三七总皂苷抑制小鼠星形胶质细胞活化缓解完全弗氏佐剂所致炎性痛

doi:10.16098/j.issn.0529-1356.2024.01.004

解剖学报 ›› 2024, Vol. 55 ›› Issue (1): 25-31.doi: 10.16098/j.issn.0529-1356.2024.01.004

三七总皂苷抑制小鼠星形胶质细胞活化缓解完全弗氏佐剂所致炎性痛

袁蕾¹ 杨智伟¹ 杨惠² 刘政海³ 何洁⁴ 万炜^1*

1.海南省热带脑科学研究与转化重点实验室，海口 571199; 2.赣州市人民医院病理科，江西赣州 341000; 3.南华大学衡阳医学院应用解剖学与生殖医学研究所，湖南衡阳 421001; 4.海南医学院病理学教研室，海口 571199

收稿日期:2023-07-06 修回日期:2023-09-01 出版日期:2024-02-06 发布日期:2024-02-06
通讯作者: 万炜 E-mail:david-wan@163.com
基金资助:
热量限制在辐射小鼠认知障碍中的机制研究;VDAC1通过调节神经发生在电离辐射致小鼠认知障碍中的作用

Panax notoginseng saponin relieving the inflammatory pain caused by complete Freund’s adjuvant by inhibiting the activation of astrocytes in mice

YUAN Lei¹ YANG Zhi-wei¹ YANG Hui² LIU Zheng-hai³ HE Jie⁴ WAN Wei^1*

1.Key Laboratory of Brain Science Research & Transformation in Tropical Environment of Hainan Province, Haikou 571199,China；
2.Department of Pathology, Ganzhou People’s Hospital, Jiangxi Ganzhou 341000,China； 3.Clinical Anatomy and Reproductive Medicine Application Institute，School of Medicine，University of South China，Hu’nan Hengyang 421001,China； 4.Department of Pathology，Hainan Medical University, Haikou 571199,China

Received:2023-07-06 Revised:2023-09-01 Online:2024-02-06 Published:2024-02-06
Contact: WAN Wei E-mail:david-wan@163.com

摘要/Abstract

摘要：

目的探讨三七总皂苷（PNS）对完全弗氏佐剂（CFA）所致慢性炎性疼痛小鼠模型的镇痛作用及可能机制。方法 48只C57BL/6J雄性小鼠随机分成生理盐水对照组（Ctrl）、CFA组（CFA）、CFA+ PNS组、CFA +地塞米松（DEX）组（CFA+DEX）。采用Von Frey纤维丝检测小鼠机械疼痛；采用免疫组织化学法检测脊髓后角胶质纤维酸性蛋白（GFAP）阳性星形胶质细胞数目和形态结构变化；采用Western blotting 检测各组小鼠相应节段脊髓GFAP、核苷酸结合寡聚化结构域（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）、Caspase-1、白细胞介素（IL）-1β和IL-18的表达。结果与Ctrl组相比，CFA组小鼠机械痛阈值在第1、3、5、7、14天明显下降，小鼠脊髓NLRP3、ASC、Caspase-1、IL-1β和IL-18的表达均显著增加。PNS干预可缓解模型小鼠机械痛觉，下调小鼠脊髓NLRP3、ASC、Caspase-1、IL-1β和IL-18的表达，且与CFA+DEX组表达差异无显著性。免疫组织化学结果显示，与Ctrl组相比，CFA组小鼠脊髓后角GFAP阳性细胞数目显著增多；PNS干预后CFA+ PNS组小鼠脊髓后角 GFAP阳性细胞数目减少；DEX干预后对小鼠脊髓后角GFAP阳性细胞数目没有明显影响。Western blotting 结果显示，与Ctrl组相比，CFA组小鼠脊髓GFAP表达明显增加；PNS干预后CFA+ PNS组小鼠脊髓GFAP表达明显减少，DEX干预后对小鼠脊髓后角GFAP表达没有明显影响。结论 PNS对炎性痛具有较好的缓解作用，其机制可能与抑制星形胶质细胞活化以及减少NLRP3炎性小体激活有关。

关键词: 三七总皂苷, 星形胶质细胞, 炎性痛, 核苷酸结合寡聚化结构域样受体热蛋白结构域相关蛋白3, 免疫组织化学, 免疫印迹法, 小鼠

Abstract:

Objective To analyse the analgesic effect and possible mechanism of panax notoginseng saponin (PNS) on mouse models of chronic inflammatory pain caused by complete Freund’s adjuvant (CFA). Methods A total of 48 male C57BL/6J mice were divided randomly into four groups: normal saline control group (Ctrl), CFA group (CFA), CFA + PNS group (CFA+PNS), CFA + dexamethasone (DEX) group (CFA+DEX). Von Frey filaments were used to detect mechanical pain in mice. Immunohistochemistry was used to detect the number and morphological changes of glial fibrillary acidic protein (GFAP) positive astrocytes. Western blotting was used to detect the expressions of GFAP, nucleotide-binding and oligomerization domain(NOD)-like receptor thermal protein domain associated protein 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), Caspase-1, interleukin (IL)-1β, and IL-18 in mice’s spinal cord segments in each group. Results Compared with the Ctrl group, mice in the CFA group showed a significant decrease in mechanical pain thresholds at day 1, day 3, day 5, day 7, and day 14. Additionally, there was a significant decrease in NLRP3, ASC, Caspase-1, IL-1β and IL-18 in the spinal cord of the mice. PNS intervention could relieve mechanical pain and down-regulate the expressions of NLRP3, ASC, Caspase-1, IL-1β and IL-18 in the spinal cord of mice, with no significant difference compared with the CFA+DEX group. CFA group mice had significantly more GFAP positive cells in their posterior horns than Ctrl group mice, as measured by immunohistochemistry; PNS intervention decreased the number of GFAP positive cells in the posterior horn of the spinal cord in model mice；DEX had no effect on the number of GFAP positive cells in the dorsal horn of spinal cord. According to Western blotting results, GFAP expression in the spinal cord of the CFA group was significantly more than that of the Ctrl group; PNS intervention significantly reduced GFAP expression in the spinal cord of CFA group mice；DEX had no effect on the expression of GFAP in the posterior horn of spinal cord. Conclusion PNS has a good alleviating effect on inflammatory pain, and its mechanism may be related to inhibition of astrocyte activation and NLRP3 inflammasome activation.

Key words:

Panax notoginseng saponins, Astrocyte, Inflammatory pain, Nucleotide-binding and oligomerization domain-like receptor thermal protein domain associated protein 3, Immunohistochemistry, Western blotting, Mouse

中图分类号:

R322.81

袁蕾杨智伟杨惠刘政海何洁万炜.

三七总皂苷抑制小鼠星形胶质细胞活化缓解完全弗氏佐剂所致炎性痛 [J]. 解剖学报, 2024, 55(1): 25-31.

YUAN Lei YANG Zhi-wei YANG Hui LIU Zheng-hai HE Jie WAN Wei.

Panax notoginseng saponin relieving the inflammatory pain caused by complete Freund’s adjuvant by inhibiting the activation of astrocytes in mice [J]. Acta Anatomica Sinica, 2024, 55(1): 25-31.

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三七总皂苷抑制小鼠星形胶质细胞活化缓解完全弗氏佐剂所致炎性痛

Panax notoginseng saponin relieving the inflammatory pain caused by complete Freund’s adjuvant by inhibiting the activation of astrocytes in mice

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