不同浓度地塞米松在体外定向诱导分化兔骨髓间充质干细胞为成骨细胞的实验研究

目的探讨兔骨髓间充质干细胞(MSCs)体外定向诱导分化为成骨细胞所需的地塞米松的最佳浓度。方法抽取兔骨髓,采用全骨髓贴壁培养法体外培养,贴壁细胞进行传代。取第3代细胞,随机分为5组,在培养基中添加成骨分化诱导剂β-甘油磷酸钠、L-抗坏血酸,5组添加的地塞米松的浓度分别为1×10-6mol/L、1×10-7mol/L、1×10-8mol/L、1×10-9mol/L、1×10-10mol/L。培养10 d后,在倒置显微镜观察细胞形态,计算细胞浓度,采用5-溴-4-氯-3-吲哚磷酸盐(BCIP)/氯化硝基四氮唑蓝(NBT)底物显色试剂检测碱性磷酸酶(ALP),茜素红染色检测钙结节。结果较高浓度(1×10-8mol/L、1×10-7mol/L、1×10-6mol/L)的地塞米松相对于较低浓度(1×10-9mol/L、1×10-10mol/L)的地塞米松可显著增加ALP表达(P均<0.05),并可增加钙结节的形成;1×10-9mol/L及其以下浓度的地塞米松对细胞增殖无明显影响(P均>0.05),1×10-7mol/L以上浓度的地塞米松可明显抑制细胞的增殖(P<0.01)。结论地塞米松在体外定向诱导分化兔骨髓间充质干细胞为成骨细胞,最佳的浓度为1×10-8mol/L。     

姜黄素对人癌细胞BCH、MGC-803、BEL-7402细胞增殖的影响

目的观察姜黄素对人癌细胞系BCH、MGC-83、BEL-7402细胞增殖的影响.方法采用Mosman MTT细胞增殖检测法.结果姜黄素在4×10-5mol/L与BCH细胞接触,在不同时相的药物细胞毒效应不同,其与BCH细胞接触16h是药物细胞毒作用最佳时期.姜黄素在(1×10-5～1×10-4)mol/L范围对上述三种体外培养癌细胞有明显的抑制活性.其IC50分别为3.58×10-5mol/L(BCH)、3.83×-5mol/L(MGC-803)、7.3×10-5mol/L(BEL-7402),并且药效与药物浓度呈相关性.结论姜黄素对三种癌细胞的增殖有明显抑制作用.其药效浓度达到国内抗肿瘤药体外药效评价标准.     

丹皮酚的电化学性质研究及其应用

目的建立丹皮酚电化学检测方法,并应用于丹皮中丹皮酚含量的测定。方法循环伏安(CV)、差分脉冲(DPV)。结果在0.1 mol/L的NaOH-NaH2PO4(pH=11)底液中1、00 mv/s的扫描速度下丹皮酚在铂电极上有一灵敏的氧化峰,峰电流与其浓度在5.0×10-6~1×10-3mol/L的范围内呈良好的线性关系,其线性回归方程为:A=0.23-4 082.64×C(r=0.998 4),检出限为1×10-6mol/L。结论该方法准确可靠,操作简便、快速,重现性好,可用于在线和野外检测。     

艾纳香二氢黄酮对脂质过氧化损伤大鼠原代培养肝细胞的保护作用

研究 5种艾纳香二氢黄酮类化合物对过氧化损伤的原代培养肝细胞的保护作用。胶原酶灌流法分离大鼠肝细胞。以 CCl4或 Fe SO4 Cys损伤原代培养肝细胞 ,1× 10 - 4 和 1× 10 - 5 mol/ L 的 5种艾纳香二氢黄酮均能明显抑制受损伤细胞的转氨酶逸出、MDA产生及 GSH耗竭。其中以 2 ,5 -二羟基艾纳香二氢黄酮的作用最强 ,在 CCl4所致肝细胞损伤实验中 ,其抑制 MDA产生的 ED5 0 约为 2 .6× 10 - 6 mol/ L ,保护 GSH的 ED5 0 约为 1.0 8× 10 - 5 mol/ L ,抑制 AL T逸出细胞的 ED5 0 约为 9.39× 10 - 6 mol/ L     

染料木素对大鼠成骨细胞增殖与分化的影响

目的:探讨染料木素体外对大鼠成骨细胞增殖与分化的影响。方法:用改良的组织块法分离培养新生大鼠颅骨成骨细胞,染料木素以不同浓度加入细胞培养体系,作用不同时间后,用MTT法检测成骨细胞的增殖情况;用对硝基苯二钠基质动力学法测定细胞内碱性磷酸酶的活性,用改良的Lowry法测蛋白含量。结果:染料木素在1×10~(-5)～1×10~(-9)mol/L浓度范围内作用24h、48h均能促进成骨细胞增殖,在1×10~(-7)～1×10~(-5)mol/L范围内作用72 h可提高成骨细胞内碱性磷酸酶的活性。结论:染料木素体外能促进成骨细胞的增殖与分化。     

问荆碱对大鼠脑囊泡膜Mg2+-ATPase,Ca2+-ATPase抑制作用的动力学研究

问荆碱对大鼠脑囊泡膜Mg²⁺-ATPase,Ca²⁺-ATPase有显著的抑制作用,对Mg²⁺-ATPase的抑制机制分别为竞争性抑制(Mg²⁺,Ki=4.93×10^-5mol/L)、非竞争性抑制(ATP,Ki=1.06×10^-4mol/L),对Ca²⁺-ATPase的抑制机制分别为混合型抑制(Ca²⁺,Ki=5.07×10^-5mol/L)、非竞争性抑制(ATP,Ki=3.27×10-4mol/L)。     

大黄素对白三烯B_4和前列腺素E_2生物合成的影响

本文研究了中药大黄的主要有效成份之一——大黄素对白三烯B_4(LTB_4)和前列腺素E_2(PGE_2)生物合成的影响。结果表明:它抑制人多形核白细胞LTB_4合成的IC_(50)值为6.4×10~(-6)mol/L(无外源性花生四烯酸)和2.6×10~(-5)mol/L(有外源性花生四烯酸),并可显著抑制全血中LTB_4的合成(兔的半体内实验),在本文各实验中,它对PGE_2的合成均未呈抑制作用,表明大黄素是一个选择性5—脂氧酶抑制剂。     

基于P糖蛋白研究仙茅对顺铂耐药非小细胞肺癌细胞的增敏机制

目的：探究中药仙茅水煎液及其主要入血成分苔黑酚葡萄糖苷(OG)对非小细胞肺癌(NSCLC)顺铂耐药细胞(A549/cis)耐药性的影响及其潜在机制。方法：用AutoDock Vina分子对接软件评估P糖蛋白(P-gp)与OG的结合能力；CCK-8法检测顺铂、仙茅水煎液、OG及其联用对A549/cis细胞活力的抑制率；蛋白质免疫印迹法检测A549/cis中P-gp的表达水平。结果：P-gp与OG模拟分子对接的结合自由能为-7.6 kcal/mol（1 cal=4.184 J），二者具有良好的结合活性；顺铂对A549/cis的半抑制浓度(IC50)约为5.5 μg/mL，仙茅水煎液与OG在梯度浓度下对A549/cis活力均无明显影响；顺铂低剂量(1.5 μg/mL)与仙茅水煎液高、低剂量(1 000 μg/mL、500 μg/mL)或OG高剂量(5×10-5mol/L)联用组、顺铂中剂量(4 μg/mL)与仙茅水煎液高、低剂量(1 000 μg/mL、500 μg/mL)或OG高、低剂量(5×10-5mol/L、1×10-5mol/L)联用组较顺铂单用组细胞抑制率均显著升高(P<0.05或P<0.01)；顺铂(1.5 μg/mL)与仙茅水煎液(1 000 μg/mL)或OG(5×10-5 mol/L)联用组A549/cis细胞的P-gp表达水平均低于顺铂单用组，其中顺铂与仙茅水煎液联用组降低显著(P<0.05)。结论：中药仙茅水煎液及OG均可以降低A549/cis细胞的耐药性，其增敏机制可能与抑制P-gp表达有关。     

金丝桃甙对坐骨神经钙含量的影响

目的:研究金丝桃甙(Hyp)对神经组织Ca~（2+）含量的影响。方法:游离的蛙坐骨神经Ca~（2+）含量用原予分光光度仪测定。结果;Hypl×l0~(-5),1×10~(-6)和1×10~(-7)mol.L~(-1)显著并呈浓度依赖性抑制50mmol,L~(-1)K~+诱导Ca~(2+)含量增加,1×10~(-4)mol.L~(-1)Hyp显著抑制10~(-5)mo1.L~(-1)NE诱导的Ca~(2+)含量增加。结论:Hyp对神经组织Ca~(2+)内流有阻滞作用。     

舒络对LPC致基底动脉环收缩反应的影响

目的研究溶血磷脂酰胆碱(LPC)对狗脑基底动脉环收缩的影响及舒络的保护作用.方法通过描记犬基底动脉环的张力变化,测定LPC对基底动脉环收缩的影响.结果LPC能剂量依赖性(1×10-9～1×10-3mol/L)促进基底动脉环收缩,在1×10-5mol/L时收缩达最大,为(133.0±11)%.舒络能剂量依赖性地减弱LPC诱导的狗脑基底动脉环收缩.结论舒络能拮抗LPC诱导的狗脑基底动脉环收缩.     

辛夷挥发油体外干预大鼠胸腔炎性白细胞5-LO活性的研究

目的:观察辛夷挥发油体外对大鼠胸腔炎性白细胞脂氧酶(5-LO)活性的影响,探讨其干预花生四烯酸代谢的抗炎机制。方法:以角叉菜胶所致急性胸膜炎模型大鼠的胸腔炎性白细胞为基本反应体系,分别加入CaCl2、MgCl2、花生四烯酸(AA)、A23187以及辛夷挥发油(0.011、0.044、0.088、0.132mg/ml)或齐留通(0.625μg/ml),提取完整白细胞的5-LO代谢产物白三烯B4(LTB4)和5-羟基二十碳四烯酸(5-HETE),运用反相高效液相色谱(RP-HPLC)紫外检测分离测定LTB4和5-HETE的合成水平,来反映5-LO的活性。结果:辛夷挥发油在0.011~0.132mg/ml浓度范围内,对大鼠胸腔白细胞LTB4与5-HETE的生物合成呈现抑制趋势。结论:辛夷挥发油体外可抑制大鼠胸腔白细胞花生四烯酸代谢酶5-LO的活性,降低LTB4与5-HETE合成水平,提示其抗炎作用可能与抑制5-LO活性,减少致炎代谢产物的生成有关。     

RP—HPLC法测定白细胞中LTB4和5-HETE的含量

目的：建立了5-脂氧酶抑制剂筛选模型的RP—HPLC测定方法。方法：采用BravaBDSC18柱（250mm×4．6mm，5μm），流动相为甲醇-0．02％乙酸水溶液，流速为0．8mL·min^-1，5-LOX的主要产物ⅡB4和5-HETE的检测波长分别为275nm和235nm。结果：LTB4和5-HETE分别在2．0～50．0μg·mL^-1（r=0．9999）和5．0～125．0μg·mL^-1（r=1．0000）范围内呈良好的线性关系；回收率分别为100．0％和97．7％（RSD分别为1．1％和1．5％）；日内、日间精密度的RSD分别为0．7％，1．1％和0．3％，1．3％。结论：该法分离效果和重复性均良好，且结果准确可靠，可用于LTB4和5-HETE的检测，为筛选新的具有抗炎活性的中药成分奠定了基础。     

Studies on the activities of tannins and related compounds, X. Effects of caffeetannins and related compounds on arachidonate metabolism in human polymorphonuclear leukocytes

As part of a series of biological examinations of various tannins and related compounds, the present paper reports the effects of caffeetannins and related compounds isolated from medicinal plants on arachidonate metabolism in human peripheral polymorphonuclear leukocytes (PMN-L). The formation of leukotriene B4 (LTB4) induced by calcium ionophore A 23187 (A 23187) in human PMN-L was inhibited by 3,5-, 4,5-, and 3,4-di-O-caffeoylquinic acid, caffeoylmalic acid, caffeoyltartric acid, rosmarinic acid, and caffeic acid. Rosmarinic acid strongly inhibited the formation of 5-hydroxy-6,8,11,14-eicosatetraenoic acid (5-HETE) and LTB4 (5-lipoxygenase products) at concentrations of 10(-5)-10(-3) M. On the other hand, the formation of prostaglandin E2 (PGE2) was enhanced in a concentration-dependent fashion by caffeic acid, caffeoylmalic acid, caffeoyltartaric acid, and 3,4-di-O-caffeoylquinic acid. On the basis of experimental results, it seems likely that caffeoyl derivatives could be developed as therapeutic drugs for treatment of allergic inflammation such as asthma.     

Effect of ganhuangenin obtained from Scutellaria radix on the chemical mediator production of peritoneal exudate cells and immunoglobulin E level of mesenteric lymph node lymphocytes in Sprague-Dawley rats

We previously showed that ganhuangenin (GHG) has beneficial antioxidative properties against lipid peroxidation in tissues of Sprague-Dawley (SD) rats. In this study, the effect of GHG on immunoglobulin E (IgE), histamine and leukotriene B(4) (LTB(4)) levels of cells isolated from SD rats was examined with regard to the manifestation of the type I allergic reaction. We showed that GHG inhibited the IgE production of mesenteric lymph node (MLN) lymphocytes. In the presence or absence of concanavalin A (ConA), the concentration used did not exert a toxic effect against MLN lymphocytes. Interestingly, the increase in the IgE content and lipid peroxidation induced by ConA was alleviated in the presence of GHG. Moreover, GHG also inhibited histamine release from the peritoneal exudate cells stimulated with a calcium ionophore, A23187, or with the histamine releaser, compound 48/80. In the case of LTB(4), GHG markedly inhibited its release at a concentration of 100 microM. Thus, it is concluded that GHG may block the common pathway for the release of histamine and LTB(4), and that the IgE level is responsible for the lipid peroxidation induced by ConA.     

喘宁软胶囊对多形核粒细胞生成和释放白三烯的抑制作用

目的：探讨喘宁软胶囊（简称喘宁）平喘作用机制。方法：分别采用生物检定法、HPLC法、免疫ELISA法离体测试大鼠多形核粒细胞（PMNs)生成和释放慢反应物质（SRS－A）、白三烯B4（LTB4）、白三烯C4（LTC4）。结果：在离体试验中，喘宁0．1，0．4，1．6，6．4g/L浓度及喘宁4g/kg ig 20d所得血清能明显抑制大鼠PMNs生成和释放SRS－A；喘宁0．025，0．05，0．1g/L及0．1，0．4，1．6g/L浓度呈量效关系分别抑制大鼠PMNs生成和释放LTB4，LTC4。结论：初步证明喘宁有平喘作用，其机制与抑制花生四烯酸（AA）生成和释放白三烯有关。     

The activity against leukocyte eicosanoid generation of essential oil and polar fractions of Adesmia boronioides Hook.f

The polar fractions (methanol and aqueous extract) and essential oil of Adesmia boronioides aerial parts were tested in vitro at concentrations of 15 and 50 microg/mL, for their effects on the COX and 5-LOX pathways of eicosanoid generation (TXB2, PGE2 and LTB4) in stimulated rat peritoneal leukocytes. Potent inhibition of LTB4 generation was displayed by the methanol extract and the essential oil, whereas the aqueous extract was essentially inactive. The methanol extract also caused potent inhibition of TXB2 generation but the essential oil and the aqueous extract were much less active. The effects on PGE2 production were much less striking, implying that the main effect is on thromboxane synthetase rather than on COX. Although the essential oil caused LDH release in leukocytes treated concurrently with ionophore, suggesting substantial toxicity to the cells, this extract did not affect cell viability according to the MTT test when incubated with the cells in the absence of ionophore. The two other extracts did not affect cell viability at the concentrations tested. It is concluded that Adesmia boronioides contains substance(s) that preferentially inhibit the 5-lipoxygenase activity of arachidonic acid metabolism and suggest that this may contribute to the antiinflammatory actions of extracts of this plant.     

Effects of wogonin,wogonoside, and 3,5,7,2',6'-pentahydroxyflavone on chemical mediator production in peritoneal exduate cells and immunoglobulin E of rat mesenteric lymph node lymphocytes

Wogonin (WG), wogonoside (WGS), and 3,5,7,2',6'-pentahydroxyl flavanone (PHF) were isolated from Scutellaria baicalensis, and their effects on histamine, leukotriene B(4) (LTB(4)), and immunoglobulin E (IgE) were examined in rats, observing for a manifestation of a type I allergic reaction. WG and WGS in the amounts of 10 and 100 microM were shown to markedly inhibit histamine release in cells stimulated with calcium ionophore A23187 or compound 48/80. PHF exerted inhibitory activity only at 100 microM. In the case of LTB(4), WG, WGS and PHF markedly inhibited LTB(4) production at the concentration of 100 microM. We also find that the increase in the IgE content induced by concanavalin A (ConA) was alleviated in the presence WG and WGS, while the inhibitory effect of PHF was much weaker. However, the magnitude of inhibitory effect observed on the content of lipid peroxidation induced by ConA was in order of PHF > WG > WGS, with PHF being the strongest. Interestingly, WG and WGS with the methoxyl group strongly inhibited histamine and IgE production, whereas PHF with the hydroxyl group in the B ring was much stronger than WG and WGS against lipid peroxidation. Based on data, it was concluded that the flavonoid components, WG, WGS, and PHF, may block a common pathway for the release of histamine and LTB(4), and that the IgE level is responsible for the lipid peroxidation induced by ConA.     

Lichen metabolites. 1. Inhibitory action against leukotriene B4 biosynthesis by a non-redox mechanism.

Of several lichen metabolites isolated from Parmelia nepalensis and Parmelia tinctorum, the didepsides atranorin (4) and diffractaic acid (5), as well as (+)-protolichesterinic acid (7), inhibited LTB4 biosynthesis in polymorphonuclear leukocytes. Ethyl hematommate (3) and (+)-usnic acid (1) were only weak inhibitors, while methyl beta-orcinolcarboxylate (2) and gyrophoric acid (6) were inactive at concentrations up to 60 microM. Redox properties of the compounds were evaluated in terms of inhibition of nonenzymatic lipid peroxidation in model membranes, reactivity against the stable free radical 2,2-diphenyl-1-picrylhydrazyl, and deoxyribose degradation as a measure of hydroxyl-radical generation. The results revealed that lichen metabolites neither acted as antioxidants against the peroxidation process in model membranes nor did they scavenge or produce free radicals, suggesting that the inhibitory effects on LTB4 biosynthesis was due to specific enzyme interaction rather than a nonspecific redox mechanism.     

六种苦豆子生物碱对炎症介质白三烯的影响

目的：研究苦参碱(matrine MT)、氧化苦参碱(oxymatrine OMT)、槐果碱(sophocarpine SC)、氧化槐果碱(oxysophocarpine OSC)、槐定碱(sophoridine SRI)、氧化槐定碱(oxysophoridine OSRI)6种苦豆子类生物碱对炎症介质白三烯巴和白三烯134的影响，来研究其抗炎作用机制。方法：以大鼠腹腔白细胞为材料，采用高效液相色谱法(HPLC)测定白细胞内白三烯C4和白三烯B4水平。结果：SRI对LTC4的生物合成影响很小，但对LTB4的生物合成有明显的抑制作用。其余5种生物碱对LTC4和LTB4的生物合成均有明显的抑制作用，且有良好的剂量-效应关系。结论：6种苦豆子生物碱对白三烯(LTC4和LTB4)的生物合成均具有不同程度的抑制作用，这可能是它们抗炎作用的机制之一。     

平喘宁对哮喘豚鼠白三烯B4、可溶性细胞间粘附分子-1含量的影响

目的观察平喘宁对哮喘豚鼠白三烯B4(LTB4)、可溶性细胞间粘附分子-1(sICAM-1)含量的影响,探讨其治疗哮喘的相关生物学机制。方法以卵蛋白致敏复制豚鼠哮喘病模型,观察豚鼠引喘潜伏期的变化;运用酶联免疫吸附反应技术(ELISA)测定豚鼠血清中的LTB4和支气肺泡灌洗液(BALF)sICAM-1的含量。结果平喘宁给药后,豚鼠的引喘潜伏期显著延长;平喘宁能显著降低哮喘豚鼠血清中LTB4和支气管肺泡灌洗液(BALF)中sICAM-1含量,与哮喘模型组比较,P<0.05或P<0.01。结论平喘宁具有明显平喘作用,其作用机制与降低哮喘豚鼠血清中LTB4和BALF中sICAM-1的含量有关。