RP-HPLC法测定红花注射液中腺苷的含量

目的 :建立HPLC法测定红花注射液中腺苷的含量。方法 :采用色谱柱 :DiamonsilTM C1 8柱 ,流动相 :甲醇 1 %醋酸(梯度洗脱 ) ,以腺苷为对照品 ,外标法定量 ,紫外检测波长 2 70nm。结果 :腺苷在 5～ 50 μg·mL- 1 范围内呈良好的线性关系。平均回收率 1 0 1 .86 % ,RSD为 1 .34 %。结论 :该方法简便、快速准确 ,可作为该制剂的质量控制方法。     

苦碟子注射液中腺苷的含量测定

目的确证苦碟子注射液中腺苷 ,并测定其含量。方法采用HPLC MS法鉴别苦碟子注射液中的腺苷 ;采用HPLC法测定腺苷含量 ,色谱柱为DimonsilTM ODSC18柱 ,流动相为甲醇 乙腈 水(体积比 5∶4∶91 ) ;流速 1mL·min-1;检测波长 2 5 9nm。结果HPLC MS确证了腺苷的存在 ,HPLC测定结果表明腺苷在 0 4～ 3 0 0g·L-1内 ,线形关系良好 (r=0 9999) ,平均回收率为 1 0 0 4% (n=5 )。结论此方法可用于苦碟子注射液中腺苷的含量测定     

吗啡对PC12细胞嘌呤核苷酸代谢相关酶基因表达的影响

目的：研究吗啡对神经细胞模型PC12嘌呤核苷酸补救合成酶与分解代谢关键酶基因表达的影响。方法：采用逆转录聚合酶链反应（RT-PCR）方法检测暴露于吗啡不同时间的PC12细胞次黄嘌呤-鸟嘌呤磷酸核糖转移酶（HGPRT）、腺苷激酶（AK）和腺苷脱氨酶（AD A）mRNA的表达水平。结果：与相应时段对照组相比，PC12细胞暴露于吗啡12及24 h时，HGPRT mRNA水平均明显增高(P<0.05)，作用48 h时HGPRT mRNA水平显著降低(P<0.05)，72 h后HGPRT mRNA水平再次升高(P<0.05)；吗啡处理的PC12细胞于12和24 h时，AK mRNA水平均明显降低(P<0.05)，作用48 h时AK mRNA水平升高(P<0.05)，72 h后AK mRNA水平恢复正常；ADA mRNA水平均表现为轻度降低，12 h差异有显著性(P<0.05)。结论：吗啡影响神经细胞嘌呤核 苷酸代谢，使细胞内腺苷酸及腺苷水平增高，这可能是吗啡依赖和耐受形成的机理之一 。     

滋阴潜阳方对高血压大鼠淋巴细胞内PKA蛋白表达和cAMP水平的影响

目的:探讨高血压的神经免疫调节机制和滋阴潜阳方对高血压大鼠淋巴细胞内信号转导作用的可能环节.方法:采用免疫组织化学染色法和DP801图像分析系统检测"二肾一夹"高血压大鼠淋巴细胞内蛋白激酶A(PKA)蛋白表达.采用放射免疫法检测环磷酸腺苷(cAMP)水平的改变.结果:模型组大鼠淋巴细胞内PKA蛋白表达和cAMP水平明显增强,滋阴潜阳组对此有抑制作用.结论:滋阴潜阳方抑制淋巴细胞内PKA蛋白表达和cAMP水平可能是其发挥神经免疫调节作用的机制之一.     

Effect of atorvastatin upon platelet activation in hypercholesterolemia, evaluated by flow cytometry

Hyperlipidemia is a well established risk factor for cardiovascular disease and atherothrombotic events, in which platelet activation also plays a significant role. However, very few studies have addressed platelet activation in hypercholesterolemia, the potential effect of lipid lowering drugs upon platelet hyperfunction, and the question of whether changes in the latter are correlated to normalization of plasma lipids. This study used whole blood flow cytometry to assess in vivo and in vitro platelet activation in a group of 33 patients with hypercholesterolemia, and also the ex vivo effect of atorvastatin (20 mg/day) upon such activation. A control group of 40 normolipidemic volunteers matched in terms of age, sex and added risk factors to the patient group was used. The results showed that hypercholesterolemic patients had in vivo a significantly greater percentage of GPIIb/IIIa- and phosphatidylserine-positive platelets compared with the control group (4.62+/-3.51% and 2.58+/-1.19% versus 2.73+/-1.08% and 1.54+/-0.68%, respectively). In vitro response of CD62 expression to thrombin was also greater in the patients than in the controls (92.51+/-6.00% versus 89.63+/-10.72%, p<0.05). Atorvastatin therapy normalized platelet hyperfunction in the patients studied and reduced GPIIb/IIIa response to ADP (from 82.65+/-6.43% to 75.84+/-4.89%, p<0.01). A significant correlation can be seen between such normalization and the decrease in plasma levels of total and LDL cholesterol.     

百令胶囊中腺苷的含量测定

采用薄层色谱紫外分光光度测定百令胶囊中腺苷的含量，样品中腺苷浓测定波长为２５６ｎｍ，相关系数ｒ＝０．９９８３，平均回收率为９９．４４％，ＣＶ％为３．８２％，方法准确，简便，重现性好，可作为虫草制剂的质量控制指标。     

Attenuation of Non‐neuronal Adenosin Triphosphate Release from Human Bladder Mucosa by Antimuscarinic Agents

Objectives: To clarify the contribution of mucosal muscarinic receptors to bladder function, we investigated the effects of various antimuscarinic drugs on stretch‐induced non‐neuronal adenosin triphosphate (ATP) release in human bladder. Methods: Human bladders were obtained from 17 patients. Bladder strips with and without mucosa were suspended in organ baths. A microdialysis probe was inserted into the strip, Ringer's solution was perfused into the probe, and dialysate was collected under tetrodotoxin pretreatment. The amount of ATP released in dialysate was measured by luciferin‐luciferase assay. The effects of various antimuscarinic drugs on non‐neuronal ATP release were evaluated. Results: Non‐neuronal ATP release from bladder strips without mucosa was approximately 10% of that from strips with mucosa. Non‐neuronal ATP release was significantly inhibited by pretreatment with nifedipine or in Ca²⁺‐free medium. Both methoctramine (M₂ receptor selective antagonist) and 4‐diphenyl‐acetoxy‐N‐methylpiperidine methiodide (4‐DAMP [M₃ receptor selective antagonist]) significantly inhibited release. However, M₁ receptor selective antagonist (pirenzepine) did not have a significant effect on release. Oxybutynin, propiverine, tolterodine and solifenacin caused concentration‐dependent inhibition in non‐neuronal ATP release. The rank order of the maximum inhibition rate was propiverine ≥ solifenacin ≥ tolterodine ≥ oxybutynin. Solifenacin showed an inhibitory effect at a lower concentration compared to other drugs. Conclusion: The data suggest that human bladder mucosa is a main source of stretch‐induced non‐neuronal ATP release, and that stimulation of M₂ and M₃ receptor subtypes of mucosa partly contributes to non‐neuronal ATP release. Various antimuscarinic drugs used for the treatment of overactive bladder may have different inhibitory effects on non‐neuronal ATP release.     

Effects of coffee and its chemopreventive components kahweol and cafestol on cytochrome P450 and sulfotransferase in rat liver.

Coffee drinking appears to reduce cancer risk in liver and colon. Such chemoprevention may be caused by the diterpenes kahweol and cafestol (K/C) contained in unfiltered beverage. In animals, K/C treatment inhibited the mutagenicity/tumorigenicity of several carcinogens, likely explicable by beneficial modifications of xenobiotic metabolism, particularly by stimulation of carcinogen-detoxifying phase II mechanisms. In the present study, we investigated the influence of K/C on potentially carcinogen-activating hepatic cytochrome P450 (CYP450) and sulfotransferase (SULT). Male F344 rats received 0.2% K/C (1:1) in the diet for 10 days or unfiltered and/or filtered coffee as drinking fluid. Consequently, K/C decreased the metabolism of four resorufin derivatives representing CYP1A1, CYP1A2, CYP2B1, and CYP2B2 activities by approximately 50%. For CYP1A2, inhibition was confirmed at the mRNA level, accompanied by decreased CYP3A9. In contrast to K/C, coffee increased the metabolism of the resorufin derivatives up to 7-fold which was only marginally influenced by filtering. CYP2E1 activity and mRNA remained unchanged by K/C and coffee. K/C but not coffee decreased SULT by approximately 25%. In summary, K/C inhibited CYP450s by tendency but not universally. Inhibition of CYP450 and SULT may contribute to chemoprevention with K/C but involvement in the protection of coffee drinkers is unlikely. The data confirm that the effects of complex mixtures may deviate from those of their putatively active components.     

反相高效液相色谱法同时测定天麻中天麻素和3种核苷

目的:建立同时测定天麻中天麻素和3种核苷含量的RP-HPLC方法。方法:采用ZORBAX-SB-C18色谱柱(4.6mm×150 mm,5μm);流动相0.1%磷酸水溶液-甲醇梯度洗脱,柱温25℃,流速0.6 mL·min-1,检测波长270 nm。结果:天麻素、尿苷、鸟苷和腺苷分别在0.286~4.29μg(r=0.999 9),0.028 9~0.433 5μg(r=0.999 7),0.066 8~1.002μg(r=1.000 0),0.135 6~2.034μg(r=1.000 0)与峰面积线性关系良好。结论:该测定方法简便、快速、准确,可以作为天麻的质量控制方法之一。