RP—HPLC法同时测定白头翁汤中盐酸小檗碱、盐酸药根碱、盐酸巴马汀的含量

目的：建立反向高效液相色谱方法同时测定白头翁汤中盐酸小檗碱、盐酸药根碱、盐酸巴马汀的含量。方法：色谱柱为XPODS—AC。。（250minx4．6mm,5I．Lm）;采用线性梯度洗脱方法,流动相：溶液A为乙腈,溶液B为O．05mol／L磷酸二氢钾（磷酸调pH值至3．0）。0—40min溶液A：溶液B为5：95—45：55;流速：1．0mL／min;柱温：室温;检测器：DAD检测器;检测波长：340nm;进样量：10μL。结果：盐酸小檗碱、盐酸药根碱和盐酸巴马汀分别在62．5-l000μg／mL、46．875—750μg／mL和37．5—600μg／mL范围内线性良好;平均加样回收率分别为99．22％（RSD为3．25％）、98．46％（RSD为1．99％）和99．20％（RSD为1．69％）。结论：本方法简便、灵敏、准确,适用于白头翁汤的质量控制。     

不同产地初加工方法对延胡索5种生物碱含量的影响刘珍珍,白桥,徐皓

以延胡索药材中五种生物碱的含量作为评价指标分析水煮、水蒸、醋煮和盐煮四种初加工方法的效果。采用甲醇加热回流法提取,高效液相色谱法检测含量。以乙腈-0.01%磷酸水溶液为流动相,按22:78的比例进行等度洗脱,流速1.0mL/min;色谱柱(Inertsil ODS-3)温度：30℃;检测波长为283nm。结果显示,原阿片碱、延胡索乙素、掌叶防己碱、小檗碱、脱氢紫堇碱5种生物碱线性范围分别为1.568～78.4、3.444～172.2、0.72～36.0、0.78～39.0、2.632～131.6 μg/mL(r≥0.9993);平均回收率(n=6)为99.26%、98.85%、102.39%、98.06%、100.23%,RSD≤2.10%;四种延胡索加工方法中有效成分含量由高到低依次为水蒸＞水煮＞醋煮＞盐煮,其中以蒸制5min最佳。该方法简单、准确性高,可为延胡索产地初加工提供科学依据与技术支持。     

黄藤素对山羊子宫内膜上皮细胞的体外毒性研究

为了探究黄藤素的体外细胞毒性,采用不同浓度的黄藤素作用山羊子宫内膜上皮细胞（EECs）后,采用MTT法绘制细胞生长曲线;倒置显微镜下观察细胞生长状态;瑞氏-姬姆萨染色法观察细胞形态学变化并通过测定乳酸脱氢酶（LDH）含量检测细胞膜的完整性。结果显示,当黄藤素的浓度为10~227 μg/mL时,黄藤素对EECs有促增殖的作用;当浓度大于227 μg/mL时出现细胞增殖抑制现象,药物对细胞的半数增殖抑制率（IC₅₀）为360 μg/mL;瑞氏-姬姆萨染色结果显示,100、200 μg/mL的黄藤素作用于EECs时均未对其细胞核、细胞浆的形态产生影响;LDH结果显示,药物浓度≥250 μg/mL时细胞LDH的释放量极显著高于正常细胞组（P<0.01）。综上所述,黄藤素作用于山羊EECs后呈现一定的毒性作用,且呈现显著的剂量依赖效应,作用于EECs的安全剂量范围为≤250 μg/mL。     

接种丛枝菌根菌对黄檗苗木主要3种生物碱含量的影响

在盆栽条件下,研究了接种丛枝菌根菌对黄檗实生苗主要药用成分含量的影响。结果表明,接种丛枝菌根菌显著增加了黄檗重要的药用成分小檗碱、药根碱及掌叶防己碱的含量,其中尤以接种摩西球囊霉和透光球囊霉的处理效果显著。     

The Study on the Cytotoxicity of Palmatine on Goat Endometrial Epithelial Cells in vitro

To explore the cytotoxicity of palmatine in vitro,goat endometrial epithelial cells (EECs) were stimulated by different concentrations of palmatine,then the growth curve of cell was drew by MTT assay which could detect the cell proliferation. The cell growth state was observed by inverted microscope,and Wright-Giemsa staining was used to observe morphological changes of the cells while LDH test was used to check the membrane permeability of EECs. The results showed that when the concentration was 10 to 227 μg/mL,palmatine had a role in promoting proliferation of the EECs,while when the concentration was more than 227 μg/mL,cell proliferation was inhibited,and 50% inhibitory rate of proliferation (IC₅₀) of this drug on EECs was 360 μg/mL.Wright-Giemsa staining results showed that themorphology of the nucleus and cytolymph were not significantly changed when 100 and 200 μg/mL palmatine effected EECs. LDH results showed that when the palmatine concentration was more than 250 μg/mL,LDH content was extremely significantly higher than that of normal cell group(P<0.01).In conclusion,palmatine presented certain toxic effect on EECs with a significant dose-response effect,and the safe dose range of palmatine was less than 250 μg/mL.     

滤光膜对黄檗幼苗三种生物碱含量的影响

以滤光膜遮光处理1年生黄檗幼苗,测定了幼苗生物量及根、茎外皮中的小檗碱、药根碱和掌叶防己碱含量.红色、黄色、蓝色和绿色滤光膜对黄檗幼苗的根和茎生物量都有不同程度的抑制作用,其中黄膜的抑制作用最小,而蓝膜和绿膜的抑制作用最强.滤光膜处理也不同程度地抑制了小檗碱、药根碱和掌叶防己碱的合成和积累,红膜的抑制作用最小,蓝膜和绿膜的抑制作用最强.滤光膜处理后,3种生物碱的单株产量都低于对照,红膜处理小檗碱的产量显著高于黄膜处理的,但红膜和黄膜处理下的药根碱和掌叶防己碱的产量差异不显著,蓝膜和绿膜处理的3种生物碱的产量都始终最低.滤光膜处理不利于黄檗幼苗的生长和上述3种生物碱的积累.     

天然黄檗不同季节主要生物碱含量差异的研究

次生代谢产物小檗碱、掌叶防己碱和药根碱是黄檗中重要的药用生物碱,本研究在吉林省5 个林业局选取了65 株不同年龄的天然黄檗,利用反相高效液相色谱法研究了黄檗多器官中3 种生物碱含量的季节差异。结果表明:小檗碱在根皮、多年生枝、当年生枝和叶中的季节差异显著,各器官中小檗碱含量夏季最低(叶除外);叶中小檗碱含量,在各个龄组均表现为从春到秋含量依次升高,变化幅度在0.017 ~0.124 mg/ g 之间;小檗碱在根皮中最高, 平均值达到24.27 mg/ g,其余依次是茎皮、多年生枝皮、当年生枝和叶片。掌叶防己碱在根皮、茎皮、当年生枝、叶和幼龄阶段的多年生枝皮中的季节差异显著,秋季含量最高(叶除外);叶中掌叶防己碱含量,在各个龄组均表现为春季最高;掌叶防己碱在茎皮中含量最多,平均值为7.02 mg/ g,其余含量由大到小依次是多年生枝皮、根皮、当年 生枝和叶。药根碱在茎皮、多年生枝皮、当年生枝、叶和成熟阶段的根皮中的季节差异显著,多数器官夏季含量最 低;叶中药根碱含量,在各个龄组均表现为春季最高;药根碱在各个器官中的分布规律与小檗碱一致,根皮中含量最高,平均值为0.84 mg/ g,其余含量由大到小依次是茎皮、多年生枝、当年生枝叶。      

Palmatine attenuates LPS-induced inflammatory response in mouse mammary epithelial cells through inhibiting ERK1/2, P38 and Akt/NF-кB signalling pathways

Palmatine has a wide range of pharmacological effects and anti-inflammatory function. However, the effect of palmatine on LPS-induced inflammatory response of mammary epithelial cells has not been reported. In this research, we studied the anti-inflammatory mechanism of palmatine in EpH4-Ev (mouse mammary epithelial cells). EpH4-Ev cells were pre-treated with palmatine and then incubated with LPS. Cells were collected for examining production of pro-inflammatory mediators by qRT-PCR, and the related inflammatory signalling pathway was detected through immunofluorescence and Western blot. The results found that palmatine could significantly reduce the expression of IL-6, TNF-α, IL-1β and COX-2 in EpH4-Ev cells. Research on mechanisms found that palmatine could significantly inhibit the protein levels of p-Akt, p-P65, p-ERK1/2 and p-P38 in EpH4-Ev cells. In conclusion, these data suggested that palmatine inhibits inflammatory response in LPS-induced EpH4-Ev cells via down-regulating Akt/ NF-кB, ERK1/2 and P38 signalling pathways.     

黄藤素柔性纳米脂质体水凝胶贴剂的制备及体外透皮给药研究

为奶牛乳房炎的治疗提供了一种新型高效的透皮给药制剂,并考察其体外透皮给药的行为特点。采用薄膜分散法制备黄藤素柔性纳米脂质体(PFL),高效液相色谱仪(HPLC)测定包封率,透射电镜(TEM)观察PFL的结构、激光粒度仪(LPSA)测定PFL的粒径和表面Zeta电位;然后将PFL制备成水凝胶贴剂(PFL-HP),并采用立式双室扩散池考察PFL-HP体外透皮给药的行为特点,并同药物水凝胶贴剂(P-HP)与加入5%氮酮的水凝胶贴剂(P-A-HP)进行对照。结果表明,PFL对黄藤素的包封率为(79±2.23)%,PFL为圆球形层状囊泡结构,粒径为(190±24)nm,Zeta电位为(-54±2.65)m V;PFL-HP膏体柔软、膏面平整光洁,黏附性良好可48 h黏附于奶牛乳房而不脱落;24 h时,PFL-HP的药物累积透过量是P-HP与P-A-HP的2.12倍(t-test,P0.05)与1.13倍(t-test,P0.05),同时,PFL-HP组中药物在皮肤组织的滞留量(即"储库效应")也高于P-HP与P-A-HP(t-test,P0.05)。黄藤素柔性纳米脂质体水凝胶贴剂(PFL-HP)药物透皮吸收效率高、黏附性良好、使用方便,奶牛对其具有良好的依从性。PFL-HP可作为治疗奶牛乳房炎的一种新型高效透皮给药制剂。