加味人参乌梅汤通过TLR4/MyD88/pNF⁃κBp65信号通路对腹泻大鼠肠黏膜屏障的影响

doi:10.3969/j.issn.1006-5725.2025.07.003

Abstract

Abstract:

Objective To investigate the effect of Modified Renshen Wumei Decoction on the TLR4/MyD88/pNF?κBp65 signaling pathway and elucidate the potential mechanism by which this formula repairs the intestinal mucosal barrier in diarrheal rats. Methods Twelve rats were randomly selected from a total of 48 rats to serve as the blank control group （CK）， while the remaining 36 rats were used to establish a disease model via a compound method. After 14 days of model preparation， the rats were randomly divided into three groups： the model group （MC）， the western medicine group （MV）， and the traditional Chinese medicine group （MRWD）. Each of the four groups （including CK） received corresponding interventions for 7 days. The concentrations of serum diamine oxidase （DAO）， D?lactic acid （D?Lac）， interleukin?1β （IL?1β）， IL?6， IL?10， tumor necrosis factor?α （TNF?α）， mucin 2 （MUC2）， MUC4， MUC6， and colonic homogenate secretory immunoglobulin A （SIgA） were measured using ELISA. Additionally， the protein and gene expressions of colonic toll?like receptor 4 （TLR4）， myeloid differentiation primary response 88 （MyD88）， phosphorylated nuclear factor kappa?light?chain?enhancer of activated B cells p65 （pNF?κBp65）， occludin， claudin?1， and zonula occludens?1 （ZO?1） were analyzed by Western blot and RT?PCR. Results （1） Intestinal mucosal injury markers： Compared with the blank group， the serum levels of DAO and D?Lac in the model group were significantly increased （P < 0.05）. Compared with the model group， both the Chinese medicine group and Western medicine group significantly decreased the serum levels of DAO （P < 0.001）， while the traditional Chinese medicine group also significantly reduced the serum levels of D?Lac （P < 0.05）. There was no significant difference in the changes of DAO and D?Lac serum levels between the Chinese medicine group and Western medicine group （P > 0.05）. （2） Inflammatory indicators： Compared with the blank group， the model group exhibited significant upregulation of TLR4， MyD88， pNF?κBp65 protein and gene expression， as well as serum levels of IL?1β， IL?6， and TNF?α （P < 0.05）， along with a significant decrease in IL?10 serum levels （P < 0.05）. Compared with the model group， both the Chinese medicine group and Western medicine group significantly downregulated TLR4， MyD88， pNF?κBp65 protein and gene expression， as well as serum levels of IL?1β， IL?6， and TNF?α （P < 0.05）， and significantly upregulated IL?10 serum levels （P < 0.05）. There was no significant difference in serum levels of TLR4， MyD88， pNF?κBp65 protein， gene expression， and IL?1β， IL?6， IL?10， and TNF?α between the Chinese medicine group and Western medicine group （P > 0.05）. （3） Intestinal mucosal barrier factors： Compared with the blank group， the model group exhibited significant downregulation in MUC2， MUC6， SIgA content， as well as Claudin?1， ZO?1 protein and gene expression， and Occludin protein expression （P < 0.05）. Compared with the model group， both Chinese and Western medicine groups significantly upregulated the content of MUC2 and SIgA， as well as the protein and gene expression of Claudin?1 and ZO?1 （P < 0.05）. The traditional Chinese medicine group also significantly increased the content of MUC6 and Occludin protein expression （P < 0.05）. No significant differences were observed between the Chinese and Western medicine groups in terms of MUC2， MUC6， SIgA serum content， and Claudin?1 and ZO?1 gene expression （P > 0.05）. However， the Western medicine group showed better Claudin?1 protein expression than the Chinese medicine group （P < 0.05）， while the ZO?1 protein expression was higher in the traditional Chinese medicine group compared to the Western medicine group （P < 0.05）. Conclusion Modified Renshen Wumei Decoction exerts an intestinal mucosal barrier repair effect in diarrhea rats by modulating the TLR4/MyD88/pNF?κBp65 signaling pathway.

Key words: modified Renshen Wumei decoction, diarrhea, TLR4, MyD88, NF-κB, intestinal mucosal barrier

CLC Number:

R285.5

Zhiwei GUAN,Qiong ZHAO,Jianli QIU,Yan XU,Qinwan HUANG,Hongyun ZHOU,Junqi ZHAO,Yinghui WU. Exploring the effect of modified Renshen Wumei Decoction on intestinal mucosal barrier in diarrhea rats based on TLR4/MyD88/pNF⁃κBp65 signaling pathway[J]. The Journal of Practical Medicine, 2025, 41(7): 944-952.

Figures/Tables 11

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References 40

1	李潘孝，冯桂玲，许天兴，等. 温脾升阳安肠汤联合匹维溴铵对腹泻型肠易激综合征患者肠黏膜免疫屏障功能恢复的促进作用研究［J］. 四川中医，2022，40（10）：102-104.
2	中西医结合防治小儿腹泻专家共识［J］. 世界中医药，2022，17（21）：2979-2984+2991.
3	叶青，刘清. 枯草杆菌二联活菌颗粒治疗小儿腹泻病的临床疗效研究［J］. 中国继续医学教育，2016，8（22）：194-195.
4	刘彬媛. 酸甘化阴法治疗小儿气阴两虚型腹泻60例临床观察［D］. 成都：成都中医药大学，2012.
5	张肖瑾，刘彬媛，赵琼，等. 加味人参乌梅汤治疗小儿气阴两虚证泄泻临床观察［J］. 实用中医药杂志，2022，38（2）：171-173.
6	曹昌霞，张国铭，王宏斌，等. 基于肠道菌群和脑-肠轴探讨益气健脾饮联合益生菌对腹泻型肠易激综合征小鼠的影响［J］.中成药，2024，46（10）：3461-3467.
7	何志辉，陈雅璐，陈俊杰，等. 基于肠道黏膜屏障探讨中医药防治腹泻型肠易激综合征的机制［J/OL］. 辽宁中医药大学学报，1-13［2024-11-21］. .
8	杨艳玲，向黎莉，陈鹏飞，等. 加味葛根芩连汤对重度湿热型溃疡性结肠炎患者免疫功能及肠黏膜屏障功能的影响［J］. 辽宁中医杂志，2023，50（3）：92-95.
9	HOU Q， HUANG Y， ZHU Z，et al. Tong-Xie-Yao-Fang improves intestinalpermeability in diarrhoea-predominant irritable bowel syndrome rats by inhibiting the NF-κB and notch signalling pathways［J］. BMC Complement Altern Med，2019，19（1）：337. doi:10.1186/s12906-019-2749-4 doi: 10.1186/s12906-019-2749-4
10	马融. 中医儿科学［M］. 4版.北京：中国中医药出版社，2016：109.
11	韩新民，汪受传，虞舜，等. 小儿泄泻中医诊疗指南［J］. 中医儿科杂志，2008，4（3）：1-3.
12	李珊珊，赵琼，管志伟，等. 加味人参乌梅汤调控腹泻大鼠GABA能信号通路的系统效应［J］. 中国实验方剂学杂志，2022，28（9）：59-65.
13	乔翠霞. 溃愈散调控TLR4/MyD88/NF-κBp65信号通路治疗溃疡性结肠炎的机制研究［D］. 南京：南京中医药大学，2019.
14	高蕊，郑晖，岳灵，等. 用不同浓度的番泻叶水煎剂建立功能性腹泻大鼠模型的稳定性观察［J］. 当代医药论丛，2018，16（10）：201.
15	吴天石，张会永，张哲，等. 脾虚证动物模型造模方法述评［J］.中医杂志，2015，56（11）：978-983.
16	黄晶晶，陈炜，赵琼，等. 腹泻病气阴两虚证病证结合模型构建及以方测证研究［J］. 成都中医药大学学报，2016，39（1）：34-38.
17	管志伟，赵琼，赵中和，等. 番泻叶致幼龄大鼠腹泻模型量-时-效关系研究［J］. 时珍国医国药，2021，32（8）：1806-1809.
18	张声生，胡玲，李茹柳. 脾虚证中医诊疗专家共识意见（2017）［J］. 中医杂志，2017，58（17）：1525-1530.
19	任士庞，赵琼，赵兴，等. 加味人参乌梅汤及其拆方对腹泻大鼠结肠黏膜病理形态学的影响［J］. 山西中医，2015，31（12）：47-49.
20	黄玉珍. 参苓白术散对脾虚泄泻幼鼠模型肠黏膜屏障的影响［D］ .南京：南京中医药大学，2016.
21	LIN Y， YANG X， YUE W，et al. Chemerin aggravates DSS-induced colitis by suppressing M2 macrophage Polarization［J］.Cell Mol Immunol，2014，11（4）：355-366. doi:10.1038/cmi.2014.15 doi: 10.1038/cmi.2014.15
22	CHEN Q， XIAO Z， HE Q Y，et al. Effect of Shenling Baizhu powder on immunity to diarrheal disease：A systematic review and meta-analysis［J］. Front Pharmacol，2022，13：938932. doi:10.3389/fphar.2022.938932 doi: 10.3389/fphar.2022.938932
23	XU J， LIU C， SHI K，et al. Atractyloside-A ameliorates spleen deficiency diarrhea by interfering with TLR4/MyD88/NF-κB signaling activation and regulatingintestinal flora homeostasis［J］. Int Immunopharmacol，2022，107：108679. doi:10.1016/j.intimp.2022.108679 doi: 10.1016/j.intimp.2022.108679
24	朱卫，曾婷婷. 基于TLR4/MyD88/NF-κB信号通路研究参苓白术散对IBS-D大鼠的干预机制［J］. 湖南中医杂志，2023，39（12）：128-134.
25	庞占菊. 双红秦榆汤对溃疡性结肠炎大鼠IL-1β及NF-KB的影响［D］. 石家庄：河北医科大学，2017.
26	朱涛，冀来喜，任佳，等. 麦粒灸调控TLR4/MyD88/NF-κB信号通路减轻溃疡性结肠炎小鼠结肠损伤的机制研究［J］. 针刺研究，2024，49（8）：836-844.
27	周方，孙波，于志丹，等. 基于TLR4/NF-κB信号通路探讨益生菌对肝内胆汁淤积大鼠的保护作用［J］. 实用医学杂志，2022，38（3）：295-299.
28	张孟爽，杜艳. 基于NF-κB信号通路的益生菌治疗溃疡性结肠炎的研究进展［J］. 中国微生态学杂志， 2019，31（1）：104-107.
29	王倬，陈强，孔斌，等. 葛根芩连裁减方结合益生菌对溃疡性结肠炎血清IL-6、IL-12、IL-10和TGF-β水平的影响［J］. 中华中医药学刊，2022，40（3）：248-251.
30	窦鑫，贺昌辉，梅笑，等. 基于“短链脂肪酸-肠屏障”途径探讨中药在腹泻型肠易激综合征中的干预研究进展［J］. 实用医学杂志，2024，40（15）：2177-2182.
31	管志伟. 加味人参乌梅汤调控腹泻大鼠肠道菌群结构和功能的宏基因组学研究［D］. 成都：成都中医药大学，2021.
32	徐锋，朱磊，李亚楠，等. 基于肠道菌群及代谢组学探讨白芷多糖对溃疡性结肠炎小鼠的治疗及其作用机制［J/OL］. 中国中药杂志，1-13［2024-12-26］. .
33	吴东升，曹晖，张彧，等. 芍药汤对溃疡性结肠炎大鼠肠黏膜免疫屏障的干预作用［J］. 中国实验方剂学杂志， 2019， 25（9）： 6-11.
34	YANG R， HU X， XIE X， et al. Propionic Acid Targets the TLR4/NF-κB Signaling Pathway and Inhibits LPS-Induced Intestinal Barrier Dysfunction：In Vitro and In Vivo Studies［J］. Front Pharmacol，2020，11：573475. doi:10.3389/fphar.2020.573475 doi: 10.3389/fphar.2020.573475
35	龙健，智慧，涂星，等. 基于NF-κB通路探讨阔叶十大功劳对急性溃疡性结肠炎小鼠的作用［J］. 中成药，2024，46（6）：2026-2031.
36	成宁宁. 黄芩素提取工艺的优化及其对溃疡性结肠炎小鼠的作用机制研究［D］.长春：长春中医药大学，2023.
37	王佳佳，迟莉，王文娟，等. 参苓白术散对溃疡性结肠炎大鼠NLRP3、NF-κB、MUC2、TFF3基因表达的影响［J］. 世界中西医结合杂志，2019，14（12）：1638-1641.
38	江勇，朱大侠，刘礼剑. 白术多糖通过调控TLR4/NF-κB信号通路对重症急性胰腺炎大鼠肠黏膜免疫屏障的影响［J］. 中成药，2021，43（3）：624-629.
39	罗晓平. 基于黏膜屏障/肠道微生态探讨黄柏酮减轻小鼠溃疡性结肠炎的作用机制［D］. 上海：上海中医药大学，2020.
40	SEMIN I， NINNEMANN J， BONDAREVA M，et al. Interplay Between Microbiota，Toll-Like Receptors and Cytokines for the Maintenance of Epithelial Barrier Integrity［J］. Front Med （Lausanne）， 2021， 8： 644333. doi:10.3389/fmed.2021.644333 doi: 10.3389/fmed.2021.644333

引物名称	引物序列（5′-3′）	长度/bp	引物编号
TLR4	上游 CCCTGGTGTTGGATTTTACG 下游 GACCACCGATACACTGATGATAG	68	NM_019178.1
MyD88	上游ACAAAGGAACTGGGAGGCA 下游TGGGGCAGTAGCAGATGAA	88	NM_198130.1
pNF-κBp65	上游CCGTTACAAGTGCGAGGGC 下游 GTGCGTCTTAGTGGTATCTGTGCT	73	NM_199267.2
Occludin	上游AGGGGAAAGCAGGGAAGG 下游CATCGCAGGACTCGCCAC	81	NM_031329.2
Claudin-1	上游 CCCTATCAATGCCAGGTATGA 下游GCAGCCCAGCCAGTAAAGA	54	NM_031699.2
ZO-1	上游CACCCAGACCCTGACCCA 下游TGCTTGCGCTTCTATCCCT	136	NM_001106266.1
GAPDH	上游GGCAAGTTCAACGGCACA 下游TCTCGCTCCTGGAAGATGG	83	NM_017008.4

项目	空白组	模型组	西药组	中药组	F值	P值
DAO/IU/L	110.53 ± 8.33	137.52 ± 9.75^??	112.76 ± 5.93^##	110.98 ± 16.22^##	6.894	0.001
D-Lac/μg/L	759.98 ± 51.46	913.96 ± 80.61^?	861.28 ± 107.70	799.49 ± 106.04^#	3.171	0.031

项目	空白组	模型组	西药组	中药组	F值	P值
TLR4	1.00 ± 0.05	1.48 ± 0.10^??	0.95 ± 0.12^##	0.96 ± 0.10^##	22.329	0.000
MyD88	1.00 ± 0.11	1.27 ± 0.08^?	0.64 ± 0.08^##	0.41 ± 0.05^##	62.125	0.000
pNF-κBp65	1.00 ± 0.13	1.35 ± 0.15^?	0.91 ± 0.03^#	0.95 ± 0.03^#	12.328	0.002

项目	空白组	模型组	西药组	中药组	F值	P值
TLR4	1.00 ± 0.09	1.52 ± 0.24^??	0.79 ± 0.16^##	0.71 ± 0.14^##	28.483	0.000
MyD88	1.01 ± 0.16	1.35 ± 0.19^?	1.04 ± 0.14^#	1.08 ± 0.17^#	5.343	0.007
pNF-κBp65	1.00 ± 0.13	1.36 ± 0.22^?	0.93 ± 0.16^##	0.86 ± 0.14^##	10.744	0.000

项目	空白组	模型组	西药组	中药组	F值	P值
IL-1β/（ng/L）	157.88 ± 14.58	203.94 ± 12.78^??	180.40 ± 10.41^#	167.06 ± 7.47^##	17.790	0.000
IL-6/（pg/mL）	9.18 ± 0.83	12.61 ± 0.45^??	10.58 ± 0.62^##	10.39 ± 1.25^##	17.065	0.000
IL-10/（pg/mL）	14.21 ± 1.69	10.90 ± 0.78^??	12.75 ± 0.92^#	13.37 ± 0.91^#	9.210	0.000
TNF-α/（ng/L）	265.87 ± 23.92	365.83 ± 41.18^??	286.13 ± 48.01^#	299.02 ± 43.18^#	6.986	0.002

Exploring the effect of modified Renshen Wumei Decoction on intestinal mucosal barrier in diarrhea rats based on TLR4/MyD88/pNF⁃κBp65 signaling pathway

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项目	空白组	模型组	西药组	中药组	F值	P值
MUC2/（ng/mL）	69.34 ± 7.99	39.73 ± 8.11^??	67.16 ± 12.82^##	65.08 ± 14.00^#	9.382	0.000
MUC4/（ng/mL）	44.29 ± 4.53	41.03 ± 5.19	40.39 ± 8.59	39.35 ± 8.19	0.580	0.635
MUC6/（ng/mL）	57.37 ± 10.90	44.51 ± 6.47^?	49.21 ± 13.28	64.02 ± 12.42^#	3.656	0.030
SIgA/（μg/mL）	7.70 ± 0.36	6.63 ± 0.45^??	7.53 ± 0.49^#	7.38 ± 0.25^#	8.578	0.001

项目	空白组	模型组	西药组	中药组	F值
Occludin	1.00 ± 0.01	0.69 ± 0.04^??	0.60 ± 0.03	0.74 ± 0.02^#	82.455
Claudin-1	1.00 ± 0.07	0.47 ± 0.11^??	1.07 ± 0.08^##	0.91 ± 0.02^##▲	40.732
ZO-1	1.00 ± 0.04	0.63 ± 0.06^??	1.11 ± 0.02^##	1.27 ± 0.09^##▲	66.861

项目	空白组	模型组	西药组	中药组	F值	P值
Occludin	1.04 ± 0.11	0.98 ± 0.13	0.98 ± 0.18	1.02 ± 0.20	0.216	0.884
Claudin-1	1.01 ± 0.12	0.59 ± 0.12^??	1.21 ± 0.25^##	1.06 ± 0.19^##	12.944	0.000
ZO-1	1.01 ± 0.16	0.70 ± 0.13^?	0.92 ± 0.12^#	1.03 ± 0.21^#	5.250	0.008

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[15]	Xue AO,Xing SU,Yu HOU,Tianyi MA,Chao. DENG. P38MAPK/NF⁃κB⁃based study of the molecular mechanism of miR⁃146a intervention in septic cardiomyopathy [J]. The Journal of Practical Medicine, 2023, 39(24): 3188-3194.