Pharmacokinetics and blood-brain barrier penetration of a novel salidroside derivative pOBz in rats北大核心CSCD

Aim To develop a ultra-high performance liquid chromatography electrospray-ionization tandem mass spectrometry ( UPLC-MS/MS ) method for the simultaneous determination of salidroside derivative pOBz in rat plasma and brain tissue, and to study the pharmacokinetic profile and penetration of the blood-brain barrier in rats after a single dose intravenous administration of pOBz. Methods SD rats were administered pOBz at a dose of 50 mg • kg -1via tail vein, and plasma and brain tissue samples were collected at different time points. UPLC-MS/MS method was used to determine the concentration of pOBz in plasma and brain tissue. The plasma concentration-time curve was fitted by DAS 2. 1. 1 software to calculate the pharmacokinetic parameters of pOBz. Results The linear regression equation of pOBz in plasma was: Y = 0.062 3X + 0.024 8 , r2=0.999, and the linear range was 0. 2 ~ 200 mg • L-1; the linearity regression equation in brain homogenate was : Y = 0.065 1X + 0.0317 , r2= 0.997, and the linear range was 0.2 ~ 200 mg • L -1. The half-life T1/2 of pOBz in plasma was 1.03 h,the area under the drug-time curve AUC0_twas 198.4 h • mg • L -1the mean residence time MRT0_twas 0.87 h, and the clearance rate CL was 0. 246 L • h-1• kg-1. 5 min after administration, pOBz could be detected in brain tissue at a concentration of 1.21 mg • L-1; it was not completely eliminated 3 h after administration. Conclusions The UPLC-MS/MS method is accurate and applicable for the pharmacokinetic study of pOBz in plasma and brain homogenate. pOBz can penetrate the blood-brain barrier and exert pharmacological effects in central nervous system. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

大黄对MCAO模型大鼠的神经保护作用

目的研究大黄提取物对MCAO模型大鼠的神经保护作用,并探讨其作用机制。方法SPF级雄性SD大鼠45只,随机分为sham组、MCAO组、MCAO+大黄组,线栓法制备MCAO模型,大黄提取物给药浓度为200 mg·kg^-1·d^-1,连续给药6 d,尼氏染色法检测尼氏体的表达,免疫荧光法检测大鼠缺血侧NeuN、NF200的表达,RT-qPCR法检测Egr1、Egr2、Egr4 mRNA表达,Western blot检测NGF及BDNF蛋白的表达。结果与MCAO组相比,大黄提取物作用后,尼氏染色法检测结果表明大黄提取物可以减少MCAO模型大鼠脑梗死面积,免疫荧光检测结果表明大黄提取物可以升高MCAO模型大鼠缺血侧NeuN、NF200的表达;经RT-qPCR检测,大黄提取物能够提高Egr1、Egr2、Egr4 mRNA基因表达;经Western blot检测,大黄提取物能够提高NGF及BDNF蛋白的表达。结论大黄提取物能够改善MCAO模型大鼠缺血侧神经元存活环境,具有神经保护作用。     

红景天苷调控PI3K/Akt信号通路对永久性脑缺血大鼠的保护作用

目的研究红景天苷(salidroside,Sal)通过调控PI3K/AKT信号通路对永久性大脑中动脉闭塞模型(pMCAO)大鼠的神经保护作用及其机制。方法健康成年雄性SPF级雄性SD大鼠70只,其中30只大鼠随机分为假手术组(sham组)、模型组(pMCAO组)、红景天苷给药组(pMCAO+Sal组),线栓法造模成功后,对sham组和pMCAO组大鼠腹腔注射生理盐水,pMCAO+Sal组按照50 mg·kg^-1·d^-1腹腔注射红景天苷(50 mg·kg^-1·d^-1),给药1 d。40只大鼠随机分为假手术组(sham组)、模型组(pMCAO组)、红景天苷给药组(pMCAO+Sal组)、抑制剂组(pMCAO+Sal+LY组);侧脑室注射PI3K抑制剂30 min后,制备pMCAO模型,1 h后腹腔注射红景天苷,给药1 d。免疫组织化学染色测定脑组织中NeuN表达,Western blot分析脑组织中NeuN蛋白、NF-κBp50核蛋白的表达,RT-qPCR检测IL-6和TNF-α的表达。结果与pMCAO组比较,红景天苷作用后,NeuN免疫荧光染色表达量明显增加,且能够促进NeuN蛋白,抑制NF-κB p50核蛋白及IL-6和TNF-αmRNA表达。经PI3K抑制剂LY294002作用后,红景天苷对上述指标作用不明显。结论红景天苷能够通过调控PI3K/AKT信号通路,抑制NF-κB p50核转录水平,进而抑制炎症因子,对pMCAO模型大鼠具有神经保护的作用。     

红景天苷衍生物对苯甲酰红景天苷对MCAO大鼠的神经保护作用

目的研究红景天苷衍生物对苯甲酰红景天苷(p-benzoyl salidroside, pOBz)对大脑中动脉闭塞模型(MCAO)大鼠的神经保护作用。方法 (1)30只健康成年♂SD大鼠,随机分为Sham组、MCAO组、MCAO+pOBz低中高剂量组(25、50、100 mg·kg^-1),线栓法造模,进行神经功能损伤评分,连续给药2 d。采用核磁共振成像检测MCAO大鼠脑梗死体积。(2)24只健康成年♂SD大鼠,随机分为Sham组、MCAO组、MCAO+pOBz组(50 mg·kg^-1)和MCAO+Sal组(50 mg·kg^-1),线栓法造模,给药1 d。Western blot检测NeuN、EGR1、Bcl-2和Bax蛋白表达。(3)18只健康成年♂SD大鼠,随机分成Sham组、MCAO组和MCAO+pOBz组(50 mg·kg^-1),连续给药2 d。免疫荧光染色观察NeuN的表达。结果 pOBz给药2 d可以降低MCAO大鼠的脑梗死体积,改善神经功能损伤,且升高NeuN和EGR1表达程度优于Sal, pOBz对MCAO大鼠脑组织Bcl-2/Bax比值的改善程度与Sal相当。结论 pOBz可改善MCAO大鼠脑梗死体积,具有优于红景天苷的神经保护作用。     

The neuroprotective effect of salidroside prophylactic administration on MCAO model rats北大核心CSCD

Aim To investigate the neuroprotective effect of prophylactic administration of salidroside (Sal) on MCAO rats. Methods A total of 52 SD adult male rats were randomly divided into sham operation group (Sham), model group (MCAO) and salidroside pre-administration group (MCAO + Sal). The dose of Sal was 50 mg·kg-1. MCAO model was administered by continuous intraperitoneal injection for 7 days, followed by the wire embolization 0.5 h after the 7th day of administration, and the materials were taken 24 h after anesthesia. TTC staining was used to detect the volume of cerebral infarction in rats, TUNEL staining was used to detect the number of nerve cell apoptosis, Western blot was used to detect the expression of Bax, Bcl-2, cleaved Caspase-3 and total Caspase-3 proteins, naphthol AS-D chloroacetate was used to detect the expression of neutrophils in brain tissues, RT-qPCR was used to detect the mRNA expression of IL-1β, IL-6 and TNF-α, and immunofluorescence staining was used to determine the expression of NeuN in brain tissues. Results Pre-administration of salidroside for seven days significantly reduced cerebral infarct volume and nerve cell apoptosis in MCAO rats, promoted the expression of Bcl-2 protein, inhibited the expression of Bax and cleaved Caspase-3 proteins, but it had no significant effect on total Caspase-3 protein, and it reduced the recruitment of neutrophils in ischemic brain tissues, decreased the mRNA expression of inflammatory factors IL-1β, IL-6, TNF-α, and up-regulated the expression of NeuN. Conclusions Pre-administration of salidroside for seven days can significantly reduce the volume of cerebral infarction in MCAO rats, inhibit nerve cell apoptosis, reduce inflammation and promote the expression of NeuN, then playing a neuroprotective role. © 2023 Publication Centre of Anhui Medical University. All rights reserved.