Intestinal anti-inflammatory activity of Ground Cherry (Physalis angulata L.) standardized CO2 phytopharmaceutical preparation

Table 1 Details of primers used for real-time PCR analysis

Target	Sequence	Annealing, °C	Oligo concentration, nmol/L	Fragment size, bp	NCBI reference sequence	cDNA concentration, μL
GAPDH	F TGACTCTACCCACGGCAAGTTCAA	60	200	141	NM_017008.3	1.000
	R ACGACATACTCAGCACCAGCATCA
β-actin	F TTGCTGACAGGATGCAGAAGGAGA	60	100	159	NM_031144.2	1.000
	R ACTCCTGCTTGCTGATCCACATCT
HPRT	F AGGGAAGTGACAATCTACCTGACG	60	100	81	AA900579.1	0.125
	R GAAATGTCTGTTGCTGCGTCCCTT
HSP70	F ACTCCTTCGTTCGGTCTGCAATCA	60	200	92	NM_031971.2	0.125
	R CTGGGAATGCAAAGCACACGTGAA
Heparanase	F TGTCAAGAGTGAAAGGCCCAGACA	60	200	141	NM_022605.1	0.125
	R GCAGCTTCAAGTGCTTGGTGACAT
NF-κB	F AAACCAAAGCCCTGAAAGGCCATC	60	200	120	XM_342346.4	0.125
	R TCGGAAGGCCTCGAATGACATCAA
MAPK1	F AACAGGTTGTTCCCAAACGCTGAC	60	200	187	NM_053842.1	0.500
	R AGTCGTCCAGCTCCATGTCAAACT
MAPK3	F TACCTGGACCAGCTCAACCACATT	60	200	173	NM_017347.2	0.500
	R AGCAGGTCAAGAGCTTTGGAGTCA
MAPK6	F AACTGAGCCAGTGGAAGAAGGGAA	60	200	164	NM_031622.2	1.000
	R TTAACGTGGCCTGGATGGACTTGA
MAPK9	F TCATGGGAGAGCTGGTGAAAGGTT	60	200	106	NM_017322.2	1.000
	R ATGAACTCTGCGGATGGTGTTCCT				NM_001270544.1
					NM_001270545.1
MUC1	F CCGCTACTACCAAGAACTGAAG	60	200	102	NM_012602.1	0.500
	R GAGCCTGACCTGAACTTGATAG
MUC2	F GGCTCTGCTCTCTGTGTTATAG	60	200	123	U68172.1	1.000
	R CAGTTTGGGAAGAAGGTAGGG
MUC3	F GGGAAATAGACCCTGCAGTTAG	60	200	107	U76551.1	0.125
	R GATCATCGCTTGCCGTCATA
MUC4	F ATGTGGAGGTGGGAGAAATG	60	200	122	AF240632.1	0.500
	R CCCTGGAACTGGAATTAGAGAC

Table 2 Macroscopic effects of the PACO2 standardized extract (25, 50 or 100 mg/kg) in acute phase response of intestinal inflammation induced by trinitrobenzenesulphonic acid

Group	Damage score, 0-101	Extension of lesion, cm2	Colonic weight, mg/cm2	Diarrhea3	Adherence3
Non-colitic	0a	0a	82.7 ± 2.10a	0%a	0%a
TNBS-control	7 (2-8)	2.55 ± 0.28	136.0 ± 5.87	87.50%	81.25%
PACO2 - 25 mg/kg	7 (5-8)	2.44 ± 0.18	135.0 ± 6.86	100.00%	71.42%
PACO2 - 50 mg/kg	7 (2-9)	2.11 ± 0.34	124.0 ± 7.72	85.71%	71.42%
PACO2 - 100 mg/kg	7 (4-9)	2.93 ± 0.20	162.0 ± 5.04	100.00%	57.14%

¹Score data are expressed as the median (range);

²Extension of lesion and colonic weight data are expressed as the mean ± SE of the mean;

³Diarrhea and adherence were analyzed by Fisher's exact test.

^aP ≤ 0.01.

Table 3 Effects of the PACO₂ standardized extract (25, 50 or 100 mg/kg) on glutathione content, myeloperoxidase and alkaline phosphatase activities in acute phase response of intestinal inflammation induced by trinitrobenzenesulphonic acid

Group	GSH, nmol/g tissue	MPO, U/g tissue	ALP, mU/mg protein
Non-colitic	1687 ± 54.2a	104 ± 13.7a	8.91 ± 2.17a
TNBS control	1295 ± 60	1703 ± 390	28.9 ± 3.07
PACO2, 25 mg/kg	1208 ± 33.8	1360 ± 286	21.0 ± 2.44
PACO2, 50 mg/kg	1350 ± 85.1	921 ± 181	18.0 ± 1.62c
PACO2, 100 mg/kg	1428 ± 50.6	831 ± 108c	19.5 ± 1.50c

Data are expressed as mean ± standard error of the mean.

^cP ≤ 0.05 and

^aP ≤ 0.01. GSH: Glutathione; MPO: Myeloperoxidase; ALP: Alkaline phosphatase.

Table 4 Effects of the PACO₂ standardized extract (25, 50 or 100 mg/kg) on cytokine levels in acute phase response of intestinal inflammation induced by trinitrobenzenesulphonic acid

Group	IFN-γ	TNF-α	IL1-β	IL-6	IL-10
Non-colitic	78.2 ± 7.9c	121.2 ± 7.0a	1395.0 ± 136.9a	421.1 ± 45.3a	467.1 ± 31.1a
TNBS control	125 ± 20.4	240.3 ± 34.3	2205.0 ± 147.4	893.8 ± 105.5	675.5 ± 38.6
PACO2, 25 mg/kg	92.1 ± 8.6	246.6 ± 34.3	1867.0 ± 221.4	891.9 ± 87.09	674.5 ± 40.4
PACO2, 50 mg/kg	67 ± 11.7c	167.9 ± 23.6	2106.0 ± 128.9	563.4 ± 55.5c	626.9 ± 66.5
PACO2, 100 mg/kg	76.4 ± 12.7c	205.5 ± 34.5	2323.0 ± 154.1	649.5 ± 86.9	537.4 ± 41.1

Data are expressed as mean ± SE of the mean.

^cP ≤ 0.05 and

^aP ≤ 0.01.