银杏内酯A、B对高胆固醇诱导的PC12细胞胆固醇代谢的影响

[目的] 基于体外血脑屏障(BBB)环境, 观察高胆固醇环境下PC12细胞胆固醇的代谢情况及银杏内酯A、B对其的影响。[方法] 利用Transwell装置共培养星形胶质细胞和脑微血管内皮细胞建立体外BBB, 并与PC12细胞共培养。在Transwell 内皮细胞侧添加40 μmol/L胆固醇, 并分别给予30 μmol/L 银杏内酯A和25 μmol/L 银杏内酯B进行干预。干预结束后, 采用反转录聚合酶链式反应(RT-PCR)技术, 检测胆固醇代谢关键酶低密度脂蛋白受体相关蛋白-1(LRP-1)、胆固醇酰基转移酶(ACAT)、ATP结合盒转运子A1(ABCA1)及胆固醇24S-羟化酶基因(CYP46)在mRNA水平的表达。[结果] 高胆固醇环境下, LRP-1表达下调(P<0.01), ACAT 及ABCA1表达上调(P<0.05), CYP46的表达有上调趋势;药物干预后, 除LRP-1有不同程度的升高(P<0.05), 其余各指标均无明显变化。[结论] GKA、B对高胆固醇环境下PC12细胞胆固醇代谢关键酶LRP-1的表达有一定影响。     

高效液相色谱法测定通腑宁颗粒去氢木香内酯含量

[目的]建立通腹宁颗粒中去氢木香内酯的含量测定方法。[方法]采用高效液相色谱法,色谱柱：Welchrom-C18（250mm×4．6μm）;流动相：甲醇．水（65：35）;流速：1mL／min;检测波长：225nm;柱温：常温。[结果]回收率RSD=1．11％。检测3批样品通腹宁颗粒中去氢木香内酯含量,限定通腹宁颗粒中去氢木香内酯含量不得低于2．5mg。[结论]高效液相色谱法测定通腹宁颗粒中去氢木香内酯含量,结果准确、可靠,可用于通腹宁颗粒质量控制。     

青蒿不同部位薄层鉴别比较研究

目的:通过收集青蒿植株不同部位的药材,对其主要化学成分进行定性研究,为黄花蒿药材资源的开发与利用奠定基础。方法:采用薄层鉴别法。结果:所建立的方法适用于青蒿不同部位的化学成分鉴别。结论:青蒿子、青蒿地上部分以及青蒿叶成分最为相似;花所含的挥发油种类最多,茎最少;茎可能不含或是含有极微量的青蒿素。其中,青蒿子、花、茎的薄层鉴别属首次报道。     

Anti-proliferative activity of aguerin B and a new rare nor-guaianolide lactone isolated from the aerial parts of Centaurea deflexa

The phytochemical investigation of the aerial parts of Centaurea deflexa led to the identification of 21 compounds, among which three phenolic acids, one sterol, ten flavonoids, one phenylpropanoid derivative, two lignans and four sesquiterpene lactones. One of the latter compounds was a new, rare active principle (1) having an uncommon 15-nor-guaianolide skeleton. The biological investigation was carried out through a bio-guided assay fractionation of C. deflexa extracts and highlighted an anti-proliferative activity of two sesquiterpene lactones, aguerin B and the newly identified 15-nor-guaianolide (1) against human pancreatic and colonic cancer cells. Of the two compounds, only aguerin B showed to induce apoptotic cell death, confirming the role as pro-apoptotic moiety of the α-methylene-γ-lactone ring present in aguerin B but not in 1.     

Inhibition of calcium-independent phospholipase A2 prevents inflammatory mediator production in pulmonary microvascular endothelium

Inhalation of allergens can result in mast cell degranulation and release of granule contents, including tryptase, in the lung. Injury to human pulmonary microvascular endothelial cells (HMVEC-L) can also result in activation of the coagulation cascade and thrombin generation. We hypothesize that these proteases activate calcium-independent phospholipase A₂ (iPLA₂), in HMVEC-L, leading to the production of membrane phospholipids-derived inflammatory mediators. Both thrombin and tryptase stimulation of HMVEC-L increased iPLA₂ activity that was inhibited by pretreatment with the iPLA₂ selective inhibitor bromoenol lactone (BEL). Arachidonic acid and prostaglandin I₂ (PGI₂) release were also increased in tryptase and thrombin stimulated cells and inhibited by BEL pretreatment. Pretreating the endothelial cells with AACOCF₃ a cytosolic PLA₂ inhibitor did not inhibit tryptase or thrombin induced arachidonic acid and PGI₂ release. In addition thrombin and tryptase also increased HMVEC-L platelet activating factor (PAF) production that significantly contributes to the recruitment and initial adherence of polymorphonuclear neutrophils (PMN) to the endothelium. Tryptase or thrombin stimulated increase in PMN adherence to the endothelium was inhibited by pretreatment of HMVEC-L with BEL or pretreatment of PMN with CV3988, a PAF receptor specific antagonist. Collectively, these data support our hypothesis that iPLA₂ activity is responsible for membrane phospholipid hydrolysis in response to tryptase or thrombin stimulation in HMVEC-L. Therefore selective inhibition of iPLA₂ may be a pharmacological target to inhibit the early inflammation in pulmonary vasculature that occurs as a consequence of mast cell degranulation or acute lung injury.     

基于色差原理分析木香有效成分含量与颜色值的相关性

目的:通过测定木香粉末色差值及有效成分木香烃内酯、去氢木香内酯和挥发油的含量,将代表颜色的指标值与代表质量的指标值相关联,探讨有效成分含量与颜色值之间的相关性,为木香的质量评价提供依据。方法:利用色差计对木香粉末色差值进行测量,以流动相甲醇-水(65∶35)等度洗脱,检测波长225 nm,采用HPLC测定木香烃内酯、去氢木香内酯的含量,运用水蒸气蒸馏法测定木香挥发油的含量,通过SPSS 21.0软件进行相关性分析。结果:木香烃内酯、去氢木香内酯、挥发油含量与L*(代表颜色深浅),b*(代表颜色红绿方向),E*ab(代表总色差)存在负相关关系,且效果极显著(P0.01),但与a*(代表颜色红绿方向)不显著相关。结论:L*,b*,E*ab越小,3种有效成分的含量越高。木香颜色值与3种有效成分含量都有关,且颜色深的木香有效成分含量较高,质量较好。通过利用色差计对木香粉末的色差值进行测定,可以快速预测木香中3种有效成分的含量,可为该药材质量评价体系的建立提供参考。     

天然生物活性N-酰基高丝氨酸内酯小分子抗原的化学合成及其抗体生成初探

目的人工合成N-酰基高丝氨酸内酯(N-acylated homoserine lactones,N-aHSLs)完全抗原并制备相应单抗,建立N-aHSLs免疫检测的新方法及海洋弧菌N-aHSLs动态监测。方法以月桂酸和高丝氨酸内酯为主要原料,通过多步系列化学反应人工合成N 十二酰高丝氨酸内酯(N-lauroyl-homoserine lactone,N-C12-HSL)及含羧基活性交联基团的半抗原衍生物12-羧基-N-十二酰高丝氨酸内酯（12-carboxyl-N-decanoyl-L-homoserine lactone,12-CD-LHL）,用1H-磁共振、高压/效液相色谱仪和液相色谱 质谱联用方法确证合成产物的结构,并用生物感应法检测合成N-aHSLs的天然生物活性;用N-羟基琥珀酰亚胺活泼酯法将12-CD-LHL半抗原衍生物连接到载体蛋白;将N-C12-HSL人工完全抗原通过传统单抗制备技术,获得抗N-aHSLs杂交瘤细胞株,用酶联免疫吸附测定法鉴定其特异性和效价。结果质谱、磁共振氢谱分析结果与理论预测相同,成功合成目标产物N-C12-HSL和12-CD-LHL半抗原衍生物;所合成的N-aHSLs能够分别激活生物感应菌株大肠杆菌MG4和紫色色杆菌026并使之显色;紫外吸收光谱扫描结果显示,半抗原已分别与牛血清白蛋白和卵清蛋白发生偶联,成功制备完全抗原;共获得3株抗N-C12-HSL单抗,均具有良好的免疫反应性。结论N-aHSLs完全抗原和相应的单抗制备为N-aHSLs动态监测和后续研究奠定了基础。     

Screening for Compositae sensitization with pure allergens: implications of molecular structure, strength of reaction, and time of testing

Background. The sesquiterpene lactone (SL) mix is the only commercial Compositae allergy screening agent that consists of pure allergens; its detection rate is lower than that of Compositae plant extracts. Objective(s). To report a 15‐year experience with routine screening with SL mix and another allergen, parthenolide, the optimal reading times when testing with SL mix constituents, and the advantages and drawbacks of using pure allergens. Materials and methods. The SL mix 0.1% and parthenolide 0.1% petrolatum were included in the baseline series. Results. Of individuals undergoing routine testing, 157/7163 (2.19%) tested positive to SL mix, 161/7162 (2.25%) to parthenolide, and 141 to both in the 15‐year period. The overall detection rate between the two was 177 persons, and 10% would have been missed without parthenolide testing. Two cases of possible active sensitization to parthenolide were recorded. Costunolide elicited positive reactions most frequently, followed by dehydrocostus lactone, and alantolactone. An important observation was that the prevalence of positive reactions to SL mix constituents was lower if they were applied 3–5 days after application of SL mix in those with 1+ or 2+ reactions. Conclusions. The benefit of detecting 10% more Compositae‐sensitive patients with parthenolide in the baseline series must be weighed against a small risk of active sensitization, which may be reduced by lowering the test concentration.