，还测定了血液溶血产物中促氧化剂/抗氧化剂的比例以及血浆和肝脏中脂质过氧化产物丙二醛（MDA）的含量。通过生物物理方法（差示扫描量热法和荧光光谱法）分析了血浆蛋白（BPP）的熔解热谱图和血清白蛋白的分子构象变化。结果表明，与AC2吸附剂相比，CCl诱导的实验动物血液和肝脏中的氧化损伤程度在AC1组中表现较轻。然而大鼠接触CCl后，以次级颗粒形式使用的AC2对白蛋白分子的一些生物物理性质（熔解温度、熔解吸热形状和固有荧光）产生了积极影响。总体而言，给予AC1和AC2均导致氧化应激表现的减轻以及血清白蛋白天然分子构象的部分恢复。这些观察结果对于严重肝损伤后机体解毒潜能的恢复具有重要意义。
