Efficacy of Wumei Baijiang prescription (乌梅败酱方) on regulatory T cells / helper T cells Immune balance in mice with ulcerative colitiss

doi:10.19852/j.cnki.jtcm.2022.01.003

Abstract

Abstract:

OBJECTIVE: To observe the therapeutic effect of Wumei Baijiang prescription (乌梅败酱方), empirical prescription of Lu Zhizheng, on experimental ulcerative colitis (UC) mice, and to investigate the mechanism of the pres-cription in UC from the perspective of the immune bal-ance of regulatory T cells (Treg) and helper T cells (Th17).

METHODS: Sixty C57BL/6 mice were randomly divided into 6 groups: normal group, model group, Chinese medicine group (high, medium and low dose group of Wumei Baijiang prescription) and control group (mesalazine sustained-release granules). Except for the normal group, the other groups used 2.5% dextran sulfate sodium to induce UC mice model. At the end of the model, the Chinese medicine group was given high, medium and low dose administration of Wumei Baijiang prescription, the control group was given slow-release granules of mesalazine, and the model group was given equal volume saline for 10 d. The changes of food intake, body weight, disease activity index (DAI) score, length of large intestine and histopathology were observed. The number of Treg, Th17, CD4+, CD8+ cells in spleen was detected by flow cytometry. The expression of tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10) and C-reactive protein (CRP) in serum was detected by enzyme-linked immunosorbent assay.

RESULTS: The middle and high-dose groups of Wumei Baijiang prescription were superior to the model group in terms of increasing food intake and body weight of colitis mice, restoring colon morphology, improving pathological damage, and reducing DAI (P < 0.05). There was no statistical difference with the mesalazine group (P > 0.05). Compared with the model group, the spleen Treg and CD4+ of the mice in the high and middle dose groups of Wumei Baijiang prescription were higher, while Th17 and CD8+ were lower (P < 0.05), and there was no statistical difference compared with the mesalazine group (P > 0.05). In addition, compared with the model group, the serum levels of TNF-α and CRP in mice with high and middle doses of Wumei Baijiang prescription and mesalazine group were lower (P < 0.05), and IL-10 content was higher ( P < 0.05).

CONCLUSIONS: Wumei Baijiang prescription can improve the general conditions of colitis mice, such as diarrhea, hematochezia, weight loss, and mucosal damage. The mechanism may be related to the regulation of Treg/Th17 immune balance.

Key words: colitis, ulcerative, mesalamine, T-lymphocytes, regulatory, Th17 cells, Wumei Baijiang prescription

Xiaolan SU, Tao ZHANG, Song GUO, Xin WANG, Lin CHEN, Wei WEI. Efficacy of Wumei Baijiang prescription (乌梅败酱方) on regulatory T cells / helper T cells Immune balance in mice with ulcerative colitiss[J]. Journal of Traditional Chinese Medicine, 2022, 42(1): 30-38.

Figures/Tables 10

Table 1 Comparison of food intake of mice in each group ($bar{x}$ ± s)

Group	n	0 d	5 d	7 d	10 d	13 d	17 d
NG	6	3.9±0.4	3.4±0.7	3.7±0.6	3.4±0.6	3.5±0.6	3.8±0.6
MG	6	3.7±0.8	2.7±0.5^a	1.3±0.4^a	1.7±0.6^a	2.3±0.6^a	2.9±0.6^a
CG	6	3.8±0.7	2.5±0.7^a	1.3±0.5^a	2.3±0.6^a	3.1±0.5^b	3.7±0.4^b
WHG	6	4.1±0.7	2.3±0.7^a	1.4±0.8^a	2.3±0.5^a	3.2±0.7^b	3.6±0.5^b
WMG	6	3.8±0.5	2.4±0.6^a	1.4±0.6^a	2.2±0.5^a	3.2±0.6^b	3.6±0.7b
WLG	6	3.7±0.6	2.3±0.7^a	1.4±0.8^a	1.9±0.5^a	2.9±0.5	3.1±0.4^a

Table 2 Comparison of weight change of mice in each group ($bar{x}$ ± s)

Group	n	0 d	5 d	7 d	10 d	13 d	17 d
NG	6	23.1±0.8	23.9±0.9	24.1±0.5	24.3±0.9	24.8±1.1	24.9±1.2
MG	6	23.3±0.5	22.4±0.8	21.3±0.7^a	18.7±1.2^a	20.9±1.5^a	22.0±1.2^a
CG	6	23.4±0.7	22.5±0.8	21.7±1.1^a	19.7±1.5^a	22.5±1.2^ab	23.8±1.3^b
WHG	6	23.2±0.6	22.7±0.6	21.9±1.2^a	19.6±2.1^a	22.7±2.9^ab	23.9±2.1^ab
WMG	6	23.9±0.4	22.2±0.7	21.4±1.7^a	19.5±1.2^a	22.5±1.8^ab	23.7±1.5^ab
WLG	6	23.2±0.7	22.5±0.9	21.1±0.8^a	19.0±0.7^a	20.5±1.7^a	21.7±1.4^a

Table 3 Comparison of DAI score of mice in each group ($bar{x}$ ± s)

Group	n	0 d	5 d	7 d	10 d	13 d	17 d
NG	6	0.00±0.00	0.06±0.13	0.00±0.00	0.03±0.09	0.06±0.13	0.03±0.09
MG	6	0.00±0.00	1.51±0.78^a	3.10±0.75^a	2.63±0.40^a	1.94±0.53^a	1.46±0.41^a
CG	6	0.00±0.00	1.53±0.87^a	2.77±1.11^a	2.17±0.43^ab	1.03±0.46^ab	0.52±0.41^b
WHG	6	0.00±0.00	1.43±0.76^a	2.50±0.95^a	2.47±1.11^a	1.01±0.98^ab	0.56±0.68^ab
WMG	6	0.03±0.09	1.95±0.86^a	3.32±0.61^a	2.61±0.78^a	1.10±0.10^ab	0.61±0.54^ab
WLG	6	0.00±0.00	1.92±0.83^a	2.80±0.98^a	2.32±0.25^a	1.72±0.83^a	1.21±0.59^a

Figure 1 Colon tissue’s HE staining results of mice in each group (× 200) A: NG: normal group (saline, 0.5 mL/d); B: MG: model group (saline, 0.5 mL/d); C: CG: mesalazine group (0.52 g·kg-1·d-1); D: WHG: high dose group of Wumei Baijiang prescription (0.736 g·kg-1·d-1); E: WMG: dose group of Wumei Baijiang prescription (0.368 g·kg-1·d-1); F: WLG: low dose group of Wumei Baijiang prescription (0.184 g·kg-1·d-1). Treatment course: 10 d.

Table 4 T lymphocyte and ratio of mice in each group (%,$bar{x}$ ± s)

Group	Treg	Th17	CD4⁺	CD8⁺	CD4/CD8
NG	5.23±0.32	1.55±0.42	18.42±1.44	9.69±1.23	1.98±0.13
MG	2.27±0.78^a	4.00±0.62^a	12.73±1.89^a	15.91±1.32^a	0.79±0.76
CG	4.75±1.32^b	2.20±0.56^b	17.53±1.61^b	10.31±1.34^b	1.68±0.98
WHG	4.28±0.50^b	2.38±0.62^b	16.33±1.93^b	10.19±1.56^b	1.59±0.87
WMG	4.26±1.53^b	2.56±0.78^b	16.17±1.95^b	11.55±2.01^b	1.39±0.96
WLG	2.48±1.01^a	3.67±1.47^a	13.5±1.26^a	13.21±1.78^a	1.02±0.88

Figure 2 Comparison of Trge cells in the spleen of mice in each group A: NG: normal group; B: MG: model group; C: CG: mesalazine group (0.52 g·kg-1·d-1); D: WHG: high dose group of Wumei Baijiang prescription (0.736 g·kg-1·d-1); E: WMG: dose group of Wumei Baijiang prescription (0.368 g·k-g1·d-1); F: WLG: low dose group of Wumei Baijiang prescription (0.184 g·kg-1·d-1). Treatment course: 10 d.

Figure 3 Comparison of Th17 cells in the spleen of mice in each group. A: NG: normal group; B: MG: model group; C: CG: mesalazine group (0.52 g·kg-1·d-1); D: WHG: high dose group of Wumei Baijiang prescription (0.736 g·kg-1·d-1); E: WMG: dose group of Wumei Baijiang prescription (0.368 g·kg-1·d-1); F: WLG: low dose group of Wumei Baijiang prescription (0.184 g·kg-1·d-1). Treatment course: 10 d.

Figure 4 Comparison of CD4+T cells in the spleen of mice in each grou A: NG: normal group; B: MG: model group; C: CG: mesalazine group (0.52 g·kg-1·d-1); D: WHG: high dose group of Wumei Baijiang prescription (0.736 g·kg-1·d-1); E: WMG: dose group of Wumei Baijiang prescription (0.368 g·kg-1·d-1); F: WLG: low dose group of Wumei Baijiang prescription (0.184 g·kg-1·d-1) . Treatment course: 10 d.

Figure 5 Comparison of CD8+T cells in the spleen of mice in each group A: NG: normal group; B: MG: model group; C: CG: mesalazine group (0.52 g·kg-1·d-1); D: WHG: high dose group of Wumei Baijiang prescription (0.736 g·kg-1·d-1); E: WMG: dose group of Wumei Baijiang prescription (0.368 g·kg-1·d-1); F: WLG: low dose group of Wumei Baijiang prescription (0.184 g·kg-1·d-1) ;Treatment course: 10 d.

Table 5 Comparison of TNF-α, IL-10 and CRP of mice in each group ($bar{x}$ ± s)

Groups	n	TNF-α (pg/mL)	IL-10 (pg/mL)	CRP (mg/L)
NG	6	126.41±4.56	10.50±0.94	1.10±0.11
MG	6	275.51±5.457^a	4.48±0.87^a	5.21±0.36^a
CG	6	188.72±5.36^ab	9.27±0.68^ab	2.00±0.11^ab
WHG	6	186.75±5.91^ab	9.43±0.34^ab	1.99±0.12^ab
WMG	6	223.79±8.25^ab	8.52±0.57^ab	2.43±0.20^ab
WLG	6	241.59±5.48^ab	5.63±0.49^a	3.45±0.12^ab

References 11

[1]	Ng SC, Shi HY, Hamidi N, et al. Worldwide incidence and prevalence of inflammatory bowel disease in the 21st century: a systematic review of population-based studies. Lancet 2018;390:2769-78.
[2]	Kessler S, Rho H, West G, Fiocchi C, Drazba J, de la Motte C. Hyaluronan (HA) deposition precedes and promotes leukocyte recruitment in intestinal inflammation. Clin Transl Sci 2006;1:57-61.
[3]	Murano. M, Maemura K, Hirata I, et a1. Therapeutic effect of in tracolonIcally administered nuclear factorkappa B (p65) Antisenseolig on ucleotide on mouse dextran sulphate sodium (DSS) inducedcolitis Clin Exp Immuno 2000;120:51-8.
[4]	Hirata Y, Ihara S, Koike K. Targeting the complex interactions between microbiota, host epithelial and immune cells in inflam-matory bowel disease. Pharmacol Re. 2016;113(Pt A):574-84.
[5]	Eastaff-Leung N, Mabarrack N, Barbour A, Cummins A, Barry S. Foxp3+ regulatory T cells, Th17 effector cells, and cytokine environment in inflammatory bowel disease. J Clin Immunol 2010;30:80-9.
[6]	Ma HQ, Zhao XJ, Zhang HJ. The expression of serum inflammatory markers and Th17/Treg cells in patients with ulcerative colitis and the clinical significance. Wei Chang Bing Xue Yu Gan Bing Xue 2018;27:885-90.
[7]	He ZC, Lin N, Yin PF. The relationship between the imbalance of Th17/Treg cells and the degree of disease activity and inflammation in patients with ulcerative colitis. Zhong Guo Yi Yao Dao Bao 2019;16:130-3.
[8]	Liu ZY, Cao H. Meta-analysis of Th17, Treg and Th17/Treg Cells in Peripheral Blood of Patients with Ulcerative Colitis. Shi Yong Zhong Yi Nei Ke Za Zhi 2020;34:109-13.
[9]	Song YF, Pei LX, Zhao TT, Sun JH. Research progress on the pathogenesis of immune factors in ulcerative colitis. Yi Xue Yan Jiu Sheng Xue Bao 2019;32:432-6.
[10]	Li PR, Wei R, Wu ZP, Zhang DK. Function and research progress of Th17/Treg cells and their related cytokines in pathogenesis of ulcerative colitis. Shi Jie Ke Ji Yan Jiu Yu Fa Zhan 2017;39:194-7.
[11]	Chen Q. Methodology for pharmacological research of Traditional Chinese Medicine. Beijing: People's Medical Publishing House, 2006: 1103

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