去氢骆驼蓬碱诱导人胃腺癌SGC-7901细胞凋亡

目的：探讨去氢骆驼蓬碱（Harmine，HM）诱导人胃腺癌SGC-7901细胞凋亡的作用。方法：MTT法检测HM对该细胞系的生存抑制率，光镜下观察凋亡细胞，流式细胞仪（FCM）检测凋亡率，基因组DNA琼脂糖凝胶电泳（DNALadder）检测凋亡梯状条带。结果：HM处理SGC-7901细胞后，细胞的存活率明显降低；细胞形态观察到有核固缩与核断裂的凋亡现象；流式细胞仪测定细胞周期的G1期前有亚二倍体的凋亡峰；基因组DNA琼脂糖凝胶电泳检测到明显的凋亡梯状条带。结论：HM能诱导人胃腺癌SGC-7901细胞的凋亡。     

The effects of harmine on the transmembrane action potentials of guinea-pig left atria as compared to those of quinidine

Summary The actions of quinidine (7.7 and 25.5 M) and harmine (10,33 and 66 M) on the action potential of isolated guinea-pig atria were studied. Both substances prolonged repolarization time dose-dependently. The depolarization velocity was considerably reduced by quinidine, while it was comparatively little affected by harmine. Antiarrhythmic compounds of the harmine type are rare.The effects on the action potential explain the differences between quinidine and harmine with regard to effects on the ECG and antagonism of aconitine-induced arrhythmia.     

Cytotoxicity of the β-carboline alkaloids harmine and harmaline in human cell assays in vitro

beta-Carboline alkaloids are natural products widely distributed in plants and also found in alcoholic beverages, well-cooked foods and tobacco smoke. Various authors have reported genotoxic activities of several carboline in prokaryotic and eukaryotic cells that have been attributed to their abilities to intercalate into DNA. But studies on the genotoxic and on the cytotoxic potencies in human cells in vitro are not found in the literature. In the present study the toxicities of one full aromatic beta-carboline alkaloid (harmine) and one dihydro-beta-carboline alkaloid (harmaline) were evaluated by means of two in vitro human cell assays: the cytochalasin-B blocked micronucleus (CBMN) assay and the viability/colony formation assay with four different human cultured non-transformed (CCD18Lu) and transformed (HeLa, C33A and SW480) cells. Neither alkaloid was able to induce micronuclei levels above that of control levels in a wide range of doses tested; although, harmine at the highest concentrations assayed induced apoptotic as well as necrotic cells. Harmine produced a good viability of all cell lines assayed (control and tumor) while harmaline significantly reduced the viability of transformed and non-transformed cell lines in a dose-dependent manner. Harmine displayed a dose-dependent inhibitory effect on cell proliferation against all human carcinoma cells, but the SW480 transformed cell line showed a higher sensitivity. These results suggested that harmine was identified as a useful inhibitor of tumor development.     

Effects of the Natural β-Carboline Alkaloid Harmine,a Main Constituent of Ayahuasca,in Memory and in the Hippocampus: A Systematic Literature Review of Preclinical Studies

Harmine is a natural β-carboline alkaloid found in several botanical species, such as the Banisteriopsis caapi vine used in the preparation of the hallucinogenic beverage ayahuasca and the seeds of Syrian rue (Peganum harmala). Preclinical studies suggest that harmine may have neuroprotective and cognitive-enhancing effects, and retrospective/observational investigations of the mental health of long-term ayahuasca users suggest that prolonged use of this harmine-rich hallucinogen is associated with better neuropsychological functioning. Thus, in order to better investigate these possibilities, we performed a systematic literature review of preclinical studies analyzing the effects of harmine on hippocampal neurons and in memory-related behavioral tasks in animal models. We found two studies involving hippocampal cell cultures and nine studies using animal models. Harmine administration was associated with neuroprotective effects such as reduced excitotoxicity, inflammation, and oxidative stress, and increased brain-derived neurotrophic factor (BDNF) levels. Harmine also improved memory/learning in several animal models. These effects seem be mediated by monoamine oxidase or acetylcholinesterase inhibition, upregulation of glutamate transporters, decreases in reactive oxygen species, increases in neurotrophic factors, and anti-inflammatory effects. The neuroprotective and cognitive-enhancing effects of harmine should be further investigated in both preclinical and human studies.     

γ-去氢骆驼蓬碱等咔啉类生物碱的辐射防护作用

γ- Harmine(Ⅰ),’harmine(Ⅱ )and harmaline(Ⅲ )were isolated from PeganumHarmala L.( Zygophylaceae).Tests were conducted with mice to detect whether γ- harmine( a newcompound), harmine,harmof(Ⅳ)and harmalol(Ⅴ) are effective radioprotective compounds againstγ-ray irradiation, Intraperitoneal injection of the hydrochlorides of the four alkaloids 50～80mg·kg^-1×1 in NIH male mice 30～45 minutes before 8.6～9. 7 Gy whole body ⁶⁰Co irradiationsignificantly increased the survival effects(1. 33～2. 61)and 30-day survivai rate in comparison withcontrol mice.The results indicatethat γ- harmine exhibited relatively good radioprotective effect.γ-harmine is the first alkaloid isolated from a plant having ptotective effects against whole-body lethal irradiation in mice.     

HPLC法同时测定哈医祛风膏中去氢骆驼蓬碱和骆驼蓬碱的含量及疗效观察

目的:建立高效液相色谱法(HPLC)测定哈医祛风膏中去氢骆驼蓬碱和骆驼蓬碱的含量及临床疗效观察。方法:采用高效液相法测定含量,色谱柱为Agilent TC-C18(4.6 mm×250 mm,5 μm),以甲醇:0.01 moL/L硫酸铵溶液(37:63,用三乙胺调pH至4.0)为流动相,流速为1 mL/min,柱温:25 ℃,检测波长在320 nm。用完全随机法分实验组和对照组进行比较哈医祛风膏疗效。结果:去氢骆驼蓬碱的线性范围为12~83 μg(r=0.999 8,n=6)平均回收率为100.14%,RSD值为1.33%、骆驼蓬碱的线性范围为12~83 μg(r=0.999 8,n=6)平均回收率为98.89%,RSD值为1.77%。疗效比较治疗组的总有效率为97.3%、对照组的总有效率为90.1%,2组有显著性差异,P<0.01。表明治疗组总体疗效明显优于对照组。结论:高效液相色谱法方法简便,准确,可靠,可以用于哈医祛风膏中去氢骆驼蓬碱和骆驼蓬碱的含量测定。并哈医祛风膏治疗颈肩腰腿痛的效果较好、疗效确切、安全性良好,可降低患者复发率,可考虑在临床上推广。     

盐酸去氢骆驼蓬碱脉冲式释药胶囊的研制

目的：制备盐酸去氢骆驼蓬减脉冲释药胶囊，并研究其体内药物动力学。方法：以pH敏感型机制为理论基础，用均匀设计对脉冲释药微丸的包衣处方进行筛选。将速释微丸和脉冲释药微丸按一定比例混合装入胶囊中即得脉冲释药胶囊。采用模拟人体胃肠道pH变化的溶出介质对脉冲释药胶囊的释药行为进行评价。结果：盐酸去氢骆驼蓬碱脉冲释药胶囊在0一2h内释放其中50％的药量，在2—6h内释放其余50％的药量。兔体内药物动力学实验结果表明，口服脉冲释药胶囊后分别在体内0．5h和6．0h有2次药物吸收高峰，脉冲释药胶囊(80mg)的生物利用度为普通胶囊（40mg)的2．08倍。结论：盐酸去氢骆驼蓬减脉冲释药胶囊可减少给药次数，提高临床用药的依从性。     

Effect of harmine and brain lesions on apomorphine induced motor activity.

Application of harmine (10 mg/kg IP) 30 min before apomorphine decreased the motoric effects of the latter. Following harmine an increase in 5-HT and a decrease in 5-HIAA in different brain regions have been found. Injection of 5,6-DHT into nucleus medianus raphe 7 days before the experiment caused a significant increase of the apomorphine effect. Harmine pretreatment reduced this escessive motility as well as additional lesion of the substantia nigra with 6-OH-DA. Lesion induced by 6-OH-DA alone was without significant effect on the hypermotility following apomorphine. Application of PCPA 3 days before testing elicited an increase of apomorphine-induced hypermotolity which could be abolished by preceding harmine application. The experiments demonstrate the inhibitory effect of the central serotoninergic system on the apomorphine syndrome as well as the serotoninergic-dopaminergic interaction in hypermotility.     

去氢骆驼蓬碱脂质体冻干剂粉的制备工艺优选

目的:制备含有不同冻干保护剂的N-三甲基壳聚糖(TMC)包衣去氢骆驼蓬碱脂质体(TMC-HM-LP)的冻干粉,并筛选其最佳制备工艺。方法:用"薄膜分散-pH梯度法"制备去氢骆驼蓬碱脂质体,并采用孵育包衣法、低温高速离心法和结合高效液相色谱(HPLC)定量方法测定其包衣脂质体的包封率;以其冻干粉的外观在冻干前和复溶后脂质体的粒径、包封率作为对比指标,优选出最佳的冻干工艺以及冻干保护剂的种类及比例。结果:以葡萄糖-乳糖-甘露醇(2:1:0.5)作为冻干保护剂,通过"分步预冻"的方法和-80℃冷冻干燥技术得到的TMC-HM-LP外观良好,冻干前后粒径和包封率变化较小。结论:采用冷冻干燥技术并结合冻干保护剂的优选,可显著提高包衣脂质体的稳定性。     

Harmine: Evaluation of its Antileishmanial Properties in Various Vesicular Delivery Systems

Harmine, a beta-carboline amine alkaloid isolated from Peganum harmala, was tested for its antileishmanial properties both in vitro and in vivo. In vitro antileishmanial activity of harmine was encouraging and prompted us to confirm the activity in vivo in hamster models. Harmine was tested both in free form and in different vesicular forms viz. liposomes, niosomes and nanoparticles. The different vesicles were prepared by the published protocols. The percent intercalation of harmine in liposomes, niosomes and nanoparticles was found to be 65, 60 and 20, respectively, when determined at 325 nm (∈_M=2.33 × 10₄ M_-1 cm_-1). At an equivalent dose of 1.5 mg/kg body weight, injected subcutaneously (SC) for a total of six doses in 15 days, harmine was found to reduce spleen parasite load by approximately 40, 60, 70 and 80%, respectively in free, liposomal, niosomal and nanoparticular forms. An inverse relationship could be established between the efficacy in the lowering of spleen parasite load and the size of the vesicles. Specific biochemical tests related to normal liver and kidney functions revealed that the toxicity of the drug was reduced in the vesicular forms in the same order as their efficacy and the same was confirmed by the histopathological studies of splenic sections. Cell cycle analysis studies using flow cytometry suggested that although harmine interferes in the cell division stage, it does not induce apoptosis in Leishmania donovani promastigotes. The results using Confocal Microscopy supported that the cell death could be attributed to necrosis due to non-specific membrane damage. Even then, because of its appreciable efficacy in destroying intracellular parasites as well as non-hepatotoxic and non-nephrotoxic nature, harmine, in the vesicular forms, may be considered for clinical application in humans.