In Vitro Nasal Transport Across Ovine Mucosa: Effects of Ammonium Glycyrrhizinate on Electrical Properties and Permeability of Growth Hormone Releasing Peptide,Mannitol, and Lucifer Yellow

Transport of growth hormone releasing peptide across ovine nasal mucosa in the absence or presence of ammonium glycyrrhizinate (AMGZ) was studied in vitro. Ovine nasal mucosa was stripped from underlying cartilage and mounted in Ussing chambers. Transepithelial conductance (G _t) and short-circuit current (I _sc) were monitored during experiments to assess tissue viability and integrity. Radiolabeled mannitol (Man; MW 182) and growth hormone releasing peptide (GHRP, SK&F 110679; MW 873) were employed to measure transport rates across the epithelium, and fluorescence spectroscopy was employed to measure rates of lucifer yellow (LY; MW 521) transport. Effects of AMGZ on ovine nasal mucosal viability and transport were determined from changes in electrical properties or fluxes of [³H]GHRP, [³H]Man, and LY. Results demonstrate that electrical properties of ovine nasal mucosa are stable over the time course of the experiments (G _t = 8.3 ± 0.5 mS/cm² and I _sc = 3.7 ± 0.2 µEq/hr · cm²; n = 21). Man fluxes were comparable in the mucosal (m)-to-serosal (s) and s-to-m directions [0.10 ± 0.01 (n = 17) and 0.10 ± 0.01 (n = 4) %/hr · cm², respectively]. Transport of GHRP and LY in the m-s direction was similar to that of Man [0.08 ± 0.01 (n = 11) and 0.09 ± 0.01 (n = 3) %/hr · cm², respectively]. GHRP flux was equivalent in the m-s and s-m directions. GHRP did not significantly alter ion transport processes as indicated by the lack of any change in G _t or I _sc. Luminal addition of AMGZ (2%, 24 mM) increases G _t and transport of both LY and [³H]Man approximately fourfold without altering transport of [³H]GHRP. No changes in transport or G _t were seen with luminal addition of AMGZ (1%, 12 mM). These studies suggest that transport of the hexapeptide GHRP occurs by a passive process and that AMGZ selectively increases the permeability of the mucosa to the low molecular weight molecules, Man and LY, but not to GHRP in vitro.     

反相高效液相色谱法测定大鼠血浆中的甘草酸铵

目的 :建立反相高效液相色谱法测定甘草甜素粉剂和四君子汤灌胃大鼠血浆中的甘草酸铵。方法 :以联苯为内标 ,甲醇 -水 -醋酸 (80∶19∶1)为流动相 ,采用ODS - 3色谱柱 (15 0mm× 4 6mm ,5 μm) ,检测波长 :2 5 0nm ,流速 :1 0mL·min-1。结果 :最低检测限为 2ng ,最低检测浓度为 0 2 2 8μg·mL-1,日间和日内RSD <10 % ,回收率为 92 46 %～10 3 2 % ,RSD为 3 8%～ 6 0 % ,在 1 2 2 4～ 38 6 μg·mL-1和 40 8～ 146 8 8μg·mL-1范围内均呈良好的线性关系。结论 :本法简便、快速、灵敏而准确 ,适用于大鼠灌胃不同甘草酸铵制剂后血浆中甘草酸铵的测定 ,为“复方效应成分动力学”研究奠定了方法学基础。     

UV法测定复方甘草酸铵注射液中甘草酸铵含量的不确定度分析

目的:对紫外分光光度法测定复方甘草酸铵注射液含量的测量的不确定度评定分析,以期找到影响不确定度的因素。方法:按照测量步骤,根据《测量不确定度评定与表示》、《化学分析中不确定度的评估指南》中有关规定评估其不确定度。结果:量化各不确定度分量,提出了该法的合成不确定度评估结果。结论:计算出扩展不确定度U=1.94%(k=2)。     

高效液相色谱法测定八珍益母丸中甘草酸铵的含量

目的:采用高效液相色谱法测定八珍益母丸中甘草酸铵的含量。方法:色谱条件:Kromasil C18柱(4.6×250mm,5μm);流动相:乙腈-0.017mol?L-1磷酸(36︰64);流速:1mL?min-1;柱温:30℃;检测波长:250nm。结果:甘草酸铵在进样量0.086~0.52μg范围内线性关系良好,相关系数r=0.9998,回收率(n=6)为99.1%,RSD为1.81%。结论:本法准确可靠,重现性好,可用于测定八珍益母丸中甘草酸铵的含量。     

The Effects of Permeation Enhancers on the Surface Morphology of the Rat Nasal Mucosa: A Scanning Electron Microscopy Study

A rat model has been developed to compare relative morphological changes in the nasal mucosa after exposure to potential membrane permeation enhancers. Scanning electron microscopy was used to characterize gross structural and specific cellular changes following exposure. Micrographs of the rat nasal mucosa were scored in four categories: (1) mucosal surface integrity, (2) ciliary morphology, (3) mucus/extracellular debris, and (4) presence of red blood cells. The order of increasing morphological damage resulting from a 5-min exposure to each surfactant was 0.5% Solulan C-24 0.5% Solulan C-24/0.5% sodium tauro-24,25-dihydrofusidate (STDHF) < 0.5% STDHF < 1.0% STDHF 1.0% Laureth-9 < 1.0% sodium taurodeoxycholate 1.0% sodium deoxycholate. The changes observed in the mucosal morphology after exposure to the various surfactants are in general agreement with data in the literature. This model is able to compare rapidly the relative morphological effects on the mucosal membrane of different nasal formulations.     

复方甘草酸铵凝胶剂制备

目的:制备复方甘草酸铵凝胶剂。方法:以甘草酸铵、苦参碱为主药,卡泊姆为基质制备凝胶剂。用紫外分光光度法分别测定甘草酸铵和苦参碱的含量并进行稳定性试验。结果:甘草酸铵的线性范围为10～70μg·ml~(-1)平均回收率为100.38%;苦参碱的线性范围为10～100μg·ml~(-1)平均回收率为100.6%。结论:该制剂制备工艺可行,凝胶性质稳定,质控方法简单、可靠。     

高效液相色谱法同时测定养血当归糖浆中芍药苷和甘草酸铵含量

目的建立同时测定养血当归糖浆中芍药苷、甘草酸铵含量的高效液相色谱法。方法色谱柱为Kromasil-C18柱(250 mm×4.6 mm,5μm),以乙腈(A)-0.085%磷酸溶液(B)为流动相(梯度洗脱),流速为1.0 mL/min,柱温为30℃,进样量为10μL,检测波长为230 nm。结果芍药苷和甘草酸铵质量浓度分别在5.54~55.35μg/mL和9.63~115.56μg/mL范围内与峰面积线性关系良好(r≥0.9990);平均加样回收率分别为88.42%和87.08%,RSD分别为3.66%和3.52%(n=6)。结论该方法简便、准确、专属性强、重复性好,可用于养血当归糖浆制剂的质量控制。     

探讨甘利欣对体外诱导的酒精性脂肪肝细胞的影响

目的 探讨甘利欣对体外诱导的酒精性脂肪肝影响.方法 以人正常肝细胞株L02为实验材料,采用乙醇诱导建立酒精性脂肪肝的离体细胞模型,后用甘利欣处理.流式细胞仪检测作用前后细胞周期的改变;分别用ALT、AST、GGT试剂盒测定谷丙转氨酶(ALT)、谷草转氨酶(AST)以及γ-谷氨酰转肽酶(GGT)的泄露量.结果 25 μg/ml甘利欣可使酒精性脂肪肝细胞的S期细胞明显减少(P＜0.05),并且使ALT、AST、GGT的泄露量明显降低(P＜0.05).结论 实验结果证明,甘利欣能减缓酒精性脂肪肝细胞的增殖生长速度,并能降低酒精性脂肪肝细胞ALT、AST及GGT的泄露量,从而减轻酒精性脂肪肝的肝功能损害.     

甘草酸单铵盐中有关物质的LC-MS~n研究

采用LC-MS~n法分析了甘草酸单铵盐原药中的主成分及其有关物质,通过液相色谱、多级质谱和紫外光谱信息确证主成分甘草酸单铵盐的结构,推测3个主要有关物质的可能结构为甘草皂苷G_2、乌拉尔甘草皂苷A的单铵盐、乌拉尔甘草皂苷B的单铵盐或18α-甘草酸单铵盐.     

Drug Metabolism in the Nasal Mucosa

Nasal delivery is a potential alternative for systemic availability of drugs restricted to intravenous administration, such as peptide and protein drugs. Although nasal delivery avoids the hepatic first-pass effect, the enzymatic barrier of the nasal mucosa creates a pseudo-first-pass effect. The xenobiotic metabolic activity in the nasal epithelium has been investigated in several species including humans. The Phase I, cytochrome P-450 enzymes have been studied extensively for their toxicological significance, since these enzymes metabolize inhaled pollutants into reactive metabolites which may induce nasal tumors. The cytochrome P-450 activity in the olfactory region of the nasal epithelium is higher even than in the liver, mainly because of a three- to fourfold higher NADPH–cytochrome P-450 reductase content. Phase II activity has also been found in the nasal epithelium. The delivery of peptides and proteins has been hindered by the peptidase and protease activity in the nasal mucosa. The predominant enzyme appears to be aminopeptidase among other exopeptidases and endopeptidases. The absorption of peptide drugs can be improved by using aminoboronic acid derivatives, amastatin, and other enzyme inhibitors as absorption enhancers. It is possible that some of the surfactants, e.g., bile salts, increase absorption by inhibiting the proteolytic enzymes. Thus, in addition to the permeation barriers, there also exists an enzymatic barrier to nasal drug delivery, which is created by metabolic enzymes in the nasal epithelium.     

鼻腔给药系统的鼻粘膜毒性及解决途径

就鼻腔给药系统可能导致的鼻粘膜毒性的评价方法、易产生毒性的制剂组分及解决毒性的方法进行简要的综述。     

Nasal Absorption in the Rat. III. Effect of Lysophospholipids on Insulin Absorption and Nasal Histology

The intranasal absorption enhancing and histological effects of a range of lysophospholipids has been investigated in the rat. Blood glucose levels fell rapidly following the administration of insulin (8 IU/kg) in combination with lysophosphatidylcholines (LPC; 0.625% w/v) which had ten or more carbon groups in their fatty acid chain. The effect of the LPC-caproyl (C6) was comparable to that of an unenhanced insulin formulation; the enhancing effect of LPC-decanoyl (C10) was similar to that of an LPC-palmitoyl/stearoyl (C16/C18) for similar concentrations. The effect of LPC-decanoyl was reduced with concentration but was still significant at 0.2% w/v (5mM). Lysophosphatidylglycerol (LPG) had a marked insulin absorption enhancing effect even at 0.0625% w/v. The histological effects of LPC-caproyl were similar to those of an unenhanced insulin formulation, while co-administration of LPC-decanoyl resulted in evidence of epithelial interaction. LPG (0.5% w/v) resulted in similar histological changes as LPC (0.625% w/v) (1), but at 0.0625% w/v no significant changes in epithelial integrity were observed. The length of the fatty acid residue of lysophospholipids was identified as an important factor for intranasal absorption enhancing activity. The nature of the polar head group may also have an influence. Increased insulin absorption was not necessarily accompanied by severe disruption of the nasal epithelium. Careful selection of lysophospholipid type and concentration may enable therapeutic drug levels to be achieved via the nasal route without prohibitive toxic effects.     

HPLC法同时测定八珍益母丸中芍药苷和甘草酸铵的含量

目的:建立HPLC法同时测定八珍益母丸中芍药苷、甘草酸铵的方法。方法:Agilent-ODS C₁₈色谱柱（250mm×4.6mm,5μm）;流动相:乙腈-0.08%磷酸梯度洗脱;流速1ml·min^-1;检测波长分别为230nm和237nm;柱温35℃。结果:芍药苷进样量在0.034～0.344μg范围内线性关系良好,r=0.999 2,平均回收率为98.83%（RSD=1.56%）;甘草酸铵在0.027～0.266μg范围内线性关系良好,r=0.9998,平均回收率为98.20%（RSD=1.49%）。结论:本法便于操作,结果准确,适用于八珍益母丸中芍药苷、甘草酸铵的同时测定。     

石杉碱甲的大鼠鼻黏膜吸收研究

目的研究治疗早老性痴呆症药物石杉碱甲的鼻黏膜吸收特性。方法采用石杉碱甲进行大鼠鼻腔灌流实验,测定循环液中药物浓度。结果随着石杉碱甲的浓度增加(0.125~0.75mg/mL),其吸收速度呈线性增加(r=0.995)。在pH值5.5~7.5内,随pH值的升高,其吸收速度增加。结论石杉碱甲的鼻黏膜吸收良好,其吸收程度与药液的浓度和pH值有较好的相关性。     

甘草酸二铵对关木通所致大鼠肾脏纤维化的影响

目的探讨甘草酸二铵对关木通所致大鼠肾脏纤维化的影响，为临床防治肾脏纤维化提供理论依据。方法将18只大白鼠随机分成3组，正常对照组、关木通组、甘草酸二铵与关木通联合组各6只。在实验前及实验第2，3，4，5，6周分别采血，测定血肌酐(SCr)、血尿素氮(BUN)、血钾、血钠、血钙等生化指标以及Ⅲ型前胶原(PCⅢ)、血清透明质酸(HA)、层粘蛋白(LN)等放射免疫指标。第6周时处死动物取肾脏作组织形态学特殊染色检查。结果关木通组与联合组用药后血生化、放免指标均明显上升(P＜0.05或P＜0.01)。联合组与关木通组肾脏纤维化程度差异无显著性(P＞0.05)。结论甘草酸二铵对关木通所致的急、慢性肾损伤无明确的防治作用；血放射免疫指标(PCⅢ、HA、LN)可作为早期慢性肾间质纤维化的重要临床观察指标。     

Uptake of Ovalbumin-conjugated Starch Microparticles by Pig Respiratory Nasal Mucosa In Vitro

The uptake of ovalbumin-conjugated starch microparticles (OVA-MP) was studied after application to porcine respiratory nasal mucosa in vitro. Nasal mucosa from freshly slaughtered pigs was mounted in horizontal Ussing chambers, which permit monitoring of the viability of the tissue exposed to microparticles and ensure that the microparticles are deposited on the mucosa. The antigen-conjugated starch microparticles have previously been shown to produce strong mucosal, cellular and systemic immune responses to conjugated model antigens following oral administration. Intranasal administration of vaccines for mucosal immunisation is an interesting alternative to oral administration, since nasal delivery systems generally require lower doses of antigen and the site of application is better suited for protection against air-borne antigens. Most of a nasally administered dose is deposited on the surface of the respiratory area of the nasal mucosa. It is therefore important to examine whether the microparticles are taken up in this area and, if so, by which cell type.

Confocal laser scanning microscopy and transmission electron microscopy (TEM) of the nasal tissue both showed intracellular OVA-MP in non-ciliated epithelial cells after 45 min' incubation. The morphology of the cells in the TEM preparations did not support the presence of either M cells (specialised antigen sampling cells) or adjacent lymphocytes. Anticytokeratin-18 (Ac18) was used as a potential M cell marker. However, there was no indication of Ac18 binding to M cells, but it did bind to mucus-producing cells in the respiratory nasal mucosa. In conclusion, OVA-MP were taken up intracellularly by non-ciliated epithelial cells in the nasal respiratory mucosa of pigs, in vitro.     

高效液相色谱法测定甘草远志合剂中甘草酸铵的含量

目的建立高效液相色谱法测定甘草远志合剂中甘草酸铵的含量。方法色谱柱：Inertsil ODS．SP（4．6mm×250mm,5μm）,流动相：乙腈-0．1％磷酸溶液（42：58）,检测波长：252nm,流速：1．0ml／min,进样量：10μl,柱温：30℃。结果甘草酸铵在4．008～160．3μg／ml（r=0．9999）范围内,峰面积与其浓度呈良好的线性关系,平均回收率为100．79％,相对标准偏差为0．98％（n=6）。结论本方法准确、灵敏度高、重复性好、简便可行,可作为甘草远志合剂的质量控制方法。     

Metabolism of Progesterone by the Nasal Mucosa in Mice and Rats

Abstract: The metabolism of 4-¹⁴C-progesterone by the nasal mucosa and lung from mice and rats was investigated in vitro. Using thin-layer radiochromatography at least 10 and 7, yet unidentified, metabolites were found at incubation with slices from the nasal mucosa and lung, respectively. The results further indicated that the rate of progesterone metabolism was higher in the nasal mucosa than in the lung. Autoradiography of lung slices incubated with ¹⁴C-progesterone showed that a selective localization of radioactivity occurred in the bronchial mucosa.     

鼻内镜下鼻中隔黏膜下部分切除术65例分析

目的探讨鼻内镜下鼻中隔黏膜下部分切除术治疗鼻中隔偏曲的临床疗效。方法本组患者65例全部进行鼻内镜下鼻中隔黏膜下部分切除术治疗,术后观察其鼻中隔畸形矫正程度及并发症发生情况。结果本组65例患者鼻中隔偏曲都一次性成功矫正,除切口黏膜外,无其他黏膜破裂。结论鼻内镜下鼻中隔黏膜下部分切除术具有切除范围小,对正常组织损伤小,术后恢复快及并发症少等优点,值得在临床上进一步推广应用。     

HPLC法测定甘草酸二铵原料中的有关物质

目的:建立HPLC法测定甘草酸二铵原料中的有关物质。方法:色谱柱:Agilent Extend-C₁₈柱（250 mm×4.6 mm,5μm）;流动相:乙腈-0.01 mol·L^-1磷酸溶液（38:62）;检测波长:252 nm;流速:1.0 ml·min^-1。结果:降解产物与主峰均分离良好;甘草酸二铵最低检测限为5.2 ng。结论:该法专属性强,准确度高,可用于甘草酸二铵原料中有关物质的检查。