硝普钠与异丙肾上腺素松弛犬气管平滑肌的协同作用

目的：研究硝普钠（ＳＮＰ）与异丙肾上腺素（ＩＳＯ）对犬气管平滑肌松弛作用是否存在协同关系，并分析协同的机制。方法：采用放射免疫分析法测定犬气管平滑肌环核苷酸量，并用磷酸二酯酶（ＰＤＥ）同工酶抑制剂为工具分析ＳＮＰ与ＩＳＯ相互作用的机制。结果：ＳＮＰ（１～１００μｍｏｌ·Ｌ－１）增强ＩＳＯ（３０μｍｏｌ·Ｌ－１）对３μｍｏｌ·Ｌ－１乙酰甲胆碱预收缩的犬气管平滑肌的松弛作用。ＳＮＰ（１００μｍｏｌ·Ｌ－１）与ＩＳＯ（３０μｍｏｌ·Ｌ－１）对犬气管平滑肌松弛的协同作用伴有气管平滑肌中ｃＡＭＰ积聚比ＳＮＰ与ＩＳＯ单独时分别增加１．２倍与０．４倍。ＰＤＥⅤ抑制剂扎普司特（３μｍｏｌ·Ｌ－１）使ＳＮＰ的气管平滑肌ｃＧＭＰ积聚由２．４倍增加到４．６倍，可进一步增加气管松弛作用１．２倍。合并ＰＤＥⅢ抑制剂氰胍哒嗪（３０μｍｏｌ·Ｌ－１）和ＰＤＥⅣ抑制剂咯利普兰（３０μｍｏｌ·Ｌ－１）或合并ＳＮＰ（１００μｍｏｌ·Ｌ－１）和咯利普兰（３０μｍｏｌ·Ｌ－１）均能进一步增强ＩＳＯ的气管ｃＡＭＰ积聚与松弛作用。结论：ＳＮＰ是通过气管平滑肌ｃＧＭＰ含量增加而抑制ＰＤＥⅢ，导致ＳＮＰ与ＩＳＯ对气管松弛的协同作用。     

磷酸二酯酶同工酶Ⅲ和Ⅳ调节气管平滑肌环腺鞋酸的区…

磷酸二酯酶同工酶Ⅳ（ＰＤＥⅣ）抑制剂咯利普兰（Ｒｏｌ，３０μｍｏｌ·Ｌ＾－１）对３０μｍｏｌ·Ｌ＾－１异丙肾上腺素所致的犬气管平滑肌环腺苷酸（ｃＡＭＰ）积聚的增强作用比ＰＤＥⅢ抑制剂氰腺哒嗪（ＳＫＦ９４８３６，３０μｍｏｌ·Ｌ－１）要强，但对异丙肾上腺素所致的犬气管平滑肌松弛的增强作用比ＳＫＦ９４８３６要弱。３０μｍｏｌ·Ｌ＾－１Ｒｏｌ或ＳＫＦ９４８３６对３０μｍｏｌ·Ｌ＾－１福斯科林所致的犬气管     

磷酸二酯酶同工酶Ⅲ和Ⅳ调节气管平滑肌环腺苷酸的区域化分布

磷酸二酯酶同工酶Ⅳ（ＰＤＥⅣ）抑制剂咯利普兰（Ｒｏｌ，３０μｍｏｌ·Ｌ－１）对３０μｍｏｌ·Ｌ－１异丙肾上腺素所致的犬气管平滑肌环腺苷酸（ｃＡＭＰ）积聚的增强作用比ＰＤＥⅢ抑制剂氰胍哒嗪（ＳＫＦ９４８３６，３０μｍｏｌ·Ｌ－１）要强，但对异丙肾上腺素所致的犬气管平滑肌松弛的增强作用比ＳＫＦ９４８３６要弱．３０μｍｏｌ·Ｌ－１Ｒｏｌ或ＳＫＦ９４８３６对３０μｍｏｌ·Ｌ－１福斯科林所致的犬气管平滑肌松弛与ｃＡＭＰ积聚的增强作用程度几乎相等．异丙肾上腺素，福斯科林单独或与ＳＫＦ９４８３６，Ｒｏｌ合用都仅引起犬气管平滑肌的可溶部ｃＡＭＰ积聚，对颗粒部ｃＡＭＰ含量无明显影响．上述结果提示犬气管平滑肌存在着ｃＡＭＰ的区域化分布，它受ＰＤＥⅢ，ＰＤＥⅣ的调节，而与平滑肌细胞的亚细胞结构无关     

普鲁托品对家兔主动脉扩血管作用的机理

应用血管平滑肌等张收缩和放射免疫测定环核苷酸水平等方法研究了普鲁托品 ( Pro)的扩血管作用及其机理 .结果表明 ,Pro浓度依赖性地松弛去氧肾上腺素 ( PE,1 μmol· L-1)和 KCl( 30 mmol·L-1)预收缩的兔胸主动脉螺旋条 ,EC50 分别为 ( 37± 2 0 )和 ( 4 0± 1 1 ) μmol·L-1.Pro30 μmol·L-1使PE和 KCl收缩兔胸主动脉的量效曲线非平行右移 ,最大反应压低 ,p D2 ′分别为 4.0 7± 0 .1 7和 3.86± 0 .1 8.格列本脲 ,N- L-硝基精氨酸 ,普萘洛尔和吲哚美辛不影响 Pro的舒血管作用 .Pro1 0和 1 0 0μmol· L-1显著增加兔胸主动脉血管平滑肌内c AMP和 c GMP水平 .结果提示 ,Pro的扩血管作用可能与增高平滑肌细胞内 c AMP和 c GMP水平有关 .     

防风及其栽培品饮片质量控制与分析

目的 对外观差异较大的防风及其栽培品进行分析比较, 探讨防风栽培品及饮片的质量标准. 方法 参照《中华人民共和国药典》2010年版一部防风项下性状、显微、薄层色谱、含量测定等方法 进行实验. 结果 防风与其栽培品在外观性状和内在质量上都有较大差别, 所含的主成分基本一致, 故薄层色谱、高效液相色谱区别很小. 结论 建议修订性状、显微鉴别, 拟订检查、含量测定限度, 完善用法用量等内容, 以完善《中华人民共和国药典》防风的质量控制方法 .     

HPLC法测定含当归药妆品中阿魏酸的含量

摘 要 目的:建立HPLC法测定含当归药妆品中阿魏酸的含量。方法: 采用HPLC法测定,色谱柱为依利特Sinochrom ODS BP(250 mm×4.6 mm,5 μm),甲醇-1%冰醋酸(45∶55)为流动相,流速为1.0 ml·min^-1,检测波长为324 nm,柱温25℃。结果:阿魏酸浓度在0.055～2.064 μg·ml^-1范围内与峰面积呈良好线性关系（r=0.999 8）;样品中阿魏酸的平均加样回收率为99.47%,RSD为1.53%（n=6）。结论:该法简便、快速准确、灵敏度高,重复性好。     

地塞米松磷酸钠对照品量值不确定问题的探讨

采用高效注相色谱的不同测定方法,对4个批号的地塞米松磷酸钠对照品的量值、降解程度进行了比较,考虑了不同批号对照品对供试品含量测定结果的影响,验证了该对照品量值的不确定性并提出了改进意见。     

普罗托品对家兔血小板功能的影响

普罗托品(protopine,Pro)体外(1—1000μmol·L~(-1))和体内(10和20mg·kg~(-1))均抑制ADP,胶原,花生四烯酸(AA)和烙铁头蛇毒血小板聚集素(TMVA)诱导的兔血小板聚集及血小板5-HT释放。Pro不抑制AA诱导的免血小板TXA_2生成。也不升高血小板内cAMP水平,但升高cGMP水平。提示其抗血小板作用的机制与升高血小板内cGMP水平,抑制血小板释放活性物质有关。     

普洛托品对兔胸主动脉平滑肌细胞增殖的影响

目的　观察普洛托品 (Pro)对正常和内皮素 (ET)诱导的血管平滑肌细胞 (VSMC)增殖的影响。方法　采用组织块贴壁法培养兔胸主动脉VSMC ,应用细胞计数、[3 H] TdR参入量、微量MTT比色测定及电镜法 ,观察Pro对VSMC增殖、DNA合成和细胞超微结构的变化。结果　ET对VSMC的增殖和DNA合成有显著的促进作用 ;1、10、10 0μmol·L-1浓度Pro能够逆转ET所致的 [3 H] TdR参入量、MTT比色的吸光度、细胞计数的增多及电镜下细胞超微结构的改变 ,且随着浓度的增加及时间的延长 ,抑制作用愈显著 ;10、10 0 μmol·L-1浓度Pro对未经ET处理的VSMC有抑制其增殖作用 ,但其抑制作用弱于ET处理组。结论　Pro可显著地抑制正常及ET诱导的VSMC增殖 ,并且对ET诱导的VSMC增殖抑制作用更明显 ,提示Pro可作为临床防治动脉粥样硬化的有效药物。     

高速逆流色谱法分离纯化延胡索乙素和原阿片碱

目的确定高速逆流色谱法分离纯化延胡索乙素和原阿片碱的条件。方法采用TBE300A型高速逆流色谱仪,以分配系数和分相时间为依据设计一组溶剂体系,初步确定适宜的溶剂体系;根据高速逆流色谱出峰数目及分离度确定较佳的溶剂体系和工作条件,并测定所收集峰的各组分的纯度。结果石油醚-乙酸乙酯-甲醇-水(22∶25∶23∶17)为溶剂体系,流速为2.0 mL/min,仪器转速800 r/min,温度25℃,从延胡索总碱中可一步纯化得到原阿片碱和延胡索乙素,得率分别为16.9%和14.7%,纯度分别为99.3%和98.8%。结论该溶剂体系分离结果可靠,可作为延胡索乙素、原阿片碱化学对照品的制备分离方法。     

脑梗死患者血清磷酸二酯酶活性的变化及意义

目的研究脑梗死患者外周血磷酸二酯酶(PDE)活性的变化及意义。方法应用高效液相色谱法(HPLC)检测298例脑梗死患者和181例对照组外周血白细胞的PDE活性,同时检测他们的血生化指标,收集临床资料,进行统计分析。结果脑梗死组外周血PDE活性为(13.78±3.12),对照组为(7.36±2.34),差异有统计学意义(P<0.01),但外周血PDE活性与病情的严重程度无关。Logistic回归分析表明,外周血PDE活性水平与脑梗死有关(P<0.01)。结论脑梗死患者PDE活性增高,HPLC能简单易行、精密可靠地测定外周血白细胞的PDE活性,PDE在脑梗死的发病中可能起重要作用。     

Tamoxifen影响人垂体腺瘤细胞增殖及其机制

目的研究tamoxifen对人垂体腺瘤细胞的增殖代谢和DNA合成的影响,并深入探讨tamoxifen影响垂体腺瘤细胞增殖作用的机制,观察tamoxifen作用后垂体腺瘤细胞周期、细胞内蛋白激酶C(PKC)及cAMP/cGMP的变化。方法采用MTT和[3H]TdR参入实验检测tamoxifen对人垂体腺瘤细胞增殖和DNA合成的影响;用流式细胞技术检测tamoxifen对垂体腺瘤细胞周期的影响;通过PKC活性及cAMP和cGMP含量测定,深入探讨tamoxifen影响垂体腺瘤细胞增殖分化的分子机制。结果①tamoxifen抑制垂体腺瘤细胞的增殖和DNA合成,并呈剂量依赖性;②tamoxifen处理的垂体腺瘤细胞G1期DNA含量升高,S期和G2期DNA含量降低;③与空白处理组相比,使用PKC的激动剂PMA处理培养的人垂体腺瘤细胞时可使胞膜和细胞总PKC活性浓度均升高,但tamoxifen作用15min后,胞浆、胞膜和细胞总PKC活性均下降;④tamoxifen作用于人垂体腺瘤细胞15min后,胞内cAMP水平升高,而cGMP没有明显改变。结论实验结果为探讨tamoxifen抑制垂体腺瘤细胞增殖的分子机制提供了重要线索,同时提示,tamoxifen对垂体腺瘤细胞增殖分化的调控作用是细胞内多信息系统相互整合的结果。     

正常人血浆中同型半胱氨酸水平及其与叶酸和维生素B12的相互关系

目的 :测定正常人血浆同型半胱氨酸 (HCY)、血清叶酸和维生素B12 水平 ,以探讨不同年龄组 3者之间的相互关系。方法 :用HPLC法测定血浆HCY水平 ,放射免疫法测定血清叶酸及维生素B12 水平。结果 :2 4 7例正常人血浆HCY水平、血清叶酸和维生素B12 水平分别为 (8 8± 2 4 ) μmol·L-1,(5 93± 2 31)ng·mL-1和 (9 3± 3 2 )ng·mL-1。血浆HCY水平随年龄增加呈明显升高 (r=0 5 4 2 0 ,P <0 0 1) ,6 1～ 70a正常老年人的血浆HCY水平为 (11 5± 2 2 ) μmol·L-1,较其他各年龄组有非常显著差别(P均 <0 0 1)。同时 ,血清叶酸和维生素B12 水平随年龄增加呈明显降低 ,与HCY水平呈负相关 (r=- 0 3891和 - 0 2 80 1,P<0 0 1)。结论 :正常人血浆HCY水平随年龄增加而明显升高 ,并与叶酸和维生素B12 水平呈负相关。     

Stereospecific effects of ketamine enantiomers on canine tracheal smooth muscle

1. Ketamine is a potent bronchodilator which relaxes airway smooth muscle (ASM). Clinically, ketamine is used as a 1:1 racemic mixture of enantiomers that differ in their analgesic and anaesthetic effects. The aim of this study was to determine whether there was a difference between the enantiomers in their ability to relax isolated ASM and to explore mechanisms responsible for any observed differences.
2. Canine tracheal smooth muscle strips were loaded with fura-2 and mounted in a photometric system to measure simultaneously force and [Ca²⁺]_i. Calcium influx was estimated by use of a manganese quenching technique.
3. In strips stimulated with 0.1 μM ACh (EC₅₀) R(−)-ketamine (1–100 μM) caused a significantly greater concentration-dependent decrease in force (P<0.0001) and [Ca²⁺]_i than S(+)-ketamine (1–100 μM) (P<0.0005). In contrast, there was no significant difference between the enantiomers in their ability to inhibit calcium influx (45% decrease in influx rate for R(−)-ketamine and 44% for S(+)-ketamine, P=0.782). In strips contracted with 24 mM isotonic KCl (which activates voltage-operated calcium channels), the enantiomers modestly decreased force and [Ca²⁺]_i; there was no significant difference between the enantiomers in their effects on force (P=0.425) or [Ca²⁺]_i (P=0.604).
4. The R(−)-enantiomer of ketamine is a more potent relaxant of ACh-induced ASM contraction than the S(+)-enantiomer. This difference appears to be caused by differential actions on receptor-operated calcium channels.

     

Characterization of the muscarinic receptor in human tracheal smooth muscle

Summary Muscarinic receptors in human tracheal smooth muscle were characterized by radioligand binding and functional studies. Specific [³H]-(–)-quinuclidinylbenzilate ([³H]-(–)-QNB) binding to tracheal smooth muscle membranes was reversible, stereoselective and of high affinity (K _d=47±4 pmol/l;R _T=920±120 fmol/g tissue). Inhibition of specific [³H]-(–)-QNB binding by the M-1 selective antagonist pirenzepine was found to occur at relative high concentrations classifying the muscarinic receptor population as belonging to the M-2 subclass.Inhibition of specific [³H]-(–)-QNB binding by muscarinic agonists revealed the presence of high and low affinity sites in nearly equal proportions. 5-Guanylylimidodiphosphate converted high affinity sites into low affinity sites although its effect was minimal. Log dose-contraction curves of methacholine had Hill coefficients of 1.10±0.04 with pD₂-values of 6.75±0.02.Inhibition of specific [³H]-(–)-QNB binding by methacholine, however, was best described by a two binding site model with pK _i-values considerably lower. The difference between these affinity values points to the presence of substantial receptor reserve.     

去上皮对离体气管平滑肌反应性的影响(英文)

去上皮致敏豚鼠气管标本对乙酰胆碱。组胺,P物质和氯化钡的敏感性比未去上皮标本为高。上述药物在去上皮标本上得到的EC_(50)仅为未去上皮的1/6—1/30。由抗原抗体反应或电场刺激所引起的去上皮标本的收缩幅度和对照组相比升高了50％—100％。这些实验结果提示,气管上皮对呼吸道,尤其是对其过敏性收缩具有重要调节作用。     

高效液相色谱法测定复方夏天无片中原阿片碱和苯甲酰乌头原碱含量

目的建立同时测定复方夏天无片中原阿片碱和苯甲酰乌头原碱含量的高效液相色谱(HPLC)法。方法色谱柱为Agilent Zorbax Eclipse SB-C18柱(250 mm×4.6 mm,3.5μm),流动相为1%冰乙酸溶液(A)-乙腈(B),梯度洗脱(0~5 min时94%A,5~27 min时94%A→58%A,27~30 min时58%A→94%A,30~35 min时94%A),柱温为30℃,测定波长为265 nm,流速为1.0 m L/min,进样量为25μL,外标法定量。结果原阿片碱和苯甲酰乌头原碱质量浓度在0.25~10.0μg/m L范围内与峰面积线性关系良好(r>0.995,n=6);平均加样回收率分别为97.47%和100.50%,RSD分别为4.37%和4.31%(n=6)。结论该方法操作简便、快速、准确、重复性好,可用于同时测定复方夏天无片中原阿片碱和苯甲酰乌头原碱的含量。     

Effect of Montelukast on bradykinin-induced contraction of isolated tracheal smooth muscle of guinea pig

Aim:

To explore the effect of montelukast on bradykinin-induced tracheal smooth muscle contraction of isolated guinea pig trachea.

Study Design:

To study the effect of bradykinin in the absence and in the presence of montelukast on the isolated tracheal smooth muscle of a guinea pig pretreated with indomethacin (10^-6M), phentolamine (10^-5M), and propranalol (10^-6M), to eliminate the effect of endogenous prostaglandins and catecholamines. The trachealis smooth muscle activity was recorded through the Isometric Force Displacement Transducer on a Four Channel Oscillograph. A cumulative dose-response relationship was demonstrated by adding successive doses of bradykinin on the tracheal strips, starting with 11 μg to 77 μg of 10^-4 concentration. A similar procedure was repeated in the presence of montelukast 0.5 μg/ml, which, was equal to approximate C_max achieved in vivo with a 10 mg oral dose of montelukast, and in the presence of 1 μg/ml of montelukast.

Statistical Analysis:

Data was expressed as mean ± standard error (SEM), and was analyzed using the SPSS version 15. A P value of less than 0.05 was considered significant.

Results:

Bradykinin produced a dose-dependent, reversible contraction of isolated tracheal smooth muscle. Montelukast significantly reduced the bradykinin-induced tracheal smooth muscle reactivity and shifted the bradykinin curve to the right and downwards, in the presence of both concentrations of montelukast. The mean magnitude of response achieved with 77 μg of bradykinin in the absence of montelukast was 39 mm ± 6.26, in the presence of 0.5 μg/ml of montelukast it was 24.17 mm ± 4.11, and in the presence of 1 μg/ml of montelukast it was 13 mm ± 2.6.

Conclusion:

It is concluded that montelukast significantly inhibits, in a dose-dependent manner, the bradykinin-induced contraction of the guinea pig tracheal smooth muscle, and alludes to an interaction between the bradykinin and leukotriene mediators.