色甘酸钠灌胃对葡聚糖硫酸钠诱导的BALB/c小鼠溃疡性结肠炎模型的影响

doi:10.3969/j.issn.1674-8115.2022.10.002

上海交通大学学报（医学版） ›› 2022, Vol. 42 ›› Issue (10): 1375-1382.doi: 10.3969/j.issn.1674-8115.2022.10.002

• 论著 · 基础研究 • 上一篇

色甘酸钠灌胃对葡聚糖硫酸钠诱导的BALB/c小鼠溃疡性结肠炎模型的影响

戈伊芹(), 黄雨霁, 李玮泽, 李延宁, 李莉()

上海交通大学医学院附属第一人民医院检验科，上海 200080

收稿日期:2022-05-15 接受日期:2022-09-13 出版日期:2022-10-28 发布日期:2022-12-02
通讯作者: 李莉 E-mail:976098334@qq.com;annylish@126.com
作者简介:戈伊芹（1997—），女，硕士生；电子信箱：976098334@qq.com。
基金资助:
国家自然科学基金(81871267);上海市科委重点生物医药项目(19441904300)

Effect of intragastric treatment of sodium cromoglycate on dextran sulfate sodium-induced ulcerative colitis in BALB/c mice

GE Yiqin(), HUANG Yuji, LI Weize, LI Yanning, LI Li()

Department of Laboratory Medicine, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200080, China

Received:2022-05-15 Accepted:2022-09-13 Online:2022-10-28 Published:2022-12-02
Contact: LI Li E-mail:976098334@qq.com;annylish@126.com
Supported by:
National Natural Science Foundation of China(81871267);Science and Technology Commission of Shanghai Municipality of Scientific and Technological Research(19441904300)

摘要/Abstract

摘要：

目的·通过使用肥大细胞膜稳定剂色甘酸钠（sodium cromoglycate，Crom）稳定肥大细胞来探究其在小鼠溃疡性结肠炎（ulcerative colitis，UC）造模过程中的作用。方法·将15只小鼠分为空白组（n=5）、模型组（n=5）和Crom组（n=5）共3组。空白组小鼠自由饮用双蒸水7 d；模型组小鼠自由饮用3%葡聚糖硫酸钠（dextran sulfate sodium，DSS）溶液7 d，构建溃疡性结肠炎模型；Crom组小鼠在饮用3% DSS溶液的第3日（即造模中期）灌胃服用Crom溶液。比较空白组、模型组和Crom组小鼠的体质量改变和疾病活动指数（disease activity index，DAI）的变化，利用苏木精-伊红染色和甲苯胺蓝染色观察小鼠肠组织的病理情况和肥大细胞数量，并利用流式细胞仪检测小鼠脾脏免疫细胞的浸润情况。3组间差异比较采用单因素方差分析（one-way ANOVA），进一步组间比较采用最小显著性差异法（least-significant difference，LSD）。结果·与空白组相比，模型组小鼠重度便血，肠道病理显示有大量炎症性浸润，体质量下降严重（P=0.005），DAI评分升高明显（P=0.001），结肠长度明显缩短（P=0.003），且甲苯胺蓝染色显示肠道中肥大细胞数量增多（P=0.000）。与模型组相比，Crom组小鼠的便血减少，结肠长度有所增加（P=0.013）；且肠道病理显示，其炎症情况较模型组有所改善，肥大细胞数量较模型组减少（P=0.000）。结论·色甘酸钠灌胃可减轻小鼠溃疡性结肠炎炎症反应，间接表明肥大细胞参与了溃疡性结肠炎的发病。

关键词: BALB/c小鼠, 溃疡性结肠炎, 色甘酸钠, 肥大细胞

Abstract:

Objective ·To explore the role of mast cell membrane stabilizer sodium cromoglycate (Crom) in the modeling of ulcerative colitis (UC) in mice. Methods ·Mice were divided into 3 groups, i.e.control group (n=5), dextran sulfate sodium (DSS) group (n=5) and DSS+Crom group (n=5). Mice in the control group drank water for 7 d, and mice in the DSS group drank 3% DSS for 7 d to establish UC model. Meanwhile, mice in the DSS+Crom group were treated with Crom since the 3^rd day during the establishment of UC. The changes of body mass and disease activity index (DAI) scores among the three groups were compared. Hematoxylin-eosin staining (H-E staining) and toluidine blue staining were used to observe the histopathology injury and the number of mast cells in colon tissues. Flow cytometry was used to detect the immune cell infiltration of spleen. Differences among the three groups were compared by using one-way ANOVA. Least significant difference (LSD) was used to further compare the differences between each two groups. Results ·Compared with the control group, the mice in the DSS group had severe hematochezia and intestinal barrier injury, the body weight of the mice in the DSS group decreased significantly (P=0.005), the DAI score increased significantly (P=0.001), the colon length shortened significantly (P=0.003), and toluidine blue staining showed that the number of mast cells in the intestine increased (P=0.000). Moreover, compared with the DSS group, the blood in the stool of the mice in the DSS+Crom group decreased, and the colon length increased (P=0.013); the intestinal pathology showed that the inflammation was improved, and the number of mast cells was reduced (P=0.000). Conclusion ·Stabilizing mast cells with sodium cromoglycate could effectively reduce colon inflammation of mice in UC, which indirectly indicates that mast cells participate in the pathogenesis of UC.

Key words: BALB/c mice, ulcerative colitis (UC), sodium cromoglycate (Crom), mast cell

中图分类号:

Q95-33

戈伊芹, 黄雨霁, 李玮泽, 李延宁, 李莉. 色甘酸钠灌胃对葡聚糖硫酸钠诱导的BALB/c小鼠溃疡性结肠炎模型的影响[J]. 上海交通大学学报（医学版）, 2022, 42(10): 1375-1382.

GE Yiqin, HUANG Yuji, LI Weize, LI Yanning, LI Li. Effect of intragastric treatment of sodium cromoglycate on dextran sulfate sodium-induced ulcerative colitis in BALB/c mice[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2022, 42(10): 1375-1382.

图/表 7

表1 DAI评分标准

Tab 1 Criteria for DAI evaluation

Score	Weight loss/%	Stool consistency	Bloody
0	0	Normal	-
1	≤5	Loose	+
2	≤10	Soft， but shaped	++
3	≤20	Loose stools	+++
4	>20	Liquid	Blood to the naked eye

表2 肠道组织病理评分标准

Tab 2 Criteria for histopathological evaluation

Score	Epithelial cell structure	Inflammatory cell infiltration	Goblet cell count	Areas of inflammatory cell infiltration
0	Normal	Normal	Normal	No
1	Slight loss	Locally， superficial	Part of the loss	Local
2	The crypt is severely deformed	Diffuse， base	The number of missing is over 50%	More than 50%
3	Crypt disappear	Full thickness	No	More than 75%

图1 3组小鼠第7日的便血情况

Fig 1 Comparison of hematochezia in the mice on the 7thday among the three groups

图2 3组小鼠第7日炎症状态评估Note： A. Comparison of body mass changes in the three groups. B. Comparison of DAIs in the three groups. C/D.Qualitative (C) and quantitative (D) analysis of colon lengths in the three groups. ①P=0.005, compared with the DSS group; ②P=0.001, ③P=0.007, ④P=0.003, compared with the control group; ⑤P=0.013, compared with the DSS+Crom group.

Fig 2 Inflammatory status of mice in the three groups evaluated on the 7th day

图3 3组小鼠结肠的H-E染色结果（×10）

Fig 3 H-E staining of colons of mice in the three groups（×10）

图4 3组小鼠肠道甲苯胺蓝染色结果（×40）Note： A.Images of toluidine blue staining of colon tissues. B. Quantitive analysis of mast cells in colon tissues of the mice. ①P=0.000, compared with the control group; ②P=0.000, compared with the DSS+Crom group.

Fig 4 Results of toluidine blue staining in intestinal tracts of the mice in the three groups（×40）

图5 小鼠脾脏淋巴细胞浸润变化

Fig5 Changes of lymphocyte infiltration in mouse spleen

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色甘酸钠灌胃对葡聚糖硫酸钠诱导的BALB/c小鼠溃疡性结肠炎模型的影响

Effect of intragastric treatment of sodium cromoglycate on dextran sulfate sodium-induced ulcerative colitis in BALB/c mice

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