| 摘要: |
| [目的] 基于NOD样受体（NLR）家族含有吡林结构域3（NLRP3）炎症小体介导巨噬细胞极化探究金匮温经汤（JGWJD）对寒冷刺激所致的大鼠子宫炎症和血管损伤的影响并阐明其分子机制。[方法] 按照随机数字表法将大鼠分为对照组（Control组）、模型组（Model组）、JGWJD高剂量组（JGWJD-H组）、JGWJD中剂量组（JGWJD-M）、JGWJD低剂量组（JGWJD-L组），每组12只。通过冰水浴＋冰箱冷冻法制备模型，采用不同浓度[16.4、8.2、4.1 g/（kg·d）]的JGWJD进行干预，通过阴道涂片检测各组大鼠动情阶段变化情况；激光散斑血流成像系统采集各组大鼠脚部与子宫表面血液灌注量；B超采集技术检测各组大鼠子宫组织结构；酶联免疫吸附法（ELISA）检测血清白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）、白细胞介素-10（IL-10）、雌二醇（E2）、孕激素（P）、促黄体生成素（LH）、促卵泡生成素（FSH）水平；苏木精-伊红（HE）染色、透射电镜观察子宫形态学变化；免疫荧光染色检测诱导型一氧化氮合酶（iNOS）、精氨酸酶-1（Arg-1）定位及强度；蛋白免疫印迹法（Western blot）检测子宫组织NLRP3、半胱氨酸天冬氨酸蛋白酶-1（caspase-1）、IL-1β、iNOS、白细胞介素-6（IL-6）、TNF-α、Arg-1、转化生长因子-β（TGF-β）、IL-10蛋白表达。[结果] 虚寒证模型大鼠精神萎靡、动情周期紊乱，脚部及子宫表面血液灌注量明显降低；子宫可见不规则低回声区，子宫体肿胀变形；内膜变薄，腺体减少，炎性细胞浸润，微绒毛稀疏，线粒体肿胀，内质网扩张水肿；IL-1β、TNF-α含量升高同时IL-10与生殖激素水平显著下降；上调NLRP3、caspase-1、IL-1β、iNOS、IL-6、TNF-α表达，抑制Arg-1、TGF-β、IL-10表达。与模型组比较，JGWJD能够恢复大鼠一般状态；升高腹温及脚部与子宫表面血液灌注量；子宫形态明显改善，内膜厚度增加，炎症减轻，局部损伤得以修复；IL-1β、TNF-α含量降低，IL-10与生殖激素水平显著升高；抑制炎症小体及相关促炎因子表达并促进抗炎修复因子释放。[结论] JGWJD能够通过拮抗炎症小体的启动与激活，降低其下游效应蛋白与炎症因子的生成与释放，同时调控与巨噬细胞极化相关的蛋白表达，抑制M1型表型相关因子，促进M2型表型相关因子表达，进而减少下游促炎因子表达，同时增加抗炎因子生成，从而恢复M1/M2免疫稳态缓解子宫炎症反应，修复血管损伤。 |
| 关键词: 吡林结构域3炎症小体 巨噬细胞极化 金匮温经汤 寒冷刺激 子宫损伤 |
| DOI：10.11656/j.issn.1673-9043.2026.09.09 |
| 分类号:R711.74 |
| 基金项目:国家自然科学基金项目（82174212）；国家自然科学基金青年基金项目（81503451）。 |
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| Mechanism of Jingui Wenjing Decoction in improving uterine injury in rats with deficiency-cold syndrome based on NLRP3 inflammasome-mediated macrophage polarization |
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ER Jiali, ZHANG Jiayu, BING Xiangnan, ZHAO Shu, DU Chenguang, QI Feng, ZHANG Biwei, XU Dingjie
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Traditional Chinese Medicial College, North China University of Science and Technology, Tangshan 063210, China
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| Abstract: |
| [Objective] To investigate the effects of Jingui Wenjing Decoction（JGWJD）on cold stress-induced uterine inflammation and vascular injury in rats，and to elucidate its molecular mechanism based on NOD-like receptor family pyrin domain-containing 3（NLRP3）inflammasome-mediated macrophage polarization. [Methods] A rat model of uterine inflammatory injury was established by combined ice-water immersion and freezing exposure. Rats were randomly divided into control group，model group，JGWJD-H group，JGWJD-M group，and JGWJD-L group，with 12 rats in each group. Rats were treated with JGWJD at doses of 16.4，8.2，and 4.1 g/（kg·d），respectively. Vaginal smears were performed to evaluate estrous cycle stages. Laser speckle contrast imaging was used to assess blood perfusion in the plantar skin and uterine surface. Transabdominal ultrasonography was employed to detect uterine structural changes. Serum levels of IL-1β，TNF-α，IL-10，E2，P，LH，and FSH were determined by ELISA. Uterine morphological alterations were observed using HE staining and TEM. Immunofluorescence staining was performed to detect the localization and expression intensity of iNOS and Arg-1. Protein expression levels of NLRP3，caspase 1，IL-1β，iNOS，IL-6，TNF-α，Arg-1，TGF-β，and IL-10 in uterine tissues were measured by Western blot. [Results] In the model group，rats exhibited mental depression，disrupted estrous cycles，and significantly reduced blood perfusion in the plantar skin and uterine surface. Ultrasonography revealed irregular hypoechoic areas，with uterine swelling and deformation. Histological and ultrastructural examination showed endometrial thinning，reduced glandular counts，inflammatory cell infiltration，sparse microvilli，mitochondrial swelling，and dilated endoplasmic reticulum. Serum levels of IL-1β and TNF-α were markedly elevated，whereas IL-10 and reproductive hormone levels were significantly decreased. Western blot analysis demonstrated upregulated expression of NLRP3，caspase-1，IL-1β，iNOS，IL-6，and TNF-α，along with downregulated expression of Arg-1，TGF-β，and IL-10 in uterine tissues. Compared with the model group，JGWJD treatment dose-dependently restored general behavioral status，increased abdominal temperature as well as plantar and uterine blood perfusion，and ameliorated uterine morphological damage，as evidenced by increased endometrial thickness，attenuated inflammation，and repaired local injury. Moreover，JGWJD significantly reduced serum IL-1β and TNF-α levels，elevated IL-10 and reproductive hormone levels，suppressed NLRP3 inflammasome activation and downstream pro-inflammatory cytokine production，while promoting the release of anti-inflammatory and tissue-repair factors. [Conclusion] JGWJD attenuates cold stress-induced uterine inflammation and vascular injury by inhibiting NLRP3 inflammasome priming and activation，thereby reducing the generation and release of downstream effector proteins and pro-inflammatory cytokines. Concurrently，it modulates macrophage polarization-related protein expression by suppressing M1 phenotype-associated factors and enhancing M2 phenotype associated factors，which restores the M1/M2 immune homeostasis，alleviates uterine inflammatory responses，and repairs vascular damage. |
| Key words: NLRP3 inflammasome macrophage polarization Jingui Wenjing Decoction cold stress uterine injury |
