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Extraction and Partial Purification of a Hepatic Stimulatory Substance in Rats, Mice, and Dogs¹

Department of Gastroenterology, University of Bari, Bari, Italy [A. F., L. P.], and Departments of Anatomy and Cell Biology [P. O., M. C.], Surgery [L. M., T. E. S.], and Gastroenterology [D. H. V. T.], University Health Center of Pittsburgh, University of Pittsburgh, Pittsburgh 15261, and Department of Crystallography, Veterans Administration Medical Center [J. R.], Pittsburgh, Pennsylvania 15240

A factor has been isolated from weanling rat liver which stimulates in vivo hepatic DNA synthesis in a dose dependent manner when injected into 40% hepatectomized rats. The factor has been partially purified by successive steps, involving ethanol precipitation, ultrafiltration through an Amicon PM 30 membrane, and finally fast protein liquid chromatography, resulting in a 38,000-fold increase in specific activity over that in the original cytosol. The factor contains a few bands in the molecular weight range of 14,000–50,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Active fractions from fast protein liquid chromatography (F₁₅₀), when injected into 40% hepatectomized rats, increased hepatic DNA synthesis 3-fold over the background stimulation due to the hepatectomy. The response was dose dependent over a range from 1.76 µg to 6.8 µg per 200-g (body weight) rat. Mitotic and labeling indexes confirmed that F₁₅₀ stimulates both replicative DNA synthesis and cell proliferation. The factor is heat and neuraminidase resistant, trypsin sensitive, organ specific, but not species specific.

¹ This study was supported by a research project grant from the Veterans Administration, Project Grant AM-29961 from the NIH, Bethesda, MD, and Grant 885/02 16544 from Consiglio Nazionale delle Ricerche, Italy.

² To whom requests for reprints should be addressed, at Veterans Administration Hospital, Building 6, Circle Drive, Pittsburgh, PA 15240.

Received 11/19/86. Revised 4/ 1/87. Revised 7/21/87. Accepted 7/31/87.

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