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慢病毒介导的RNA干扰对大鼠神经元PKCγ mRNA及蛋白表达的影响

Effect of lentivirus-mediated RNA interference on expression of
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摘要 目的 探讨慢病毒介导的RNA干扰（RNAi）对大鼠神经元PKCymRNA及蛋白表达的影响.方法 孕16d的SD大鼠,原代培养大鼠胚胎皮层神经元,随机分为3组,每组6孔,对照组（C组）：不行任何处理;阴性对照组（NC组）：每孔加入阴性对照慢病毒3×10^5个;RNAi组：每孔中加入编码PKCγ shRNA的慢病毒载体（LV-PKCγ shRNA）颗粒3×10^5个.5 d后采用RT-PCR和Western blot法检测PKCγ mRNA和蛋白的表达水平,并计算干扰效率.结果 与C组比较,RNAi组大鼠神经元PKCγ mRNA及其蛋白表达水平均降低（P〈0.05）,NC组上述指标差异无统计学意义（P〉0.05）.基因干扰效率99.3%,蛋白干扰效率85.2%.结论 慢病毒介导的RNAi可有效地下调大鼠原代神经元PKCγ基因及其蛋白的表达. Objective To investigate the effect of lentivirus-mediated RNA interference（RNAi） on the expression of PKCγ mRNA and protein in rat neurons.Methods Primary cultured cortical neurons from SD rat （16 days of pregnency） embryos were randomly divided into 3 groups with 6 wells in each group：control group （group C）,negative group（group NC）and RNAi group.Group C received no treatment.Each well in group NC was given negative lentivirus 3 × 105.Each well in group RNAi was given the recombinant lentiviral vector containing PKCγ shRNA（LV-PKC7 shRNA）.The expression of PKCT mRNA and protein in rat neurons was detected by RT-PCR and Western blot 5 days later.The interference efficiency was ealculated.Results Compared with group NC,the expression of PKCγ mRNA and protein was down-regulatedin group RNAi（P〈0.05）,but no significant change Was found in group NC（P〉0.05）.The interference efficiency of gene and protein were 99.3%and 85.2%respectively.Conclusion Lentivirus-mediated RNAi can down restate the expression of PKC7 gene and protein in rat neurons.
作者 邹望远 郭曲练 宋宗斌 刘畅 潘韫丹
机构地区 中南大学湘雅医院麻醉科
出处 《中华麻醉学杂志》 CAS CSCD 北大核心 2010年第4期430-432,共3页 Chinese Journal of Anesthesiology
基金 国家自然科学基金（30772080,30801074）
关键词 RNA干扰 蛋白激酶C 慢病毒属 神经元 RNA interference Protein kinase C Lentivirus Neurons
分类号 R3 [医药卫生—基础医学]
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参考文献11

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二级参考文献11

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中华麻醉学杂志
2010年 第4期

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