摘要
目的 建立精密肺切片方法学,探讨中性红比色法检测肺切片活力的有效性。方法 1%低熔琼脂糖充盈肺组织,振荡切片机切片,厚度分别为 400、500、600、700μm, 培养液pH分别为 6 8、7 0、7 2、7 4,预培养 1h,于 24孔培养板持续浸润培养 0、2、4、6h,以中性红比色法,MTT比色法乳酸脱氢酶漏出率,超氧化物歧化酶活力为检测手段确定肺切片最适制备培养条件。结果 肺切片厚 600μm,培养液pH7 0时,活力保持最佳,可稳定维持 6h;不同厚度肺切片中性红摄取率与MTT还原量呈正相关 (r1 = 0 91,P0 05),不同pH培养液培养的肺切片中性红摄取率与MT还原量呈正相关(r2 =0 98,P<0 01 )。结论 精密肺切片是简便可行、有待发展的新型体外模型,中性红比色法操作简便,结果稳定,灵敏准确,可作为肺切片活力判定指标。
Aim To establish the metho d of precise lung slicing and examine the validity of neutral red(NR) assay for measuring lung slices viability.Method Inflated with 1% low melting agaro se solution, lung lobes were cut into slices by shaking microtome. With the thic kness of 400,500,600,700 μm,lung slices were cultured in medium pH 6.8,7 .0,7.2, 7.4, respectively. After 1 h pre-incubation, lung slices were cont inuously submerged in 24-well plate, incubated for 0,2,4,6 h,respectively. NR assay, MTT assay, LDH leakage and SOD activity were used to assess the slices viability under different slices thickness, medium pH and culturing time. Result When the slice thickness was 600 μm and medium pH was 7.0, t he viability of slices maintained best and steady for 6 h. There were positive c orrelations between NR uptake and MTT reduction in slices under different thickn ess(r 1 = 0.91, P<0.05) and medium pH(r 2 = 0.98, P<0.01). Conclusion PCLS is a simple, easy, feasible newly designed i n vitro model, which is worth further developing; NR assay is a convenient and sensitive index in determing lung slices viability.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2005年第2期249-252,共4页
Chinese Pharmacological Bulletin
关键词
肺切片
中性红
体外模型
lung slice
neutral central
in vitro model
