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样本ＲＮＡ加样量的差异是影响Ｎｏｒｔｈｅｒｎ杂交ＲＮＡ分析和反转录ＰＣＲ（ＲＴ－ＰＣＲ）半定量分析的重要因素。因５－磷酸甘油醛脱氢酶（ＧＡＰＤＨ）和β－肌动蛋白（β－ａｃｔｉｎ）ｍＲＮＡ水平一般相对恒定，常用于基因表达研究时样本加样量的校正。但有报道表明某些因素对ＧＡＰＤＨ和β－ａｃｔｉｎ的ｍＲＮＡ水平有一定影响。脑星状细胞（ＡＣ）是近年来神经科学的研究热点之一，有关其基因表达的研究得到广泛的重视。本研究的目的是观察ＧＡＰＤＨ和β－ａｃｔｉｎ的ｍＲＮＡ水平是否作为培养ＡＣ缺氧状态下基因表达研究的标准参照。方法：ＡＣ取自新生ＮＭＩＲ小鼠大脑。培养５天后转至不含血清的培养基中，在缺氧箱（０３％氧）或正常氧供下培养２４小时。提取细胞总ＲＮＡ后，在甲醛／０１％琼脂糖凝胶上电泳使其变性和分离，并转移至尼龙膜上。样本加样ＲＮＡ量用溴化乙锭染色法评估。ＧＡＰＤＨ和β－ａｃｔｉｎｃＤＮＡ探针用磷３２－ｄＣＴＰ标记。Ｎｏｒｔｈｅｒｎ杂交放射自显影后的ｍＲＮＡ水平用计算机进行分析和统计学处理。结果：２４小时缺氧后，ＡＣ的ＧＡＰＤＨｍＲＮＡ水平（ｎ＝９）显著增高，达正常组（ｎ＝１０）５０３．０％，其Ｐ＜０００１。与此相反?
The differences in sample loading amount are important factors affecting the semi-quantitative analysis of Northern hybridization RNA and reverse transcription PCR (RT-PCR). Because GAPDH and β-actin mRNA levels are generally constant, they are commonly used for sample size correction in gene expression studies. However, it has been reported that some factors have some influence on the mRNA level of GAPDH and β-actin. Brain stellate cells (ACs) are one of the hot topics in neuroscience in recent years. Their research on gene expression has drawn extensive attention. The purpose of this study was to investigate whether mRNA levels of GAPDH and β-actin are used as standard references for the study of gene expression in AC hypoxia. Methods: AC was taken from the brain of neonatal NMIR mice. After 5 days of cultivation, the cells were transferred to serum-free medium and incubated for 24 hours in an anoxic chamber (0.3% oxygen) or normal oxygen. Total cellular RNA was extracted, denatured and separated on a formaldehyde / 0.1% agarose gel and transferred to a nylon membrane. Sample loading The amount of RNA was assessed by ethidium bromide staining. GAPDH and beta-actinc DNA probes are labeled with phospho-32-dCTP. Northern hybridizations after autoradiography mRNA levels were analyzed by computer and statistically processed. Results: After 24 hours of hypoxia, GAPDH mRNA level in AC increased significantly (n = 9) to 503.0% in normal group (n = 10), P <0001. opposite of this?