青藤碱对人脑胶质瘤的抑制作用及肿瘤内药代动力学特征

Inhibitory Effect of Sinomenine on Human Glioblastoma and Its Pharmacokinetic Characteristics

摘要:

目的 观察青藤碱（sinomenine）对人脑胶质瘤的抑制作用及在脑胶质瘤内的药代动力学特征。方法 构建稳定表达荧光素酶的脑胶质瘤U87细胞系，脑内接种建立同时用于药效学和药代动力学研究的小鼠胶质瘤模型。药效学部分：模型小鼠随机分为模型组和青藤碱低、中、高剂量组（50、100、150 mg·kg^-1），腹腔注射给药14 d，观察脑内肿瘤荧光值，分析其对脑胶质瘤增殖的抑制作用；取脑肿瘤及肿瘤周围脑组织，蛋白免疫印迹法（Western blot）检测血管内皮生长因子A（VEGFA）、药物转运体P-糖蛋白（P-gp）和乳腺癌耐药蛋白（BCRP）及闭合蛋白（Occludin）的表达水平。药代动力学部分：小鼠分为正常给药组（50 mg·kg^-1）和模型给药组（50、100、150 mg·kg^-1），青藤碱单次腹腔注射后，微透析技术活体采集脑肿瘤细胞外液，每15 min收集1次，共收集至给药后6 h。高效液相色谱-质谱联用法（HPLC-MS/MS）检测透析液中青藤碱浓度，计算药代动力学参数，分析青藤碱在脑和脑胶质瘤内的药代动力学特征。结果 与模型组比较，青藤碱给药14 d后脑内肿瘤荧光值明显降低（P<0.05），并呈一定的剂量依赖性。青藤碱可抑制肿瘤周围VEGF的升高和Occludin的降解，抑制胶质瘤VEGF、P-gp、BCRP蛋白表达。青藤碱单次给药后，在7.5 min即可到达脑组织和肿瘤组织。和正常组比较，肿瘤内C_max、AUC均明显升高，T_max缩短（从1.63 h缩短至0.71 h），CLz/F降低。在50~150 mg·kg^-1剂量范围内，脑胶质瘤内的青藤碱呈现一定的线性药代动力学过程。结论 青藤碱对于脑胶质瘤具有明显的抑制作用，抑制VEGF的增高和药物转运蛋白的外排，降低肿瘤侵袭，维持血脑屏障完整。青藤碱可以快速通过血肿瘤屏障并达峰，在肿瘤内具有线性药代动力学特征。

Abstract:

Objective To observe the inhibitory effect of sinomenine on human glioblastoma and its pharmacokinetic characteristics in glioblastoma.Method A human glioblastoma U87 cell line stably expressing luciferase was constructed， and a mouse glioma model was established for use in both pharmacodynamic and pharmacokinetic studies. Pharmacodynamics： Model mice were randomly divided into model group and sinomenine low-， medium-， and high-dose groups （50， 100， 150 mg·kg^-1）. Sinomenine was administered intraperitoneally for 14 days. The fluorescence value of brain tumors was observed to analyze its inhibitory effect on glioblastoma proliferation. Brain tumors and the surrounding brain tissue were collected， and the expression levels of vascular endothelial growth factor A （VEGFA）， P-glycoprotein （P-gp）， breast cancer resistance protein （BCRP）， and Occludin were detected by Western blot. Pharmacokinetics： Mice were divided into a normal group （50 mg·kg^-1） and model groups （50， 100， 150 mg·kg^-1）. After a single intraperitoneal injection of sinomenine， extracellular fluid from brain tumors was collected in vivo by microdialysis every 15 min for 6 h. Sinomenine concentrations in the dialysate were detected by HPLC-MS/MS， and pharmacokinetic parameters were calculated to analyze pharmacokinetic characteristics of sinomenine in the brain and glioblastoma.Result Compared with model group， after 14 days of sinomenine administration， the fluorescence value of brain tumors significantly decreased （P<0.05） in a dose-dependent manner. Sinomenine inhibited the increase in VEGF and the degradation of Occludin in the tissue surrounding the tumor and inhibited the expression of VEGF， P-gp， and BCRP in glioblastoma. After a single administration， sinomenine was detected in brain and tumor tissues within 7.5 min. Compared with normal group， the C_max and AUC in the tumor significantly increased， T_max shortened （from 1.63 h to 0.71 h）， and CLz/F decreased. In the dose range of 50-150 mg·kg^-1， sinomenine exhibited a linear pharmacokinetic process in glioblastoma.Conclusion Sinomenine has a significant inhibitory effect on glioblastoma， which can inhibit VEGF elevation and drug transporter efflux， reduce tumor invasion， and maintain the integrity of the blood-brain barrier. Sinomenine can rapidly cross the blood-tumor barrier， reach peak concentration， and exhibit linear pharmacokinetic characteristics in the tumor.