Substance P as a Novel Anti-obesity Target Iordanes Karagiannides, Daniel Torres, Yu–Hua Tseng, Collin Bowe, Eugenia Carvalho, Daniel Espinoza, Charalabos Pothoulakis, Efi Kokkotou  Gastroenterology  Volume 134, Issue 3, Pages 747-755.e1 (March 2008) DOI: 10.1053/j.gastro.2007.12.032 Copyright © 2008 AGA Institute Terms and Conditions

Figure 1 CJ, a neurokinin-1 receptor antagonist, prevents weight gain in lean mice fed a high-fat diet. (A) CJ (300 μg)-treated mice gained significantly less weight on a high-fat diet compared to control mice. (B) Body weight recovery after discontinuation of CJ treatments. Mice received daily IP CJ injections (150 μg for 7 days followed by 300 μg for the next 3 days) and body weight monitoring continued for a week after cessation of treatment. (C) Similar results were obtained in food intake. Gastroenterology 2008 134, 747-755.e1DOI: (10.1053/j.gastro.2007.12.032) Copyright © 2008 AGA Institute Terms and Conditions

Figure 2 CJ-treated mice develop less adiposity when fed a high fat diet. (A) Epididymal fat pad weight in control, CJ-treated and pair-fed mice. (B) CJ treatment affects fat, but not lean, mass in mice, as assessed by DEXA scanning. (C) Reduced serum leptin levels in CJ-treated mice. Gastroenterology 2008 134, 747-755.e1DOI: (10.1053/j.gastro.2007.12.032) Copyright © 2008 AGA Institute Terms and Conditions

Figure 3 Weight loss in CJ-treated (300 μg) mice is associated with a significant reduction in food intake. Gastroenterology 2008 134, 747-755.e1DOI: (10.1053/j.gastro.2007.12.032) Copyright © 2008 AGA Institute Terms and Conditions

Figure 4 CJ-treated (300 μg) mice showed significantly improved insulin sensitivity at the end of their treatment. (A) Serum insulin levels. (B) Glucose tolerance test. (C) Insulin tolerance test. Gastroenterology 2008 134, 747-755.e1DOI: (10.1053/j.gastro.2007.12.032) Copyright © 2008 AGA Institute Terms and Conditions

Figure 5 Acute effects of CJ on appetite. (A) In an acute feeding paradigm, mice were treated IP with a single dose of CJ (300 μg) at 30 minutes prior to the start of their feeding cycle (dark cycle), and food intake was monitored at intervals up to 24 hours. (B) CJ also inhibits re-feeding. Mice were food deprived overnight and then treated as above. (C) Mice were treated with a single low (36 μg) or high (72 μg) dose of SP as in (A) and their food intake was monitored acutely. (D) Mice were injected IP with SP (100 μg) at the start of their light cycle and food was removed from the cages. Glucose levels were measured at various time points in tail vein blood by a glucometer. (E) Mice were treated IP with a single dose of SP (100 μg) at the start of the light cycle in the absence of food, and hypothalamic expression of the appetite regulating neuropeptides POMC and NPY was measured after 3 hrs by real-time RT-PCR. Gastroenterology 2008 134, 747-755.e1DOI: (10.1053/j.gastro.2007.12.032) Copyright © 2008 AGA Institute Terms and Conditions

Figure 6 CJ treatment reduces food intake and weight gain in leptin-deficient mice. (A) There was a significant reduction in the weight of leptin-deficient (ob/ob) mice treated with CJ (300 μg for 7 days followed by 600 μg for 10 additional days). (B) Epididymal fat pad weight and (C) food intake in mice treated as in (A). (D) Hypothalamic expression of SP and NPY (as a positive control) was evaluated in fed and overnight fasted wild-type mice (n=6/group) by real time RT-PCR. Gastroenterology 2008 134, 747-755.e1DOI: (10.1053/j.gastro.2007.12.032) Copyright © 2008 AGA Institute Terms and Conditions

Figure 7 CJ treatment results in a dose dependent weight loss and improved insulin sensitivity in obese (DIO) mice. Mice were fed a high-fat/hypercaloric diet for 12 weeks in order to become obese. Mice were treated with 150 μg (group J) or 300 μg (group JJ) of CJ or saline (group C: control; group PF: pair-fed) for 18 days. (A) Body weight changes, (B) epididymal fat pad weight changes, and (C) serum leptin levels in response to the various treatments. (D) Weight loss during CJ treatment was associated with reduced food intake in the same mice. (E) Serum insulin levels and (F) changes in a glucose tolerance test performed just before and at the completion of CJ treatments. Gastroenterology 2008 134, 747-755.e1DOI: (10.1053/j.gastro.2007.12.032) Copyright © 2008 AGA Institute Terms and Conditions

Supplemental Figure 1 Saccharin preference in mice during a conditioned taste aversion test. LiCl treatment, but not CJ treatment, significantly reduced saccharin intake. ***P < .0001. Gastroenterology 2008 134, 747-755.e1DOI: (10.1053/j.gastro.2007.12.032) Copyright © 2008 AGA Institute Terms and Conditions