目的 通过转录组测序（RNA-Seq）、微小 RNA 测序（miRNA-Seq）及实时荧光定量 PCR（RT-qPCR）分 析，探究全蝎-蜈蚣药对对脑胶质瘤小鼠的作用及机制。方法 将 18 只雄性 C57BL/6J 小鼠随机分为模型组、 全蝎-蜈蚣组（1 g·kg-1 ）、替莫唑胺组（50 mg·kg-1 ），每组 6 只。通过颅内注射表达荧光素酶报告基因的小鼠胶 质瘤细胞（GL261-Luc）构建原位脑胶质瘤小鼠模型。注射 GL261-Luc 细胞 1 周后，各组按照上述剂量灌胃给药 （10 mL·kg-1 ），每日 1 次，连续 2 周。采用活体成像测定原位胶质瘤体积；分析小鼠外周血中白细胞、淋巴细 胞数量及单核细胞、粒细胞比例；检测小鼠血清中谷丙转氨酶（ALT）、谷草转氨酶（AST）、总胆红素（TBIL）及 直接胆红素（DBIL）的水平；采用 HE 染色法观察脑组织病理变化；免疫组化法检测原位胶质瘤组织 Ki67 蛋白 表达水平；采用 RNA-Seq 及 miRNA-Seq 对模型组、全蝎-蜈蚣组小鼠原位胶质瘤组织进行系统性分析和联合 分析；RT-qPCR 法检测胶质瘤组织中 miR-206-3p 及 XC 趋化因子受体 1（XCR1）mRNA 表达水平。结果 与 模型组比较，全蝎-蜈蚣组小鼠的肿瘤体积显著缩小（P＜0.01），体质量无明显变化（P＞0.05）；外周血白细胞 及淋巴细胞数量显著升高（P＜0.05，P＜0.01），单核细胞及粒细胞比例显著降低（P＜0.01）；血清 ALT 水平明 显降低（P＜0.05），AST、TBIL、DBIL 水平无明显变化（P＞0.05）；肿瘤组织可见细胞凋亡后留下的局部空泡， 同时细胞密度降低，原位瘤内新生血管减少；原位胶质瘤组织的 Ki67 表达显著下调（P＜0.01），miR-206-3p 表达量明显降低（P＜0.05），XCR1 mRNA 表达量显著升高（P＜0.01）。mRNA-Seq、miRNA-Seq 分析及二者联 合分析均提示，全蝎-蜈蚣具有显著的免疫调节作用，可能通过调节细胞黏附分子、促进免疫细胞的活化和迁 移、改善肿瘤微环境等多途径发挥抑制胶质瘤生长的作用。结论 全蝎-蜈蚣药对能够抑制小鼠原位脑胶质瘤 的进展，可能与通过 miR-206-3p/XCR1 信号改善脑胶质瘤免疫微环境有关。

Objective To explore the effects and mechanisms of the Scorpio-Scolopendra drug pair on glioma in mice through transcriptome sequencing （RNA-Seq）， microRNA sequencing （miRNA-Seq）， and real-time quantitative PCR （RT-qPCR） analysis. Methods Eighteen male C57BL/6J mice were randomly divided into model group，ScorpioScolopendra group （1 g·kg-1 ），and temozolomide group （50 mg·kg-1 ），with 6 mice in each group. An orthotopic glioma mouse model was established by intracranial injection of luciferase reporter gene-expressed mouse glioma cells （GL261- Luc）. One week after GL261-Luc cell injection，each group was administered the corresponding dose via intragastric gavage （10 mL·kg-1 ），once daily for 2 consecutive weeks. The counts of white blood cells and lymphocytes，and the proportions of monocytes and granulocytes in peripheral blood were analyzed. Serum levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， total bilirubin （TBIL）， and direct bilirubin （DBIL） were measured. Histopathological changes in brain tissue were observed via HE staining. Ki67 protein expression in orthotopic glioma tissue was detected by immunohistochemistry. RNA-Seq and miRNA-Seq were systematically and jointly performed on orthotopic glioma tissues from the model and Scorpio-Scolopendra groups. mRNA expression levels of miR-206-3p and XC chemokine receptor 1（XCR1） in glioma tissues were determined by RT-qPCR. Results Compared with the model group，the Scorpio-Scolopendra group showed significantly reduced tumor volume （P＜0.01） with no significant change in body mass（P＞0.05）. The counts of white blood cells and lymphocytes in peripheral blood were significantly increased （P＜0.05，P＜0.01），while the proportions of monocytes and granulocytes were significantly decreased （P＜ 0.01）. Serum ALT levels were significantly decreased （P＜0.05）， while AST，TBIL，and DBIL levels showed no significant changes （P＞0.05）. Local vacuoles left after cell apoptosis were observed in tumor tissue， along with reduced cell density and decreased neovascularization within the orthotopic tumor. Ki67 expression in orthotopic glioma tissue was significantly downregulated （P＜0.01）. The expression level of miR-206-3p was markedly decreased （P＜ 0.05），while the mRNA expression level of XCR1 was significantly increased （P＜0.01）. Analyses from mRNA-Seq， miRNA-Seq， and their integration all indicated that the Scorpio-Scolopendra drug pair exhibits significant immunomodulatory effects. It likely inhibits glioma growth through multiple pathways by modulating cell adhesion molecules， promoting the activation and migration of immune cells， and improving the tumor microenvironment. Conclusion The Scorpio-Scolopendra drug pair can inhibit the progression of orthotopic glioma in mice，which may be related to its ability to improve the glioma immune microenvironment via the miR-206-3p/XCR1 signaling axis.

R285.5

国家自然科学基金项目（82374534）；广州市科技计划项目（202201011634）；深圳市自然科学基金面上项目（JCYJ20240813180803005）； 深圳市宝安区“医疗卫生三名工程”项目（SZZYSM202311015）；深圳市宝安区中医药临床研究专项（2023ZYYLCZX）。