Orally administered extract from Prunella vulgaris attenuates spontaneous colitis in mdr1a-/- mice

Table 1 Histopathological scores of cecal tissue

Microscopic parameter	FVBWT vehicle (n = 6)	mdr1a-/- vehicle (n = 10)	mdr1a-/-P. vulgaris (n = 7)
Mucosal height (μm)	3.5 ± 0.2a	4.6 ± 0.2	3.9 ± 0.2
Ulceration	0.2 ± 0.2	1.9 ± 0.3	1.2 ± 0.4
Inflammation	1.3 ± 0.2b	3.6 ± 0.2	2.5 ± 0.4c
Edema	0.7 ± 0.5a	2.4 ± 0.3	0.9 ± 0.3c
Stromal collapse (necrosis)	0.0 ± 0.0a	1.7 ± 0.4	0.5 ± 0.4
Gland hyperplasia	1.2 ± 0.2b	2.8 ± 0.2	2.1 ± 0.3c
Additive cecal score	6.9 ± 0.9b	17.0 ± 1.2	11.1 ±1.5
Mice exhibiting cecal neutrophil infiltrate	17%b	100%	38%c

Average values are shown here ± standard error of the mean except where noted.

^aP < 0.05,

^bP < 0.01 compared to mdr1a^-/- vehicle with Kruskal-Wallis test with Dunn’s multiple comparisons test.

^cP < 0.05 compared to mdr1a^-/- vehicle with Mann-Whitney test.

Table 2 Assessment of selected cytokines in the serum of mice

Cytokine/chemokine	FVBWT vehicle (n = 6)	mdr1a-/- vehicle (n = 16)	mdr1a-/-P. vulgaris (n = 13)
G-CSF	140.1 ± 15b	9694.0 ± 2563	4597.0 ± 1931
GM-CSF	ND	21.1 ± 11	1.5 ± 1
IL-9	77.9 ± 27a	200.1 ± 38	117.8 ± 14
IL-10	NDb	19.4 ± 4	4.6 ± 2ad
IL-17	4 ± 2	16 ± 4	1 ± 1
CXCL10	8.4 ± 5b	724.3 ± 136	397.2 ± 85
KC	69.5 ± 24b	527.1 ± 119	253.4 ± 58
CXCL9	108.7 ± 37b	3901.0 ± 858	1319.0 ± 277c
TNFα	9.3 ± 0.1b	14.8 ± 1	9.9 ± 3c

Serum samples were collected at the time mice were euthanized and analyzed as described in Materials and Methods. Average values (pg/mL serum) are shown here ± standard error of the mean except where noted.

^aP < 0.05,

^bP < 0.01 compared to mdr1a^-/- vehicle with Kruskal-Wallis test with Dunn’s multiple comparisons test.

^cP < 0.05,

^dP < 0.01 compared to mdr1a-/- vehicle with unpaired t-test with Welch’s correction. ND: Not detectable; P. vulgaris: Prunella vulgaris.

Table 3 Attenuation of inflammatory gene expression in mdr1a^-/- mice treated orally with an extract from Prunella vulgaris

		Fold change compared to vehicle-treated mdr1a-/- micea
Gene	Pathway affiliation	FVBWT vehicle healthy	mdr1a-/-P. vulgaris healthy	mdr1a-/-P. vulgaris colitic
Ccl2	NF-κB, LDL	-7.3	-3.4	-1.1
Cxcl1	NF-κB	-11.3	-7.2	-1.0
Cxcl9	NF-κB, Jak/Stat	-23.9	-4.2	-1.1
Icam1	NF-κB, Phospholipase C	-3.2	-1.2	1.8
Il1a	NF-κB	-13.4	-8.0	1.3
Mmp10	NF-κB, Jak/Stat	-6.6	-8.5	1.2
TNFα	NF-κB	-10.2	-2.6	1.2
Vcam1	NF-κB, Phospholipase C, LDL	-3.8	-2.0	1.2
Ccl20	NF-κB	-3.6	-2.0	1.4
Il2	NF-κB, NFAT, Calcium, PKC	-3.7	-2.2	-1.5

^aA negative value indicates that there was a lower level of gene expression when compared to the level of gene expression in vehicle-treated mdr1a^-/- mice that developed severe colitis. P. vulgaris: Prunella vulgaris.