大鼠骨骼肌挫伤后ASCT2 mRNA的表达变化

Expression of ASCT2 mRNA in rat skeletal muscle after contusion

目的应用实时定量PCR技术检测大鼠骨骼肌挫伤后细胞中性氨基酸载体ASCT2 mRNA表达,分析其与挫伤时间的关系。方法建立大鼠骨骼肌挫伤模型,分别取伤后4、8、12、16、20、24、28、32 h及对照组检材。提取总RNA,逆转录合成第一链的cDNA,以RPL13为内参基因进行荧光定量检测,采用2-△△Ct法比较其与对照组肌肉组织中ASCT2 mRNA的相对表达量。结果损伤组肌肉组织中ASCT2 mRNA在挫伤后12、16h表达量分别为对照组的196.40%和189.15%,损伤后4、8、20、24、28、32h ASCT2 mRNA表达量与对照组相比无统计学差异(P>0.05),说明ASCT2 mRNA在损伤12～16h其表达量较正常组及其他损伤时间组要高,而在损伤20h以后ASCT2 mRNA表达量又降至正常水平。结论大鼠骨骼肌挫伤后32h之内ASCT2 mRNA的表达有一定的时间规律性,可望作为推断骨骼肌损伤时间的指标之一。

Objective To investigate the expression of ASCT2 mRNA in the contused muscle of rats and the relationship between the ASCT2 expression and the wound age. Methods The samples were taken at 4, 8, 12, 16, 20, 24, 28 and 32h and from the control group after contusion of rats, respectively. Total RNA was extracted from the muscle samples of each group and reverse transcription polymerase chain reaction （RT- PCR）was conducted to synthesize the 1st strand eDNA. The amount of ASCT2 mRNA in each sample was quantified using RPL13 intrinsic fluorescent assay and compared by the 2 - a a ct method with that from control samples. Results In the contusion group, the expression of ASCT2 mRNA increased rapidly and peaked at 12h, at 20h degraded to the amount that was as much as the control groups, then degraded stably. Conclusion It is suggested that ASCT2 mRNA analysis may be useful for estimation of wound age because of its timerelated expression after contusion in muscle.