梓醇促进大鼠骨髓间充质干细胞增殖过程中Wnt信号通路的变化

 目的：观察梓醇促进大鼠骨髓间充质干细胞（bone marrow mesenchymal stem cells, BMSCs）增殖过程中Wnt信号通路的变化。方法：（1）采用机械分离及差速贴壁法分离培养SD大鼠BMSCs,将细胞分为空白对照组与梓醇（1.0 mg/L）处理组,流式细胞术检测各组细胞细胞周期分布情况,计算其增殖指数。（2）实时荧光定量PCR法检测各组细胞中Wnt3a、Wnt5a、Wnt11及β-catenin mRNA的表达水平;Western blotting技术检测各组细胞β-catenin蛋白的表达变化。结果：（1）空白对照组与梓醇处理组BMSCs增殖指数分别为8.90%±0.46%和17.93%±1.68%（P<0.01）。(2）与空白对照组相比,梓醇处理组Wnt5a、Wnt11、β-catenin mRNA及β-catenin蛋白表达均明显提高,但Wnt3a mRNA表达无显著变化。结论：梓醇在促进BMSCs增殖过程中可同时激活经典与非经典Wnt信号通路。     

HPLC测定清热达郁口服液中梓醇的含量

目的 建立测定清热达郁口服液中梓醇含量的方法.方法 采用HPLC法,测定清热达郁口服液中的梓醇含量,Diamonsil C18(250 mm ×4.6 mm,5μm)柱,流动相乙腈-0.1%磷酸溶液(1:99),流速1.0 ml·min-1,检测波长210 nm.结果 梓醇在1.1～8.8 μg范围内线性关系良好(r=0.9999),其平均回收率为98.80%(RSD=1.2%,n=5).结论 所建方法简便、准确、可靠,可用于清热达郁口服液的质量控制.     

正交实验优选地黄叶中梓醇的提取工艺

目的对干地黄叶中梓醇的提取方法进行优选，确定最佳提取工艺。方法采用正交实验表L8（2）^7安排实验。结果提取方法对梓醇提取率影响最大。结论确定地黄叶中梓醇的最佳提取工艺，即加水煎煮提取，提取次数为2次，1h／次。     

反相高效液相色谱法检测咽喉康口服液中梓醇的含量

用反相ＨＰＬＣ以水－乙腈（９６：４）为流动相，检测波长２１０ｎｍ，对咽喉康口服液中梓醇含量进行了检测，其保留时间为５．９６０分，平均加样回收率为９９．１２％，ＲＳＤ为１．４０％。本检测方法简便快速，准确可靠。     

Catalpol improves cholinergic function and reduces inflammatory cytokines in the senescent mice induced by d-galactose

The neuroprotective effects of catalpol, an iridoid glycoside isolated from the fresh rehmannia roots, on the cholinergic system and inflammatory cytokines in the senescent mice brain induced by d-galactose were assessed. The results showed that acetylcholinesterase (AChE) activity increased in senescent mice brain and choline acetyltransferase (ChAT) positive neurons, detected by immunohistochemical staining, decreased remarkably in the basal forebrain of senescent mice. Simultaneously, muscarinic acetylcholine receptor M1 (mAChR1) expression declined in senescent mice brain by western blotting method. We also found that the contents of tumor necrosis factor (TNF-α), interleukin-1β (IL-1β) and advanced glycation endproducts (AGEs) increased in senescent mice brain by ELISA method. However, administration of catalpol for 2-weeks significantly reversed the biochemical markers mentioned above. These results suggest that catalpol can exert protective effects on senescent mice brain induced by d-galactose and this effect may be due to its protective effects on cholinergic and immune impairment in mice brain. Thus catalpol is worth testing for further preclinical study aimed for senescence or neurodegenerative diseases such as Alzheimer’s disease.     

RP-HPLC法测定天王补心丸中梓醇的含量

目的:建立天王补心丸中梓醇含量的测定方法。方法:采用反相高效液相色谱法。色谱柱为Agilent-ZORBX SB-C18柱,流动相为乙腈-0.1%磷酸溶液(V/V)=1:99,流速为1.0mL.min-1,检测波长为210nm,进样量为20μL,柱温为40℃。结果:梓醇检测浓度在2.004～20.046μg.mL-1范围内与其峰面积积分值呈良好的线性关系(r=0.9993);平均加样回收率为99.9%,RSD=2.2%(n=9)。结论:该法简便、准确、重复性好,适用于天王补心丸的质量控制。     

梓醇在大鼠体内的药代动力学和生物利用度研究

目的:研究梓醇在大鼠体内的药代动力学和生物利用度。方法:建立LC-MS/MS分析方法,应用Xcalibur 1.4数据处理工作站,以待测物(梓醇)与内标物(桃叶珊瑚苷)的色谱峰面积比计算大鼠灌胃50、100和200mg/kg和静脉注射50mg/kg梓醇后,各时间的血浆药物浓度。选用DAS 2.0软件计算药动学参数,t检验法统计实验数据,计算生物利用度。结果:梓醇在大鼠体内的药动学过程符合二室模型,大鼠灌胃50、100和200mg/kg梓醇后,AUC0→∞与剂量呈正比,t1/2与剂量无关;大鼠灌胃与静注50mg/kg梓醇后,AUC0→∞分别为(70±23)、(104±11)μg.h.mL-1,该剂量下梓醇的绝对生物利用度为66.7%。结论:梓醇的吸收和消除呈一级动力学特征,在大鼠体内的绝对生物利用度较高。     

基于谱效相关的生地黄多指标成分定量分析方法的研究

目的建立谱效相关的生地黄多指标定量分析方法。方法采用高效液相色谱法建立生地黄止血作用指纹图谱,以梓醇为对照品,对达到基线分离的色谱峰进行定量,计算出各指纹峰相对于梓醇的含量。结果确定出13个指纹峰,计算其相对于梓醇的含量。结论所建方法简便、准确、可靠,可作为生地黄止血作用的质量控制方法。     

乌梅散颗粒的质量标准研究

目的建立乌梅散颗粒（乌梅、熟地、麦冬、炙甘草）的质量标准。方法采用薄层色谱法（TLC）对乌梅、熟地、麦冬、炙甘草进行鉴别;采用高效液相色谱法测定乌梅散中梓醇的含量。色谱条件为Synergi（4.6 mm×250mm,5μm）谱柱;流动相为乙腈-水（2∶98）;检测波长为206nm;流速为1.0ml/min;柱温为室温。结果建立了乌梅、熟地、麦冬、炙甘草的薄层定性鉴别的方法;建立了HPLC测定乌梅散中梓醇含量的方法,梓醇在1.0-40μg范围内线性关系良好,梓醇的平均回收率为99.48%,RSD为2.57%。结论建立的乌梅散颗粒鉴别及梓醇含量测定方法简单、易于操作、准确度高、重现性好,可用于乌梅散颗粒的质量控制。     

Catalpol protects primary cultured astrocytes from in vitro ischemia-induced damage

Catalpol, an iridoid glycoside abundant in the roots of Rehmannia glutinosa, has been previously found to prevent the loss of CA1 hippocampal neurons and to reduce working errors in gerbils after ischemia-reperfusion injury. In the present study, we investigated the effects of catalpol on astrocytes in an ischemic model to further characterize its neuroprotective mechanisms. Primary cultured astrocytes exposed to oxygen-glucose deprivation (OGD) followed by reperfusion (adding back oxygen and glucose, OGD-R), were used as an in vitro ischemic model. Treatment of the astrocytes with catalpol during ischemia-reperfusion increased astrocyte survival significantly in a concentration-dependent manner, as demonstrated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, lactate dehydrogenase (LDH) release and morphological observation. In addition, catalpol prevented the decrease in mitochondrial membrane potential, inhibited the formation of reactive oxygen species (ROS) and the production of nitric oxide (NO), decreased the level of lipid peroxide and the activity of inducible nitric oxide synthase (iNOS), and elevated the activities of superoxide dismutase (SOD), glutathione peroxidase (GPx) and the content of glutathione (GSH). Our results suggest that catalpol exerts the most significant cytoprotective effect on astrocytes by suppressing the production of free radicals and elevating antioxidant capacity.