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基于NLRP3炎症小体探讨降糖3号方缓解糖尿病小鼠炎症反应的分子机制
The molecular mechanisms of
Jiang Tang San Hao Formula alleviating inflammatory responses in diabetic mice via the NLPR3 inflammasome- 北京中医药大学学报 2024年47卷第11期 页码：1541-1549
- 作者机构：
1.北京中医药大学中医学院 北京 100029
2.北京大学医学部
- 作者简介：
赵祎，女，在读硕士生
#赵丹丹，女，博士，副研究员，硕士生导师，主要研究方向：脏腑相关理论防治糖尿病等代谢性疾病的基础与临床研究，E-mail: bucmzhaodandan@163.com
- 基金信息：国家自然科学基金项目(82174329);北京中医药大学基本科研业务费揭榜挂帅项目(2023-JYB-JBZD-010);中医药传承与创新“百千万”人才工程（岐黄工程）岐黄学者项目(1040063321005)
DOI：10.3969/j.issn.1006-2157.2024.11.009
中图分类号： R285.5收稿日期：2024-05-23，
网络出版日期：2024-10-12，
纸质出版日期：2024-11-30
- 稿件说明：
移动端阅览
赵祎, 李润琪, 徐冰蕊, 等. 基于NLRP3炎症小体探讨降糖3号方缓解糖尿病小鼠炎症反应的分子机制[J]. 北京中医药大学学报, 2024,47(11):1541-1549.
ZHAO Yi, LI Runqi, XU Bingrui, et al. The molecular mechanisms of
Jiang Tang San Hao Formula alleviating inflammatory responses in diabetic mice via the NLPR3 inflammasome[J]. Journal of beijing university of traditional chinese medicine, 2024, 47(11): 1541-1549.赵祎, 李润琪, 徐冰蕊, 等. 基于NLRP3炎症小体探讨降糖3号方缓解糖尿病小鼠炎症反应的分子机制[J]. 北京中医药大学学报, 2024,47(11):1541-1549. DOI： 10.3969/j.issn.1006-2157.2024.11.009.
ZHAO Yi, LI Runqi, XU Bingrui, et al. The molecular mechanisms of
Jiang Tang San Hao Formula alleviating inflammatory responses in diabetic mice via the NLPR3 inflammasome[J]. Journal of beijing university of traditional chinese medicine, 2024, 47(11): 1541-1549. DOI： 10.3969/j.issn.1006-2157.2024.11.009.
摘要
目的
2
观察降糖3号方对2型糖尿病(T2DM)小鼠机体与肠道炎症反应及NLRP3炎症小体的影响，揭示其抗糖尿病的分子机制。
方法
2
选取4周龄C57BL/6 N雄性小鼠，高脂饲料联合链脲佐菌素腹腔注射构建T2DM小鼠模型，将成模小鼠随机数字表法分成模型组、二甲双胍组、降糖3号方组，同时设置正常组。各组小鼠每日灌胃相应药液，二甲双胍组灌胃二甲双胍0.20 g/kg，降糖3号方组灌胃降糖3号方4.26 g/kg，正常组、模型组灌胃等量灭菌水，连续给药8周。药物干预结束后，酶联免疫吸附法检测血清及结肠组织中脂多糖(LPS)、肿瘤坏死因子－α(TNF-α)、白细胞介素－1β(IL-1β)、白细胞介素－6(IL-6)含量，苏木素－伊红染色观察结肠组织病理形态，免疫组织化学法检测结肠组织中NOD样受体热蛋白结构域相关蛋白3(NLRP3)、凋亡相关斑点样蛋白(ASC)、半胱氨酸蛋白酶－1(Caspase-1)、G蛋白偶联受体43(GPR43)、闭锁小带蛋白－1(ZO-1)、闭合蛋白(Occludin)蛋白表达，实时荧光PCR法检测结肠组织中NLRP3、ASC、Caspase-1、GPR43、ZO-1、Occludin mRNA表达。
结果
2
各组间血清、结肠炎性因子表达差异具有统计学意义，与模型组比较，降糖3号方组LPS、TNF-α、
IL-1β、IL-6表达降低(均
P
<
0.05)；降糖3号方可上调结肠中ZO-1、Occludin、GPR43的蛋白及mRNA表达，降低NLRP3、ASC、Caspase-1的蛋白及mRNA表达(均
P
<
0.05)，同时改善结肠组织病理损伤。
结论
2
T2DM小鼠呈现慢性炎症反应状态，降糖3号方能够通过抑制NLRP3炎症小体，缓解T2DM小鼠机体与肠道炎症反应，修复肠黏膜屏障，这可能是其抗糖尿病的分子机制之一。
Abstract
Objective
2
This study aimed to observe the effect of
Jiang Tang San Hao
Formula (JTSHF) on systemic and intestinal inflammation
as well as on the NLRP3 inflammasome in type 2 diabetic mice (T2DM)
and to elucidate its anti-diabetic molecular mechanisms.
Methods
2
Four-week-old male C57BL/6 N mice were used to establish the T2DM model using a high-fat diet combined with streptozotocin injection. The diabetic mice were randomly divided into the model
metformin
and JTSHF groups. A control group was also set to provide baseline comparisons. Each group of mice was orally administered with the corresponding medication daily. The metformin group was orally administered with 0.20 g/kg metformin
the JTSHF group was orally administered with 4.26 g/kg JTSHF
and the control group and model group were orally administered with an equal amount of sterile water continuously for 8 weeks.After an 8-week drug intervention via gavage
the lipopolysaccharide (LPS)
tumor necrosis factor-alpha (TNF-α)
interleukin 1 beta (IL-1β)
and interleukin 6 (IL-6) serum and colon levels were quantified using an enzyme-linked immunosorbent assay (ELISA). The pathological morphology of the colon was observed using hematoxylin and eosin staining. NOD-like receptor protein 3 (NLRP3)
apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC)
caspase-1
zonula occludens-1 (ZO-1)
occludin
and G-protein coupled receptor 43 (GPR43) protein expression in the colon were assessed using immunohistochemistry. The mRNA expression levels of NLRP3
ASC
caspase-1
ZO-1
Occludin
and GPR43 in the colon were detected using Real-time PCR.
Results
2
The ELISA data revealed significant differences in inflammatory markers among the groups. Compared with the model group
the JTSHF group exhibited notably reduced LPS
TNF-α
IL-1β
and IL-6 levels (
P
<
0.05). Moreover
compared with the model group
JTSHF treatment upregulated ZO-1
occludin
and GPR43 protein and mRNA expression in the colon and downregulated NLRP3
ASC
and Caspase-1 protein and mRNA expression (
P
<
0.05).
Conclusion
2
The inflammatory reaction of T2DM mice is apparent. JTSHF effectively alleviates the systemic and intestinal inflammatory response of T2DM mice by inhibiting the NLRP3 inflammasome and repairing the intestinal mucosal barrier
highlighting the potential molecular mechanisms of the anti-diabetes effects of JTSHF.
关键词
Keywords
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