At 7 weeks of age, Leprflox/flox AlbCre ob/ob mice were treated with low dose leptin so as to maintain obesity and hyperinsulinemia (Supporting Fig. 1). This dose of leptin lowered plasma cholesterol and triglycerides in ob/ob mice with and without hepatic leptin signaling (Fig. 2A-C). However, plasma triglyceride levels in Leprflox/flox AlbCre+ ob/ob mice did not decrease as much as in their Leprflox/flox AlbCre- ob/ob controls (Fig. 2C). By the last day of leptin treatment, the Leprflox/flox AlbCre+ ob/ob mice had GSK1120212 cost 36% higher plasma triglycerides

than their littermate controls (Fig. 2C). The effects of leptin treatment persisted even after leptin therapy ceased, with plasma triglyceride levels in both groups only returning to near pre-leptin levels 50 days after the leptin pump was removed (Fig. 2C), indicating leptin treatment in ob/ob mice has long-term effects on lipid metabolism. Because the effect on plasma triglycerides in Leprflox/flox AlbCre ob/ob mice was subtle, we sought to reproduce these results in a complementary mouse model. We treated leptin receptor-deficient db/db mice with Ad-Lepr-b, which confers liver-selective expression18 and restores phospho-STAT3 signaling in the liver.16 Upon treatment with Ad-Lepr-b, the db/db mice remained

obese and hyperinsulinemic (Supporting Fig. 2). Also, db/db mice treated with Ad-Lepr-b and control db/db mice treated with Ad-β-gal both had a response to the virus itself independent of the Lepr-b or see more β-gal constructs. We attribute this to an acute phase immune response to the virus, which has been shown to have effects on lipid metabolism.19 Nonetheless, we observed no differences in plasma cholesterol and free fatty acids between Ad-Lepr-b– and Ad-β-gal–treated db/db mice (Fig. 2D-E). Although both virus-treated groups had an increase in plasma triglycerides,

db/db mice treated with Ad-Lepr-b had lower fasting plasma triglycerides than the Ad-β-gal–treated controls between 1 and 3 weeks postinfection, with Ad-Lepr-b treated mice reaching 31% lower plasma triglycerides 12 days postinfection (Fig. 2F). These data are similar to those of Lee et al.,14 who treated fa/fa rats with an adenovirus Florfenicol expressing β-gal or Lepr-b and also saw a marked increase in plasma triglycerides in the β-gal–treated animals compared with the Lepr-b–treated animals. Collectively, the data show that under obese, hyperinsulinemic conditions, hepatic leptin signaling is required for maintaining normal plasma triglyceride levels. Because leptin has been implicated in regulating the amount of triglyceride incorporation into VLDL,17 we evaluated lipoprotein profiles in Leprflox/flox AlbCre+ mice and their littermate controls. Mice lacking hepatic leptin signaling had no alterations in the distribution or amount of cholesterol (Fig. 3A). Interestingly, Leprflox/flox AlbCre+ mice had elevated triglycerides in fractions consistent in size with VLDL particles (Fig. 3B).