Effects of L-Tetrahydropalmatine on Energy Metabolism,Endothelin-1 and NO during Acute Cerebral Ischemia-reperfusion of Rats

To investigate the effects of L-tetrahydropalmatine (L-THP) on ener-gy metabolism, endothelin-1 (ET-1 ) and NO during acute cerebral ischemia-reperfusion of rats, 24 Wistar rats were randomly divided into four groups, with 6rats in each group: sham-operation group, simple ischemia group, ischemia-reperfusion group and treatment group (L-THP group). Cerebral ATP, lactate,ET-1 and NO levels were measured in all groups. Our results showed that treat-ment with L-THP could increase cerebral ATP levels, but decrease cerebral lac-tate, ET-1 and NO concentrations during ischemia-reperfusion in the treatmentgroup. It is concluded that L-THP could improve cerebral energy metabolism and protect the injured brain tissue, the mechanism of which might be related to suppression of overproduction of ET-1 and NO.

Effects of L-Tetrahydropalmatine on Neuron Apoptosis during Acute Cerebral Ischemia-Reperfusion of Rats

To investigate the effects of L-Tetrahydropalmatine (L-THP ) on neuron apoptosis during acute cerebral ischemia-reperfusion of rats and explore the effects of heat shock protein (HSP) on neuron apoptosis, Wistar rats were randomly divided into 3 groups: normal group, ischemia- reperfusion group and treatment group. The condition of neuron apoptosis, the survival state of neuron, pathological changes under an electron microscope and the number of HSP70 positive cells were measured in all groups. Results showed that the apoptosis neuron number was increased obviously at the 24th h during reperfusion and was further increased at the 48th h, the 72th h. While the number of survival neurons was decreased gradually with the prolongation of reperfusion time. Treatment with L-THP could decrease the apoptosis neuron number but increase the survival neuron number and the HSP70 positive cell number. Our study suggested that L-THP could decrease apoptosis and necrosis of neuron, up-regulate the expression of HSP70 and protect the cerebral ischemic injury.

Effects of L-Tetrahydropalmatine on the Expressions of bcl-2 and bax in Rat after Acute Global Cerebral Ischemia and Reperfusion

To investigate the effects of L-Tetrahydropalmatine (L-THP) on the expressions of bcl-2, bax and neuronal apoptosis after cerebral ischemia and reperfusion, 60 Wistars rats were randomly divided into 3 groups: sham-operation group (group S, n = 20), ischemic-reperfusion group treated with saline (group I, n=20) and ischemia-reperfusion group treated with L-THP (group T, n=20) .The rat model of global cerebral ischemia and reperfusion was induced by Pulsinelli's four-vessel occlusion method. The expression of bcl-2 and bax mRNA was detected by in situ hybridization and reverse transcriptional polymerase chain reaction (RT-PCR). The number of apoptotic neurons was examined by terminal deoxynucleotidyl-transferase (TdT)-mediated dUTP nick end-labeling (TUNEL) method. Compared with group S, the expression of bcl-2 and bax mRNA in group I was increased significantly (P<0.01), and the number of apoptotic neurons increased either (P< 0.01). After L-THP treatment, the expression of bcl-2 mRNA was up-regulated (P<0.01) and that of bax mRNA was down-regulated (P<0.01); the number of apoptotic neurons was decreased (P<0.01). Our results indicated that bcl-2 may suppress apoptosis and bax promote apoptosis after cerebral ischemia and reperfusion. L-THP could ameliorate cerebral ischemia and reperfusion damage by reducing the apoptosis through regulating bcl-2 and bax.

ELECTROPHYSIOLOGICAL EFFECTS OF TETRAHYDROPALMATINE ON CANINE PURKINJE FIBER AND VENTRICULAR MUSCLE IN VITRO

The cellular electrophysiologic properties of a new antiarrkythmic substance,tetrahydropalmatine(TRP),were studied in isolated canine cardiac Purldnje fibers and ventricular muscles.In lharkinjc fibers,the shortening of APD50 produced by THP was concentration dependent,APD90was shortened only at higher concentrations of THP,APA and Vmax were decreased by increasing theTHP concentration to 100 and 300 μmol/L. In Veutricular muscles,THP,at higher Concentration,shortened APDs,and APD90 The reductions of Vmax and APA were significant only at highest concentration,300 μmol/L,ERP was relatively prolonged in both Purldnje fibers and ventricular muscles.These studies suggest that THP may inhibit Ca2+ and Na+ movement across the membrane of myocardlal cells.

Effect of tetrahydropalmatine analogs on Fos expression induced by formalin-pain

目的：研究四氢巴马汀（ＴＨＰ）同类物对福尔马林致痛诱导的Ｆｏｓ蛋白表达的影响，以阐明ＴＨＰ同类物的镇痛机制．方法：在右后肢脚掌皮下注射５％福尔马林５０μＬ，诱发炎性疼痛，用免疫组织化学方法观察Ｆｏｓ蛋白表达．结果：腹腔注射ＴＨＰ同类物和Ｄ２受体拮抗剂螺哌隆诱导的Ｆｏｓ蛋白表达主要位于纹状体和伏膈核．Ｄ２受体激动剂喹吡罗可阻滞ｌＴＨＰ和螺哌隆诱导的Ｆｏｓ蛋白表达．ＴＨＰ同类物明显增加脑干下行痛觉调制系统的Ｆｏｓ蛋白表达，并能明显抑制福尔马林诱导的脊髓背角浅层和深层的Ｆｏｓ蛋白表达．结论：ＴＨＰ同类物通过阻滞纹状体和伏膈核的Ｄ２受体，加强脑干下行痛觉调制系统的功能，抑制外周痛觉信息在脊髓水平的传入，达到它们的镇痛作用．

Network Pharmacology and Experimental Validation to Explore Mechanism of Tetrahydropalmatine on Acute Myocardial Ischemia

Objective:To explore the potential molecular mechanism of tetrahydropalmatine(THP)on acute myocardial ischemia(AMI).Methods:First,the target genes of THP and AMI were collected from SymMap Database,Traditional Chinese Medicine Database and Analysis Platform,and Swiss Target Prediction,respectively.Then,the overlapping target genes between THP and AMI were evaluated for Grene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis and protein-protein interaction network analysis.The binding affinity between the protein and THP was assessed by molecular docking.Finally,the protective effects of THP on AMI model and oxygen and glucose deprivation(OGD)model of H9C2 cardiomyocyte were explored and the expression levels of target genes were detected by RT-qP CR in vivo and in vitro.Results:MMP9,PPARG,PTGS2,SLC6A4,ESR1,JAK2,GSK3B,NOS2 and AR were recognized as hub genes.The KEGG enrichment analysis results revealed that the potential target genes of THP were involved in the regulation of PPAR and hormone pathways.THP improved the cardiac function,as well as alleviated myocardial cell damage.Furthermore,THP significantly decreased the RNA expression levels of MMP9,PTGS2,SLC6A4,GSK3B and ESR1(P<0.05,P<0.01)after AMI.In vitro,THP significantly increased H9C2 cardiomyocyte viability(P<0.05,P<0.01)and inhibited the RNA expression levels of PPARG,ESR1 and AR(P<0.05,P<0.01)in OGD model.Conclusions:THP could improve cardiac function and alleviate myocardial injury in AMI.The underlying mechanism may be inhibition of inflammation,the improvement of energy metabolism and the regulation of hormones.

Tetrahydropalmatine protects against methamphetamine-induced spatial learning and memory impairment in mice

Objective The purpose of this study was to investigate the effect of methamphetamine （MA） on spatial learning and memory and the role of tetrahydropalmatine （THP） in MA-induced changes in these phenomena in mice. Methods Male C57BL/6 mice were randomly divided into eight groups, according to different doses of MA, different doses of THP, treatment with both MA and THP, and saline controls. Spatial learning and memory were assessed using the Morris water maze. Western blot was used to detect the expression of extracellular signal-regulated protein kinase （ERK） in the mouse prefrontal cortex （PFC） and hippocampus. Results Repeated MA treatment significantly increased the escape latency in the learning phase and decreased the number of platform site crossings in the memory-test phase. ERK1/2 expression was decreased in the PFC but not in the hippocampus of the MA-treated mice. Repeated THP treatment alone did not affect the escape latency, the number of platform site crossings or the total ERK1/2 expression in the brain. Statistically significantly shorter escape latencies and more platform site crossings occurred in MA＋THP-treated mice than in MA-treated mice. Conclusion Repeated MA administration impairs spatial learning and memory in mice, and its co-administration with THP prevents this impairment, which is probably attributable to changed ERK1/2 expression in the PFC. This study contributes to uncovering the mechanism underlying MA abuse, and to exploring potential therapies.

白芥子穴位给药配伍对延胡索乙素药效动力学影响的研究

目的:探讨“冬病夏治”全方配伍和无白芥子配伍延胡索乙素在模型家兔“肺俞”穴皮下药代动力学特征及药代动力学-药效动力学(PK-PD)模型的相关性。方法:支气管哮喘模型家兔随机分成延胡索单方组、缺白芥子组、全方组,微透析技术收集14 h穴位皮下透析液,液相色谱-质谱法(Liquid Chromatography Mass Spectrometry,LCMS)法检测方中君药延胡索主要成分延胡索乙素浓度,获得药代动力学参数;酶联免疫吸附试验(ELISA)法检测对应时间点模型动物血清中IgE水平,获得药效学参数;对药动学、药效学参数进行PK-PD模型拟合。结果:白芥子配伍后的药峰浓度(C_(max))、药时曲线下面积(AUC_(0-t))、平均滞留时间(MRT_(0-t))均显著增加(P<0.01,P<0.01,P<0.05),达峰时间(T_(max))提前(P<0.01);“浓度-时间-效应”三维曲线表明,方中有白芥子配伍时,药效出现更快、消退更慢,起效时间晚于峰浓度,具有一定滞后性。结论:动力学参数、PK-PD模型结果表明,白芥子配伍能够改变“方中君药”——延胡索的主要成分延胡索乙素穴位局部的皮下分布,促进方中君药有效成分快速吸收,延长滞留时间,在方剂中起到主药、改善其他药物分布的“双重”作用。

延胡索叶片愈伤组织诱导及其2种活性成分含量测定

目的:以叶片为外植体诱导延胡索(Corydalis yanhusuo)形成愈伤组织,测定愈伤组织生长指标,检测生物碱类成分含量,为研究延胡索生物碱代谢调控机制和培育优良品系提供依据。方法:运用2,4-二氯苯氧乙酸(2,4-D)与6-苄基氨基嘌呤(6-BA)诱导形成愈伤组织,采用重量法测定生物量,组织化学染色法定位生物碱,高效液相色谱法测定延胡索乙素和脱氢紫堇碱含量。结果:延胡索叶片经MS+2 mg/L 2,4-D+1 mg/L 6-BA处理,愈伤组织诱导率可达75%;外观呈黄白色,表面呈颗粒状,生长迅速;继代培养的愈伤组织鲜重(2.7153±0.4398)g,增殖倍数4.43;干重(0.1552±0.0207)g,含水量94.16%。愈伤组织薄壁细胞内含能被碘化铋钾染色的橘黄色斑块,提示生物碱的存在;愈伤组织中延胡索乙素含量达(0.8600±0.3502)mg/g DW(干重),脱氢紫堇碱含量为(0.1744±0.0372)mg/g DW。结论:利用叶片作为外植体获得的延胡索愈伤组织诱导率高、生长迅速,是快速、稳定获取延胡索乙素等生物碱类活性成分的适宜材料。

HPLC法同时测定冠心安口服液中蒙花苷、延胡索乙素和2,3,5,4’-四羟基二苯乙烯-2-O-β-D-葡萄糖苷的含量

目的建立能同时测定冠心安口服液中蒙花苷、延胡索乙素和2,3,5,4’-四羟基二苯乙烯-2-O-β-D-葡萄糖苷含量的HPLC分析方法。方法Capcell Pak UG C_(18)色谱柱分离;蒙花苷、延胡索乙素和2,3,5,4’-四羟基二苯乙烯-2-O-β-D-葡萄糖苷检测波长分别设定为334、281、320 nm;柱温为室温;进样体积为10μL;以乙腈为流动相A,1 mL·L^(−1)磷酸溶液(三乙胺调节pH至6.0)为流动相B,梯度洗脱。结果蒙花苷、延胡索乙素和2,3,5,4’-四羟基二苯乙烯-2-O-β-D-葡萄糖苷的质量浓度分别在0.0245~2.4500、0.0457~4.5700、0.0474~4.7400μg·mL^(−1)范围内线性良好;平均回收率分别为100.8%、99.9%、101.4%;精密度和重复性RSD值(n=6)均在5.0%以内;供试品溶液在24 h内稳定。结论此方法具有前处理简单、分析时间短和检测结果准确等优点,适用于冠心安口服液制剂的质量控制。

延胡索乙素纳米混悬剂制备及体外评价

目的制备延胡索乙素纳米混悬剂(THP-NS)及其冻干粉,并对其进行体外评价。方法通过沉淀-均质法制备延胡索乙素纳米混悬剂,在单因素实验基础上,以粒径与多分散系数(PDI)的归一值作为考察指标,考察延胡索乙素用量、透明质酸钠用量、泊洛沙姆407用量对纳米混悬剂制备的影响,通过Box-Behnken设计响应面法优化THP-NS的处方工艺,并制备冻干粉末。采用透射电镜法(TEM)、差示扫描量热法(DSC)、X射线衍射(XRD)、傅里叶变换红外光谱法(FT-IR)对THP-NS进行表征,透析袋法评价其体外溶出度。结果延胡索乙素纳米混悬剂最佳处方为延胡索乙素用量51.0 mg,透明质酸钠用量3.1 mg,泊洛沙姆407用量18.4 mg。平均粒径为(195.57±1.80)nm,PDI为0.23±0.019,与Box-Behnken响应面法预测值接近。Zeta电位值为-31.0 mV,延胡索乙素纳米混悬剂外貌为球形,THP-NS中延胡索乙素以结晶状态存在,THP-NS在24 h累积溶出度达到97.43%。结论将延胡索乙素制备成纳米混悬剂药物处方设计合理、工艺可行,显著提高了其体外溶出率。

参连肠炎丸中4种活性成分含量测定

目的利用高效液相色谱切换波长的方法同时测定参连肠炎丸中甘草苷、延胡索乙素、橙皮苷和盐酸小檗碱含量。方法色谱柱为Agilent Eclipse XDB-C_(18)(5μm,4.6 mm×250 mm),流动相为乙腈-0.2%磷酸,进行梯度洗脱,流速为1.0 ml/min,甘草苷、延胡索乙素、橙皮苷和盐酸小檗碱的检测波长和切换时间分别为237 nm(0~7 min)、283 nm(7~12 min)、283 nm(7~12 min)、345 nm(12~20 min)。结果甘草苷、延胡索乙素、橙皮苷和盐酸小檗碱4种成分良好分离,分离度均在1.5以上。4种成分线性关系良好,平均回收率分别为100.19%,99.32%,99.64%,100.08%,RSD分别为1.78%,1.69%,1.89%,2.04%(n=6)。结论本法操作简单、结果准确,回收率高,重复性好,可为参连肠炎丸质量的有效控制提供一定的技术支撑。

一测多评法同时测定蒲元和胃胶囊中7种成分含量

目的建立同时测定蒲元和胃胶囊中延胡索乙素、盐酸巴马汀、α-香附酮、咖啡酸、木犀草素、甘草苷、甘草酸铵的含量的一测多评法。方法采用高效液相色谱法测定,色谱柱为Diamonsil C_(18)柱(250 mm×4.6 mm,5µm),流动相为乙腈-0.1%磷酸溶液(梯度洗脱),流速为1.0 mL/min,柱温为30℃,检测波长为237 nm,进样量为10µL。以木犀草素为内参物,建立与延胡索乙素、盐酸巴马汀、α-香附酮、咖啡酸、甘草苷、甘草酸铵的相对校正因子;采用一测多评法和外标法检测7种成分的含量,比较计算值与实测值的差异。结果延胡索乙素、盐酸巴马汀、α-香附酮、咖啡酸、木犀草素、甘草苷、甘草酸铵的质量浓度分别在2.67~133.41µg/mL、0.89~44.60µg/mL、1.36~67.85µg/mL、1.53~76.65µg/mL、2.00~100.12µg/mL、3.00~150.20µg/mL、8.51~425.61µg/mL范围内与峰面积线性关系良好(r≥0.9995,n=6);精密度、稳定性、重复性试验结果的RSD均小于2.0%(n=6);平均加样回收率分别为99.09%,97.34%,97.42%,96.85%,98.60%,99.20%,99.09%,RSD分别为0.72%,1.68%,1.75%,1.29%,0.69%,0.79%,0.97%(n=9)。延胡索乙素、盐酸巴马汀、α-香附酮、咖啡酸、木犀草素、甘草苷、甘草酸铵的相对校正因子分别为1.3159,0.7059,1.1805,1.2241,1.2385,6.7178;在不同色谱柱、色谱仪、流速、柱温下的重复性均良好,结果的RSD均低于1.50%。一测多评法与外标法的含量计算结果相对误差的绝对值均小于2.0%。结论该方法重复性好、结果准确可靠,可用于蒲元和胃胶囊的质量控制。

延胡索乙素改善PDGF-BB诱导的VSMCs氧化应激损伤的机制

目的研究延胡索乙素(Thp)对血小板衍生生长因子-BB(PDGF-BB)诱导的大鼠主动脉血管平滑肌细胞(VSMCs)氧化应激损伤的保护作用,并基于核因子红系2相关因子2(Nrf2)/血红素加氧酶1(HO-1)信号通路探究其可能机制。方法在Thp抑制PDGF-BB诱导的VSMCs氧化应激损伤效应研究中,将VSMCs分为对照组、PDGF-BB组(25 ng/mL)及Thp低、中、高浓度组(5、10、20 mg/mL)。在Thp作用机制研究(沉默Nrf2)中,将VSMCs分为PDGF-BB+阴性对照siRNA(NC-siNrf2)组(25 ng/mL PDGFBB+NC-siNrf2),PDGF-BB+Thp+NC-siNrf2组(25 ng/mL PDGF-BB+10 mg/mL Thp+NC-siNrf2),PDGF-BB+Nrf2小干扰RNA(siNrf2)组(25 ng/mL PDGF-BB+siNrf2),PDGF-BB+Thp+siNrf2组(25 ng/mL PDGF-BB+10.0 mg/mL Thp+siNrf2)。2个实验均检测VSMCs的增殖、迁移能力,活性氧(ROS)水平,超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性以及Nrf2和HO-1蛋白表达。结果与对照组比较,PDGF-BB组VSMCs的增殖、迁移能力显著增强(P<0.01),ROS水平显著升高(P<0.01),SOD、CAT活性及Nrf2、HO-1蛋白的相对表达量均显著降低(P<0.01);与PDGF-BB组比较,Thp不同浓度组VSMCs的增殖、迁移能力均显著下降(P<0.01),ROS水平均显著降低(P<0.01),SOD、CAT活性及Nrf2、HO-1蛋白的相对表达量均显著升高(P<0.01)。沉默Nrf2可显著逆转Thp对PDGF-BB诱导VSMCs氧化应激损伤的改善作用(P<0.01)。结论Thp可以通过激活Nrf2介导的抗氧化防御途径来降低VSMCs的氧化应激水平,从而抑制VSMCs的增殖、迁移。

延胡索乙素的药理作用研究进展

延胡索是一种常见的中药材,具有活血散瘀、理气止痛的功效。在临床治疗上,延胡索作为一种常见的镇痛药,具有镇痛及催眠作用,常用于内科病痛(如消化性溃疡痛)、产前阵痛、产后子宫缩痛、月经痛、头痛及失眠等症。延胡索的化学成分主要包括生物碱类、萜类、氨基酸类、多糖类、有机酸类及挥发油类等。延胡索乙素是延胡索的主要有效化学成分,对中枢神经系统、心血管系统以及消化系统具有显著作用,可用于止痛镇静、抗药物成瘾、抗心律失常、抗肿瘤、改善血流动力学及降血脂等。本文对延胡索乙素的药理进行总结,为进一步研究此药材提供参考。

复方延胡索乙素止痛凝胶成型性工艺优化研究

目的:制备复方延胡索乙素止痛凝胶(Compound Tetrahydropalmatine Pain Relief Gel,CTPRG),并对其进行质量评价。方法:CTPRG以卡波姆940为基质,将外观、黏度、涂展性、离心稳定性、低温稳定性、高温稳定性作为综合考察指标,采用单因素试验结合正交实验优化处方。结果:所得CTPRG最佳处方为1.00 g卡波姆940,20 g丙二醇,8 g甘油,0.50 g三乙醇胺。产品外观如果冻,表面光滑、均匀、细腻;离心、高温、低温稳定性均良好,无分层、析出药液现象。结论:CTPRG外观良好、质量稳定,研究成果可为元胡止痛方的外用制剂开发利用提供数据基础。

夏天无总碱中抗胆碱酯酶活性成分的研究

夏天无为罂粟科植物伏生紫堇Corydalis decumbens (Thunb.) Pers.的干燥块茎,是我国常用中草药,有活血通络、行气止痛之功效,用于中风偏瘫、跌打损伤、风湿性关节炎和坐骨神经痛等[1,2],现代药学研究认为其主要有效部位为生物碱[2,3].

延胡索乙素在大鼠体内的立体选择性药代动力学

目的研究dl延胡索乙素在大鼠体内的立体选择性动力学过程。方法血浆样品经液液萃取后,用非手性手性HPLC联用法测定血浆中延胡索乙素对映体的浓度,配对t检验比较两对映体的血药浓度和药代动力学参数。结果大鼠血浆中l延胡索乙素的浓度均明显高于d延胡索乙素的浓度,两对映体的AUC,MRT和Cmax均有显著性差异。结论大鼠灌胃延胡索乙素消旋体后,两对映体在体内的药代动力学过程具有显著的立体选择性。

不同产地延胡索饮片炮制前后有效成分含量的测定

目的：比较不同产地延胡索生品和炮制品中延胡索乙素、含水量及醇溶性浸出物的含量。方法：将14批不同产地的延胡索生品，按照中国药典醋煮法进行炮制。采用高效液相色谱法测定生品和炮制品中延胡索乙素的含量，并对含水量及醇溶性浸出物的含量进行考察。结果：14批延胡索饮片生品的水分含量、醇溶性浸出物及延胡索乙素的含量分别在12．8％～14．8％、16．5％-19．6％和0．043％～0．075％；延胡索经醋煮后，水分含量、醇溶性浸出物及延胡索乙素的含量分别在7．58％-11．7％、17．8％-23．5％和0．046％～0．079％。结论：由于各地栽培延胡索土壤、气候等差异，不同产地延胡索炮制前后延胡索乙素、含水量及醇溶性浸出物的含量有所不同。

苏元胡的生物碱成分研究

应用多种色谱方法从苏元胡Corydalis humosa Migo的块茎的乙醇提取物中分离得到6个化合物,它们的结构分别被鉴定为:四氢巴马汀(1)、药根碱(2)、小檗碱(3)、四氢小檗碱(4)、巴马汀(5)及(-)-四氢黄连碱(6).化合物1～3为从本植物中首次分离得到.