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Cell Biology International (2005) 29, 1047–1049 (Printed in Great Britain)
Possible role of endogenous growth inhibitors in regeneration of organs: Searching for new approaches
N. Giorgobiania*, D. Dzidziguria, M. Rukhadzeb, L. Rusishvilia and G. Tumanishvilia
aLaboratory of Developmental Biology, Tbilisi State University, 1 Chavchavadze Avenue, 0108 Tbilisi, Georgia, USA
bLaboratory of Neurobiology, Tbilisi State University, Tbilisi, Georgia, USA Abstract A hydrophilic protein component (12–17 Keywords: Endogenic growth inhibitor, Growth regulation. *Corresponding author. 1 Introduction Endogenous regulators of proliferation are protein molecules, encoded by regulating genes. They send specific signals to target cells to induce proliferation. In our opinion, one of the topical trends in the field of regulation of cell proliferation is investigation of endogenous growth inhibitory factors. Growth regulatory protein factors have been obtained from different organs of adult animals (Balazs and Blazsek, 1979; Kusen and Stoika, 1985; Amano and Iseki, 2001); at those times, comparatively scanty data existed about growth inhibitory factors of the myocardium. Most investigations included non-specific factors that affect: (a) the expression of synthesis of the different myocardium proteins; (b) the hypertrophy of cardiomyocytes; and (c) a directed action against death of cardiomyocytes after infarction (Kardami, 1990; Detillieux et al., 2003; Nishida et al., 2003; Kardami et al., 2004). In general, our interest is mainly in specific growth inhibitory factors of myocardium and identification of similar factors in other organs. The objective of the present work was growth inhibitory factors obtained from: (i) the myocardium of animals of different phylogenetic stages; and (ii) different organs of the same animal. 2 Materials and methods Myocardium, liver, kidney, and brain of white adult rats, as well as myocardium of adult snail, pigeon, and pig, were used as a material for investigation. Thermostable protein fractions were obtained by the method of alcohol precipitation of Balazs and Blazsek (1979), with modification. Animals were decapitated under diethyl ether. Organs were removed quickly, separated from capsules of connective tissues and vessels, rinsed with the physiological solution, and crushed. Aqueous homogenates were prepared in a tissue/distilled water ratio of 1:8. The homogenates were saturated step-wise with 96% ethanol to obtain 81% ethanol fraction, which was heated in a water bath (100 Hydrophobic interaction chromatography (HIC) was used for comparative analysis of TSPC (Queiroz et al., 2001). A hydrophilic polymeric sorbent, HEMA BIO Phenyl-1000 (particle size 10 3 Results and discussion The chromatograms in Fig. 1 show that TSPC extracted from the hearts of the animals (snail, pigeon, rat, pig) contains a hydrophilic component with a retention time of 5.5 
Fig. 1 Chromatograms of TSPC extracted from the heart of snail, pigeon, pig and rat. 
Fig. 2 The hydrophilic (1) and hydrophobic (2) components of TSPC. In previous work, we found that TSPC extracted from heart, liver, kidney, and brain of adult white rats inhibits both proliferation and transcription on average by 40–50% in cells of homologous organs of 7-day-old rats (Salakaya et al., 2000; Giorgobiani et al., 2002, 2003; Rusishvili et al., 2003; Rukhadze et al., 2005). We had also determined the molecular weights of TSPC components. In particular, electrophoresis in polyacrylamide gel showed the presence of a low molecular weight subfraction of 12–17 Focusing our attention here on the hydrophilic component of TSPC, the problem is whether this hydrophilic component and the low molecular weight fraction described earlier (Giorgobiani et al., 2002) are the same fraction. Chromatography of the low molecular weight TSPC component (Fig. 3) shows that the 12–17 
Fig. 3 Chromatogram of low molecular weight TSPC component. 
Fig. 4 Spectrum of hydrophilic component in the UV region (190–360 Chromatograms in Fig. 5 show hydrophilic and hydrophobic components of TSPC extracted from different organs (heart, liver, kidney, brain) of the same animal (adult white rat). According to the results, hydrophilic fractions of TSPC of all investigated organs seemed identical. The presence of the hydrophilic fraction in the myocardium of different animals indicates that a phylogenetically conservative group of thermostable proteins inhibiting proliferation processes has been found. 
Fig. 5 Chromatograms of hydrophilic and hydrophobic components of TSPC extracted from different organs (heart, liver, kidney, brain) of rat. In the light of these results, we suggest two approaches for the use of hydrophilic (12–17 References Amano O, Iseki, S. Expression and localization of cell growth factors in the salivary gland. Kaibogaku Zasshi 2001:76:1, 2:201-12 Balazs A, Blazsek, I. Control of cell proliferation by endogenous inhibitors. Akademia Kiado (Budapest) 1979:302: Detillieux KA, Sheikh, F, Kardami, E, Cattini, PA. Biological activities of fibroblast factor-2 in the adult myocardium. Cardiovasc Res 2003:57:8-19 Giorgobiani N, Chkhobadze, M, Rusishvili, L, Dzidziguri, D, Tumanishvili, G. Study of endogenic growth inhibitors in different organs of white rats. Proc Georgian Acad Sci Ser B 2003:9:46-9 Giorgobiani N, Rusishvili, L, Dzidziguri, D, Tumanishvili, G. The study of nucleolar parameter in evaluation of the rat myocardium functional state after the treatment with specific growth-inhibitory cardiomyocyte factor. Proc Georgian Acad Sci 2002:28:Suppl:27-31 Kardami E. Stimulation and inhibition of cardiac myocyte proliferation in vitro. Mol Cell Biochem 1990:92:129-35 Kardami E, Jiang, ZS, Jimenes, SK, Hirst, CJ, Sheikh, F, Zahradka, P. Fibroblast growth factor 2 isoforms end cardiac hypertrophy. Cardiovasc Res 2004:63:3:458-66 Kusen SI, Stoika, PS. Molecular mechanisms in action of polypeptide growth-factors. 1985: Lowry DH, Rosebrough, NJ, Farr, AL, Randell, RJ. Protein measurement with the folin phenol reagent. J Biol Chem 1951:193:265-75 Nishida S, Nagamine, H, Tanaka, Y, Watanabe, G. Protective effect of basic fibroblast growth factor against myocyte death and arrhythmias in acute myocardial infarction in rats. Circ J 2003:67:334-9 Queiroz JA, Tomaz, CT, Cabral, JMS. Hydrophobic interaction chromatography of proteins. J Chromatogr 2001:87:143-59 Rukhadze M, Dzidziguri, D, Giorgobiani, N, Kerkenjia, S. The study of growth inhibitive protein factor by various mode of HPLC and estimation of its binding with drugs. Biomed Chromatogr 2005:19:36-42 Rusishvili L, Giorgobiani, N, Dzidziguri, D, Tumanishvili, G. Comparative analysis of cardiomyocyte growth inhibitive factor in animals of different classes. Proc Georgian Acad Sci Ser B 2003:1:1–2:42-5 Salakaya T, Giorgobiani, N, Dzidziguri, D, Tumanisvili, G. Further study of the growth inhibiting factor (GIF) isolated from rat ventricular myocardium. Bull Georgian Acad Sci 2000:162:175-7 doi:10.1016/j.cellbi.2005.10.012 |
ISSN Print: 1065-6995
ISSN Electronic: 1095-8355 Published by Portland Press Limited on behalf of the International Federation for Cell Biology (IFCB) |
Abstract