Emergency Department of Traumatology, Shanghai Pudong Hospital, Shanghai, China (mainland).
Med Sci Monit. 2018 Nov 7;24:7980-7987. doi: 10.12659/MSM.910730.
BACKGROUND Paraquat (PQ) can over-accumulate in alveolar epithelial cells. Anthocyanin (An) can exert anti-oxidative properties. The role of An in PQ-induced toxicity is unclear, so we aimed to explore whether An could inhibit epithelial mesenchymal transition (EMT) induced by PQ in alveolar cells. MATERIAL AND METHODS lveolar epithelial cells were treated with PQ and An with concentration gradient for 12, 24, and 48 h. The cell viability, ROS level, and apoptosis rate were determined using the Cell Counting Kit-8 (CCK-8) and flow cytometry, respectively. The lactate dehydrogenase (LDH) leakage, methane dicarboxylic aldehyde (MDA) level, glutathione peroxidase (GPx), and superoxide dismutase (SOD) activities were determined by spectrophotometric method. The mRNA and protein expressions were detected using quantitative real-time PCR (qPCR) and Western blot, respectively. RESULTS An reduced the PQ-induced apoptosis in a dose-dependent manner. Moreover, An reduced the ratio of Bax/Bcl-2 to ROS level. We found that An suppressed the activity of LDH and MDA and improved SOD and GPX levels. Additionally, the level of PQ-induced E-cadherin was decreased by An while the expressions of vimentin, α-smooth muscle actin (α-SMA), and collagens type I (col-I) were increased. Furthermore, An inhibited the levels of transforming growth factor β1 (TGF-β1) and activin receptor-like kinase 5 (ALK5) and reduced the phosphorylation of smad2. CONCLUSIONS Our study shows newly discovered effects of anthocyanidins on EMT and supports their chemopreventive effects in paraquat-induced apoptosis in alveolar type II cells.
百草枯(PQ)可在肺泡上皮细胞中过度蓄积。花色苷(An)可发挥抗氧化作用。An 在 PQ 诱导的毒性中的作用尚不清楚,因此我们旨在探讨 An 是否可以抑制 PQ 在肺泡细胞中诱导的上皮间质转化(EMT)。
用 PQ 和 An 以浓度梯度处理肺泡上皮细胞 12、24 和 48 h。分别用细胞计数试剂盒-8(CCK-8)和流式细胞术测定细胞活力、ROS 水平和细胞凋亡率。用分光光度法测定乳酸脱氢酶(LDH)漏出、甲烷二羧酸醛(MDA)水平、谷胱甘肽过氧化物酶(GPx)和超氧化物歧化酶(SOD)活性。用实时荧光定量 PCR(qPCR)和 Western blot 分别检测 mRNA 和蛋白表达。
An 呈剂量依赖性降低 PQ 诱导的细胞凋亡。此外,An 降低 Bax/Bcl-2 比值和 ROS 水平。发现 An 抑制 LDH 和 MDA 的活性,提高 SOD 和 GPX 水平。此外,An 降低 PQ 诱导的 E-钙粘蛋白水平,增加波形蛋白、α-平滑肌肌动蛋白(α-SMA)和 I 型胶原蛋白(col-I)的表达。此外,An 抑制转化生长因子β1(TGF-β1)和激活素受体样激酶 5(ALK5)的水平,降低 smad2 的磷酸化。
本研究显示花色苷对 EMT 的新发现作用,并支持其在肺泡型 II 细胞 PQ 诱导的凋亡中的化学预防作用。