Changes in the electrical and viscoelastic parameters of erythrocytes in patients with manifestations of metabolic syndrome, COVID-19 convalescents, when exposed to citicoline in an in vitro experiment

M. Kruchinina, Andrei A. Gromov, Elina V. Kruchinina, Yulia A. Shishakina
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Abstract

Aim. To study changes in the electrical and viscoelastic parameters of erythrocytes using the method of dielectrophoresis in patients with manifestations of the metabolic syndrome who underwent COVID-19, when exposed to the drug citicoline in an in vitro experiment to reduce the severity of microcirculatory disorders. Materials and methods. 31 men were examined with manifestations of metabolic syndrome (50.6±9.9 years), COVID-19 convalescences, within 8 to 12 months after the disease, the diagnosis was confirmed by PCR, ELISA. The electrical and viscoelastic parameters of erythrocytes were studied by dielectrophoresis twice: the initial levels of indicators were determined and after 30 minutes of exposure with after 30 minutes of exposure to a solution of the drug Ronocyte (oral solution with the active substance citicoline sodium – 104.50 mg., which is equivalent to 100.00 mg citicoline at a concentration of 0.01 µl per 0.3 µl of red blood cell suspension in 0.3M sucrose solution (pH 7.36). Results. Exposure of erythrocyte suspension of patients with Ronocyte solution led to change in the levels of electrical, viscoelastic parameters: an increase in the average cell diameter (p=0.0003), the proportion of discocytes (p=0.0004), the amplitude of cell deformation at high frequencies of the electric field (p=0.000002), cell capacity (p=0.000007), the velocity of erythrocytes to the electrodes (p=0.003), dipole moment (p=0.002), polarizability at 106 and 0.5×106 Hz (p=0.000019 and p=0.0015, respectively), relative polarizability (p0.05) and, conversely, to reduce summarized rigidity (p=0.000003), viscosity (p=0.000002), electrical conductivity (p0.000001), aggregation indices (p=0.00003), destruction at frequencies of 106 Hz (p=0.003), 0.5x106 Hz (p=0.00002), 0.1×106 Hz (p0.00001), polarizability at low frequencies of the electric field (p=0.02). Under the influence of the drug, the equilibrium frequency of erythrocytes shifted to the low-frequency range compared to the initial values (p0.0000001). The revealed changes indicate an increase in the surface charge of erythrocytes, their ability to deform, and cell resistance under the action of citicoline. Conclusion. For the first time, the effect of the drug with the active substance citicoline, improving the rheological properties of erythrocytes, was discovered. Preparations with the active substance - citicoline should be considered promising for conducting a full-fledged clinical study to study the reduction of circulatory disorders at the microcirculatory level in patients with manifestations of metabolic syndrome who have undergone COVID-19 coronavirus infection.
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在体外实验中,代谢综合征表现患者(COVID-19 康复者)暴露于柠檬胆碱后红细胞的电参数和粘弹性参数的变化
目的利用模电泳法研究接受 COVID-19 治疗的代谢综合征患者在体外实验中接触药物柠檬胆碱以减轻微循环障碍严重程度时红细胞电参数和粘弹性参数的变化。材料和方法对 31 名有代谢综合征表现(50.6±9.9 岁)的男性进行了检查,他们在病后 8 至 12 个月内接受了 COVID-19 康复治疗,并通过 PCR 和 ELISA 进行了确诊。红细胞的电参数和粘弹性参数通过模电泳法进行了两次研究:分别测定了初始指标水平和接触药物 Ronocyte(活性物质柠檬胆碱钠口服溶液-104.50 毫克,相当于 100.00 毫克柠檬胆碱,浓度为每 0.3 微升红细胞悬液 0.01 微升,0.3M 蔗糖溶液(pH 值 7.36))30 分钟后的指标水平。结果患者的红细胞悬液接触罗诺细胞溶液后,电学、粘弹性参数水平发生了变化:细胞平均直径增加(p=0.0003)、盘状细胞比例增加(p=0.0004)、电场高频下细胞变形幅度增加(p=0.000002)、细胞容量增加(p=0.000007)、红细胞到电极的速度增加(p=0.003)、偶极矩增加(p=0.002)、106 和 0.5×106 Hz 时的极化性(分别为 p=0.000019 和 p=0.0015)、相对极化性(p0.05),反之,则会降低总结刚性(p=0.000003)、粘度(p=0.000002)、导电性(p0.000001)、聚集指数(p=0.00003)、106 Hz(p=0.003)、0.5x106 Hz(p=0.00002)、0.1×106 Hz(p0.00001)频率下的破坏性、电场低频下的极化性(p=0.02)。在药物影响下,红细胞的平衡频率与初始值相比向低频范围移动(p0.0000001)。这些变化表明,在柠檬胆碱的作用下,红细胞的表面电荷、变形能力和细胞抵抗力都有所增加。结论首次发现含有活性物质柠檬胆碱的药物具有改善红细胞流变特性的作用。含有活性物质柠檬胆碱的制剂被认为有望开展全面的临床研究,以研究减少 COVID-19 冠状病毒感染的代谢综合征患者微循环障碍的情况。
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