Upregulated adenosine 2A receptor accelerates post-infectious irritable bowel syndrome by promoting CD4+ T cells’ T helper 17 polarization

Table 1 The dilution rate of primary antibodies

Primary antibodies	Dilution rate
β-actin	1:1000
A2AR	1:500
RORγt	1:2000
STAT3	1:1000
p-STAT3	1:2000
JAK2	1:1000
p-JAK2	1:1000

Table 2 Primers used in the present study

Name	Primer	Sequence	Size
mus β-actin	Forward	5‘-CACGATGGAGGGGCCGGACTCATC-3’	240 bp
	Reverse	5‘-TAAAGACCTCTATGCCAACACAGT-3’
musRORγt	Forward	5‘-CGCACCAACCTCTTTTCA-3’	274 bp
	Reverse	5‘-GCTCCACACCACCGTATTT-3’
mus JAK	Forward	5‘-AGTGGCGGCATGATTTTGTT-3’	181 bp
	Reverse	5‘-GCTCGAACGCACTTTGGTAA-3’
mus STAT3	Forward	5‘-GTAGAGCCATACACCAAGCAGCAG-3’	123 bp
	Reverse	5‘-AATGTCGGGGTAGAGGTAGACAAGT-3’
mus 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 the 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 ± 0.53	2.56 ± 0.62	3.67 ± 0.57
PI-IBS (n = 6)	0.00 ± 0.00	2.11 ± 0.49a	3.22 ± 0.58	3.89 ± 0.46
IBS + Sch58261 (n = 6)	0.00 ± 0.00	1.89 ± 0.40c	2.89 ± 0.45	3.67 ± 0.52
IBS + CGS21680 (n = 6)	0.00 ± 0.00	2.00 ± 0.41	3.11 ± 0.54	3.78 ± 0.57

^aP < 0.05 vs control group.

^cP < 0.05 vs post-infectious irritable bowel syndrome group. AWR: Abdominal withdrawal reflex; PI-IBS: Post-infectious irritable bowel syndrome.

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

Group	Time to first black stool (min)	Bristol stool grade
Control (n = 6)	105.39 ± 15.16	1.01 ± 0.12
PI-IBS (n = 6)	69.45 ± 8.72a	2.54 ± 0.48a
IBS + Sch58261 (n = 6)	94.70 ± 14.56c	1.29 ± 0.35c
IBS + CGS21680 (n = 6)	78.91 ± 9.74	3.11 ± 0.54

^aP < 0.05 vs control group.

^cP < 0.05 vs post-infectious irritable bowel syndrome group. PI-IBS: Post-infectious irritable bowel syndrome.