长链非编码RNA HOTAIR在急性肝损伤中的表达及其对细胞铁死亡及炎症发生的干预作用

Expression of long non-coding RNA HOTAIR in acute liver injury and its intervention on cell ferroptosis and inflammation

目的探讨长链非编码RNA HOTAIR在急性肝损伤中的表达及其对细胞铁死亡及炎症发生的干预作用。方法选取无特定病原体级C57BL/6小鼠,将小鼠分为对照组、急性肝损伤组、急性肝损伤+HOTAIR对照质粒腺病毒干预组(NC组)、急性肝损伤+HOTAIR过表达腺病毒干预组(OV组)、急性肝损伤+HOTAIR短发夹RNA干预组(KD组)、急性肝损伤+HOTAIR过表达腺病毒+Fer-1(一种铁死亡抑制剂)干预组(OV+Fer-1组),急性肝损伤模型通过1%四氯化碳溶液5 ml/(kg·d)的量灌胃得到,对照组小鼠灌胃用等量0.9%氯化钠溶液。比较各组小鼠血清和肝组织中HOTAIR、炎症因子[肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)]、肝损伤标志物(丙氨酸转氨酶、天冬氨酸转氨酶)、铁死亡标志物(Fe^(3+)、谷胱甘肽、丙二醛)水平,观察小鼠肝组织病理变化。结果急性肝损伤组小鼠血清HOTAIR表达水平高于对照组[(2.9±0.6)比(1.1±0.3)](P<0.001)。OV组小鼠血清和肝组织中TNF-α、IL-1β及丙氨酸转氨酶、天冬氨酸转氨酶水平均高于NC组,而KD组上述指标均低于NC组(均P<0.05)。OV组小鼠血清和肝组织中丙二醛和Fe^(3+)水平均高于NC组、谷胱甘肽水平低于NC组,而KD组小鼠血清和肝组织中丙二醛和Fe^(3+)水平低于NC组、谷胱甘肽水平高于NC组(均P<0.05)。OV+Fer-1组小鼠血清和肝组织中TNF-α、IL-1β及丙氨酸转氨酶、天冬氨酸转氨酶水平均低于OV组(均P<0.05)。与NC组相比,OV组小鼠肝组织中炎性细胞浸润程度显著增加,而这种作用在OV+Fer-1组中被显著回调。结论HOTAIR可上调急性肝损伤引起的炎症反应,铁死亡信号通路可能介导了这一作用机制。

Objective To investigate the expression of long non-coding RNA HOTAIR in acute liver injury and its intervention on cell ferroptosis and inflammation.Methods C57BL/6 mice with specific pathogen free were selected and divided into control group,acute liver injury group,acute liver injury+HOTAIR control plasmid adenovirus intervention group(NC group),acute liver injury+HOTAIR overexpression adenovirus intervention group(OV group),acute liver injury+HOTAIR short hairpin RNA(shRNA)intervention group(KD group),and acute liver injury+HOTAIR overexpression adenovirus+Fer-1(a ferroptosis inhibitor)intervention group(OV+Fer-1 group).The acute liver injury model was obtained by gavage of 5 ml/(kg·d)of 1%carbon tetrachloride solution,while the control group mice were gavaged with an equal amount of 0.9%sodium chloride solution.The levels of HOTAIR and inflammatory factors[tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)],liver injury markers(alanine transaminase,aspartate transaminase)and ferroptosis markers(Fe^(3+),glutathione,malondialdehyde)in serum and liver tissue of mice in each group were compared.The pathological changes in liver tissue of mice in each group were observed.Results The expression level of serum HOTAIR in mice with acute liver injury group was higher than that in the control group[(2.9±0.6)vs(1.1±0.3)](P<0.001).The levels of TNF-α,IL-1β,alanine transaminase and aspartate transaminase in serum and liver tissue of OV group were higher than those of NC group,while the levels of KD group were lower than those of NC group(all P<0.05).The levels of malondialdehyde and Fe^(3+)in the serum and liver tissue of OV group were higher than those of NC group,and the level of glutathione was lower than that of NC group;however,the levels of malondialdehyde and Fe^(3+)in the serum and liver tissue of KD group were lower than those of NC group,and the level of glutathione was higher than that of NC group(all P<0.05).The levels of TNF-α,IL-1β,alanine transaminase and aspartate transaminase in serum and liver tissue of OV+Fer-1 group were lower than those of OV group(all P<0.05).Compared with the NC group,the degree of inflammatory cell infiltration of liver tissue in the OV group mice was significantly increased,and this effect was significantly reversed in the OV+Fer-1 group.Conclusion HOTAIR can up-regulate the inflammatory response caused by acute liver injury,which may be mediated by ferroptosis signaling pathway.