Attenuation of TGF-beta-induced apoptosis in primary cultures of hepatocytes by calpain inhibitors

Apoptosis induced in primary cultures of rat hepatocytes by transforming growth factor beta(1) (TGF-beta(1)) was greatly attenuated by inhibitors (5 mu M) of calpain I and calpain II, respectively. Both inhibitors prevented the TGF-beta-elicited increase of nucleosomal DNA fragments and the occurrence of DNA-breaks in the TUNEL reaction. The detrimental effect of TGF-beta on cell viability measured by the WST-1 test was strongly reduced by calpain inhibitors. Calpain II > I inhibitors suppressed spontaneous DNA cleavage in hepatocytes during culture and prevented the appearance of immunocytochemically visible TGF-beta (APAAP staining), which occurs in untreated parenchymal cell cultures. The data show that inactivation of calpains attenuates both the TGF-beta-elicited and the spontaneous apoptosis of cultured hepatocytes; the latter effect is likely due to the suppression of endogenous TGF-beta activation. It is suggested that calpains participate in these processes. (C) 1997 Academic Press.