Adenosine 2A receptor contributes to the facilitation of post-infectious irritable bowel syndrome by γδ T cells via the PKA/CREB/NF-κB signaling pathway

Table 1 The dilution ratio for primary antibodies

Primary antibodies	Dilution ratio
β-actin	1:1000
A2AR	1:500
NF-κB	1:2000
PKA	1:1000
p-PKA	1:2000
CREB	1:1000
p-CREB	1:1000
ZO-1	1:1000
Occludin	1:1000
Claudin-1	1:1000
IL-1β	1:1000
IL-6	1:1000
IL-17A	1:500
IFN-α	1:1000

A2AR: Adenosine 2A receptor; IL: Interleukin; IFN-α: Interferon-α.

Table 2 Primers used in this study

Name	Primer	Sequence	Size
β-actin	Forward	5’-CACGATGGAGGGGCCGGACTCATC-3’	240 bp
	Reverse	5’-TAAAGACCTCTATGCCAACACAGT-3’
NF-κB	Forward	5’-CACCGGATTGAAGAGAAGCG-3’	194 bp
	Reverse	5’-AAGTTGATGGTGCTGAGGGA-3’
CREB	Forward	5’-GCTGGCTAACAATGGTACGG-3’	230 bp
	Reverse	5’-CCATAACAACTCCAGGGGCA-3’
PKA	Forward	5’-GGGCGTGCTGATCTATGAGA -3’	169 bp
	Reverse	5’-TCGCTTTGTCAGATCCACCT-3’
A2AR	Forward	5’-GCCTCTTCTTCGCCTGCTTTGTCC-3’	140 bp
	Reverse	5’-GCCCTTCGCCCTCATACCCGTCAC-3’

Table 3 The effect of adenosine 2A receptor on the abdominal withdrawal reflex score in post-infectious irritable bowel syndrome mouse

Distending air volume (mL)	AWR
	0.25	0.35	0.5	0.65
Control (n = 6)	0.00 ± 0.00	1.67 ± 0.05	2.52 ± 0.18	3.65 ± 0.08
PI-IBS (n = 6)	0.00 ± 0.00	2.13 ± 0.12a	3.15 ± 0.14	3.90 ± 0.15
IBS + SCH58261 (n = 6)	0.00 ± 0.00	1.79 ± 0.12c	2.90 ± 0.25	3.57 ± 0.05
IBS + CGS21680 (n = 6)	0.00 ± 0.00	2.01 ± 0.42	3.09 ± 0.53	3.62 ± 0.11

^aP < 0.05 vs the control group.

^cP < 0.05 vs the PI-IBS group.

AWR: Abdominal withdrawal reflex; PI-IBS: Post-infectious irritable bowel syndrome.

Table 4 The effect of adenosine 2A receptor on the intestinal mobility in post-infectious irritable bowel syndrome mouse

Group	First black stool time (min)	Bristol stool grade
Control (n = 6)	407 ± 10.33	1.00 ± 0.00
PI-IBS (n = 6)	132 ± 12.59a	2.67 ± 0.52a
IBS + SCH58261 (n = 6)	246 ± 14.20c	1.33 ± 0.51c
IBS + CGS21680 (n = 6)	168 ± 11.38	2.66 ± 0.54

^aP < 0.05 vs the control group.

^cP < 0.05 vs the PI-IBS group.

PI-IBS: Post-infectious irritable bowel syndrome.