Impact and mechanism of NOD-like receptor thermal protein domain-associated protein 3 on pathological features and inflammatory response in a rat model of hemorrhoids
-
摘要:目的
探讨NOD样受体热蛋白结构域相关蛋白3(NLRP3)对痔疮模型大鼠创面病理特征、炎症反应及环磷酸腺苷(cAMP)/瞬时受体电位香草酸亚型1(TRPV1)信号通路的影响。
方法将40只6周龄健康雄性SD大鼠随机分为对照组、模型组、NLRP3激动剂组和NLRP3抑制剂组，每组10只。对照组不进行干预，其余3组均构建急性痔疮模型。NLRP3激动剂组、NLRP3抑制剂组在造模成功后分别给予创面局部皮下注射NLRP3激动剂、NLRP3抑制剂，模型组、对照组则给予等量生理盐水皮下注射。观察各组大鼠创面病理特征并评估创面积分，采用酶联免疫吸附试验(ELISA)检测血清肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6水平，采用蛋白质印迹法(Western blot)检测创面组织中NLRP3、cAMP和TRPV1蛋白表达量，采用实时荧光定量聚合酶链反应(qRT-PCR)检测创面组织中NLRP3、cAMP、TRPV1基因mRNA表达量。
结果与对照组相比，模型组创面组织炎性细胞浸润增多，水肿明显，组织结构紊乱，结缔组织增生; 与模型组相比, LRP3激动剂组炎性细胞浸润增多，而NLRP3抑制剂组炎性细胞浸润减少。干预后，与对照组相比，模型组创面积分和血清TNF-α、IL-6水平升高，创面组织中NLRP3、cAMP、TRPV1蛋白相对表达量和NLRP3、cAMP、TRPV1基因mRNA表达量升高，差异有统计学意义(P < 0.05); 干预后，与模型组相比, NLRP3激动剂组上述指标均升高, NLRP3抑制剂组上述指标均降低，差异有统计学意义(P < 0.05)。
结论NLRP3炎症小体可能通过激活cAMP/TRPV1信号通路，参与调控大鼠急性痔疮的发生与发展。
-
关键词:
- 痔疮 /
- NOD样受体热蛋白结构域相关蛋白3 /
- 环磷酸腺苷 /
- 瞬时受体电位香草酸亚型1 /
- 炎症因子
Abstract:ObjectiveTo investigate the effects of NOD-like receptor thermal protein domain-associated protein 3 (NLRP3) on wound pathological characteristics, inflammatory responses, and cyclic adenosine monophosphate (cAMP)/transient receptor potential vanilloid subtype 1 (TRPV1) signaling pathways in a rat model of hemorrhoids.
MethodsForty healthy male SD rats aged 6 weeks were randomly divided into control group, model group, NLRP3 agonist group, and NLRP3 inhibitor group, with 10 rats in each group. The control group received no intervention, while acute hemorrhoid models were established in the other three groups. After successful modeling, the NLRP3 agonist group and the NLRP3 inhibitor group were administered local subcutaneous injections of NLRP3 agonist and NLRP3 inhibitor at the wound site, respectively. The model group and the control group received subcutaneous injections of equal volume of saline. The pathological characteristics of the wounds in each group were observed and wound scores were assessed. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum levels of tumor necrosis factor (TNF)-α and interleukin (IL)-6. Western blot analysis was employed to detect protein expression levels of NLRP3, cAMP, and TRPV1 in the wound tissue. Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect mRNA expression levels of NLRP3, cAMP, and TRPV1 in the wound tissue.
ResultsCompared with the control group, the model group exhibited increased inflammatory cell infiltration, pronounced edema, disrupted tissue structure, and connective tissue hyperplasia in the wound tissue. When compared to the model group, the NLRP3 agonist group showed an increase in inflammatory cell infiltration, whereas the NLRP3 inhibitor group demonstrated a reduction in inflammatory cell infiltration. After intervention, compared with the control group, the model group had elevated wound scores and serum levels of TNF-α and IL-6. Additionally, the relative protein expression levels of NLRP3, cAMP, and TRPV1, as well as the mRNA expression levels of NLRP3, cAMP, and TRPV1 in the wound tissue, were increased in the model group (P < 0.05). Following intervention, when compared to the model group, the NLRP3 agonist group showed an increase in all aforementioned indicators, whereas the NLRP3 inhibitor group exhibited a decrease in these indicators (P < 0.05).
ConclusionThe NLRP3 inflammasome may participate in the development and progression of acute hemorrhoids in rats by activating the cAMP/TRPV1 signaling pathway.
-
-
![]()
图 2 各组大鼠创面组织中NLRP3、cAMP和TRPV1蛋白表达量比较
与对照组比较, #P < y0.05; 与模型组比较, *P < 0.05; 与NLRP3激动剂组比较, △P < 0.05。
![]()
图 4 各组大鼠创面组织中NLRP3、cAMP、TRPV1基因的mRNA表达量比较
与对照组比较, #P < 0.05; 与模型组比较, *P < 0.05; 与NLRP3激动剂组比较, △P < 0.05。
表 1 各组大鼠创面积分比较($\bar{x} \pm s$)
分 组别 n 干预前 干预7 d后 对照组 10 0.3±0.1 0.3±0.1 模型组 10 7.8±0.5* 7.9±0.5* NLRP3激动剂组 10 7.7±0.4* 8.5±0.3*#▲ NLRP3抑制剂组 10 7.6±0.4* 5.4±0.3*#△▲ 与对照组比较, *P < 0.05; 与模型组比较, #P < 0.05;
与NLRP3激动剂组比较, △P < 0.05;
与干预前比较, ▲P < 0.05。
下载: 导出CSV
表 2 各组大鼠血清TNF-α和IL-6水平比较($\bar{x} \pm s$)
mg/L 组别 n TNF-α IL-6 干预前 干预7 d后 干预前 干预7 d后 对照组 10 0.5±0.1 0.6±0.1 0.9±0.2 0.9±0.3 模型组 10 8.6±2.3* 8.7±2.4* 12.5±3.4* 12.7±3.5* NLRP3激动剂组 10 8.4±2.2* 12.3±3.6*#▲ 12.2±3.1* 15.5±4.5*#▲ NLRP3抑制剂组 10 8.5±2.3* 4.8±0.9*#△▲ 12.6±3.4* 9.9±2.5*#△▲ TNF-α: 肿瘤坏死因子-α; IL-6: 白细胞介素-6。与对照组比较, *P < 0.05; 与模型组比较, #P < 0.05;
与NLRP3激动剂组比较, △P < 0.05; 与干预前比较, ▲P < 0.05。
下载: 导出CSV
-
[1] PATA F, SGRÓ A, FERRARA F, et al. Anatomy, physiology and pathophysiology of haemorrhoids[J]. Rev Recent Clin Trials, 2021, 16(1): 75-80.
[2] PORWAL A, KUNDU G C, BHAGWAT G, et al. Herbal medicine AnoSpray suppresses proinflammatory cytokines COX-2 and RANTES in the management of hemorrhoids, acute anal fissures and perineal wounds[J]. Exp Ther Med, 2022, 23(1): 86.
[3] 徐珊, 窦燕, 李小军, 等. 内镜下负压套扎术对Ⅱ、Ⅲ度痔疮患者肛肠动力学、炎症反应及免疫功能的影响[J]. 现代生物医学进展, 2024, 24(6): 1187-1190, 1170. [4] 李萌. PKA介导骨癌诱导的大鼠特异性背根神经节神经元兴奋性的机制研究[J]. 全科口腔医学杂志: 电子版, 2019, 6(21): 158-159. [5] 何裕智, 李俊. 自动痔疮套扎术对直肠内脱垂合并便秘患者肠黏膜屏障功能及黏膜TRPV1、5-HT表达的影响[J]. 现代消化及介入诊疗, 2021, 26(5): 607-610. doi: 10.3969/j.issn.1672-2159.2021.05.013 [6] 中华中医药学会中药实验药理专业委员会. 中药痔疮动物模型制备规范(草案)[J]. 中药药理与临床, 2017, 33(5): 211-212. [7] ZHANG Y L, SUN S W, HAN Z G. Establishment and validation of clinical prediction model for hemorrhoid recurrence after procedure for prolapse and hemorrhoids[J]. Medicine, 2023, 102(26): e34062. doi: 10.1097/MD.0000000000034062
[8] 徐珊, 窦燕, 李小军, 等. 苦参凝胶对痔疮模型大鼠的治疗效果及机制研究[J]. 中医药学报, 2024, 52(3): 34-39. [9] 袁一鸣, 王艳, 裴飞, 等. NLRP3炎性小体与周围神经损伤关系的研究进展[J]. 华中科技大学学报: 医学版, 2023, 52(5): 735-741. [10] 张艺嘉, 余清华, 王宇, 等. 菝葜通过NLRP3通路对巴豆油诱发痔疮大鼠模型的改善作用[J]. 湖北中医药大学学报, 2024, 26(2): 10-14. [11] FU J N, WU H. Structural mechanisms of NLRP3 inflammasome assembly and activation[J]. Annu Rev Immunol, 2023, 41: 301-316.
[12] SEOANE P I, LEE B L, HOYLE C, et al. The NLRP3-inflammasome as a sensor of organelle dysfunction[J]. J Cell Biol, 2020, 219(12): e202006194.
[13] 李惠雯, 严建, 宾东华, 等. 参榆洗液对痔术后大鼠痛觉敏化及cAMP/PKA信号通路和TRPV1蛋白表达的影响[J]. 现代中西医结合杂志, 2024, 33(8): 1065-1071. [14] MA Y Q, CHEN J, YU D Q, et al. cAMP-PKA signaling is involved in regulation of spinal HCN channels function in diabetic neuropathic pain[J]. Neurosci Lett, 2021, 750: 135763. http://www.sciencedirect.com/science/article/pii/S0304394021001415
[15] 梁冉冉, 刁志君, 杨晓航, 等. 穴位敏化现象与TRPV1通道的关系研究[J]. 针灸临床杂志, 2023, 39(7): 1-5. [16] YU Q, ZHAO Y, ZHANG X Y, et al. The beneficial effect of Sanhuang ointment and its active constituents on experimental hemorrhoids in rats[J]. J Ethnopharmacol, 2024, 319(Pt 2): 117173.
[17] ZHAO C Y, ZHAO W. NLRP3 inflammasome-a key player in antiviral responses[J]. Front Immunol, 2020, 11: 211.
[18] XIA C Y, GUO Y X, LIAN W W, et al. The NLRP3 inflammasome in depression: potential mechanisms and therapies[J]. Pharmacol Res, 2023, 187: 106625.
[19] 刘宁远, 刘殿龙, 郑丽华. 痔的治疗现状与展望[J]. 中国临床医生杂志, 2024, 52(1): 1-3. [20] 李彦娜, 杨阳, 刘晓娜, 等. 冷冻疗法联合马应龙麝香痔疮膏对混合痔术后水肿及疼痛程度的影响[J]. 检验医学与临床, 2023, 20(16): 2452-2455. [21] 顾菁华, 杜培欣, 吴伟, 等. 黄连解毒汤对急性痔疮模型大鼠肛周病理组织形态、免疫功能以及炎症因子的影响[J]. 中国中医急症, 2020, 29(1): 25-28. -
期刊类型引用(2)
1. 王向阳. PD-1抑制剂联合SOX方案治疗晚期胃癌患者的疗效及对外周血T淋巴细胞亚群水平的影响. 宁夏医学杂志. 2024(06): 461-464 . 
百度学术
2. 陈守华，姚卫东，徐美华，肖金章，徐薇薇. PD-1单抗和紫杉醇脂质体联合奈达铂一线治疗进展期食管癌的疗效观察. 中国肿瘤临床与康复. 2022(02): 138-141 . 百度学术
其他类型引用(0)
计量
- 文章访问数: 34
- HTML全文浏览量: 10
- PDF下载量: 1
- 被引次数: 2
苏公网安备 32100302010246号
