斑点免疫印迹法快速测定地龙炮制品中蚓激酶及其水解产物

目的测定参环毛蚓、秉氏环毛蚓、日本杜拉蚓和天锡杜拉蚓炮制品中蚓激酶及其水解产物,建立斑点免疫印迹法评价相应生药材药效成分明确的中药炮制品质量新方法。方法以兔抗蚓激酶单抗为一抗,应用斑点免疫印迹法对地龙炮制品中蚓激酶及其水解产物作定性检测,杂交信号应用Quantity One软件作定量分析。结果现行中药材参环毛蚓与山东产地方品种:秉氏环毛蚓、日本杜拉蚓、天锡杜拉蚓炮制品中蚓激酶及其水解产物的质量分数分别为125.23、118.34、81.05、72.13μg/g。结论秉氏环毛蚓炮制品中蚓激酶及其水解产物与现行地龙炮制品参环毛蚓无显著差异,理论上可作为广地龙的替代品。斑点免疫印迹法简便快捷,结果准确,无需大型设备,可用于地龙与类似中药材炮制品的品质评价。     

免疫活性地龙肽的制备及其对小鼠NK细胞活性的影响

目的:提取低分子量免疫活性地龙肽,并就其对自然杀伤(NK)细胞活性的影响进行体外研究。方法:采用粗分离、凝胶过滤层析和阴离子交换层析等方法分离纯化地龙肽;采用乳酸脱氢酶法测定NK细胞活性。结果:用凝胶过滤层析分离得到地龙肽T(分子量8000～20000,含有7种蛋白)。用离子交换层析又将地龙肽T分为A、B两部分,地龙肽A由5种蛋白组成,其中有4种分子量在13000以下;地龙肽B仅由一种蛋白构成,分子量约11000;地龙肽A和T可明显提高NK细胞的活性;地龙肽A、B和T皆可减弱环磷酰胺和地塞米松的免疫抑制作用,提高NK细胞的杀伤率。结论:地龙肽T、A单独应用即可以增强NK细胞的活性,并与IL-2具有协同作用;地龙肽T、A和B还可以拮抗地塞米松、环磷酰胺等免疫抑制剂。     

地龙胶囊对小鼠耳廓微循环障碍改善的实验研究

[目的]探讨地龙胶囊对小鼠血液微循环障碍形成的保护作用及机理。[方法]以高分子右旋糖酐建立小鼠微循环障碍模型。用WX-9型微循环显微镜和微循环检测分析系统观察小鼠耳廓微循环在给药前、给药后10min、20min、30min等不同时间,微血管的口径、流速、流态、毛细血管交叉网开放数量等项指标的变化。[结果]地龙胶囊大、小剂量组预防均使小鼠微血管细静脉、细动脉管径增大,与注射生理盐水比较具有显著性差异(P<0·01);能提高血流速度,其中以大剂量组较明显(P<0·001)。血液流态有显著的改善;毛细血管交叉网开放数目增加。[结论]地龙胶囊能对微循环障碍可发挥积极的治疗作用。     

补阳还五汤对脑出血大鼠脑内Caspase-3活化的影响

目的研究补阳还五汤对脑出血大鼠的保护作用。方法采用微量注射器向大鼠苍白球内注入Ⅶ型胶原酶0.4U建立脑出血模型,用免疫组织化学法观察脑出血后1、2、4、7d,4个时间点活化Caspase-3表达变化,以阳性细胞计数作为观察指标,并观察血肿大小的变化。结果脑出血后1d大鼠血肿周围活化Caspase-3阳性神经元细胞计数最多,2d开始下降,7d明显下降,与假手术组比较具有统计学意义(P<0.01);补阳还五汤组与同时间点模型组比较:1、2d计数无统计学意义,4、7d均明显减少(P<0.05)。造模后1d血肿最大,2d有所变小,7d明显变小;治疗组在1、2d与模型组比较血肿大小无明显差异(P>0.05),4、7d具统计学意义(P<0.05)。结论大鼠脑出血后血肿周围神经元活化Caspase-3表达明显上调;补阳还五汤能下调Caspase-3的活化,减少神经元的凋亡,同时促进血肿的吸收。     

蚓激酶的研究与临床应用

蚓激酶作为中药地龙(即蚯蚓)的主要抗凝、溶栓成分,经动物实验及临床观察证实具有良好的纤溶、抗凝、溶栓和改善血流变等药理作用,并取得了较理想的结果。与同类药物比较,蚓激酶具有不良反应小、安全性高、疗效较显著、生产成本低等优势。国内外的学者对蚓激酶进行了大量的研究,现就蚓激酶基本的理化性质、生化方面、作用机制、临床应用等的研究成果做一简要综述,总结其中存在的问题并对前景作出展望,以期能有助于今后对蚓激酶的进一步研究开发和利用。     

In vitro anthelmintic activity of Melia azedarach naturalized in Argentina

The anthelmintic activity of the drupe extracts of Melia azedarach L. (Meliaceae) growing in Argentina was tested against tapeworms, hookworms, nodular worms and earthworms, and was shown to be better than the standards piperazine phosphate and hexylresorcinol against tapeworms and hookworms, respectively.     

Effects of Norepinephrine and Epinephrine on Resting Membrane Potential in Body Wall Muscle Cells of Lumbricus Terrestris Eearthworm

Norepinephrine and to a lesser extent epinephrine increased the resting membrane potential of earthworm body wall muscle cells. Ouabain, phentolamine, propranolol, and replacement of Ca²⁺ with Mg²⁺ in the incubation medium abolished this effect. External 3'5'-cAMP in high concentration, dibutyryl cAMP, and dibutyryl cGMP did not induced hyperpolarization of muscle cell membranes. It was concluded that norepinephrine and epinephrine increased the resting membrane potential in earthworm body wall muscle cells via activation of Na⁺, K⁺ pump. This process involves Ca²⁺ and sarcolemmal adrenergic structures, similar to -adrenoceptors in vertebrate, but not the cyclic nucleotide systems.     

Two Populations of Miniature Excitatory Synaptic Ionic Currents in Somatic Muscle Cells of Lumbricus Terrestris Earthworm Body Wall

Three types of miniature excitatory synaptic currents were recorded in the same synaptic region of earthworm muscle cells: monoexponential (tau=1.2 msec) and biexponential (tau(1)=1.2 and tau(2)=8.0 msec). It was hypothesized that earthworm muscle cells contain at least two populations of acetylcholine-sensitive ionic channels, which do not belong to classical nicotinic and muscarinic acetylcholine receptors.     

中药脑泰方对大鼠脑缺血后VEGF和Angiopoietins表达的实验研究

目的 探讨中药脑泰方对大鼠脑缺血后血内皮生长因子(vascular endothelial growth factor,VEGF)和血管生成素(angiopoietins,Ang)的表达变化及其在血管生成中的作用.方法 选择40只雄性SD大鼠,采用随机区组法分为A正常对照组,B假手术组,C MCAO模型组,D脑泰方组;,观察各组大鼠神经功能缺失计分、脑梗死体积;RT-PCR、Western blot方法检测不同组大鼠脑组织中VEGF、Ang-1及Ang-2 mRNA和蛋白的表达变化.结果 神经功能缺失计分及TTC染色显示脑泰方组脑缺血情况缓解;大鼠脑缺血后VEGF、Ang-2 的表达水平增高,脑泰方组表达显著高于模型组(P<0.05);Ang-1的表达水平在大鼠脑缺血后呈下降趋势,脑泰方组表达显著低于模型组(P<0.05).结论 大鼠脑缺血后VEGF、Ang-1和Ang-2共同协调作用促进血管生成,中药脑泰方在脑缺血损伤后治疗血管新生样作用中发挥着促进作用.     

Chronic toxicity of commercial chlorpyrifos to earthworm Pheretima peguana

A chronic toxicity study was conducted in earthworms (Pheretima peguana) exposed to soil spiked with chlorpyrifos at concentrations of 0, 0.1, 1, 10, and 100 mg/kg soil dry matter for 7, 14, and 28 days. The integrity of the coelomocyte lysosomal membrane, nervous system, and male reproductive tissue was monitored using, respectively, the neutral‐red retention assay, acetylcholinesterase (AChE) enzyme assay, and histomorphology of spermatogenic cells in the seminal vesicles and cocoon production (at 28 days after 28 days’ exposure). Chlorpyrifos decreased the coelomocyte neutral‐red retention time (NRRT) significantly (p < 0.05) at concentrations > 0.1 mg/kg soil as early as day 7 of exposure and was dose‐ and time‐dependent. Chlorpyrifos inhibition of AChE activity was greater at day 7 than at day14 (p < 0.05) indicating possibly nerve recovery. Chlorpyrifos induced concentration‐dependent damage to spermatogenic cells and cytophores in premature stages. The number and size of premature, maturing, and fully mature spermatogenic stages were increased at low concentrations (<1 mg/kg) but a number of these maturation stages declined at higher concentrations (10 and100 mg/kg) on day 28. The most severe effects were observed in the maturing and fully mature stages at the highest chlorpyrifos concentration, and this had an adverse impact on cocoon production and cocoon viability. Collectively, the results suggest induction of widespread effects on multiple organ systems in P. peguana exposed to chlorpyrifos. Although NRRT and AChE activity were the most sensitive of the biomarkers, cocoon production and cocoon viability could still be considered as diagnostic tools for monitoring effects from low‐dose long‐term chlorpyrifos toxicity and for evaluating population effects. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1450–1459, 2016.