人脾脏巨噬细胞总RNA的提取和产率研究

Extraction and productivity of total RNA in macrophages isolated from human spleen

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 extract total RNA from macrophages isolated from human spleen and determine the RNA productivity.Methods Macrophages were isolated and purified by anchorage cultivation from the excisional human spleen in the patients suffering from portal hypertension.Total RNA was extracted from 5 cases of macrophages through TRIzol method respectively.Spectrophotometric analysis was used to determine the yield and purity of total RNA and agarose gel electrophoresis was used to test whether there existed RNA degeneration.The average productivity of total RNA was calculated considering the amount of macrophages in each case.Results The results of spectrophotometric analysis showed that the average value of A_ 260nm/A_ 280nm was 1.83± 0.13,indicating that the total RNA had a high purity.The results of agarose gel electrophoresis showed no RNA degeneration.The average productivity of total RNA was (41.58± 23.13) μg per 108 macrophages.Conclusion Through TRIzol method,an enough amount of total RNA with high purity and good integrity could be extracted successfully from macrophages of human spleen.