Objective: To observe the effects of point-moxibustion with Zhuang medicinal thread on differentially expressed genes (DEGs) in the dorsal root ganglion (DRG), tissue morphology, and the expression of Fc epsilon RI (FcεRI) pathway proteins spleen tyrosine kinase (Syk) and membrane spanning 4-domain A2 (MS4A2) in rat model of postherpetic neuralgia (PHN), and to explore the potential mechanism by which this therapy alleviates pain sensitization.
Methods: Thirty-nine male Sprague-Dawley (SD) rats were randomly divided into a control group, a model group, and a moxibustion group, with 13 rats in each group. The PHN model was established in the model and moxibustion groups by intraperitoneal injection of resiniferatoxin. In the moxibustion group, bilateral L4-L6 "Jiaji" (EX-B2) points were treated with point-moxibustion with Zhuang medicinal thread from day 7 post-modeling, with two cones per acupoint per session, every other day for a total of 10 sessions. Mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were measured at 1 day before modeling and on days 1, 4, 7, 13, 19, and 25 after modeling. After intervention, HE staining was used to observe DRG morphology. RNA sequencing was performed to analyze DEGs in DRG and conduct bioinformatics analysis. The expression of Syk and MS4A2 mRNA and proteins in the FcεRI pathway in DRG was detected by quantitative PCR and Western blot.
Results: Compared with the control group, the model group exhibited decreased MWT (P<0.05) and increased TWL (P<0.05); histopathological analysis revealed neuronal atrophy, nuclear displacement, and intracellular vacuoles, with a slightly loose arrangement; the RNA-Seq identified 3,207 DEGs (1,997 upregulated and 1,210 downregulated); the mRNA and protein expression levels of Syk and MS4A2 were significantly increased (P<0.01). Compared with the model group, the moxibustion group showed increased MWT (P<0.05) and decreased TWL (P<0.05), with relatively normal neuronal morphology; the RNA-Seq identified 426 DEGs (250 upregulated and 176 downregulated); the mRNA and protein expression levels of Syk and MS4A2 were significantly decreased (P<0.05). Venn diagram analysis identified 156 DEGs that showed a reversal in expression trends after treatment, including Syk and MS4A2, which are associated with pain sensitization. KEGG pathway analysis indicated that these DEGs were primarily enriched in the FcεRI pathway.
Conclusion: Point-moxibustion with Zhuang medicinal thread could alleviate pain sensitization in PHN rats, possibly by inhibiting the FcεRI signaling pathway and downregulating the expression of Syk and MS4A2.
目的：观察壮医药线点灸对带状疱疹后遗神经痛（PHN）模型大鼠背根神经节差异表达基因（DEGs）、组织形态及高亲和力免疫球蛋白ε受体（FcεRI）通路蛋白脾酪氨酸激酶（Syk）、跨膜4域A2（MS4A2）表达的影响，探讨壮医药线点灸改善痛觉敏化的可能机制。 方法：将39只雄性SD大鼠随机分为对照组、模型组和壮医药线点灸组，每组13只。模型组和壮医药线点灸组大鼠腹腔注射树脂毒素建立PHN大鼠模型。壮医药线点灸组于造模成功后（造模第7天）予双侧L4-L6“夹脊”穴壮医药线点灸，每次每穴2壮，隔日1次，共干预10次。分别于造模前1 d，造模第1、4、7、13、19天及造模第25天（干预完成）测定机械缩爪阈值（MWT）和热刺激缩爪潜伏期（TWL）；干预后，采用HE染色观察大鼠背根神经节形态，转录组测序（RNA-Seq）技术分析背根神经节DEGs并行生物信息学分析，实时荧光定量PCR和Western blot法检测背根神经节FcεRI通路蛋白Syk、MS4A2 mRNA和蛋白表达。 结果：与对照组比较，模型组大鼠MWT降低（P<0.05），TWL升高（P<0.05）；神经元萎缩，细胞核偏离中心，细胞内多空泡结构，排列稍疏；RNA-Seq总计筛到3 207个DEGs（包括1 997个上调DEGs和1 210个下调DEGs）；Syk、MS4A2 mRNA和蛋白表达均升高（P<0.01）。与模型组比较，壮医药线点灸组大鼠MWT升高（P<0.05），TWL降低（P<0.05）；神经元形态较正常；RNA-Seq总计筛到426个DEGs（包括250个上调DEGs和176个下调DEGs）；Syk、MS4A2 mRNA和蛋白表达均降低（P<0.05）。维恩图分析得到156个DEGs在壮医药线点灸后表达趋势逆转，包括与痛敏相关的Syk和MS4A2。京都基因与基因组百科全书（KEGG）分析显示，这些DEGs主要富集于FcεRI通路等。 结论：壮医药线点灸可改善PHN大鼠的痛觉敏化，其作用机制可能与抑制FcεRI信号通路，进而下调Syk和MS4A2表达有关。.
Keywords: FcεRI pathway; differentially expressed genes; point-moxibustion with Zhuang medicinal thread; postherpetic neuralgia (PHN); transcriptome sequencing.