Beneficial impact of human placenta extracts on erythrocyte membrane thermostability

Abstract

PURPOSE: To study the influence of human placenta extract (HPE) and its individual fractions on the thermal stability of human erythrocyte membrane. METHODS: HPE fractions were isolated by gel chromatography. Thermal hemolysis of erythrocytes, exposed to 55°C was measured spectrophotometrically. Cytosol microvscosity and barrier function of erythrocyte membranes at hyperthermia were investigated by EPR spin probe TEMPON. Thermal denaturation of erythrocyte membrane proteins were studied by differential scanning calorimetry. RESULTS: Pre-treatment of erythrocytes with HPE or its fractions inhibited thermal hemolysis. Low-molecular fractions (below 4 kDa and 12-20 kDa) were the most effective in thermal hemolysis inhibition ((31.7±3.3) % and (31.5±3.2) %, respectively). The latter fractions markedly reduced the hyperthermia (55°C)-induced permeability of erythrocytes for ferricyanide ions and inhibited the thermo-induced structural transitions in erythrocyte membrane between 40 and 50°C, which are associated with cytoskeletal proteins. HPE fractions reversibly increased the denaturation temperatures of erythrocyte membrane proteins, except that of spectrin, and enlarged the enthalpy of denaturation of all membrane proteins. CONCLUSIONS: HPE and its individual fractions increased the thermal stability of erythrocyte membranes and erythrocytes. This effect was attributed to the reversible binding of some low molecular ingredient of HPE to the integral proteins and consequent stabilization of their interaction with under-membrane cytoskeleton.

Influence of Freeze-Thawing of Placenta on the Biological Activity of its Extracts

“…Вплив ЕПЛ на температуро-залежну динаміку стану цитозолю (мікров'язкість) та бар'єрну функцію мембрани еритроцитів при гіпертермії досліджували методом електронного парамагнітного резонанса (ЕПР) спінових зондів, як описано нами [9], із застосуванням зонду ТЕМПОН та розширюючого агента, фериціаніда калію -K 3 Fe(CN) 6 , що не проникає в інтактні еритроцити. Для приготування зразків до 0,5 мл еритроцитарної маси контрольної чи відмитої від ЕПЛ або окремих фракцій додавали по 50 мкл водних розчинів ТЕМПОН (10 -2 М) та К 3 Fe(CN) 6 (1 М) і реєстрували спектри ЕПР на спектрометрі Bruker ER 100 D, Німеччина з термостатичним пристроєм (точність ± 0,5ºС).…”