5). In contrast to db/db mice, in db/m mice only TNF-α increased significantly with MCD feeding. However, TNF-α mRNA levels increased 14-fold in db/db mice fed the MCD diet compared to only a 2-fold increase in db/m mice. Furthermore, ICAM-1 and MCP-1 also increased to a greater extent in db/db mice; 5- and 8.5-fold in db/db mice compared to 2- and 3-fold in db/m mice, respectively (P < 0.05). Although

ICAM protein expression did increase more dramatically after MCD feeding in db/db compared to db/m mice, protein expression in db/db mice did not exceed that of db/m mice on the MCD diet. On densitometric analysis the effect of the MCD diet was only significant in db/db mice (Table 1). Db/m mice fed the MCD diet had a >50% reduction in SAM levels compared to control MAPK inhibitor diet fed animals (P < 0.01). In contrast, the MCD diet had learn more no significant effect on SAM levels, suggesting that SAM depletion may not play as prominent a role in the development of steatohepatitis or UPR activation of the UPR in db/db mice. There were no significant differences in hepatic SAH levels or SAM/SAH ratio between the groups

(Table S2). Others have demonstrated that JNK1 knockout mice are resistant to MCD-induced steatohepatitis.26 Although complete JNK inhibition in humans may not be advisable, partial inhibition with a pharmacologic inhibitor may be of benefit for the treatment NASH. We performed an experiment to assess the effect of partial JNK inhibition on MCD-induced steatohepatitis. The wildtype strain C57BLKS/J was used instead of the db/db or db/m strain in an attempt to more directly ascertain the effect of JNK inhibition 上海皓元 independent of diabetes or defective leptin signaling. Preliminary experiments in wildtype mice documented the presence of steatohepatitis after 2 weeks of MCD feeding. Therefore, MCD and control diet-fed C57BLKS/J mice were treated with SP 600125, a specific pharmacologic JNK inhibitor, for 2 weeks to assess the drug’s ability to attenuate the development of MCD induced liver injury. SP600125 decreased both JNK2/3 and JNK1

protein levels (Fig. 6). As expected, mice fed the MCD diet developed steatohepatitis; however, the severity was not affected by SP600125 did not improve histology in mice fed MCD diet. Serum ALT and hepatic triglyceride content were unchanged in MCD-fed mice treated with SP 600125, compared to MCD-fed mice given vehicle (Table 2A). Although SP 600125 failed to have a biochemical or histological effect, it did significantly reduce downstream inflammatory mediators (MCP-1, TNFα, ICAM, and iNOS). Its effects on UPR activation were less clear, and appeared to be more selective. A significant reduction in mRNA levels was appreciated for CHOP; however, there was no effect on EDEM mRNA levels (Table 2B).