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Title: [The expression and significance of TRPM8 among chronic rhinosinusitis with nasal polyps].
Author: Tong XT, Liu PQ, Zhou HQ, Jin J, Liu KY, Huang JY, Zhang W, Chen SM, Tao ZZ, Xu Y.
Journal: Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi; 2021 Oct 07; 56(10):1059-1065. PubMed ID: 34666466.
Abstract:
Objective: To compare the expression and difference of melastatin-related transient receptor potential 8(TRPM8) among chronic rhinosinusitis, nasal polyps and normal mucosa tissues. And to explore the significant expression of TRPM8 among CRSwNP. Methods: Fifty-one patients underwent endoscopic sinus surgery in the Department of Otorhinolaryngology Head and Neck Surgery of Renmin Hospital of Wuhan University from February 2019 to January 2020 were recruited, including 33 males and 18 females, aged from 14 to 65 years old (34.55±1.689).Immunohistochemistry was used to detected the expression of TRPM8 protein among CRSsNP(17),CRSwNP (17) and control tissuses(17). In addition, the correlation between the expression of TRPM8 protein in CRSwNP patients and preoperative CT Lund-Mackay scores and preoperative VAS scores and sinonasal outcome test-20 scores was analyzed, respectively. The primary human nasal epithelial cells were cultured in vitro and the expression of TRPM8 was detected by quantitative real-time PCR and western blotting. The tissue in control group, chronic rhinosinusitis without nasal polyps (CRSsNP) group and the CRSwNP group were collected and grinded into tissue homogenized. The expression of TRPM8 protein was detected by western blotting after 24 h stimulation after homogenate was added into the medium of RPMI 2650 and primary nasal epithelial cells. Results: Compared with the control, the expression of TRPM8 was significantly up-regulated in nasal polyps (t=6.852, P<0.05). TRPM8 was mainly expressed in epithelial cells. The expression of TRPM8 in the epithelial cells of CRSsNP had no difference with the control group (t=1.980, P>0.05). In addition, the expression of TRPM8 in CRSwNP patients was positively correlated with the preoperative CT Lund-Mackay scores and VAS scores and SNOT-20 scores (r=0.512, P<0.05;r=0.853, P<0.01;r=0.814, P<0.01). After cultured primary epithelial cells in vitro, the expression level of TRPM8 in epithelial cells derived from nasal polyp was significantly higher than that in control group (t=8.845, P<0.05). By adding the homogenization of control and CRSsNP and CRSwNP tissues, the expression of TRPM8 in RPMI 2650 cells and primary nasal epithelial cells was changed and that was significantly increased after adding the homogenization of the group of CRSwNP. Conclusion: TRPM8 is highly expressed in nasal polyps epithelial cells, suggesting that TRPM8 may be involved in the pathogenesis of nasal polyps regulated by nasal epithelial cells. 目的： 检测M型瞬时受体电位通道8（melastatin-related transient receptor potential 8，TRPM8）在慢性鼻窦炎、鼻息肉和对照组中鼻甲黏膜组织中表达及表达差异，探讨在慢性鼻窦炎伴鼻息肉（CRSwNP）中TRPM8表达的意义。 方法： 前瞻性纳入2019年2月至2020年1月在武汉大学人民医院耳鼻咽喉头颈外科行鼻内镜手术患者51例，分为慢性鼻窦炎伴鼻息肉（CRSwNP）组17例、慢性鼻窦炎不伴鼻息肉（CRSsNP）组17例和对照组（单纯解剖上存在鼻中隔偏曲或鼻骨骨折的患者）17例。其中男性33例，女性18例，年龄为14~65（34.55±1.689）岁。利用免疫组织化学技术检测3组患者组织（CRSwNP组取息肉组织、CRSsNP组取钩突组织和对照组取中鼻甲组织）中TRPM8的表达及分布。根据免疫组织化学结果分析TRPM8表达与CRSwNP患者术前CT Lund-Mackay评分、鼻部症状视觉模拟量表（VAS）评分和鼻腔鼻窦结局测试20（SNOT-20）量表评分的相关性；利用原代细胞培养及实时定量反转录聚合酶链反应（RT-PCR）和免疫蛋白印迹法检测鼻息肉上皮细胞及中鼻道上皮细胞中TRPM8的mRNA及蛋白表达水平；收取3组临床标本制备成组织匀浆后加入人鼻腔上皮细胞系和原代正常鼻黏膜上皮细胞中刺激24 h，随后利用蛋白免疫印迹技术检测TRPM8蛋白表达。采用SPSS 23.0及GraphPad Prism软件进行统计学分析，两组间采用独立样本t检验，相关性分析采用Spearman检验。 结果： 与对照组相比，CRSwNP组中TRPM8表达明显上调（t=6.852，P<0.05），同时蛋白主要表达在鼻黏膜上皮细胞中；CRSsNP组中TRPM8表达差异无统计学意义（t=1.980，P>0.05）。TRPM8表达与术前CT Lund-Mackay评分、VAS评分和SNOT-20均呈正相关（r=0.512，P<0.05；r=0.853，P<0.01；r=0.814，P<0.01）；体外培养对照组中鼻道上皮细胞以及息肉上皮细胞后，检测发现息肉上皮细胞中TRPM8的mRNA及蛋白表达水平明显高于对照组上皮细胞，差异有统计学意义（t=8.845，P<0.05）；相较于加入正常对照和慢性鼻窦炎组织匀浆而言，加入鼻息肉组织匀浆的人鼻腔上皮细胞系（RPMI 2650）细胞中TRPM8表达增高。 结论： TRPM8在鼻息肉上皮细胞中呈高表达且其表达量与患者VAS评分呈正相关，提示TRPM8可能参与鼻息肉发病过程中鼻黏膜上皮细胞调控的发病机制。.

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