Abstract: Objective　To investigate the feasibility of using BAF/GM cell line method to detect the biological activity of GH,and to provide referential methods for clinical detect biological activity of GH.Methods　42 children with ISS who received treatment in the Department of Pediatrics，the First Affiliated Hospital of Xinjiang Medical University from July 2011 to December 2013 were selected（ISS group）,other 52 healthy children after physical examination in the Department of Prevention and Health Care of this hospital were also selected as control group.BAF/GM cell line was established,which was routinely cultured in DMEM high glucose medium.The BAF/GM cell line after washed was transferred into the culture medium containing 10% horse serum for 16 hours cultivation.10% horse serum was adopted to culture the resuspension cells,the cell concentration was adjusted to 3×105/ml,and 100 μl after mixing was dropped into the 96-hole micro pore plate.Diluted the standard GH to a final concentration of 0.01-1 000.00 μg/L,took 10 μl standard GH and dropped it into the 96-hole micro pore plate,and cultured for 22 hours.We added 22 μl reaction liquid into each hole and further cultured for 3 hours，and detected the OD value at the wavelength of 490 nm.The standard curve of the equivalent relationship between GH biological activity equivalent and OD value was made.We used the culture medium containing 10% horse serum to make up thyroid-stimulating hormone（100 mU/L）,luteinizing hormone（10 000 U/L）,follicle-stimulating hormone（10 000 U/L）,prolactin（100 U/L）,fibroblast growth factors（1 000 μg/L）,epidermal growth factors（100 μg/L）,IGF-1（10 μg/L）,hydrocortisone（100 μg/L）,Novolin R（1.0 U/L）,and L-thyroxine sodium（643.5 nmol/L）,intervention test was made by adding 5 μl（20 μg/L） standard GH solution into each hole.We collected fasting venous blood of the two groups,and adopted IRMA method to measure the level of serum GH，inactivated serum samples,and through MTS method to obtain OD value at the wavelength of 490 nm,and then substituted it into the standard curve to calculate the biological activity equivalent of the serum GH.Results　The standard curve of GH biological activity equivalent and the OD value relation was Y=0.852 0ln（X）+0.498 8,R2=0.969 8.X was the GH biological activity equivalent,and Y was the OD value.After the addition of thyroid-stimulating hormone,luteinizing hormone,follicle-stimulating hormone,prolactin,fibroblast growth factors,epidermal growth factors,IGF-1,hydrocortisone,Novolin R,and L-thyroxine sodium,there were no significant difference between the detected OD value and the standard GH（P>0.05）.There was no significant difference in GH level between the two groups（P>0.05）.The biological activity equivalent of GH and the GH biological activity in ISS group were lower than those in the control group,which showed significant differences（P<0.05）.Conclusion　BAF/GM cell line is the potential method for detecting biological activity of GH,and has certain anti-interference ability.

Key words: Growth hormone, Biological activity, BAF/GM cell line, Idiopathic short stature

摘要： 目的探讨基于BAF/GM细胞株法检测生长激素（GH）生物活性的可行性，为临床检测GH生物活性提供参考方法。方法　选取2011年7月—2013年12月新疆医科大学第一附属医院儿科收治的特发性身材矮小（ISS）患儿42例（ISS组），另选取同期本院预防保健科体检健康儿童52例（对照组）。建立BAF/GM细胞株，常规培养于DMEM高糖培养基。培养液洗涤后转移到含10%马血清的培养基中培养16 h。采用含10%马血清的培养基重悬细胞，调整细胞浓度为3×105个/ml，混匀后取100 μl滴在96孔板中。稀释标准GH溶液至终浓度为0.01～1 000.00 μg/L，取10 μl标准GH溶液滴加到96孔板中，培养22 h。每孔加入22 μl反应液，继续培养3 h，在490 nm处测定吸光度（OD）值。绘制OD值与GH生物活性当量关系的标准曲线。采用含10%马血清的培养基配制促甲状腺激素（100 mU/L）、黄体生成素（10 000 U/L）、卵泡刺激素（10 000 U/L）、催乳素（100 U/L）、成纤维细胞生长因子（1 000 μg/L）、表皮生长因子（100 μg/L）、人类胰岛素样生长因子1（IGF-1，10 μg/L）、氢化可的松（100 μg/L）、诺和灵R（1.0 U/L）、L-甲状腺素钠（643.5 nmol/L），同时每孔加标准GH溶液5 μl（20 μg/L）进行干扰实验。采集两组受试者空腹静脉血，采用放射免疫分析法（IRMA）测定血清GH水平；灭活血清标本，以MTS法测定490 nm处OD值，代入标准曲线计算血清GH生物学活性当量。结果　GH生物活性当量与OD值关系的标准曲线为Y=0.852 0ln（X）+0.498 8，R2=0.969 8，其中X为GH生物活性当量，Y为OD值。加入促甲状腺激素、黄体生成素、卵泡刺激素、催乳素、成纤维细胞生长因子、表皮生长因子、IGF-1、氢化可的松、诺和灵R和L-甲状腺素钠后测得OD值分别与标准GH溶液比较，差异均无统计学意义（P>0.05）。两组受试者GH水平比较，差异无统计学意义（P>0.05）。ISS组GH生物活性当量、GH生物活性低于对照组，差异均有统计学意义（P<0.05）。结论　基于BAF/GM细胞株法是潜在的检测GH生物活性的良好方法，且具有一定的抗干扰能力。

关键词: 生长激素, 生物活性, BAF/GM细胞株, 特发性身材矮小