Abstract：
BACKGROUND: Formaldehyde (FA) is associated with the occurrence of leukemia, and oxidative stress is considered to be a major reason. As an endogenous biomarker of oxidative stress, few studies focus on the relationship between peroxiredoxin III (PrxIII) and FA toxicity. Our previous research observed high expression of PrxIII occurred in the process of apoptosis of bone marrow cells (BMCs) induced by FA, however the exact mechanism is unclear. Therefore, this paper aimed to explore the possible association between FA toxicity and PrxIII gene.
METHODS: We first, used a Cell Counting Kit-8 (CCK-8) to detect the viability of BMCs after they were exposed to different doses of FA (50, 100, 200 μmol/L) for different exposure time (12, 24, 48 h), then chose 24 h as an exposure time to detect the expression of PrxIII for exposing different doses of FA by Quantitative reverse transcription-PCR (qRT-PCR) and Western blot analysis. Based on our preliminary experimental results, we chose 100 μmol/L FA as an exposure dose to expose for 24 h, and used a small interfering RNA (siRNA) to silenced PrxIII to examine the cell viability by CCK-8, reactive oxygen species (ROS) level by DCFH-DA, apoptosis by Annexin V/PI double staining and cell cycle by flow cytometry (FCM) so as to explore the possible regulatory effect of PrxIII silencing on FA-induced bone marrow toxicity.
RESULTS: High expression of PrxIII occurred in the process of FA-induced oxidative stress. Silencing of PrxIII prevented FA from inducing oxidative stress, thus increasing cell viability, decreasing ROS level, rescuing G0 -G1 and G2 -M arrest, and reducing cell apoptosis.
CONCLUSIONS: PrxIII silencing might be a potential target for alleviating FA-induced oxidative damage.
METHODS: We first, used a Cell Counting Kit-8 (CCK-8) to detect the viability of BMCs after they were exposed to different doses of FA (50, 100, 200 μmol/L) for different exposure time (12, 24, 48 h), then chose 24 h as an exposure time to detect the expression of PrxIII for exposing different doses of FA by Quantitative reverse transcription-PCR (qRT-PCR) and Western blot analysis. Based on our preliminary experimental results, we chose 100 μmol/L FA as an exposure dose to expose for 24 h, and used a small interfering RNA (siRNA) to silenced PrxIII to examine the cell viability by CCK-8, reactive oxygen species (ROS) level by DCFH-DA, apoptosis by Annexin V/PI double staining and cell cycle by flow cytometry (FCM) so as to explore the possible regulatory effect of PrxIII silencing on FA-induced bone marrow toxicity.
RESULTS: High expression of PrxIII occurred in the process of FA-induced oxidative stress. Silencing of PrxIII prevented FA from inducing oxidative stress, thus increasing cell viability, decreasing ROS level, rescuing G0 -G1 and G2 -M arrest, and reducing cell apoptosis.
CONCLUSIONS: PrxIII silencing might be a potential target for alleviating FA-induced oxidative damage.
摘要：
背景：甲醛（FA）与白血病的发生有关，氧化应激被认为是一个主要原因。作为氧化应激的内源性生物标志物，很少有研究关注过氧化物氧化还原蛋白III（PrxIII）与FA毒性之间的关系。我们前期的研讨不雅察到PrxIII的高表达在FA引诱的骨髓细胞(BMCs)凋亡进程中，然而，确切的机制尚不清楚。因此,本文旨在探讨FA毒性与PrxIII基因的可能关联。
方法：我们首先，使用细胞计数试剂盒-8（CCK-8）检测BMC暴露于不同剂量的FA（50、100、200μmol/L）不同暴露时间（12、24、48h）后的活力，然后选择24小时作为暴露时间，通过定量逆转录PCR（qRT-PCR）和Westernblot分析检测暴露不同剂量FA的PrxIII的表达。根据我们的初步实验结果，我们选择100μmol/LFA作为暴露剂量，暴露24小时，并使用小干扰RNA（siRNA）沉默PrxIII，通过CCK-8检查细胞活力，通过DCFH-DA检查活性氧（ROS）水平，通过膜联蛋白V/PI双重染色和流式细胞术（FCM）检测细胞周期，以探讨PrxIII沉默对FA诱导的骨髓毒性的可能调节作用。
结果：PrxIII的高表达发生在FA诱导的氧化应激过程中。沉默PrxIII可防止FA诱导氧化应激，从而增加细胞活力，降低ROS水平，拯救G0-G1和G2-M逮捕，减少细胞凋亡。
结论：PrxIII沉默可能是减轻FA诱导的氧化损伤的潜在目标。
方法：我们首先，使用细胞计数试剂盒-8（CCK-8）检测BMC暴露于不同剂量的FA（50、100、200μmol/L）不同暴露时间（12、24、48h）后的活力，然后选择24小时作为暴露时间，通过定量逆转录PCR（qRT-PCR）和Westernblot分析检测暴露不同剂量FA的PrxIII的表达。根据我们的初步实验结果，我们选择100μmol/LFA作为暴露剂量，暴露24小时，并使用小干扰RNA（siRNA）沉默PrxIII，通过CCK-8检查细胞活力，通过DCFH-DA检查活性氧（ROS）水平，通过膜联蛋白V/PI双重染色和流式细胞术（FCM）检测细胞周期，以探讨PrxIII沉默对FA诱导的骨髓毒性的可能调节作用。
结果：PrxIII的高表达发生在FA诱导的氧化应激过程中。沉默PrxIII可防止FA诱导氧化应激，从而增加细胞活力，降低ROS水平，拯救G0-G1和G2-M逮捕，减少细胞凋亡。
结论：PrxIII沉默可能是减轻FA诱导的氧化损伤的潜在目标。
