Table_3_Tetrandrine alleviates oxaliplatin-induced mechanical allodynia via modulation of inflammation-related genes.XLS

Oxaliplatin, a platinum-based chemotherapy drug, causes neuropathic pain, yet effective pharmacological treatments are lacking. Previously, we showed that tetrandrine (TET), with anti-inflammatory properties, reduces mechanical allodynia in nerve-injured mice. This study explores the effect of TET on oxaliplatin-induced mechanical allodynia and gene changes in mice. Male C57BL/6J mice received oxaliplatin intraperitoneally to induce mechanical allodynia. Post-treatment with TET or vehicle, the mechanical withdrawal threshold (WMT) was assessed using von Frey filaments. TET alleviated oxaliplatin-induced mechanical allodynia. RNA sequencing identified 365 differentially expressed genes (DEGs) in the Control vs. Oxaliplatin group and 229 DEGs in the Oxaliplatin vs. TET group. Pearson correlation analysis of co-regulated DEGs and inflammation-related genes (IRGs) revealed 104 co-regulated inflammation-related genes (Co-IRGs) (|cor| > 0.8, P < 0.01). The top 30 genes in the PPI network were identified. Arg2, Cxcl12, H2-Q6, Kdr, and Nfkbia were highlighted based on ROC analysis. Subsequently, Arg2, Cxcl12, Kdr, and Nfkbia were further verified by qRCR. Immune infiltration analysis indicated increased follicular CD4 T cell infiltration in oxaliplatin-treated mice, reduced by TET. Molecular docking showed strong binding affinity between TET and proteins encoded by Arg2, Cxcl12, Kdr, and Nfkbia. In summary, TET may alleviate oxaliplatin-induced peripheral neuropathy in clinical conditions.

奥沙利铂（Oxaliplatin）作为一类铂类化疗药物，可诱发神经性疼痛，目前尚无有效的临床药物治疗方案。此前本研究团队已证实，具备抗炎活性的粉防己碱（tetrandrine, TET）可缓解神经损伤小鼠的机械性痛觉超敏（mechanical allodynia）。本研究旨在探讨粉防己碱对奥沙利铂诱导的小鼠机械性痛觉超敏及体内基因表达变化的调控作用。实验选用雄性C57BL/6J小鼠，通过腹腔注射奥沙利铂构建机械性痛觉超敏模型；给予粉防己碱或溶剂对照处理后，采用冯·弗雷细丝（von Frey filaments）检测小鼠的机械撤足阈值（mechanical withdrawal threshold, WMT）。结果表明，粉防己碱可显著缓解奥沙利铂诱导的机械性痛觉超敏。RNA测序（RNA sequencing）分析显示，对照组与奥沙利铂组间共鉴定出365个差异表达基因（differentially expressed genes, DEGs），奥沙利铂组与粉防己碱组间则鉴定出229个DEGs。对共调控差异表达基因与炎症相关基因（inflammation-related genes, IRGs）开展皮尔逊相关分析，最终筛选得到104个共调控炎症相关基因（co-regulated inflammation-related genes, Co-IRGs）（|相关系数|>0.8，P<0.01）。进一步通过蛋白质相互作用网络（protein-protein interaction network, PPI网络）筛选出排名前30的核心基因，并结合受试者工作特征曲线（receiver operating characteristic curve, ROC）分析，锁定Arg2、Cxcl12、H2-Q6、Kdr及Nfkbia为关键靶基因。随后通过qRCR对上述核心基因的表达水平进行了验证。免疫浸润分析显示，奥沙利铂处理组小鼠的滤泡CD4+T细胞浸润水平显著升高，而粉防己碱可逆转这一异常免疫浸润状态。分子对接实验证实，粉防己碱与Arg2、Cxcl12、Kdr及Nfkbia编码的蛋白具有较强的结合亲和力。综上，粉防己碱有望在临床场景中缓解奥沙利铂诱导的周围神经病变。