毛蜂窝菌提取物体外抗肿瘤活性的研究

以人肺癌细胞NCI-H460、中枢神经系统癌细胞SF-268及乳腺癌细胞MCF-7为供试细胞株,采用MTT(四甲基偶氮唑盐)法对毛蜂窝菌(Hexagona apiaria)野生子实体、栽培子实体、培养废料、发酵菌丝体的醇提物和发酵液乙酸乙酯萃取物、发酵液水相部分等各提取物进行了体外抗肿瘤活性研究.结果表明,毛蜂窝菌发酵液乙酸乙酯萃取物对三种肿瘤细胞株都有较明显的抑制作用,抑制率达80%以上;其它提取物也有一定的抑制作用,但抑制率在40%以下.     

猴头菌子实体醇提物的生物活性

通过对猴头菌(Hericium erinaceus)子实体的乙醇提取物进行萃取,得到了石油醚、氯仿、乙酸乙酯、正丁醇和剩余水分部共5个分部,通过定性鉴定分析了其中次生代谢物质的成分,并以保护PC12细胞损伤和抑制DPP-IV酶的活性为实验模型,寻找具有抗衰老、降血糖等生物活性组分。结果表明,猴头菌子实体的醇提取物中可能含有生物碱、酚类(或鞣质)、甾类(或萜类)、蒽醌等物质。5个分部对NaN3诱导所致的PC12细胞损伤均具有一定恢复作用,与对照(69.5%)比,剩余水分部活性最强(84.6%)。石油醚分部和剩余分部在浓度为1.0mg/mL时对DPP-IV酶的抑制率超过了阳性对照药(25.25%),而氯仿萃取分部在浓度为0.4mg/mL时活性已超过阳性对照药,说明氯仿分部具有较强的降血糖潜力。     

冬虫夏草菌丝体多酚提取及对3种癌细胞抑制作用研究

以冬虫夏草的菌丝体为原料,提取溶剂为乙醇,采用Folin-Ciocalteu法测定冬虫夏草菌丝体中多酚的含量,目的在于研究乙醇浓度、料液比、提取温度和提取时间对多酚提取率的影响。以单因素试验为前提,以正交试验来指导优化提取参数,测定了所得多酚含量及其对Caco-2结肠癌细胞、Hep G-2肝癌细胞、MCF-7乳腺癌细胞的抑制作用。研究结果表明,在冬虫夏草菌丝体的提取工艺中,最佳的提取工艺条件为:温度为70℃,提取时间60 min,乙醇的浓度70%,料液比(g∶mL)1∶35;冬虫夏草菌丝体中提取的粗多酚对试验中3种癌细胞都表现出一定的抑制作用,当菌丝体粗多酚的浓度为100μg·mL~(-1)时,对3种癌细胞抑制率达到最高,分别为72.12%(Caco-2结肠癌细胞)、83.67%(HepG-2肝癌细胞)和58.78%(MCF-7乳腺癌细胞)。     

滇西北松口蘑菌塘的生态因子调查

以滇西北丽江市拉市乡松口蘑(Tricholoma matsutake)产地为例,进行了松口蘑菌塘生态因子的调查。结果发现:(1)松口蘑菌塘多集群分布于狭窄的山脊及西南坡,坡面坡度在30~60°之间,上层云南松荫蔽度0.4~0.6,下层高山栎、黄背栎等灌木丛荫蔽度达0.7~0.9,落叶层及腐质层1~6 cm;(2)出菇期间菌塘地表附近空气的平均温度14.1℃,平均相对湿度78.7%;(3)出菇期间菌塘的地下温度在15.0~20.5℃之间,西南坡的菌塘温度相对较高;(4)菌塘土壤的含水量比非菌塘土壤的含水量低,出菇期间平均低16.7%;(5)菌塘土pH约为5.5,比非菌塘土低7.9%;(6)菌塘土除全钾的含量比非菌塘土高之外,其余营养成分均比非菌塘的低。     

八种食药用菌子实体醇提物对乳腺癌细胞MCF-7的体外影响

以乳腺癌细胞株MCF-7为实验对象,采用细胞增殖抑制率、细胞粘附抑制率、细胞克隆数和细胞迁移距离为评价指标,研究8种食药用菌醇提物对乳腺癌的影响。刺芹侧耳(Pleurotus eryngii)、姬松茸(Agaricus blazei)和灰树花(Grifola frondosa)的醇提物(200μg/mL)处理能显著降低MCF-7细胞的克隆形成数和细胞迁移距离,较阴性对照分别降低了64.3%、64.5%、61.6%和54.8%、54.8%、48.0%,3种醇提物对MCF-7细胞粘附数和细胞增殖的抑制率分别为48.5%、55.1%、49.3%和30.0%、34.1%、29.5%;另外5种食用菌醇提物作用效果不明显。结果初步表明,刺芹侧耳、姬松茸和灰树花的醇提物(200μg/mL)具有一定的体外抑制乳腺癌活性。     

粗毛纤孔菌提取物的抗氧化和抑菌活性研究

采用福林酚法和NaNO2-Al(NO3)3比色法分别测定了粗毛纤孔菌子实体70%乙醇提取物中的多酚和黄酮,含量分别为(64.02±0.73)μg/mg和(5.74±0.13)μg/mg;并对70%乙醇提取物及主要化合物Hispidin的抗氧化和抑菌活性进行了测定。结果表明:70%乙醇提取物和Hispidin具有强烈的清除DPPH和·OH自由基活性的作用,在浓度为200mg/kg时清除率分别与BHA相当,清除率达到90%左右;采用K-B纸片扩散法测定了70%乙醇提取物及Hispidin对大肠杆菌Escherichia coli ATCC8099、金黄色葡萄球菌Staphyloccocus aureus ATCC6538、枯草芽孢杆菌Bacillus subtilis的抑制活性,结果除70%乙醇提取物高剂量(40.96mg/mL)对大肠杆菌的抑制率达到40.35%,其余均没有明显活性。     

基于ITS序列对林芝市售七种野生大型食用真菌的鉴定

以西藏林芝市售7种野生大型真菌(分别编号为XZQGJ、XZDJG、NCSBJ、NCNGJ、NCTLJ、NCSR、NCWZNGJ)为试材,采用分子生物学的方法对其进行鉴定,为进一步开发利用林芝地区大型真菌资源提供了参考依据。结果表明:XZQGJ属于腊伞属的Hygrophorus russula(红菇腊伞);XZDJG属于肉齿菌的Sarcodon cf.leucopus(裂盖肉齿菌);NCSBJ属于枝瑚菌属,与Ramaria sp.(小刺枝瑚菌)(KY774253.1)亲缘性最近,但林芝扫把菌与其它相似种均有一定的遗传距离,因此林芝扫把菌是一个独立的种;NCNGJ属于牛肝菌属,与Boletus recapitulatus(KP938967.1)亲缘性最近;NCTLJ属于钉菇属,与Gomphus clavatus(棒状钉菇)(KX008988.1)亲缘性最近;NCSR属于口蘑属,与Tricholoma matsutake(松口蘑)(MF521899.1)亲缘性最近,但林芝松茸与其它相似种均有一定的遗传距离,因此林芝松茸是一个独立的种;NCWZNGJ属于牛肝菌科的uncultured Boletaceae(未驯化牛肝菌)。     

基于气候特征的中国松口蘑分布规律分析及适生性评估

分别采用生态位因子分析(ecological niche factor analysis,ENFA)和最大熵模型(maximum entropy model,MaxEnt)对松口蘑(Tricholoma matsutake)在中国的分布规律及适生区范围进行研究。结果表明,松口蘑的气候生态位非常狭窄,其分布极易受到气候条件的制约。夏季降雨丰沛,环境昼夜温差较大,年温差相对较小是我国松口蘑分布区的基本气候特征。中国的松口蘑适生区主要可划分为西南和东北两大区域:西南区以横断山区和滇中高原区为核心,川西高原和西藏东南部也存在较大面积的松口蘑最适分布区;东北地区,松口蘑的高度适生区主要分布在辽宁、吉林两省东北部以及黑龙江东南部与朝鲜半岛毗邻区域(即长白山山区)。此外,陕西南部的秦岭山区,山西东南部、河南北部、山东北部以及内蒙古自治区境内大兴安岭地区也存在一定范围的松口蘑适生区。总体而言,松口蘑在中国的适生区沿滇中高原—横断山区—秦岭—长白山一线分布。     

鱼腥草不同溶剂提取物对豌豆根腐病菌的抑制作用

用蒸馏水、95%乙醇和石油醚为溶剂对采自鄂西北山区的鱼腥草进行全草浸提,探讨不同溶剂提取物对豌豆根腐病菌的抑制作用。试验结果,以浸膏的质量得率为指标,水浸法提取率最高,石油醚超声波法提取率次之,95%乙醇浸泡法提取物最少,分别为14.34%,9.85%,7.99%;当处理浓度不低于0.000 5 g/ml时,对豌豆根腐病菌菌丝生长的抑制率以95%乙醇提取物和石油醚提取物较高,均超过40%,且在最高处理浓度0.002 g/ml时分别达到60.32%和63.26%。而对豌豆根腐病菌孢子萌发的抑制率,在最高处理浓度为0.002 g/ml时,石油醚提取物最高,为90.00%,95%乙醇提取物和水提取物分别为85.71%和59.26%。     

短波紫外处理对口蘑采后贮藏品质的影响

以新鲜口蘑为试材,采用1.5、3.0kJ/m2短波紫外线(UV-C)照射处理后4℃贮藏,研究短波紫外对采后口蘑贮藏期间主要品质的影响。结果表明:经1.5kJ/m2和3.0kJ/m2的短波紫外线处理后,可以显著抑制口蘑失重率和PPO活性的上升,降低口蘑的呼吸高峰值;延缓维生素C、硬度及L值的下降,并使硬度维持在较高水平,同时促进了类黄酮及酚类物质的积累,从而较好地保持口蘑的感官品质和营养价值,延长口蘑的贮藏保鲜期。表明短波紫外线处理对采后口蘑的贮藏保鲜具有广泛的应用价值。     

猴头菌子实体提取物对人肺癌细胞SPCA-1的抑制作用

猴头菌(Hericium erinaceus)子实体的醇提物用不同极性有机溶剂分部提取,得到石油醚、氯仿、乙酸乙酯和正丁醇分部,其中仅石油醚分部能抑制SPCA-1细胞增殖和促进其释放ROS(reactive oxygen species),对石油醚分部的进一步研究发现石油醚分部可诱导SPCA-1细胞早期凋亡和降低G_0/G_1期细胞数量。     

樟芝发酵液和液体发酵菌丝体提取物对SPCA-1细胞的体外抑制作用

樟芝(Taiwanofungus camphoratus)发酵液和经95％乙醇浸提后的菌丝体醇提物分别用石油醚和氯仿进行分级提取,得到石油醚和氯仿提取物.对不同提取物进行体外抑制SPCA-1肺癌细胞增殖实验,发现各提取物均可抑制SPCA-1细胞的增殖,其中发酵液石油醚提取物抑制作用最好,IC5.为62.5 μg/mL,发酵液和菌丝体氯仿提取物的抑制作用较好,IC50分别为120.9和109.5 μg/mL,菌丝体石油醚提取物的抑制作用较差,在25～150 μg/mL作用浓度下,抑制率低于20％.各提取物对SPCA-1细胞凋亡和生长周期作用的测定结果表明,各提取物在肺癌细胞SPCA-1中通过诱导细胞凋亡及S期周期阻滞来表现其抑制作用.     

山茶靛牛肝菌子实体提取物的体外抗肿瘤活性

依次用95％乙醇和蒸馏水提取山茶靛牛肝菌(Boletus pseudoregius)子实体,再用不同有机溶剂萃取95％乙醇提取物,考察山茶靛牛肝菌子实体不同有机溶剂萃取物和水提物的体外抗肿瘤活性.结果表明:石油醚萃取物对白血病细胞K562抑制作用较弱,对人肠癌细胞SW620和人肝肿瘤细胞HepG-2的增殖有较强的抑制作用,IC50分别为95.7和195.4 μg/mL;正丁醇萃取物对K562、SW620和HepG-2的增殖都有较强的抑制作用,IC50分别为24.2、89.0和59.6 μg/mL;石油醚和正丁醇萃取物在实验浓度范围内对人正常前列腺基质永生化细胞WPMY-1的增殖无抑制作用;氯仿和乙酸乙酯萃取物对K562、SW620和HepG-2肿瘤细胞和WPMY-1正常细胞的增殖均有较强抑制作用,其中氯仿萃取物的抑制作用最强;水提物对以上3种肿瘤细胞抑制作用不明显,对正常细胞的增殖无抑制作用.     

Effects of propofol on invasion and migration of human gastric cancer SGC-7901 cells

AIM: To investigate the effects of propofol on invasion and migration of gastric cancer cell line SGC-7901. METHODS: Cultured gastric cancer cell line SGC-7901 was randomly divided into 4 groups, and then diffe-rent concentrations (1, 3, 5 and 7 mg/L) of propofol were added and incubated for 24 h. The cell viability was measured by MTT assay. The invasion and migration abilities of the SGC-7901 cells were detected by Transwell assay and wound-healing assay. The expression of cysteine-rich angiogenic inducer 61 (CYR61), CD44v6 and matrix metalloproteinase-7 (MMP-7) in the SGC-7901 cells were examined by immunocytochemistry and Western blot.RESULTS: Propofol at 5 mg/L does not affect the viability of SGC-7901 cells, whereas significantly suppresses the invasion and migration abilities, and down-regulates the expression of CD44v6 and MMP-7 (P<0.05). CONCLUSION: The decreased invasion and migration abilities of SGC-7901 cells were partly due to the inhibition of CD44v6 and MMP-7 expression.     

Effect of ruthenium-polypyridine complex on apoptosis of gastric cancer SGC-7901 cells

AIM:To study the effect of ruthenium-pyridine complex Ru1 on apoptosis of gastric cancer SGC-7901 cells. METHODS:MTT assay and crystal violet staining method were used to detect the viability and cell number of SGC-7901 cells treatment with Ru1. Annexin V-FITC and PI staining was performed to test the apoptosis rate of SGC-7901 cells. The protein expression of Bax and Bcl-2 was detected by Western blot. RESULTS:The results of MTT assay and crystal violet staining showed that the ruthenium-pyridine complexes significantly reduced the viability and cell number of SGC-7901 cells. Treatment with Ru1 for 24 h significantly increased the apoptotic rate of SGC-7901 cells (P<0.05). Ru1 up-regulated the expression of Bax protein and down-regulated the expression of Bcl-2 protein in the SGC-7901 cells (P<0.05). CONCLUSION:Ru1 induces apoptosis of SGC-7901 cells by affecting the expression of Bax and Bcl-2 proteins.     

Inhibitory effect of ampelopsin combined with mitomycin on proliferation of gastric carcinoma cells

AIM: To investigate the synergistic effects of ampelopsin (AMP) and a chemotherapeutic drug mitomycin (MMC) on the proliferation of gastric cancer cell line SGC-7901.METHODS: SGC-7901 cells were cultured in vitro and divided into 4 groups: control group, AMP group, MMC group and AMP+MMC group. Cell proliferation was measured by MTT method. The apoptotic index was examined by flow cytometry. The expression of apoptotic proteins, Bcl-2 and survivin, was detected by Western blotting.RESULTS: AMP at the concentrations ranging from 2.2 mg/L to 14.84 mg/L exerted inhibitory effect on the growth of SGC-7901 cells. Cell proliferation in AMP (14.84 mg/L) group was inhibitory by (60.85±1.13) %, significantly higher than that in control group (P<0.05). The inhibitory rates of cell proliferation varied from (17.40±0.30) % to (72.23±1.36) % when the concentrations of MMC increased from 1×10^-3 g/L to 1×10^-2 g/L. AMP combined with MMC showed a synergistic effect on the growth of SGC-7901 cells. The inhibitory rates of cell proliferation varied from (21.83±2.50) % to (46.70±1.45) % when the concentrations of MMC increased from 1×10^-3g/L to 5×10^-3g/L. The inhibitory effect of AMP plus MMC was higher than that of MMC or AMP alone. The protein levels of Bcl-2 and survivin were inhibited in AMP group, MMC group and AMP+MMC group, and were significantly lower in AMP+MMC group than those in AMP group or MMC group.CONCLUSION: AMP enhances the inhibitory effect of MMC on the growth of SGC-7901 cells. The mechanism is related to the inhibition of apoptotic protein expression.     

MicroRNA-146a promotes apoptosis of gastric cancer SGC-7901 cells by targeting TAK1

AIM: To explore the effect of microRNA-146a (miR-146a) on apoptosis of human gastric cancer SGC-7901 cells and the underluing mechanism. METHODS: miR-146a mimic (up-regulated miR-146a expression) and miR-146a inhibitor (down-regulated miR-146a expression) were transfected into the SGC-7901 cells by liposome method. At the same time, miRNA nonsense sequence transfection group as the negative control group (NC group) was set up. RT-qPCR was used to evaluate the levels of miR-146a in the SGC-7901 cells after transfection. The effects of miR-146a on the cell apoptosis and growth were assessed by flow cytometry analysis and CCK-8 assay, respectively. The effect of over-expression or knockdown of miR-146a on transforming growth factor-β-activated kinase 1 (TAK1)/nuclear factor-kappa B (NF-κB) signaling was evaluated by RT-qPCR and Western blot. RESULTS: miR-146a modulated apoptosis of SGC-7901 cells. Over-expression of miR-146a significantly increased apoptosis, whereas knockdown of miR-146a inhibited the apoptosis of SGC-7901 cells. The expression of TAK1 at mRNA and protein levels was significantly decreased when miR-146a mimic was transfected into the SGC-7901 cells (P<0.05). On the contrast, the expression of TAK1 at mRNA and protein were significantly higher in miR-146a inhibitor transfection group than that in NC group (P<0.05), suggesting that miR-146a negatively regulated TAK1 expression. Moreover, knockdown of TAK1 enhanced the apoptosis of SGC-7901 cells (P<0.01), while over-expression of TAK1 inhibited the apoptosis of SGC-7901 cells(P<0.01). Additionally, both over-expression of miR-146a and knockdown of TAK1 led to a prominent increase in the expression of NF-κB inhibitor protein alpha (IκBα) and a significat decrease in B cell lymphoma-2 (Bcl-2) level in the SGC-7901 cells. CONCLUSION: miR-146a significantly promotes apoptosis of SGC-7901 cells by inhibition of NF-κB pathway via targeting TAK1.     

高效液相色谱法测定食用菌中福美双残留量

建立食用菌中福美双残留量的高效液相色谱(HPLC)检测方法.新鲜食用菌子实体样品粉碎后经二氯甲烷超声(53 Hz)提取20 min,离心(3320 g、5 min),二氯甲烷经氮气吹干,甲醇定容后用HPLC测定.以等体积甲醇和0.1％甲酸水溶液为流动相,1 mL/min等度洗脱,在0.05～10 μg/mL范围内,福美双的峰面积与其浓度呈线性相关,R≥0.999,方法检出限为0.02 mg/kg,添加回收率为79.4％～90.2％,变异系数为1.15％～7.51％％.     

MicroRNA-126 enhances radiosensitivity of SGC-7901 cells via targeting inhibition of EZH2

AIM: To investigate the molecular biological mechanisms by which microRNA-126 (miR-126) enhances the radiosensitivity of gastric cancer cells. METHODS: SGC-7901 cells were cultured in vitro. In order to over-express miR-126 in SGC-7901 cells, miR-126 mimic was transfected. The mRNA and protein levels of enhancer of zeste ho-molog 2 (EZH2) were detected by RT-qPCR and Western blot, respectively. The targeting relationship between miR-126 and EZH2 was determined by dual-luciferase reporter assay. To estimate the effect of EZH2 on miR-126-enhanced radiosensitivity of the SGC-7901 cells, the pcDNA3.1-EZH2 vector was also co-transfected with miR-126 mimic, and then CCK-8 assay and flow cytometry were used to detect the viability and apoptotic rate of the cells after radiation. RESULTS: Over-expression of miR-126 significantly inhibited the expression of EZH2 in SGC-7901 cells both at protein and mRNA levels (P<0.05). A direct targeting relationship between miR-126 and EZH2 was confirmed by dual-luciferase reporter assay. Compared with the cells only transfected with miR-126 mimic, co-transfection of pcDNA3.1-EZH2 with miR-126 mimic increased the viability but reduced the apoptosis of the cells treated by radiation (P<0.05). CONCLUSION: Targeting inhibition of EZH2 may be one of the mechanisms by which miR-126 enhances the radiosensitivity of gastric cancer cells.     

林芝地区松口蘑营养成分

以西藏林芝地区林芝、波密、工布江达3个县采集的野生松口蘑子实体为材料,测定其外观大小及粗多糖、粗蛋白、游离氨基酸、铁和锌等营养成分含量,并与其它地区的松口蘑营养成分含量进行比较。结果表明:林芝县松口蘑铁含量显著高于其它区域;林芝地区松口蘑菌盖直径4.0~7.1 cm、菌柄长5.9~8.6 cm,与内地松口蘑菌盖直径5.9~21.5 cm、菌柄长5.0~18.0 cm相比,个体较小;粗多糖、粗蛋白和锌含量较低,而粗脂肪和铁的含量较高。