桑黄化学成分研究

目的 :对药用真菌桑黄进行化学成分研究。方法 :利用正相硅胶柱层析 ,Al₂O_{3柱层析 ,SephadexLH-20柱层析 ,反相HPLC ,重结晶等分离手段和IR ,MS及 1D ,2DNMR等结构鉴定光谱方法。结果 :从桑黄中分离鉴定了 5个黄酮和两个香豆素类化合物 ,分别为柚皮素、樱花亭、二氢莰非素、7 甲氧基二氢莰非素、北美圣草素、香豆素及莨菪亭。结论 :这些化合物均为首次从针层孔属真菌中发现。}     

Anti-angiogenic activity of methanol extract of Phellinus linteus and its fractions

Aim of the study

The aim of the present study was to investigate the effects of MeOH extract of PL (PLME) and its fractions on angiogenesis.

Materials and methods

PLME and its subsequent fractions (methylene chloride, ethyl acetate, n-butanol and aqueous fractions) were evaluated in vitro. Specifically, the anti-angiogenic activities of PLME and its fractions were investigated by measuring their effects on the proliferation, migration, tube formation and phosphorylation of vascular endothelial growth factor receptor (VEGFR)-2 in human umbilical vein endothelial cells (HUVECs). In addition, the in vivo Matrigel plug model was applied to evaluate new vessel formation.

Results

The results revealed that PLME and its subsequent fractions, except for the aqueous fraction, led to significant inhibition of the proliferation, migration, tube formation and VEGFR-2 phosphorylation of HUVECs as well as in vivo angiogenesis.

Conclusions

These findings indicate the potential for the use of PLME in pathological situations involving stimulated angiogenesis, such as inflammation and tumor development.     

桑黄麦角甾醇生物合成关键酶PlERG24基因克隆与表达分析

目的 克隆桑黄菌丝甾醇C14还原酶（ERG24）基因全长,并进行生物信息学及表达模式的初步分析。方法 基于桑黄菌丝转录组数据设计PlERG24基因引物,采用PCR技术获得其cDNA全长序列,并通过Ex PASy等在线分析软件对其进行生物信息学分析,同时利用实时荧光定量PCR技术分析该基因的表达模式。结果 从桑黄菌丝中克隆到1 412 bp的PlERG24基因,开放阅读框长度为1 326 bp,编码441个氨基酸,理论相对分子质量为49 358.61,等电点为5.28,为不含信号肽的疏水性蛋白,推测定位于质膜,具有6个磷酸化位点;系统进化树分析表明,该序列与暴马桑黄的ERG24基因同源性最高。表达模式分析表明,在桑黄菌丝生长的一个周期内,PlERG24基因表达在25 d时达到最高6.36。结论 获得了PlERG24基因全长序列,为进一步研究该基因的功能以及利用基因工程等手段改良麦角甾醇生物合成途径奠定基础。     

Antiproliferative and antimetastatic effects of the ethanolic extract of Phellinus igniarius (Linnearus: Fries) Quelet

AIM OF THIS STUDY: Phellinus igniarius (Linnearus: Fries) Quelet (Phellinus igniarius) has been used in oriental countries for treatment of various diseases including cancer. However, it is unclear how Phellinus igniarius exerts anticancer effects. MATERIALS AND METHODS: In this study the ethanolic extract from the fruiting body of Phellinus igniarius (EEPI) was used to evaluate the antiproliferative and antimetastatic effects in human hepatocarcinoma SK-Hep-1 cells and rat heart vascular endothelial cells (RHE cells). RESULTS: We found that EEPI inhibited the proliferation of both cell lines in a dose-dependent manner, and the IC50 values at 48 h were 72 and 103 microg/ml for SK-Hep-1 cells and RHE cells, respectively. EEPI at non- or sub-cytotoxic concentrations (25-100 microg/ml) markedly inhibited the migration and invasion of SK-Hep-1 cells. EEPI added at 25 microg/ml significantly decreased the secretion of matrix metalloproteinase-2 (MMP-2) (49%, p<0.01) and vascular endothelial growth factor (VEGF) (13%, p<0.05) in SK-Hep-1 cells. EEPI at 25 microg/ml completely inhibited matrigel-induced tube formation in RHE cells. Importantly, EEPI (25 or 50 microg/ml) in combination with oxaliplatin (Oxa) or 5-flurouracil (5-FU) synergistically inhibited the proliferation of SK-Hep-1 cells. CONCLUSION: These results demonstrate the antiproliferative and antimetastatic effects of EEPI in vitro and the potential of EEPI as an adjuvant for chemotherapy.     

桑黄等四种抗癌药物抗癌活性比较

目的:研究比较桑黄、灵芝、阿加里斯茸、PL-2.PL-5(MeSima)的抗癌活性.方法:以小鼠移植性胃癌和S180肿瘤模型观察桑黄、灵芝、阿加里斯茸、PL-2.PL-5的抗肿瘤作用.结果:桑黄、灵芝、阿加里斯茸、PL-2.PL-5对小鼠胃癌的抑制率分别为43.09%、38.99%、31.01%和35.70%;对S180肉瘤的抑制率分别为46.07%、37.22%、27.99%和10.43%.结论:桑黄在4种药物中有较好的抑瘤作用.     

鲍氏针层孔菌生产菌丝体及胞外多糖的液体发酵培养

目的：考察营养因子和环境因子对鲍氏针层孔菌液体发酵生产菌丝体和胞外多糖产量的影响。方法：通过摇瓶实验，对不同的碳源、氮源、接种量、起始pH 和温度进行筛选。结果：鲍氏针层孔菌液体发酵生产菌丝体和胞外多糖时，所需的最优碳源和氮源分别是葡萄糖和大豆蛋白胨；适宜的接种量、起始pH、温度分别为6％，6.0，28 ℃。结论：通过摇瓶实验获得的数据将为鲍氏针层孔菌工业化液体深层发酵培养提供参考。     

桑黄治疗大鼠肝纤维化实验研究

目的:研究桑黄对肝纤维化的治疗作用。方法:采用四氯化碳诱导大鼠肝纤维化模型,实验分正常对照组、模型对照组、桑黄高、中剂量组,观察血清酶学,血清胶原成分含量及肝组织病理变化。结果:桑黄高、中、低剂量对肝纤维化有明显的治疗作用,能显著降低血清氨基转移酶水平和胶原成分含量,抑制肝组织内胶原纤维增生。结论:桑黄可用于治疗肝纤维化,且疗效与剂量呈正相关。     

桑黄多糖对人TLR4信号通路的激活作用研究

目的 观察桑黄多糖（polysaccharide from Phellinus igniarius,PPI）对人Toll-样受体4（Toll like receptor 4,TLR4）通路的激活作用及作用特点。方法 采用转染了人TLR4基因、分泌型胚胎碱性磷酸酶报告基因的HEK-Blue^TM hTLR4细胞模型,快速评价PPI对TLR4通路的激活作用,并用PMA诱导的THP-1细胞模型观察药物对TLR4通路作用的特点;MTT法检测细胞活性;ELISA法检测细胞因子分泌量;Real time PCR检测细胞mRNA表达水平。结果 PPI作用24 h不影响细胞活性,但能剂量依赖性地增加HEK-Blue^TM hTLR4细胞分泌型胚胎碱性磷酸酶的表达,并使THP-1细胞TLR4通路相关细胞因子TNF-α,IP-10分泌量增加。TLR4受体阻断剂TAK-242能显著降低PPI诱导的细胞因子分泌量（P<0.01）。Real-time PCR结果显示PPI既可以增加MyD88通路TNF-α、IL-6、IL-12、IL-1β、COX-2等细胞因子的mRNA表达,又可以增加TRIF通路IP-10、IFN-β等细胞因子的mRNA表达。结论 PPI对人TLR4受体具有直接激活作用,能同时激活TLR4下游MyD88和TRIF 2条分支,对TLR4分支通路的激活没有选择性。