Cellular biological effects of matrine on K562 and K562/Vin cells

AIM: To investigate the cellular biological effects of matrine on K562 and K562/Vin cells and discuss the anticancer mechanism of matrine. METHODS: MTT assay was used to detect the IC₅₀ of matrine on these two cell lines and the reversal effect of matrine on K562/Vin cell's resistance to vincristine. In addition, the growth curve of cells was drawed. The p-glycoprotein (P-gp) expression was determined by immunohistochemistry analysis. The morphological changes of cells under light microscopy and the structural changes under transmission electron microscope were observed. RT-PCR assay was used to detect the hTERT-mRNA expression. RESULTS: The IC₅₀ of matrine was 3.4, 4.6 mmol·L ^-1 for K562 and K562/Vin cells, respectively. Matrine (4.0 mmol·L ^-1) inhibited the growth of K562, K562/Vin cells, 2.0 mmol·L ^-1 matrine inhibited expression of P-gp and with 492.4 reversal index. Matrine killed K562 cells by inducing the apoptosis and the same effect on K562/Vin cells was also observed. The hTERT-mRNA expression of K562 cells were also inhibited by matrine. CONCLUSIONS: Matrine enhanced the cytotoxicity of vincristine in K562/Vin cells, induced the apoptosis of K562 and K562/Vin cells, also inhibited the hTERT-mRNA expression in K562 cells. It shows that matrine would be an effective anticancer medicine.     

从苦豆子种子中提取生物碱的方法研究

对苦豆子种子中生物碱进行了提取、分离及苦参碱的定性和定量分析的研究。将苦豆子粉碎后,在室温下分别用0.4%盐酸、95%乙醇、1%氨水氯仿、无水乙醇溶剂冷浸7天共4次,经脱脂、酸化、碱化、萃取,得到生物碱;通过薄层层析(TLC)法进行单碱的分离,气相色谱(GC)法进行苦参碱的定性和定量分析。结果表明:以上4种提取方法出碱率分别为3.67%、2.73%、2.47%和1.60%;薄层层析(TLC)检查,以上4种方法提取的苦豆子生物碱中至少含有四种单一生物碱;气相色谱检测证明了以上4种方法提取的苦豆子生物碱中均含有苦参碱,其百分含量分别为8.460%、7.553%、5.675%、0.893%。其中0.4%盐酸冷浸提取法提取率明显高于其他溶剂提取率,为苦豆子生物碱优选提取方法。     

苦参碱、印楝素与矿物油两种复合液防治小绿叶蝉、跗线螨的药效试验研究

为了研究植物源农药苦参碱、印楝素与矿物油两种复合液防治小绿叶蝉、跗线螨的药效,应用苦参碱、印楝素乳油与矿物油两种复合液;单剂苦参碱、印楝素;两种化学农药硫丹、乐果6种处理。在广西桂北茶园第二次小绿叶蝉高峰及跗线螨高峰前期进行田间防效小区对比试验。结果表明,6种处理对小绿叶蝉校正防效:两种复合液A1药后第2和7天分别达86.54%和78.43%、B1分别达76.04%和88.57%,药后第14天A1和B1分别仍达60.29%、58.59%;单剂苦参碱、印楝素校正防效:药后第2和7天苦参碱仅分别达64.94%和57.32%、印楝素分别达53.54%和67.17%;两种化学农药硫丹、乐果校正防效:药后第2和7天硫丹仅分别为52%和42.18、乐果为41.04%和36.92%。6种处理对跗线螨的防效:两种复合液A1药后第2和7天高达91.14%和82.93%、B1高达82.34%和93.67%,药后第21天分别仍达68.03%、69.57%;单剂苦参碱、印楝素校正防效:药后第2和7天苦参碱分别达74.19%和68.34%、印楝素达61.6%和76.47%,药后第14天仅为50.12%、51.18%;两种化学农药硫丹及乐果:药后第2天分别为83.45%、70.54%,药后第21天分别仅为37.53%、4.73%。结果说明两种复合液比其他4种处理在防治小绿叶蝉及跗线螨上显示出较好的防效及较长的持效期。两种复合液不仅能有效地控制小绿叶蝉及害螨,而且对天敌安全。可在有机茶园生产基地的茶叶生产中推广使用。     

Effect of low expression of miR-21 on apoptosis of HepG2 cells induced by matrine

AIM: To explore whether miR-21 low expression enhances the effect of matrine (MAT) on the apoptosis of hepatocellular carcinoma cells.METHODS: Real-time fluorescence quantitative PCR (RT-qPCR) was used to detect the expression of miR-21 in the HepG2 cells treated with different concentrations of MAT. The effect of miR-21 on MAT-induced HepG2 cell apoptosis was analyzed by flow cytometry. The mRNA and protein expression of Bcl-2 and Bax in the HepG2 cells treated with MAT was determined by RT-qPCR and Western blot.RESULTS: The expression of miR-21 increased with the increasing concentration of MAT. Low expression of miR-21 promoted MAT-induced apoptosis, and enhanced the expression of Bax at mRNA and protein levels (P<0.05), while inhibited the expression of Bcl-2 at mRNA and protein levels (P<0.05).CONCLUSION: Low expression of miR-21 enhances MAT-induced HepG2 cell apoptosis by inhibiting the expression of Bcl-2 and promoting Bax expression.     

1%苦参碱可溶性液防治草原蝗虫田间药效试验

苦参碱是1种新型的生物防治制剂,具有对人畜安全、不污染环境、易于就地取材、节约开支、使用经济等优点。为了确定苦参碱是否对草原蝗虫具有良好的防治效果,2010～2011年连续2 a以常用药剂4.5%高效氯氰菊酯乳油为药剂对照,在天然草场上测试了1%苦参碱可溶性液使用量4.5 g/hm2、6.0 g/hm2和7.5 g/hm2对草原蝗虫的防治效果。结果表明：1%苦参碱可溶性液4.5～7.5 g/hm2处理对草原蝗虫表现出了较好的防治效果,2 a防治效果均在90%以上。本研究结果可为1%苦参碱药剂的产品登记以及为生产上选择适于防治草原蝗虫的药剂提供科学依据。     

苦参对鸡大肠杆菌的抑菌作用及其机理研究

【目的】阐明苦参的抑菌作用及机理，为生产实践提供理论依据。【方法】采用牛津杯法及液体倍比稀释法对鸡大肠杆菌等常见致病细菌进行了敏感性试验；通过电镜观察，SDS-聚丙烯酰胺凝胶电泳对蛋白表达的研究，生长曲线的绘制以及流式细胞仪对菌体细胞周期的分析，对苦参碱粗提物的抑菌作用及机理进行研究。【结果】苦参对革兰氏阳性菌、革兰氏阴性菌均有明显抑制作用，抑菌谱广；药物是通过抑制鸡大肠杆菌功能蛋白的表达，从而影响菌体的细胞周期，使I期菌数增加，R期菌数下降，最终菌体不能达到正常对数生长期而直接进入衰亡期；菌体形态结构发生明显变化，表现为菌体缢缩变形，中间凹陷呈规则元宝状，细胞质固缩，质壁分离，最终细胞壁破损，内容物泄漏而使菌体死亡。【结论】苦参碱对鸡大肠杆菌抑菌机理是抑制调控细菌生长、分裂相关蛋白的合成，阻滞其分裂和生长，并且与细胞内的蛋白质结合，细胞质固缩、解体而死亡。     

参芍洁阴洗液最佳提取工艺研究

优选参芍洁阴洗液的最佳提取工艺。采用正交试验方法,以苦参碱和氧化苦参碱总含量为指标,筛选参芍洁阴洗液最佳提取工艺条件。参芍洁阴洗液最佳提取工艺条件为水煮液密度1.12g/mL(20℃),醇沉百分数60%,pH=8。优选得到的工艺稳定可行,能显著提高苦参碱和氧化苦参碱的含量。     

Antiviral activity and underlying molecular mechanisms of Matrine against porcine reproductive and respiratory syndrome virus in vitro

Porcine reproductive and respiratory syndrome (PRRS), caused by porcine reproductive and respiratory syndrome virus (PRRSV), is an acute infectious disease. The prevalence of PRRS has made swine industry suffered huge financial losses. Matrine, a natural compound, has been demonstrated to possess anti-PRRSV activity in Marc-145 cells. However, the underlying molecular mechanisms were still unknown. The main objective of our study was to discuss the effect of Matrine on PRRSV N protein expression and PRRSV induced apoptosis. Indirect immunofluorescence assay (IFA) and Western blot were used to assess the effect of Matrine on N protein expression. Apoptosis was analyzed by fluorescence staining. In addition, the effect of Matrine on caspase-3 activation was investigated by Western blot. Indirect immunofluorescence assay and Western blot analysis demonstrated that Matrine could inhibit N protein expression in Marc-145 cells. And Matrine was found to be able to impair PRRSV-induced apoptosis by inhibiting caspase-3 activation.     

外源赖氨酸对苦参中生物碱含量的影响

以1年生苦参为材料,测定苦参根中生物总碱、苦参碱和氧化苦参碱含量,研究添加外源赖氨酸对苦参中生物碱含量的影响。结果表明,与对照相比,添加不同质量浓度的赖氨酸后,苦参总碱的含量都有所升高,添加量为600 mg/L时最有利于苦参总碱含量的增加,7、8、9、10月4个采样时间苦参总碱含量分别为对照的159.43%,168.33%,137.97%,144.52%;灌根和叶面喷施2种添加方法间没有显著性差异;添加600 mg/L外源赖氨酸后,7、8、9、10月4个时期苦参碱和氧化苦参碱的总量比对照分别提高102.38%,77.79%,48.63%,51.15%。因此,添加适当浓度的外源赖氨酸可提高苦参中生物碱的含量。     

苦参碱金属配合物的合成及抑菌活性研究

为了明确苦参碱金属配合物的抑菌活性,于无水乙醇中合成2种配合物,采用元素分析、红外光谱分析、差热热重同步分析、单晶衍射分析对配合物的结构进行表征,确定组成及结构。采用菌丝生长速率法测定配体及配合物对番茄灰霉病菌、小麦赤霉病菌、马铃薯晚疫病菌的抑菌活性。结果表明,合成的配合物经结构表征后,确定其结构为1分子苦参碱与1分子金属离子配位。药剂质量浓度为800mg/L时,苦参碱金属配合物的抑菌活性均高于配体,尤其锌配合物对番茄灰霉病菌的抑制率提高近7.1倍,铁配合物对小麦赤霉病菌以及锌配合物对马铃薯晚疫病菌的抑菌活性最强,其抑制率分别为79.8%、95.2%。苦参碱能与铁、锌金属离子形成金属配合物,且配合物的抑菌活性相对于配体有不同程度的提高。