The mechanism of Wumei pill on ulcerative colitis in mice based on Nrf2/ARE antioxidant stress pathway

doi:10.11958/20230533

Abstract

Abstract:

Objective To explore the mechanism of Wumei pill on ulcerative colitis (UC) in mice based on the anti oxidative stress pathway of nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant response element (ARE). Methods Seventy SPF male C57BL/6 mice were randomly divided into the control group, the UC group, the mesalazine group (MES group, 0.82 g/kg MES), the low dose Wumei pill group (WMW-L group, 5 g/kg crude drug), the middle dose Wumei pill group (WMW-M group, 10 g/kg crude drug), the high dose Wumei pill group (WMW-H group, 20 g/kg crude drug) and the high dose Wumei pills+Nrf2 inhibitor ML-385 group (WMW-H+ML-385 group, Wumei pills crude drug 20 g/kg+20 mg/kg ML-385), with 10 rats in each group. The disease activity index (DAI) score and colonic mucosa injury score were performed in mice after the last administration. Pathological changes of colonic mucosa in mice were observed by HE staining. The levels of interleukin (IL) -1β, tumor necrosis factor-α (TNF-α) and IL-6 in serum and colon tissue of mice were measured by enzyme-linked immunosorbent assay (ELISA). The content of malondialdehyde (MDA) in serum and colon tissue of mice was determined by thiobarbituric acid colorimetry (TBA). The activity of superoxide dismutase (SOD) in serum and colon tissue of mice was measured by xanthine oxidase method. The activity of glutathione peroxidase (GSH-px) in serum and colon tissue of mice was determined by direct method with dithiodinitrobenzoic acid (DTNB). The positive expression of Nrf2 in colon tissue of mice was observed by immunohistochemistry. The expression of heme oxygenase-1 (HO-1) and NAD(P)H:quinone oxidoreductase-1 (NQO1) proteins in colon tissue of mice were detected by Western blot assay. Results Compared with the control group, the DAI score, colonic mucosa injury score, colonic histopathology score, levels of IL-1β, TNF-α, IL-6 and MDA in serum and colonic tissue, and expression levels of Nrf2, HO-1 and NQO1 protein in colonic tissue of mice were increased in the UC group, levels of SOD and GSH-px in serum and colon tissue decreased (P<0.05), the colon mucosa of mice was seriously damaged. Compared with the UC group, changes of corresponding indexes were contrary to the above in the MES group, the WMW-M group and the WMW-H group. However, the expression levels of Nrf2, HO-1 and NQO1 proteins in colon tissue were increased (P<0.05), and the damage of colon mucosa in mice was alleviated. Changes of the above indexes were dose-dependent in the WMW-L group, the WMW-M group and the WMW-H group. There were no significant differences in the above indexes between the WMW-H group and the MES group. ML-385 attenuated the improvement effect of high dose Wumei pill on colon mucosa injury. Conclusion Wumei pill may alleviate the colon mucosal damage of UC mice by activating Nrf2/ARE antioxidant stress pathway.

Key words: colitis, ulcerative, NF-E2-related factor 2, antioxidant response elements, oxidative stress, Wumei pill

CLC Number:

R285.5

Figures/Tables 8

References 25

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组别	DAI（n=10）	结肠黏膜损伤评分（n=10）	组织病理学评分（n=5）
对照组	0	0	0
UC组	3.86±0.43^a	4.47±0.49^a	3.76±0.21^a
MES组	1.04±0.07^b	1.59±0.13^b	1.15±0.10^b
WMW-L组	3.80±0.41	4.39±0.46	3.71±0.32
WMW-M组	2.13±0.15^bc	2.73±0.22^bc	2.25±0.17^bc
WMW-H组	0.96±0.08^bcd	1.54±0.14^bcd	1.08±0.11^bcd
WMW-H+ML-385组	2.97±0.22^e	3.75±0.31^e	3.14±0.28^e
F	363.996^＊	312.249^＊	270.269^＊

组别	DAI（n=10）	结肠黏膜损伤评分（n=10）	组织病理学评分（n=5）
对照组	0	0	0
UC组	3.86±0.43^a	4.47±0.49^a	3.76±0.21^a
MES组	1.04±0.07^b	1.59±0.13^b	1.15±0.10^b
WMW-L组	3.80±0.41	4.39±0.46	3.71±0.32
WMW-M组	2.13±0.15^bc	2.73±0.22^bc	2.25±0.17^bc
WMW-H组	0.96±0.08^bcd	1.54±0.14^bcd	1.08±0.11^bcd
WMW-H+ML-385组	2.97±0.22^e	3.75±0.31^e	3.14±0.28^e
F	363.996^＊	312.249^＊	270.269^＊

组别	血清（n=10）			结肠组织（n=5）
组别	IL-1β	TNF-α	IL-6	IL-1β	TNF-α	IL-6
对照组	38.26±6.87	165.22±18.65	11.76±2.43	47.80±9.33	178.95±19.45	25.63±3.29
UC组	114.35±12.56^a	396.87±32.15^a	158.49±19.36^a	136.92±15.14^a	415.57±34.68^a	178.48±12.30^a
MES组	52.17±7.20^b	228.41±20.52^b	62.30±5.38^b	64.48±6.24^b	237.73±21.43^b	75.26±7.15^b
WMW-L组	109.54±8.82	385.47±30.26	152.79±16.22	130.68±13.34	409.50±30.47	174.34±11.92
WMW-M组	73.42±5.94^bc	297.15±23.08^bc	93.51±8.42^bc	90.27±7.49^bc	356.53±24.26^bc	118.92±10.16^bc
WMW-H组	48.79±4.85^bcd	219.84±18.57^bcd	58.37±5.06^bcd	59.82±5.56^bcd	231.46±20.36^bcd	69.44±6.39^bcd
WMW-H+ML-385组	82.06±7.69^e	324.25±25.36^e	117.54±9.30^e	104.52±8.33^e	388.24±26.84^e	142.50±11.08^e
F	137.824^＊	126.668^＊	234.576^＊	62.662^＊	70.542^＊	186.631^＊

组别	血清（n=10）			结肠组织（n=5）
组别	IL-1β	TNF-α	IL-6	IL-1β	TNF-α	IL-6
对照组	38.26±6.87	165.22±18.65	11.76±2.43	47.80±9.33	178.95±19.45	25.63±3.29
UC组	114.35±12.56^a	396.87±32.15^a	158.49±19.36^a	136.92±15.14^a	415.57±34.68^a	178.48±12.30^a
MES组	52.17±7.20^b	228.41±20.52^b	62.30±5.38^b	64.48±6.24^b	237.73±21.43^b	75.26±7.15^b
WMW-L组	109.54±8.82	385.47±30.26	152.79±16.22	130.68±13.34	409.50±30.47	174.34±11.92
WMW-M组	73.42±5.94^bc	297.15±23.08^bc	93.51±8.42^bc	90.27±7.49^bc	356.53±24.26^bc	118.92±10.16^bc
WMW-H组	48.79±4.85^bcd	219.84±18.57^bcd	58.37±5.06^bcd	59.82±5.56^bcd	231.46±20.36^bcd	69.44±6.39^bcd
WMW-H+ML-385组	82.06±7.69^e	324.25±25.36^e	117.54±9.30^e	104.52±8.33^e	388.24±26.84^e	142.50±11.08^e
F	137.824^＊	126.668^＊	234.576^＊	62.662^＊	70.542^＊	186.631^＊

组别	血清（n=10）			结肠组织（n=5）
组别	SOD/（U/mg）	MDA/（μmol/L）	GSH-px/（U/mg）	SOD/（U/mg）	MDA/（μmol/L）	GSH-px/（U/mg）
对照组	114.26±3.20	1.85±0.14	203.50±12.46	127.49±8.62	2.24±0.17	234.26±17.34
UC组	23.53±1.89^a	8.67±0.45^a	96.21±5.29^a	33.27±2.38^a	10.38±0.86^a	117.16±6.85^a
MES组	89.08±5.21^b	3.04±0.20^b	167.34±10.18^b	89.46±6.43^b	4.25±0.25^b	210.34±16.47^b
WMW-L组	24.68±2.14	8.25±0.63	98.07±8.69	34.29±3.18	10.16±0.72	119.72±8.86
WMW-M组	60.35±5.36^bc	5.74±0.30^bc	143.25±10.26^bc	66.82±5.33^bc	6.79±0.44^bc	165.83±12.45^bc
WMW-H组	96.48±8.93^bcd	2.58±0.16^bcd	173.34±14.30^bcd	92.78±7.45^bcd	3.88±0.28^bcd	217.33±16.09^bcd
WMW-H+ML-385组	43.57±5.84^e	6.25±0.44^e	126.34±10.09^e	56.28±6.18^e	7.49±0.65^e	143.15±12.27^e
F	485.133^＊	548.147^＊	145.492^＊	161.288^＊	170.028^＊	64.769^＊