L. S. Kurilova

The effect of oxidized glutathione and its pharmacological analogue glutoxim on intracellular Ca2+ concentration in macrophages Ca2+

Using Fura-2AM microfluorimetry, the effect of oxidized glutathione (GSSG) and its pharmacological analogue glutoxim on the intracellular Ca2+ concentration in rat peritoneal macrophages was investigated. It was shown that GSSG or glutoxim increase the intracellular Ca2+ concentration by inducing Ca2+ mobilization from thapsigargin-sensitive Ca2+ stores and subsequent Ca2+ entry from external medium. Dithiothreitol, which reduces S-S-bonds in proteins, completely prevents or reverses the increase of intracellular Ca2+ concentration induced by GSSG or glutoxim. This suggests that the increase of intracellular Ca2+ concentration induced by GSSG or glutoxim can be mediated by their interactions with functionally important SH-groups of proteins involved in Ca2+-signaling.Two structurally different tyrosine kinase inhibitors genistein and methyl-2,5-dihydroxycinnamate prevent or completely reverse the increase in the intracellular Ca2+ concentration induced by GSSG or glutoxim. On the contrary, tyrosine phosphatase inhibitor Na orthovanadate enhances the increase of intracellular Ca2+ concentration evoked by oxidizing agents. The data suggest that tyrosine kinases and tyrosine phosphatases are involved in the regulatory effect of GSSG and glutoxim on the intracellular Ca2+ concentration in macrophages.

Involvement of small G proteins and vesicle traffic in the glutoxim and molixan effects on the intracellular Ca(2+) concentration in macrophages.

252 Glutoxim (bisodium salt of oxidized glutathione (GSSG) containing nanoconcentrations of cissplatii num; PharmaVAM, Russia) is a pharmacological GSSG analogue used as an immunomodulating and hematopoesissstimulating agent in the complex therr apy of bacterial and viral infections, psoriasis, and in radioo and chemotherapy of malignant tumors [1]. Molixan (PharmaVAM) is a complex of glutoxim and inosine with an antiviral, immunomodulating, and hepatoprotective action used in the therapy of acute viral hepatitis B and C, mixed hepatitis, and liver cirr rhosis [1]. Glutoxim and molixan belong to the pharr macological group of thiopoetines, which affect intraa cellular redox regulation. However, the cellular and molecular mechanisms of their action are insuffii ciently understood. In our previous studies, it was first shown that GSSG, glutoxim, and molixan increased the intracell lular Ca 2+ concentration ([Ca 2+ ] i) in rat peritoneal macrophages by mobilizing calcium ions from thapsii garginnsensitive Ca 2+ stores and subsequently stimuu lating the Ca 2+ uptake [2–4]. Using a wide range of agents affecting different components of intracellular signaling systems, we first identified the principal elements of the signal cascade triggered by GSSG and glutoxim and resulting in a [Ca 2+ ] i increase in macrophages, namely, tyrosine kinases and tyrosine phosphatases [3, 5], phosphatii dylinositol kinases [6], and the key enzymes of the phosphoinositide signaling system, phospholipase C and protein kinase C [7]. It was also found that the effects of glutoxim and molixan on [Ca 2+ ] i in macc rophages were mediated by actin cytoskeleton elee ments [8] and microtubules [9]. The involvement of microtubules and the actin cytoskeleton in the glutoxim and molixan action on [Ca 2+ ] i in macrophages invites the assumption that macrophage activation induced by these agents is mediated by vesicle traffic. It is known that intracellu lar trafficking of secretory vesicles depends on microo tubules, which regulate the transport efficiency and organize the vesicle traffic by acting like cellular highh ways. Agents causing microtubule disintegration have been shown to inhibit secretion in different types of cells [10]. In addition, it was reported that glutoxim could induce vesicle exocytosis in macrophages conn taining M. tuberculosis [11]. Based on these data, we considered it worthwhile to investigate the possible involvement of vesicle transport and small G proteins, important components of the exocytosis signaling pathway, in mediating the glutoxim and molixan effects on the [Ca 2+ ] i level in macrophages. Experiments were …

Cyclooxygenase and lipoxygenase inhibitors modulate the Glutoxim and Molixan effects on the intracellular Ca 2+ concentration in macrophages

277 Glutoxim (disodium salt of GSSG with cisplatin at nano concentration, FARMA VAM, Moscow), a pharmacological analogue of oxidized glutathione (GSSG), is used as an immunomodulator and hemo stimulator in integrated treatment of bacterial and viral diseases and psoriasis, as well as in radiation ther apy and chemotherapy of cancer [1]. Another disul fide containing drug, Molixan (a complex of Glu Cyclooxygenase and Lipoxygenase Inhibitors Modulate the Glutoxim and Molixan Effects on the Intracellular Ca2+ Concentration in Macrophages Z. I. Krutetskaya, L. S. Kurilova, V. G. Antonov, and Academician A. D. Nozdrachev Received: April 1, 2013

Involvement of Microtubules in the Effects of Glutoxim and Molixan on the Intracellular Concentration of Ca 2+ in Macrophages

Currently, a great number of disulfidecontaining drugs altering the redox status and having a physiolog� ically significant effect on cells have been developed and introduced into clinical practice. The pharmaceu� tical agent Glutoxim®, disodium salt of oxidized glu� tathione (GSSG) with a platinum nanoadditive (PHARMAVAM, Moscow, Russia) is used as an immunomodulator and a hemostimulant in the inte� grated therapy of bacterial and viral diseases, psoriasis, as well as radioand chemotherapies of cancer (1). Another disulfidecontaining agent, molixan (a com� plex of glutoxim and inosine nucleoside), has a similar application. However, the mechanisms of the cellular and molecular effects of these drugs are not com� pletely understood. We were the first to find out earlier that GSSG, glu� toxim, and molixan enhance the intracellular Са 2+

[The involvement of actin cytoskeleton in glutoxim and molixan effect on intracellular Ca(2+)-concentration in macrophages].

Glutoxim and molixan belong to new generation of disulfide-containing drugs with immunomodulatory, hepatoprotective and hemopoetic effect on cells. Using Fura-2AM microfluorimetry, two structurally distinct actin filament disrupters latrunculin B and cytochalasin D, and calyculin A, which causes actin filaments condensation under plasmalemma, we have shown the involvement of actin cytoskeleton in the intracellular Ca2+-concentration increase induced by glutoxim or molixan in rat peritoneal macrophages. Morphological data obtained with the use of rhodamine-phalloidine demonstrated that glutoxim and molixan cause the actin filaments reorganization in rat peritoneal macrophages.