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目的研究超顺磁氧化铁(SPIO)标记对大鼠脂肪干细胞(ADSCs)转铁蛋白受体(TfR)和铁蛋白轻链(Fn-L)基因及蛋白表达的影响。方法实验通过医院伦理委员会的批准。标记组(实验组)采用多聚赖氨酸(PLL,终质量浓度为1.5μg/mL)介导SPIO(终质量浓度为50μg/mL)标记ADSCs。在2、4、8、16、24、96、168、336、504、672 h,分别用实时荧光定量聚合酶链反应(RT-PCR)和WesternBlot实验定量检测实验组和未标记组(对照组)TfR和Fn-L基因及蛋白表达水平。结果 PLL-SPIO标记ADSCs后,实验组Fn-L mRNA(2、4、8、16、24、96、168、336 h)及蛋白(16、72、96、168 h)(P均<0.05)表达水平会暂时性升高,至标记后一定时间,Fn-L mRNA(504、672h)及蛋白(336、504、672h)表达水平两组间差异无统计学意义(P均>0.05);实验组TfR mRNA(2、4、8、16、72、96、168、336 h)及蛋白(24、96 h)(P均<0.05)表达水平会暂时性减低,至标记后一定时间,TfRmRNA(504、672 h)及蛋白(168、336、504、672 h)表达水平两组间差异无统计学意义(P均>0.05)。结论在一定浓度内,PLL-SPIO标记ADSCs,对Fn-L和TfR基因及蛋白表达仅产生暂时性影响;从而为细胞内标记过程的安全性提供了实验依据。
Objective To investigate the effects of SPIO labeling on gene and protein expression of transferrin receptor (TfR) and ferritin light chain (Fn-L) in rat adipose derived stem cells (ADSCs). Methods The experiment was approved by the Hospital Ethics Committee. Labeling group (experimental group) ADSCs were labeled with polylysine (PLL, final concentration of 1.5μg / mL) mediated SPIO (final concentration of 50μg / mL). At 2,4,8,16,24,96,168,336,504 and 672 h respectively, the expression of GFP was detected by real-time fluorescence quantitative polymerase chain reaction (RT-PCR) and Western Blot, respectively ) TfR and Fn-L gene and protein expression levels. Results After PLL-SPIO labeled ADSCs, Fn-L mRNA (2,4,8,16,24,96,168,336 h) and protein (16,72,96,168 h) in experimental group (all P <0.05) The expression level of Fn-L mRNA (504,672h) and protein (336,504,672h) had no significant difference between the two groups (P> 0.05) The expression of TfR mRNA (2,4,8,16,72,96,168,336 h) and protein (24,96 h) (all P <0.05) decreased temporarily. At a certain time after labeling, TfR mRNA ( 504,672 h) and protein (168,336,504,672 h), there was no significant difference between the two groups (P> 0.05). CONCLUSION: PLL-SPIO-labeled ADSCs have only a temporary effect on the expression of Fn-L and TfR genes and proteins at a certain concentration, which provides an experimental basis for the safety of intracellular labeling.