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3×10~5个/ml,混匀后取100μl滴在96孔板中。稀释标准GH溶液至终浓度为0.01~1 000.00μg/L,取10μl标准GH溶液滴加到96孔板中,培养22 h。每孔加入22μl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 m U/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 000μg/L)、表皮生长因子(100μg/L)、人类胰岛素样生长因子1(IGF-1,10μg/L)、氢化可的松(100μg/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5μl(20μg/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=0.8520ln(X)+0.498 8,R~2=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。 Objective To investigate the feasibility of detecting the biological activity of growth hormone (GH) based on the method of BAF / GM cell line and to provide a reference method for the clinical detection of biological activity of GH. Methods Forty-two patients with idiopathic short stature (ISS) admitted to the First Affiliated Hospital of Xinjiang Medical University from July 2011 to December 2013 were enrolled in this study. ISS group and 52 healthy children Example (control group). BAF / GM cell lines were established and cultured in DMEM high glucose medium. The medium was washed and then transferred to medium containing 10% horse serum for 16 h. Resuspend the cells with 10% horse serum medium, adjust the cell concentration to 3 × 10 ~ 5 / ml, and mix 100μl in a 96-well plate. Dilute the standard GH solution to a final concentration of 0.01 to 1 000.00 μg / L. Add 10 μl of standard GH solution to a 96-well plate and incubate for 22 h. Add 22 μl of reaction solution to each well and continue to culture for 3 h. Measure the absorbance (OD) at 490 nm. Draw a standard curve of the OD value versus the GH bioactivity equivalent. Thyrotropin (100 mU / L), luteinizing hormone (10 000 U / L), follicle-stimulating hormone (10 000 U / L) and prolactin (100 U / L) were prepared using 10% horse serum- , Fibroblast growth factor (1 000 μg / L), epidermal growth factor (100 μg / L), human insulin-like growth factor 1 (IGF-1,10μg / L), hydrocortisone (100μg / L) (1.0 U / L), sodium L-thyroxine (643.5 nmol / L) and 5ul of standard GH solution per well (20μg / L). Fasting venous blood was collected from both groups. Serum GH levels were determined by radioimmunoassay (IRMA). Serum samples were inactivated. OD value at 490 nm was determined by MTS method. Serum GH bioequivalence was calculated by standard curve. Results The standard curve of the relationship between the bioequivalence of GH and OD was Y = 0.8520ln (X) +0.498 8 and R ~ 2 = 0.969 8, where X was the biological activity equivalent of GH and Y was the OD value. OD values ​​were measured after addition of thyrotropin, luteinizing hormone, follicle stimulating hormone, prolactin, fibroblast growth factor, epidermal growth factor, IGF-1, hydrocortisone, norepinephrine and sodium L-thyroxine Respectively, compared with the standard GH solution, the difference was not statistically significant (P> 0.05). There was no significant difference in GH level between the two groups (P> 0.05). GH biological activity equivalent and GH biological activity in ISS group were lower than those in control group, with statistical significance (P <0.05). Conclusion Based on the BAF / GM cell line method, it is a potential method to detect GH biological activity and has some anti-interference ability.