蒺藜果实化学成分及其抗氧化活性研究

目的 研究蒺藜Tribulus terrester果实的化学成分及其抗氧化活性。方法 综合利用硅胶柱色谱、Sephadex LH-20凝胶柱色谱等分离手段进行分离纯化;结合NMR、MS、IR、UV等波谱学技术来鉴定化合物的结构;通过DPPH法、ABTS法以及羟基自由基清除方法测定化合物的抗氧化活性。结果 从蒺藜果实75%甲醇提取物中分离鉴定了10个化合物,分别为(E)-4-hydroxy-N-(3-(4-hydroxy-3-methoxyphenyl)acryloyl)benzamide(1)、蒺藜酰胺（2）、海柯皂苷元（3）、25R-螺甾-4-烯-3,12-二酮（4）、槲皮素（5）、3,4’-二羟基-3’-甲氧基苯丙酮（6）、对羟基苯甲酸（7）、(9Z,11E)-13-hydroxy-9,11-octadecadienoic acid(8)、(8E,12Z)-10-hydroxy-8,12-octadecadienoic acid(9)、肉豆蔻酸（10）,其中化合物1～6对DPPH、ABTS自由基以及羟基自由基均有较显著的活性,并且化合物5清除ABTS自由基和羟基自由基半数抑制浓度（...     

Quercetin protects against cisplatin‐induced acute kidney injury by inhibiting Mincle/Syk/NF‐κB signaling maintained macrophage inflammation

Acute kidney injury (AKI) with high incidence and mortality is the main cause of chronic kidney disease. Previous studies have indicated that quercetin, an abundant flavonoid in plants, exhibited renoprotective role in AKI. However, the underlying mechanism is largely unknown. In this study, we try to explore whether quercetin protects against AKI by inhibiting macrophage inflammation via regulation of Mincle/Syk/NF‐κB signaling. The results demonstrated that quercetin can significantly inhibit expression and secretion of IL‐1β, IL‐6, and TNF‐α in LPS‐induced bone marrow‐derived macrophages (BMDMs) and reduce activity of Mincle/Syk/NF‐κB signaling in vitro. We also found that quercetin can strongly reduce the concentration of serum creatinine, BUN, IL‐1β, IL‐6, and TNF‐α in cisplatin‐induced AKI model. Furthermore, quercetin down‐regulated protein levels of Mincle, phosphorylated Syk and NF‐κB in kidney macrophages of AKI, as well as inhibited M1, up‐regulated M2 macrophage activity. Notably, the down‐regulation of LPS‐induced inflammation by quercetin was reversed after adding TDB (an agonist of Mincle) in BMDMs, suggesting that quercetin suppresses macrophage inflammation may mainly through inhibiting Mincle and its downstream signaling. In summary, these findings clarified a new mechanism of quercetin improving AKI‐induced kidney inflammation and injury, which provides a new drug option for the clinical treatment of AKI.     

基于网络药理学及实验验证探究清热利胆汤治疗胆汁瘀积性肝损伤的作用机制

目的 基于网络药理学及实验验证探究清热利胆汤治疗胆汁瘀积性肝损伤的作用机制.方法 采用网络药理学方法筛选清热利胆汤和胆汁瘀积性肝损伤相关的核心靶点,构建基于活性成分与疾病共同靶点的蛋白质-蛋白质相互作用(protein-protein interaction,PPI)网络,利用DAVID数据库对潜在作用靶点进行京都基因...     

The changes of antioxidant defense system caused by quercetin administration do not lead to DNA damage and apoptosis in the spleen and bone marrow cells of rats.

Quercetin may have the opposite effect, namely anti- as well as pro-oxidant. The aim of this study was to assess the results of quercetin anti- and/or pro-oxidant activity in the bone marrow and spleen cells of rats. The experimental rats were treated daily, with quercetin in a dose of 8 or 80mg/kg b.w. by gavage for 40 days. The intracellular redox state in cells were assessed by measuring the ferric ion reducing antioxidant power (FRAP) level and malonodialdehyde concentration. HO-1 mRNA expression was examined with real-time PCR. The extent of DNA damage was determined by the alkaline-labile comet assay. A potential pro-apoptotic quercetin action was determined using the FITC-Annexin V kit. The quercetin and isorhamnetin concentrations in serum were analyzed by HPLC-ECD. MDA concentration and FRAP values, were significantly decreased in the spleen and bone marrow cells of rats treated with quercetin, in a dose of 80mg/kg b.w. in comparison with the control rats; no significant changes were observed after quercetin was administered in a dose ten times as low. Treatment with quercetin dose-dependently upregulated the expression of HO-1 mRNA in the bone marrow cells. Quercetin administration to the rats did not induce either DNA damage or apoptosis in the examined cells. The results of our study prove that changes in the antioxidant state, caused by quercetin, do not lead to DNA damage or exert any pro-apoptotic activity in vivo.     

辽西蜂胶醇提取物与槲皮素对培养心肌细胞过氧化损伤保护作用的比较研究

目的 评价由改良的聚酰胺柱层析技术研制的辽西蜂胶醇提取物(EEP)对心肌细胞过氧化损伤的保护作用及强度.方法 参照文献方法并加以改进获得FEP样品,以1000μmol/L的Fe2 和500μmol/L的半胱氨酸造成心肌细胞过氧化损伤,通过检测培养液中乳酸脱氢酶(LDH)的漏出量、培养细胞丙二醛(MDA)和超氧化物歧化酶(SOD)含量等指标,对比研究EEP和槲皮素对心肌细胞过氧化损伤的保护作用.结果 EEP和槲皮素在25～100μg/Ml均能剂量依赖性地对抗培养介质中LDH漏出和培养细胞MDA的产生,并能对培养细胞SOD具有保护作用;抑制LDH漏出,EEP和槲皮素的IC50分别为22.9和27.4μg/Ml;抑制MDA产成,EEP和槲皮素的IC50分别为43.9和36.2μg/Ml;保护SOD活力,EEP和槲皮素对抗SOD活力降低的IC50分别为23.3和147.6μg/Ml.结论 对于Fe2 /半胱氨酸体系造成的培养乳鼠心肌细胞过氧化损伤模型,EEP对心肌细胞过氧化损伤和对SOD活性的保护作用均强于槲皮素,但对心肌细胞脂质过氧化的抑制作用,EEP略弱于槲皮素.     

高效液相色谱法测定槲皮素纳米脂质体药物含量及包封率

目的:建立测定槲皮素脂质体药物含量及包封率的 RP—HPLC 法。方法:采用 Diamonsil~(TM)C_(18)柱(4.6 mm×250 mm,5μm);流动相:甲醇-0.4%磷酸(55:45);柱温:30℃;流速:1.0 mL·min~(-1);紫外检测波长:360 nm。采用透析法分离槲皮素脂质体中的游离药物。结果:在本色谱条件下槲皮素与辅料及溶剂峰分离良好,槲皮素在1.2～50μg·mL~(-1)范围内线性关系良好(r=0.9998,n=5),回收率在99.7%～101.2%之间,日内 RSD 及日间 RSD 均小于2%(n=5)。结论:该方法准确可靠、简单快速,可用于槲皮素纳米脂质体含量及包封率的测定。     

RP—HPLC同时测定菝葜药材中总槲皮素和山柰酚含量

目的:建立同时测定菝葜药材中槲皮素和山柰酚含量的反相高效液相色谱法。方法:色谱柱:Lichrospher C_(18)柱(250 mm×4.6 mm,5 μm),流动相:甲醇-0.5%磷酸水溶液(55:45),流速:1.0 mL·min~(-1),检测波长:365 nm。结果:槲皮素在2.97～29.7μg·mL~(-1)浓度范围内线性关系良好(r=0.9999),山柰酚在3.01—30.1μg·mL~(-1)浓度范围内线性关系良好(r=0.9999),方法回收率分别为98.9%和99.7%。结论:本方法简便、准确,灵敏度高,重复性好,可为评价菝葜药材质量提供依据。     

HPLC同时测定维吾尔族药复方小艾飞中5种有效成分的含量

目的:建立HPLC同时测定复方小艾飞中槲皮素、山柰素、乔松素、高良姜素4种黄酮类成分及胡椒碱含量的方法。方法:采用Hypersil BDS C18色谱柱(4.6 mm×250 mm,5μm),以乙睛(A)-1%磷酸溶液(B)-甲醇(C)为流动相,梯度洗脱(0~12 min,28%A∶52%B∶20%C;12~13 min,28%~30%A,52%~50%B,20%C;13~45 min,30%A∶50%B∶20%C),流速1.0 m L·min-1,柱温30℃,进样量20μL,检测波长280 nm。结果:槲皮素、山柰素、乔松素、高良姜素、胡椒碱的标准曲线分别为Y=357 314X-88 066(r=0.999 9),Y=442 853X-74 204(r=0.999 9),Y=647 926X-327 436(r=0.999 9),Y=471 672 X-256 408(r=0.999 9),Y=290 000X-391 906(r=0.999 9),线性关系良好。平均加样回收率(n=9)分别为98.4%(RSD 1.6%),98.9%(RSD 1.7%),101.3%(RSD 2.2%),98.6%(RSD 2.1%),98.6%(RSD 1.8%)。结论:该方法准确、简便,具有良好的重复性,可用于维吾尔族药复方小艾飞的质量控制。     

基于UPLC指纹图谱和黄酮醇苷含量的不同来源银杏叶的质量比较

目的 比较不同产地、性别、树龄银杏Ginkgobiloba叶在同一采收期的质量,为其资源评价提供依据。方法 收集5个产地共36批的银杏单株叶样品,采用超高效液相色谱法测定芦丁、槲皮素、异鼠李素和山柰素含量。用国家药典委员会中药色谱指纹图谱相似度评价系统软件处理分析,建立银杏叶药材UPLC指纹图谱。结合主成分分析及偏最小二乘判别分析探讨不同性别银杏叶的质量差异。结果 各组样品相似度均大于0.85,雌株之间相似度低于雄株;产地、性别和树龄对银杏叶的化学成分组成和比例有一定的影响,成熟植株指纹图谱相似度变化幅度大于苗期植株;HM居群与其它居群样品的物候期不同。结论 不同产地雌雄株银杏叶的性状特征存在差异。LT居群银杏叶质量较佳且更稳定;苗期植株稳定性高于成熟植株;不同性别银杏叶质量差异标志物有所差异。为控制银杏叶内在质量建立了UPLC指纹图谱快速检测方法,并进行指纹图谱相似度评价,结合主成分分析及偏最小二乘判别分析方法用于分析不同性别银杏叶的质量,为银杏叶的质量比较和资源评价提供了科学依据。     

RP-HPLC法测定鬼箭羽中3种活性成分的含量

目的:建立同时测定鬼箭羽中降糖活性成分山柰酚、柚皮素及槲皮素含量测定的方法。方法:采用反相高效液相色谱法,以AlltimaC18(4.6mm×250mm,5μm)为色谱柱,乙腈-0.1%甲酸(38:62)为流动相,检测波长258nm,流速0.8mL/min,柱温30℃。结果:山柰酚、柚皮素及槲皮素的线性范围分别为0.02064~0.2580μg(r=0.9996),0.05592~0.699μg(r=0.9997),0.01656~0.207μg(r=0.9998);加样回收率分别为100.6%(RSD=2.5%),96.4%(RSD=1.1%),101.9%(RSD=1.7%)。结论:本法简便、准确、重现性好,适用于鬼箭羽中3种活性成分的同时测定,可用于鬼箭羽药材的质量控制。