肉苁蓉愈伤组织培养及所含有效成分量的研究

目的为扩大和提高肉苁蓉Cistanchedeserticola资源的利用效率。方法以肉苁蓉肉质茎的不同组织部位作为外植体,采用正交实验方法,筛选诱导愈伤组织产生的不同培养基和培养条件,并继代培养。应用HPLC方法对愈伤组织培养物中松果菊苷和洋丁香苷(毛蕊花糖苷)的量进行测定。结果肉苁蓉肉质茎的维管组织部分是诱导愈伤组织的最适外植体,鳞片叶次之,髓组织部分诱导效果较差。在暗培养、25~27℃条件下以B5为基本培养基附加6-BA(0.5~2mg/L)与IAA(0.5~1.5mg/L)诱导愈伤组织效果最佳;在半光照(光培养10h/d,暗培养14h/d)条件下,愈伤组织生长正常,最佳继代时间为25~30d。愈伤组织培养物中松果菊苷和洋丁香苷(毛蕊花糖苷)达4.37%。结论筛选出了适宜的肉苁蓉愈伤组织培养方法,且培养物中主要药用有效成分松果菊苷和洋丁香苷(毛蕊花糖苷)的量达到并超过了《中国药典》要求(0.3%)的标准。     

肉苁蓉提取工艺及干燥方式研究

目的选择肉苁蓉最佳提取工艺参数,研究不同干燥方式对5种苯乙醇苷类成分的影响。方法采用单因素筛选结合Box-Behnken响应面法优化提取工艺。最优条件提取后,利用HPLC法检测不同干燥方式肉苁蓉的松果菊苷、肉苁蓉苷A、毛蕊花糖苷、异毛蕊花糖苷、2′-乙酰基毛蕊花糖苷成分含量并采用单因素方差分析、聚类分析、主成分分析、密切值法分析5种苯乙醇苷类成分含量,选择最佳干燥方式。结果最佳提取工艺:甲醇体积分数55.14%,液料比46.39,提取时间38.50 min。聚类分析、主成分分析、密切值法分析表明,采用冷冻干燥法所得到的肉苁蓉品质最好,80℃烘干次之,40℃烘干最低。结论使用该工艺提取肉苁蓉,其5种苯乙醇苷类成分提取完全、充分。虽然冷冻干燥法炮制肉苁蓉所保留有效成分最高,但从生产角度考虑,采用80℃烘干法炮制能达到成本和功效的平衡。     

Therapeutic potential of phenylethanoid glycosides: A systematic review

Phenylethanoid glycosides (PhGs) are generally water-soluble phenolic compounds that occur in many medicinal plants. Until June 2020, more than 572 PhGs have been isolated and identified. PhGs possess antibacterial, anticancer, antidiabetic, anti-inflammatory, antiobesity, antioxidant, antiviral, and neuroprotective properties. Despite these promising benefits, PhGs have failed to fulfill their therapeutic applications due to their poor bioavailability. The attempts to understand their metabolic pathways to improve their bioavailability are investigated. In this review article, we will first summarize the number of PhGs compounds which is not accurate in the literature. The latest information on the biological activities, structure–activity relationships, mechanisms, and especially the clinical applications of PhGs will be reviewed. The bioavailability of PhGs will be summarized and factors leading to the low bioavailability will be analyzed. Recent advances in methods such as bioenhancers and nanotechnology to improve the bioavailability of PhGs are also summarized. The existing scientific gaps of PhGs in knowledge are also discussed, highlighting research directions in the future.     

苁蓉总苷胶囊HPLC指纹图谱研究

该文建立苁蓉总苷胶囊的HPLC指纹图谱,为其质量评价提供依据。以毛蕊花糖苷为参照物,采用高效液相色谱法,对苁蓉总苷胶囊进行了指纹图谱分析。色谱条件:Waters C18色谱柱(4.6 mm×150 mm,5μm);流动相甲醇-0.1%甲酸溶液梯度洗脱;流速1.0 m L·min-1;检测波长330 nm;柱温30℃。该方法精密度高、重复性好,所测样品中的13个成分均达到基线分离,并且15批成品指纹图谱相似度在0.95以上。该方法准确、可行,是一种简便、有效的中药质量评价手段。     

管花肉苁蓉松果菊苷次生代谢部位研究

目的：通过对管花肉苁蓉寄生生长和柽柳-管花肉苁蓉不同部位松果菊苷含量的研究,为明确管花肉苁蓉松果菊苷的次生代谢部位及调控提供理论依据。方法：高效液相系统分析了指标性成分松果菊苷含量,比较分析了管花肉苁蓉干物质积累量、松果菊苷积累量和寄生柽柳根粗的相关关系。结果：随着管花肉苁蓉干物质积累量的增加,植株内的松果菊苷积累量显著增加,两者与寄生的柽柳根粗呈极显著正相关关系。吸器韧皮部的松果菊苷质量分数为1553%,高于吸器木质部、管花肉苁蓉植株及其他部位的松果菊苷含量。结论：吸器韧皮部可能是管花肉苁蓉松果菊苷的次生代谢部位。     

松果菊苷抑制卵巢癌SKOV3细胞的增殖、侵袭和干细胞样特性

目的 探讨松果菊苷对卵巢癌SKOV3细胞的作用。方法 选择不同浓度的松果菊苷处理卵巢癌SKOV3细胞,24 h后CCK8检测细胞存活率;选择不同浓度松果菊苷浓度(0,20,40,80μmol·L^-1)进行后续实验,EdU染色检测细胞增殖;Transwell检测细胞侵袭;免疫印迹法检测VEGF、E-cadherin、Vimentin的表达及p38 MAPK的磷酸化;显微观察细胞肿瘤球形态;流式细胞仪检测CD133和CD44的含量。结果 SKOV3细胞的存活率与松果菊苷呈浓度依赖性降低,松果菊苷浓度≥40μmol·L^-1时,SKOV3细胞的存活率较松果菊苷0μmol·L^-1时均明显降低(P<0.05)。与松果菊苷0μmol·L^-1组比较,松果菊苷40μmol·L^-1组和80μmol·L^-1组EdU阳性细胞数和侵袭细胞数均减少(P<0.05);VEGF和Vimentin蛋白表达水平及p38 MAPK磷酸化水平均降低(P<0.05);E-cadherin蛋白表达水平升高(P<0.05);肿瘤干细胞成球直径减小(P<0.05),成球数减少(P<0.05);CD133和CD44表达均降低(P<0.05)。结论 松果菊苷可以抑制卵巢癌SKOV3细胞的增殖、侵袭、干细胞样特性及p38 MAPK的磷酸化。     

Identification of HPV16's E6 gene in suspected cases of cervical lesions and docking Study of its L1 protein with active components of Echinacea purpurae

BackgroundHPV 16 is the primary etiologic agent of cervical cancer and the presence of L1 and E6 oncoproteins are largely responsible for its virulence. It was the objective of this study to identify HPV16 isolates from suspected cases of cervical cancer at Specialist Hospital Sokoto and Sir Yahaya Memorail Hospiatal Birnin Kebbi, Nigeria and also to identify potent HPV16''s L1 protein inhibitor using in silico analysis.MethodsA total of 144 cervical samples consisting of 21 low grade squamous intraepithelial lesion, 6 high grade lesion and 117 negative pap smears were collected. The samples were subjected for molecular detection using PCR targeting E6 gene of the virus. Data generated for the molecular prevalence was statistically analyzed using Chi-square method. AutoDock Vina was used to carry out the molecular docking between 2hr5 and Chicoric acid, curcumin and Echinacoside.ResultsOut of the 144 samples, 24 samples were positive for the PCR representing 16.9% molecular prevalence rate. There is statistically significant association between cyto-diagnoses and presence of HPV16 (P < 0.05). Docking analysis showed that the Chicoric acid components of Echinacea purpurae have strong binding affinity (-8.7 kcal/mol) to the L1 protein of the HPV.ConclusionThis study provides data on HPV 16 epidemiology in northern Nigeria, and also provides novel evidence for consideration on certain interacting residues, when synthesizing Anti-HPV compounds in the wet lab.     

松果菊苷对高糖诱导的人视网膜毛细血管内皮细胞增殖、迁移及新生血管形成的影响

目的 探讨松果菊苷(ECH)对高糖诱导的人视网膜毛细血管内皮细胞(HRCECs)增殖、迁移及新生血管形成的影响及其作用机制.方法 将HRCECs分为5组:正常对照(Con)组、高糖组(HG,25 mmol·L-1 D-葡萄糖)、HG+1μmol·L-1 ECH组、HG+5μmol·L-1 ECH组和HG+25μmol·...     

HPLC法同时测定苁蓉总苷胶囊中松果菊苷、毛蕊花糖苷、异毛蕊花糖苷的含量

目的：建立同时测定苁蓉总苷胶囊中松果菊苷、毛蕊花糖苷、异毛蕊花糖苷三种成分含量的HPLC法。方法：Waters C18色谱柱（4.6 mm×150 mm,5 μm）,流动相：甲醇-0.1%甲酸溶液梯度洗脱;流速：1.0 mL·min-1;检测波长：330 nm;柱温30 ℃。结果：松果菊苷、毛蕊花糖苷、异毛蕊花糖苷分别在27.792-277.92 μg·mL-1 （r=0.999 6,n=6）,2.418 4-24.184 μg·mL-1 （r=0.999 6,n=6）,5.106-51.06 μg·mL-1 （r=0.999 8,n=6）范围内呈良好的线性关系,加样回收率均符合含量测定要求。结论：该方法简便、准确,可以用于苁蓉总苷胶囊中松果菊苷、毛蕊花糖苷、异毛蕊花糖苷三个成分的含量测定。     

松果菊苷对脑缺血大鼠纹状体细胞外液中氨基酸水平的影响

目的研究松果菊苷(ECH)对急性脑缺血大鼠纹状体细胞外液中4种氨基酸水平和脑梗死率的影响,以探讨ECH对脑神经保护作用的可能机制。方法 SD大鼠随机分为假手术对照组、模型组、阳性药川芎嗪组(CXQ,40 mg.kg-1)、ECH高剂量(ECH 40 mg.kg-1)组、ECH低剂量(ECH 20mg.kg-1)组和ECH配伍冰片(ECH 40 mg.kg-1,冰片400mg.kg-1)组。各组大鼠给予相应的药物或者生理盐水腹腔注射,每天1次,连续7 d。在给药d 3,脑纹状体埋置探针套管,末次给药1 h后,制作大鼠局灶性脑缺血模型(MCAO),模型成功后立刻进行微透析。将透析液注入高效液相-荧光检测器(HPLC-RF),此方法较氨基酸分析仪相比较,具有最低检测限低等特点,检测各组纹状体细胞外液中天门冬氨酸(Asp)、谷氨酸(Glu)、甘氨酸(Gly)、γ-氨基丁酸(GABA)的含量。结果与假手术对照组相比,模型组的Asp、Glu、Gly、GABA水平均明显升高;ECH给药组与模型组相比,ECH高剂量组能明显降低Asp、Glu的水平,而ECH低剂量组与ECH配伍冰片组Asp、Glu降低均不明显;ECH高、低组与配伍冰片组对Gly、GABA的影响均不明显;与模型组相比,ECH高、低剂量组能明显地缩小脑梗死面积。结论 ECH对脑神经的保护作用可能与对抗脑缺血后兴奋性氨基酸升高有关。