摘要
采用美国药典方法筛选合格的去脑垂体幼龄大鼠,并将胫骨染色法改为石蜡包埋、切片、苏木精-伊红(HE)染色,用于测定重组人生长激素(rhGH)水针剂的生物活性。结果表明,采用术后8～14 d内体重变化小于10%的标准筛选去脑垂体大鼠比中国药典法更合理,可避免大鼠体重不稳定而发生的误判;相比于中国药典2010年版收载的硝酸银染色法,胫骨HE染色法可操作性强,人为影响因素较少,切片可长期保存,染色后胫骨骨骺板即软骨带清晰可见,边界明确,胫骨骨骺板宽度易测量。与模型组相比,rhGH水针剂的各剂量组胫骨骨骺板宽度显著增宽(P<0.01),体重显著增加(P<0.01),且呈剂量相关性。本法及中国药典法测得rhGH水针剂的生物活性均大于2.5 IU/mg。
The qualified hypophysectomized rats were screened by the method in US pharmacopeia and the following improvements of tibia staining method were carried out： the tibia was embedded in paraffin, then sliced with slicer and stained with hematoxylin-eosin （HE）. This method was adopted for the bioassay of liquid injection of recombinant human growth hormone （rhGH）. The results showed that the screening standard of body weight changes of the rats during the postoperative period of 8 - 14 d within 10% was more reasonable than Chinese pharmacopeia method, because this could avoid the misscarrages of justice caused by unstable body weight of rats. Compared with the silver nitrate staining method in Chinese pharmacopeia, the HE staining method was easily operated and had less subjective effects, which slice could store for a rather long peroid of time. The staining effect with HE was more clear, the boundary of tibial epiphyseal plate could clearly define and the width could be easily measured. Compared with the model group, the tibial epiphyseal plate width of the low-, middle- and high-dose groups of liquid injection of rhGH was significantly widened （P〈0.01） and the body weight was significantly increased （P〈0.01） in a dose-dependent manner. The biological activities of the injection measured by this method and Chinese pharmacopeia method were both above 2.5 IU/mg.
出处
《中国医药工业杂志》
CAS
CSCD
北大核心
2013年第6期592-596,共5页
Chinese Journal of Pharmaceuticals
关键词
重组人生长激素
水针剂
生物活性
去脑垂体大鼠
胫骨HE染色
recombinant human growth hormone
liquid injection
biological activity
hypophysectomized rat
tibia hematoxylin-eosin staining
