苦参素注射液在犬体内的药动学过程

目的:建立用HPLC法测定血浆中苦参素浓度的方法,考察苦参素注射液犬体内的药动学过程.方法:以氯仿-正丁醇(98:2)为提取溶剂处理血样.6条beagle犬iv苦参素注射液,测各时间点血药浓度,PAPP 2.0程序处理数据.结果:苦参素注射液犬体内的各个重要药动学参数如下:α为(0.053±0.007)μg·min-1,β为(0.008 3±0.001 0)μg·min-1,t1/2β为(84.1±4.3)min,k10为(0.025±0.003)μg·min-1,k12为(0.018 8±0.002 0)μg·min-1,k21为(0.018±0.003)μg·min-1,AUC为(85.9±11.3)mg·h·L-1.结论:本法提取回收率较高,拟合结果表明苦参素体内分布属于二室开放模型.     

苦参片质量标准的研究

目的：修订完善苦参片的质量标准。方法：修订现行标准的显微鉴别和薄层鉴别;增订高效液相色谱法测定苦参片中苦参碱和氧化苦参碱的总量。结果：显微鉴别中苦参组织特征均易察见;薄层色谱中斑点清晰,专属性强;苦参碱,氧化苦参碱分别在24.2～4840μg（r=0.9999）、20.8～4160μg（r=0.9999）范围内线性关系良好,平均回收率分别为97.53%（RSD为1.17%）、96.97%（RSD为1.35%）。结论：修订后的标准简便、准确,可用于苦参片的质量控制。     

RP-HPLC 法同时测定成人血清中苦参素和阿德福韦酯的浓度方法学研究

目的：建立一种简易的测定联合服用苦参素和阿德福韦酯成人血清中苦参素和阿德福韦酯含量方法。方法采用Shim-pack VP-ODS C18色谱柱（250 mm ×4．6 mm,5μm）,流动相为甲醇∶水（70∶30）,流速为1．1 mL·min －1,检测波长为229 nm。结果苦参素保留时间4．6 min 左右,标准曲线的线性范围为0．0125～2 mg·L －1,r ＝0．9992。阿德福韦酯保留时间7．3 min 左右,标准曲线的线性范围为0．0125～0．75 mg·L －1,r ＝0．9991,苦参素和阿德福韦酯的日间、日内变异系数均在7％以下,回收率＞92％。结论该法简便,灵敏,快速,可用于苦参素和阿德福韦酯临床联用治疗的成人血药浓度的监测。     

苦参素对慢性病毒性肝炎及肝硬化患者血清胆碱酯酶影响的初步观察

观察苦参素对慢性病毒性肝炎及肝炎后肝硬化患者血清胆碱酯酶的影响。 4 8例患者分成慢性肝炎组(36例 )和肝硬化组 (12例 ) ,检测苦参素治疗前、后患者血清中胆碱酯酶及肝功能生化指标。苦参素治疗后患者血清胆碱酯酶明显下降 ,但疗程大于 3个月后 ,胆碱酯酶可稳定于一个较低的水平 ;患者均未出现明显的副反应 ;停药后胆碱酯酶能较快地恢复原水平。苦参素能明显降低患者血清中胆碱酯酶水平 ,而且这种作用为可逆性 ,可能不会引起明显的不良反应。     

苦参碱类生物碱的镇痛作用研究进展

苦参碱类生物碱（包括有苦参碱、氧化苦参碱、氧化槐定碱、槐果碱、氧化槐果碱等）具有镇痛作用,镇痛特征是以中枢性为主、无成瘾性和耐药性,综述其镇痛作用及作用机制研究进展。苦参碱类生物碱的中枢性镇痛作用机制可能是：激活大麻素受体-2和上调电压门控钙离子N-型通道表达,促进抑制性神经递质（γ-氨基丁酸、甘氨酸）合成和释放,下调GAT-1表达,使突触间隙γ-氨基丁酸浓度升高以及上调γ-氨基丁酸-A受体表达,从而增强γ-氨基丁酸能神经功能。γ-氨基丁酸能神经功能的增强可抑制兴奋性神经递质谷氨酸过度表达并下调NMDA受体和蛋白激酶Cγ表达,降低NMDA受体对谷氨酸的兴奋性,从而下调电压门控钙离子L-型通道的表达,抑制钙离子内流,阻滞CaMKⅡ/CREB通路,减轻炎症反应和产生镇痛作用。     

氧化苦参碱介导细胞自噬减轻溃疡性结肠炎小鼠结肠黏膜氧化性损伤的作用机制研究

目的 探讨氧化苦参碱介导细胞自噬减轻溃疡性结肠炎（ulcerative colitis,UC）小鼠结肠黏膜氧化性损伤的作用机制。方法 采用2,4,6-三硝基苯磺酸灌肠复制小鼠UC模型,将造模成功小鼠按体质量随机分为模型组、氧化苦参碱组（50 mg·kg^-1·d^-1,ig）、羟基氯喹组（50 mg·kg^-1·d^-1,ig）、氧化苦参碱+羟基氯喹组,另设正常组,每组10只。治疗1周后测定各组小鼠疾病活动度（disease activity,DAI）、结肠质量系数及病理形态;MitoSOX Red法测定结肠ROS含量,ELISA法测定结肠组织SOD、MDA、MPO、GSH-PX含量;透射电镜结合免疫荧光观测结肠细胞自噬程度,Western blot测定Atg5和Beclin-1蛋白表达。结果 与模型组和羟基氯喹组比较,氧化苦参碱组小鼠DAI和结肠质量系数均有显著减小（P<0.01）,结肠病理损伤明显减轻;ROS、MDA和MPO含量极显著降低（P<0.01）,SOD和GSH-PX含量极显著增加（P<0.01）;结肠黏膜细胞自噬,程度显著增强,Atg5和Beclin-1蛋白表达极显著上调（P<0.01）。结论 氧化苦参碱可促进UC小鼠细胞自噬,减轻小鼠结肠黏膜氧化性损伤。     

外翻肠囊法研究氧化苦参碱磷脂复合物的肠吸收特性

目的 通过对比研究氧化苦参碱及其磷脂复合物大鼠离体肠吸收情况,考察磷脂复合物对氧化苦参碱肠吸收的影响。方法 采用大鼠离体外翻肠囊模型研究氧化苦参碱磷脂复合物的肠吸收特性。HPLC测定不同剂量的氧化苦参碱及氧化苦参碱磷脂复合物在大鼠十二指肠、空肠、回肠和结肠内的累积吸收量。结果 氧化苦参碱磷脂复合物在不同肠段的吸收较氧化苦参碱均有提高,低剂量组氧化苦参碱磷脂复合物在空肠、结肠处的吸收较氧化苦参碱有显著提高;中剂量组氧化苦参碱磷脂复合物在十二指肠、空肠、结肠处的累积吸收量较氧化苦参碱有显著提高;高剂量组氧化苦参碱磷脂复合物在空肠的吸收较氧化苦参碱有显著提高。结论 与氧化苦参碱相比,磷脂复合物对氧化苦参碱在大鼠肠道的吸收有显著的促进作用。     

A Study on Isolation of Chemical Constituents from Sophora Flavescens Ait. and Their Anti-Glioma Effects

Background

Sophora flavescens Ait. is a traditional Chinese medicine with a long history in China. It is mainly used in the treatment of heat dysentery and similar ailments in the clinical. The objective of this paper was to isolate, purify and identify alkaloids from Sophora flavescens Ait. and to explore their inhibitory effects on C6 glioma cells.

Materials and Methods

Column chromatography, extraction and NMR spectroscopy were used to structurally identify the isolated compounds. MTT assay and flow cytometry were used to detect the inhibitory effect of matrine on C6 cells.

Results

Three compounds were isolated from Sophora flavescens Ait., namely matrine, oxymatrine and lupeol. Different concentrations of matrine solution all had inhibitory effects on growth of C6 cell lines, which showed apparent dose-effect relationship. Compared with the control group, proportion of G0/G1 phase cells increased in each matrine concentration group to a maximum of 79.8%; proportion of S phase cells reduced, and proportion of G2/M phase cells declined slightly to a minimum of 6.3%, suggesting that after the action of matrine proliferation of C6 cells was significantly inhibited and the cells were arrested in the G1 phase.

Conclusion

We concluded that Sophora flavescens Ait. has an inhibitory effect on C6 cell proliferation.     

Extraction and Isolation of Alkaloids of Sophora Alopecuroides and Their Anti-Tumor Effects in H22 Tumor-Bearing Mice

Background

Alkaloids of Sophora alopecuroides have good biological activity, and are widely used in clinical settings, which not only have pharmacological activities of anti-cancer, cancer suppression, as well as the inhibition, and killing of various microorganisms; but also possess extensive pharmacological effects on immune system, nervous system and cardiovascular system. The objective of this paper was to extract and isolate total alkaloids of Sophora alopecuroides (TASA), and to study their anti-tumor effects in H22 tumor-bearing mice.

Materials and Methods

TASA were extracted and isolated using thin-layer chromatography, and column chromatography; and the isolated compounds were analyzed using nuclear magnetic resonance. The inhibitory effects of TASA on tumor in H22-bearing mice were determined by MTT assay.

Results

Three compounds were isolated from Sophora alopecuroides L., which were matrine, oxymatrine and sophoridine, respectively. Meanwhile, mouse H22 sarcoma model was established and different doses of TASA apparently inhibited solid H22-tumor in mice; it inhibited the thymus, and spleen to some extent; the degree of inhibition was more obvious for the spleen.

Conclusion

TASA has an anti-tumor effect in H22 tumor-bearing mice.     

高效液相色谱法同时测定山豆根中苦参碱和氧化苦参碱含量

目的建立同时测定山豆根中苦参碱和氧化苦参碱含量的方法。方法用三氯甲烷超声提取，通过中性氧化铝柱，依次以三氯甲烷、三氯甲烷-甲醇洗脱。色谱柱以氨基键舍硅胶为填充剂（250mm×4．6mm，5μm），流动相为乙腈-无水乙醇-3％磷酸溶液（80：10：10），流速为1．0mL／min，检测波长205nm，柱温30℃。结果苦参碱进样量在0．1021—4．082μg范围内与峰面积值呈良好的线性关系（r=0．9999），氧化苦参碱进样量在0．1943—3．886μg范围内与峰面积值呈良好的线性关系（r=0．9998），平均回收率均在99％以上。结论方法准确、灵敏度高、重现性好，能同时测定山豆根中苦参碱和氧化苦参碱的含量，可作为山豆根药材质量控制的含量检测方法。