黄连解毒汤在阿尔茨海默病模型大鼠体内药代动力学及对脑内炎症微环境改善的研究

Pharmacodynamics of Huanglian Jiedu decoction in Alzheimer’s disease(AD) model rats and effect on improvement of inflammation microenvironment in brain

研究黄连解毒汤（HLJDD）中有效成分（生物碱类、环烯醚萜类和黄酮类）在阿尔茨海默病（AD）模型大鼠体内药代动力学过程,并探讨黄连解毒汤治疗阿尔茨海默病的相关性。实验分为正常组（n=6）,假手术组（n=4）和模型组（n=12）。假手术组大鼠连续45 d皮下注射D-半乳糖（50 mg·kg^-1）造成亚急性衰老模型,模型组大鼠在注射D-半乳糖的基础上,基底核注射Aβ25-35（4.0 g·L^-1）和鹅膏蕈氨酸（2.0 g·L^-1）。模型组大鼠分为黄连解毒汤单次给药组（n=6）和黄连解毒汤连续给药1周组（n=6）。采用LC-QQQ-MS方法测定不同时间点生物碱类、环烯醚萜类和黄酮类成分的血药浓度。并利用Bio-Plex多因子检测技术检测大鼠脑脊液中7个炎症因子（MIP-2,IL-1β,IL-2,IL-8,IL-10,IL-13,TNF-α）的水平。环烯醚萜类成分在AD模型大鼠体内吸收和消除较快,生物利用度高。原小檗碱型生物碱在血浆中含量较低,其AUC（药时曲线下面积）在给药1周组大鼠中高于单次给药组。黄酮类成分在大鼠体内的药时曲线出现了明显的双峰现象。黄连解毒汤给药后,AD模型大鼠脑脊液中的炎症因子表现出回调趋势,其中IL-1β/IL-10（P〈0.05）出现显著性差异。基本阐述了环烯醚萜类,生物碱类和黄酮类成分（共41个化合物）在AD模型大鼠体内的药代动力学行为,黄连解毒汤能够通过调节炎症因子的水平改善阿尔茨海默病模型大鼠中枢炎症状态。

To study the pharmacokinetics of active ingredients（alkaloids,iridoids and flavonoids） in Huanglian Jiedu decoction（HLJDD） in Alzheimer＇s disease（AD） model rats,and investigate its mechanism in treatment of AD. All of rats were divided into normal control group（n = 6）,shame operation group（n = 6） and model group（n = 12）. Rats in shame operation group received daily subcutaneous injection of D-galactose（D-gal 50 mg·kg^-1） for a total of 45 d to induce subacute aging model. Based on the operation of shame operation group,the rats in model group were given with an injection of Aβ25-35（4. 0 g·L^-1）-ibotenic acid（2. 0 g·L^-1）into the nucleus basalis magnocellularis. Then rats in model group were divided into HLJDD one day administration group（n = 6） and HLJDD one week group（n = 6）. The plasma concentration of alkaloids,iridoid and flavonoids was determined by liquid chromatography tandem mass spectrometry（LC-QQQ-MS） at different time points. The levels of seven inflammatory factors（MIP-2,IL-1β,IL-2,IL-8,IL-10,IL-13,TNF-α） in cerebrospinalfluid（CSF） were measured by Bio-Plex multi-factor detection technology. Iridoids in AD model rats,with high bioavailability,were easily absorbed and eliminated. The plasma concentration of alkaloid was lower,and the AUC（area under the curve） was higher in HLJDD one week group than that in HLJDD one day group. The plasma concentration-time curves of flavonoids showed obvious bimodal phenomena. After the gastric administration of HLJDD,the inflammatory factors in CSF of AD rats demonstrated a callback trend,including IL-1β/IL-10（P〈0. 05） with significant difference. The pharmacokinetic behaviors of iridoids,alkaloids and flavonoids（41 compounds） in AD model rats were fundamentally elucidated,and HLJDD can improve the central inflammatory status of AD rats by regulating the levels of inflammatory factors.