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AIM To extract and purify the transforming growth factor13(TGF β),and to demonstrate its biological activity invivo and induction of apoptosis of hepatocytes in vitro.METHODS TGF β was isolated from fresh bovine plateletsby acid/ethanol extraction method and purified with ionexchange and gel chromatography.The extracted TGF βwas injected subcutaneously to mice,and its biologicalactivity in vivo was observed 72 hrs post-injection by HEstaining.The morphological changes were observed by HEstaining and the occurrence of apoptosis was detected byTUNEL method after the human normal hepatic cell lineQZG was treated with 8μg·L ~1 TGFβ for 12 hrs in vitro.RESULTS The molecular mass 25 ku TGF β protein wassuccessfully extracted.It was able to induce localizedgranulation tissue formation in vivo.TGF β-treatedhepatocytes showed obvious apoptotic morphologicalchanges,including the pyknosis and dense-stained nucleiand cytoplasm,the fragmentary,annular or crescentnuclei,and the“bubbling”cytoplasm.Moreover,itsapoptotic rate was significantly higher than that of thecontrol group(P<0.05).CONCLUSION Biological active TGF β protein is extractedand purified successfully from bovine platelets,and it isable to induce the apoptosis of hepatocytes.
AIM To extract and purify the transforming growth factor 13 (TGF β), and to demonstrate its biological activity invivo and induction of apoptosis of hepatocytes in vitro. METHODS TGF β was isolated from fresh bovine platelets by acid / ethanol extraction method and purified with ion exchange and gel chromatography. The extracted TGF βwas injected subcutaneously to mice, and its biologicalactivity in vivo was observed 72 hrs post-injection by HE staining. The morphological changes were observed by HE stain and the occurrence of apoptosis was detected by TUNEL method after the human normal hepatic cell line QZG was treated with 8 μg · L ~ 1 TGFβ for 12 hrs in vitro .RESULTS The molecular mass 25 ku TGF β protein wassuccessfully extracted. It was able to induce localized tissue formation in vivo. TGF-β-treated hepatocytes of the highly apoptotic morphological changes, including the pyknosis and dense-stained nucleiand cytoplasm, the fragmentary, annular or crescentnuclei, and the “bubbling ” cy (P <0.05). CONCLUSION Biological active TGF β protein is extracted and purified successfully from bovine platelets, and it is able to induce the apoptosis of hepatocytes.