限定骨骼肌提取液（ME，10～50kDa）影响培养的大鼠脊髓腰段运动神经元生长的定量研究

BIOASSAY OF EFFECTION OF THE LIMITED SKELETAL MUSCLE EXTRACTS(10-50kDa)ON THE NEURONS OF RAT LUMBAR VENTRAL SPINAL CORD IN VITRO

用胎鼠的限定骨骼肌提取液（ＭＥ，１０～５０ｋＤａ）作用于无血清培养的胚胎大鼠脊髓腰段腹角细胞后，用四唑盐（ＭＴＴ）微量自动比色定量法和ＬＤＨ活性测定法检测了骨骼肌提取液对大鼠脊髓运动神经元生长存活的生物活性，结果显示：细胞培养４ｄｉｖ后，ＭＴＴ微量比色的ＯＤ值随加入骨骼肌提取液浓度的增加而升高，含骨骼肌提取液２００和４００μｇ／ｍｌ蛋白的实验孔与单纯无血清的对照孔比较有极显著性差异（Ｐ＜０．００１），说明２００μｇ／ｍｌ以上浓度的骨骼肌提取液对脊髓腹角神经元的生长有明显的促进作用。细胞培养１４ｄｉｖ后，ＬＤＨ活性测定ＯＤ值的线性回归斜率亦随加入骨骼肌提取液浓度的增加而升高，说明存活细胞数的增加。实验说明培养神经细胞的ＭＴＴ和ＬＤＨ的检测方法有可能成为评估运动神经营养因子活性的指标。

The effect of skeletal muscle extracts(ME)on the survival and growth of rat spinal cord neurons in culture was studied by the MTT colorimetric microassay and the quantitative determination of lactate dehydrogenase(LDH)efflux assay.The OD of MTT microassay was raised as ME increased in the medium,and it was statistically significant higher than that in the control which cultured with medium serum free(P<0.001)in 4 days of culture,The LDH effluxassay also showed that activity was raised as ME concentrated in 14 days of culture,The positive ChAT immunoreactive neurons showed that the majority of cultured cells which expressed MTT microassay and LDH effluxassay were motoneurons of spinal cord.The present result supported the hypothesis that the motoneuron putative trophic factors contained in skeletal muscle improved the survival and growth of motoneurons,and the MTT and LDH assay could be used for the measures for the neurotrophic factor's bioassay.