威灵仙总皂苷调控miR-26a-5p/Wnt5a信号通路对三叉神经痛大鼠的镇痛作用

doi:10.11958/20251039

天津医药 ›› 2025, Vol. 53 ›› Issue (8): 802-807.doi: 10.11958/20251039

威灵仙总皂苷调控miR-26a-5p/Wnt5a信号通路对三叉神经痛大鼠的镇痛作用

郭青玉¹(), 储成志²^,^△()

1 皖南医学院研究生学院（邮编241002）
2 安徽中医药高等专科学校中医系

收稿日期:2025-03-13 修回日期:2025-05-26 出版日期:2025-08-15 发布日期:2025-08-12
通讯作者: ^△E-mail：346686012@qq.com
作者简介:郭青玉（2000），女，硕士在读，主要从事中西医结合基础方面研究。E-mail：guoqingyu0312@163.com
基金资助:
安徽省高等学校自然科学研究项目(KJ2021A1282)

Analgesic effect of total saponins of Clematis on trigeminal neuralgia rats by regulating the miR-26a-5p/Wnt5a signaling pathway

GUO Qingyu¹(), CHU Chengzhi²^,^△()

1 Graduate School of Wannan Medical College, Wuhu 241002, China
2 Department of Traditional Chinese Medicine, Anhui College of Traditional Chinese Medicine

Received:2025-03-13 Revised:2025-05-26 Published:2025-08-15 Online:2025-08-12
Contact: ^△E-mail：346686012@qq.com

郭青玉, 储成志. 威灵仙总皂苷调控miR-26a-5p/Wnt5a信号通路对三叉神经痛大鼠的镇痛作用[J]. 天津医药, 2025, 53(8): 802-807.

GUO Qingyu, CHU Chengzhi. Analgesic effect of total saponins of Clematis on trigeminal neuralgia rats by regulating the miR-26a-5p/Wnt5a signaling pathway[J]. Tianjin Medical Journal, 2025, 53(8): 802-807.

摘要/Abstract

摘要：

目的探讨威灵仙总皂苷（TSC）调控微小RNA（miR）-26a-5p/Wnt5a信号通路对三叉神经痛（TN）大鼠的镇痛作用。方法通过眶下神经缩窄术建立TN大鼠模型，并将50只大鼠分为TN组、不同剂量（50、200 mg/kg）TSC组、TSC+NC antagomir（200 mg/kg TSC+5 nmoL NC antagomir）组、TSC+miR-26a-5p antagomir（200 mg/kg TSC+5 nmoL miR-26a-5p antagomir）组，每组10只，另取10只作为假手术组，仅暴露大鼠眶下神经但不结扎。酶联免疫吸附试验（ELISA）检测肿瘤坏死因子（TNF）-α、白细胞介素（IL）-6水平；HE染色检测组织病理学变化；实时荧光定量逆转录聚合酶链反应检测miR-26a-5p、Wnt5a mRNA表达；蛋白质印迹检测三叉神经节组织胶质源性神经营养因子（GDNF）、Wnt5a、脑源性神经营养因子（BDNF）蛋白表达；双萤光素酶报告基因系统验证miR-26a-5p与Wnt5a的靶向关系。结果与假手术组相比，TN组TNF-α、IL-6、Wnt5a蛋白及mRNA表达增加，机械痛阈值、miR-26a-5p、GDNF、BDNF表达降低（P<0.05）；与TN组相比，50、200 mg/kg TSC组TNF-α、IL-6、Wnt5a蛋白及mRNA表达显著降低，机械痛阈值、miR-26a-5p、GDNF、BDNF表达显著增加，不同剂量TSC组间有差异（P<0.05）；抑制miR-26a-5p逆转了TSC对TN大鼠的保护作用（P<0.05）。结论 TSC通过调控miR-26a-5p/Wnt5a信号通路对TN大鼠发挥镇痛作用。

关键词: 三叉神经痛, 镇痛, 威灵仙总皂苷, miR-26a-5p/Wnt5a信号通路

Abstract:

Objective To investigate the analgesic effect of total saponins of Clematis (TSC) on trigeminal neuralgia (TN) rats by regulating the miR-26a-5p/Wnt5a signaling pathway. Methods The TN rat model was established through suborbital nerve constriction surgery. Fifty rats were divided into the TN group, the different doses of (50, 200 mg/kg) TSC groups, the TSC+NC antagonist (200 mg/kg TSC+5 nmoL NC antagonist) group and the TSC+miR-26a-5p antagonist (200 mg/kg TSC+5 nmoL miR-26a-5p antagonist) group2025， with 10 mice per group. Another 10 rats were taken as the sham operation group, and only the infraorbital nerve of rats was exposed but not ligated. ELISA was used to detect the levels of tumor necrosis factor (TNF) -α and interleukin (IL) -6. HE staining was used to detect histopathological changes. Real-time fluorescence quantitative reverse transcription polymerase chain reaction was used to detect the expression levels of miR-26a-5p and Wnt5a mRNA. Western blot assay was used to detect protein expressions of glial neurotrophic factor (GDNF), Wnt5a and brain-derived neurotrophic factor (BDNF) in trigeminal ganglion tissue. Dual luciferase reporter gene system was applied to validate the targeting relationship between miR-26a-5p and Wnt5a. Results Compared with the sham operation group, the expression levels of TNF-α, IL-6, Wnt5a protein and mRNA were obviously increased in the TN group, and the mechanical pain threshold, the expression levels of miR-26a-5p, GDNF and BDNF were obviously reduced (P<0.05). Compared with the TN group, the expression of TNF-α, IL-6, Wnt5a protein and mRNA were greatly reduced in the 50 and 200 mg/kg TSC groups, and the mechanical pain threshold, the expression levels of miR-26a-5p, GDNF and BDNF were greatly increased. There were differences in different doses of TSC between the different groups (P<0.05). Inhibition of miR-26a-5p reversed the protective effect of TSC on TN rats. Conclusion TSC exerts analgesic effects on TN rats by regulating the miR-26a-5p/Wnt5a signaling pathway.

Key words: trigeminal neuralgia, analgesia, total saponins of Clematis, miR-26a-5p/Wnt5a signaling pathway

中图分类号:

R745.11

图/表 8

图1 各组大鼠机械痛阈值的比较 A—F分别为假手术组、TN组、不同剂量（50、200 mg/kg）TSC组、TSC+NC antagomir组、TSC+miR-26a-5p antagomir组；a与假手术组比较，b与TN组比较，c与50 mg/kg TSC组比较，d与TSC+NC antagomir组比较，P<0.05。

Fig.1 Comparison of mechanical pain thresholds between six groups of rats

表1 各组大鼠血清TNF-α、IL-6水平比较

Tab.1 Comparison of TNF-α and IL-6 levels between six groups of rats （n=10，ng/L，$\bar{x}±s$）

组别	TNF-α	IL-6
假手术组	86.34±8.82	53.82±5.42
TN组	183.26±18.44^a	105.24±10.82^a
50 mg/kg TSC组	138.09±13.97^b	78.24±7.99^b
200 mg/kg TSC组	92.05±9.31^bc	58.64±5.93^bc
TSC+NC antagomir组	91.52±9.22	59.34±6.08
TSC+miR-26a-5p antagomir组	136.52±13.75^d	77.82±7.88^d
F	90.175^＊＊	64.326^＊＊

图2 各组大鼠三叉神经节组织病理学变化（HE染色，×100）

Fig.2 Histopathological changes of the trigeminal ganglion in each group (HE staining, ×100)

表2 各组大鼠miR-26a-5p、Wnt5a mRNA表达水平比较

Tab.2 Comparison of miR-26a-5p and Wnt5a mRNA expressions between six groups of rats （n=10，$\bar{x}±s$）

组别	Wnt5a mRNA	miR-26a-5p
假手术组	0.98±0.10	1.02±0.11
TN组	2.01±0.21^a	0.25±0.03^a
50 mg/kg TSC组	1.51±0.16^b	0.54±0.06^b
200 mg/kg TSC组	1.15±0.12^bc	0.85±0.09^bc
TSC+NC antagomir组	1.09±0.11	0.88±0.09
TSC+miR-26a-5p antagomir组	1.49±0.16^d	0.61±0.07^d
F	65.780^＊＊	125.119^＊＊

图3 各组大鼠组织中GDNF、BDNF、Wnt5a蛋白表达 A：假手术组；B：TN组；C：50 mg/kg TSC组；D：200 mg/kg TSC组；E：TSC+NC antagomir组；F：TSC+miR-26a-5p antagomir组。

Fig.3 Expression levels of GDNF, BDNF and Wnt5a proteins in tissue of rats in each group

表3 各组大鼠组织中GDNF、BDNF、Wnt5a表达水平比较

Tab.3 Comparison of GDNF, BDNF and Wnt5a expression levels between rat tissue of six groups （n=10，$\bar{x}±s$）

组别	GDNF/β-actin	BDNF/β-actin	Wnt5a/β-actin
假手术组	0.86±0.09	1.02±0.11	0.42±0.05
TN组	0.22±0.03^a	0.35±0.04^a	1.38±0.15^a
50 mg/kg TSC组	0.58±0.06^b	0.65±0.07^b	0.95±0.10^b
200 mg/kg TSC组	0.78±0.08^bc	0.83±0.09^bc	0.72±0.08^bc
TSC+NC antagomir组	0.76±0.08	0.91±0.10	0.67±0.07
TSC+miR-26a-5p antagomir组	0.49±0.05^d	0.68±0.07^d	1.03±0.11^d
F	120.925^＊＊	80.538^＊＊	114.387^＊＊

图4 miR-26a-5p与Wnt5a预测位点

Fig.4 Predictive sites of miR-26a-5p and Wnt5a

表4 miR-26a-5p靶向调节Wnt5a表达

Tab.4 miR-26a-5p targeted regulation of Wnt5a expression （n=6，$\bar{x}±s$）

组别	萤光素酶活性
mimics NC+Wnt5a WT组	1.01±0.11
miR-26a-5p mimics+Wnt5a WT组	0.54±0.06
t	9.188^＊＊
mimics NC+Wnt5a MUT组	1.05±0.11
miR-26a-5p mimics+Wnt5a MUT组	1.02±0.11
t	0.472

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威灵仙总皂苷调控miR-26a-5p/Wnt5a信号通路对三叉神经痛大鼠的镇痛作用

Analgesic effect of total saponins of Clematis on trigeminal neuralgia rats by regulating the miR-26a-5p/Wnt5a signaling pathway

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