1. Volume 140, Issue 4, Pages 1283-1291.e2 (April 2011)
Synergistic Role of TRPV1 and TRPA1 in Pancreatic Pain and Inflammation 
Erica S. Schwartz, Julie A. Christianson, Xiaowei Chen, Jun–Ho La, Brian M. Davis, Kathryn M. Albers, G.F. Gebhart 
Gastroenterology 
Volume 140, Issue 4, Pages e2 (April 2011)
DOI: /j.gastro
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2. Figure 1
Cerulein produces pancreatitis. Pancreatic histology, examined (A) 1 day after repeated injection of vehicle (PBS) or (B) 1, (C) 3, and (D) 7 days after repeated injection of cerulein (N = 6 per group), revealed significant edema (closed arrowheads) and neutrophil infiltration (open arrowhead) as early as 1 day after cerulein treatment with recovery by 3 days. No evidence of inflammation was observed in vehicle-treated pancreas. (E) MPO activity (units/mg tissue) was also significantly increased 1 and 3 days following cerulein treatment (n = 8 each day) but was not significant from vehicle (veh, n = 6) treatment by day 7 (F3,24 = one-way ANOVA; P < .001). *P < .05 relative to vehicle treatment (Bonferoni post hoc test). caer, cerulein.
Gastroenterology  , e2DOI: ( /j.gastro )
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3. Figure 2
Pancreatic inflammation increases TRP channel mRNA expression. The expression levels of TRPV1 and TRPA1 mRNA in pancreatic (A) DRG and (B) NG neurons were also elevated 1 and 3 days after cerulein treatment. n = 6 for vehicle and n = 9 for caer-1d– and caer-3d–treated mice, and the number of cells used for mRNA quantification is given in each bar. *P < .05, 2-way ANOVA with a Holm–Sidak post hoc test to compare vehicle vs cerulein-treated groups. veh, vehicle; caer, cerulein.
Gastroenterology  , e2DOI: ( /j.gastro )
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4. Figure 3
Pancreatitis increases the percentage of pancreatic neurons responding to CAP (TRPV1) and MO (TRPA1). Ca2+ imaging was performed on pancreatic DRG (left panels) and NG (right panels) neurons from vehicle- and cerulein (caer)-treated mice in response to (A) CAP or (B) MO. In vehicle-treated mice (open circles), the percentage of neurons expressing functional TRPV1 was significantly higher in pancreatic DRG neurons than in NG neurons. In contrast, the functional expression of TRPA1 was similar between the 2 ganglia types. Cerulein treatment significantly increased the percentage of pancreatic DRG neurons that express functional TRPV1 or TRPA1. Cerulein treatment only produced a transient (1 day posttreatment) significant increase in the percentage of pancreatic NG neurons expressing TRPV1 or TRPA1. All data were analyzed by 2-way ANOVA, and in all cases F > 10.2, P < .01. *P < .05 with a Holm–Sidak post hoc test to compare groups (n = at least 8 animals and n = 38 cells per group).
Gastroenterology  , e2DOI: ( /j.gastro )
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5. Figure 4
Pharmacologic antagonism of TRP channels reduces pancreatic inflammation. Pancreatic histology, examined 1 day after repeated injections of (A) cerulein or TRP channel antagonists: (B) 100 mg TRPV1 and (C) 100 mg TRPV1 (AMG9810) plus 100 mg TRPA1 (HC ) (n = 6 per group). (D) TRPV1 (AMG9810) or TRPA1 (HC ) receptor antagonists had no effect on MPO activity (units/mg tissue) in vehicle-treated mice (n = 6) but dose-dependently attenuated pancreatic inflammation in cerulein-treated mice. Doses are expressed as milligrams per kilogram in the bars; n = 5–8 per treatment group; F9,50 = one-way ANOVA with a Bonferoni post hoc test; *P < .01 (vs vehicle); †P < .01 (vs cerulein). Combined treatment with 100 mg/kg of both antagonists was comparable to or more effective than 300 mg/kg of either antagonist given alone (**P < .05). veh, vehicle; caer, cerulein.
Gastroenterology  , e2DOI: ( /j.gastro )
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6. Figure 5
Pharmacologic antagonism of TRP channels blocks pancreatitis pain. Activity monitoring revealed that (A) the total distance traveled, (B) time spent moving in the horizontal plane, and (C) rearing events in the vertical plane were are all significantly reduced in cerulein- vs vehicle-treated mice (n = 6 per group; all data were analyzed by 2-way ANOVA and in all cases F > 10.1, P < .01). Morphine (2.5 mg/kg subcutaneously) given 30 minutes before testing significantly reversed all 3 behavioral parameters in cerulein-treated mice (all data analyzed by 2-way ANOVAs and in all cases F > 7.2, P < .01). Combined treatment with TRPV1 and TRPA1 antagonists (each at 100 mg/kg) significantly prevented cerulein-produced behaviors at all times tested. All data were analyzed by 2-way ANOVA and in all cases F > 10.2, P < .01. *P < .01 (vs cerulein); **P < .001 (vs vehicle); Holm–Sidak post hoc test. caer, cerulein.
Gastroenterology  , e2DOI: ( /j.gastro )
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7. Supplementary Figure 1
Pancreatitis increases peak response amplitude of pancreatic neurons to CAP (TRPV1) and MO (TRPA1). Ca2+ imaging of pancreatic DRG (left column) and NG (right column) neurons from vehicle- and cerulein (caer)-treated mice revealed significant increases in peak amplitude to application of (A) CAP or (B) MO. All data were analyzed by 2-way ANOVA and in all cases F1,58 = 10.1, P < .01. *P < .05, Holm–Sidak post hoc test to compare groups (n = at least 8 animals and n = 38 cells per group).
Gastroenterology  , e2DOI: ( /j.gastro )
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