He Lan-Zhi, Meng Ya-Kun, Han Yan-Zhong, Zhang Zhen-Fang, Yin Ping, Sang Xiu-Xiu, Xiao Xiao-He, Bai Zhao-Fang
School of Pharmacy, Hunan University of Traditional Chinese Medicine, Changsha 410208, China.
China Military Institute of Chinese Medicine, 302 Military Hospital, Beijing 100039, China.
Zhongguo Zhong Yao Za Zhi. 2016 Nov;41(22):4234-4239. doi: 10.4268/cjcmm20162224.
This paper was aimed to investigate the protective effects of luteolin (Lut) against acetaminophen(APAP)-induced damage in L02 liver cells. CCK-8 was used to detect the cell activation of L02 cells treated by different Lut. The concentration and time of APAP induced L02 cell damage was screened. The effect of Lut on APAP induced apoptosis of L02 cells was detected by cell morphological observation, CCK-8 assay and flow cytometry. The contents of MDA, GSH and SOD activity in cell supernatant were detected by colorimetric assay. The expression of apoptosis-related genes Bax, Bcl-2 and caspase-3 was detected by RT-PCR. The results showed that Lut in 2.5-40 μmol•L⁻¹ range does not affect the activity of L02 cells; 12 mmol•L⁻¹ APAP incubated with L02 cell 12 h to establish damage model. Compared with the model group, the cell status of Lut group was significantly improved, the cell body was increased, the adherence ability was recovered, and the apoptosis rate was obviously decreased. MDA content decreased significantly (P<0.05, P<0.01), GSH and SOD activity significantly increased (P<0.05, P<0.01), at the same time, it could up-regulate expression of Bcl-2 mRNA and down-regulate the expression of Bax and caspase-3 mRNA. In conclusion,Lut has protective effect on APAP induced L02 cell injury, and its mechanism may be related to the reduction of oxidative stress and inhibition of apoptosis.
本文旨在研究木犀草素(Lut)对乙酰氨基酚(APAP)诱导的L02肝细胞损伤的保护作用。采用CCK-8法检测不同浓度Lut处理的L02细胞的活性。筛选出APAP诱导L02细胞损伤的浓度和时间。通过细胞形态学观察、CCK-8法检测和流式细胞术检测Lut对APAP诱导的L02细胞凋亡的影响。采用比色法检测细胞上清液中丙二醛(MDA)、谷胱甘肽(GSH)含量及超氧化物歧化酶(SOD)活性。采用RT-PCR法检测凋亡相关基因Bax、Bcl-2和caspase-3的表达。结果显示,2.5-40μmol•L⁻¹范围内的Lut不影响L02细胞活性;12 mmol•L⁻¹ APAP与L02细胞孵育12 h建立损伤模型。与模型组相比,Lut组细胞状态明显改善,细胞体积增大,贴壁能力恢复,凋亡率明显降低。MDA含量显著降低(P<0.05,P<0.01),GSH和SOD活性显著升高(P<0.05,P<0.01),同时,可上调Bcl-2 mRNA表达,下调Bax和caspase-3 mRNA表达。综上所述,Lut对APAP诱导的L02细胞损伤具有保护作用,其机制可能与减轻氧化应激和抑制细胞凋亡有关。