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目的 探讨提取人脾脏巨噬细胞总RNA的方法并测算其产率。方法 收集 5例门静脉高压症脾亢患者的手术切除脾脏 ,经贴壁培养法分离纯化出巨噬细胞 (Mφ)后 ,采用TRIzol试剂一步法提取 5例脾脏Mφ样本的总RNA。分光光度法和琼脂糖凝胶电泳检测总RNA的产量和质量。最后计算出每 10 8个Mφ总RNA的产率。 结果 5例样本Mφ总RNA的A2 60nm与A2 80nm比值平均为 1.83± 0 .13 ,提示总RNA纯度较高 ,电泳结果提示总RNA无降解。每 10 8个Mφ平均可抽提 (4 1.5 8± 2 3 .13 ) μg总RNA。 结论 采用TRIzol试剂一步法提取脾脏Mφ总RNA是可行的 ,收获的人脾脏Mφ总RNA纯度高、完整性好、量较多 ,可满足后续实验要求。
Objective To investigate the method of extracting total RNA from human spleen macrophages and evaluate its yield. Methods The spleen of 5 patients with portal hypertension were collected. The macrophages (Mφ) were isolated and purified by adherent culture. Total RNA was extracted from 5 splenic Mφ samples by TRIzol reagent in one step. Spectrophotometry and agarose gel electrophoresis to measure total RNA yield and quality. Finally, the yield of total RNA per 10 8 Mφ was calculated. Results The average ratio of A2 60nm to A2 80nm of Mφ total RNA in 5 samples was 1.83 ± 0.13, which indicated that the purity of total RNA was high. The results of electrophoresis suggested that total RNA did not degrade. The average total RNA was extractable per 10 8 Mφ (4 1.5 8 ± 2 3 .13) μg. Conclusion It is feasible to extract Mφ total RNA from spleen by TRIzol reagent in one step. The purity of Mφ total RNA in harvested human spleen is high, the integrity is good and the quantity is large, which can meet the requirements of subsequent experiments.