Guo Bo, Yan Shengzhe, Zhai Lei, Cheng Yanzhen
School of Clinical Medicine, Guangzhou Health Science College, Guangzhou, 510450 Guangdong China.
Department of Endocrinology, Zhujiang Hospital, Southern Medical University, Guangzhou, 510280 Guangdong China.
Cytotechnology. 2024 Apr;76(2):259-269. doi: 10.1007/s10616-023-00614-x. Epub 2024 Feb 20.
LncRNA HOTAIR has been reported to be associated with metabolic diseases of the liver. However, the effect of HOTAIR on non-alcoholic fatty liver disease (NAFLD) inflammation and its potential mechanism have not been reported. Genes and proteins expression were detected by qRT-PCR and Western blot respectively. The level of inflammatory cytokines was assessed by ELISA. HepG2 cell viability was detected by MTT assay. TG level and lipid accumulation were measured by Assay Kit and Oil red O staining, respectively. Direct binding relationship between HOTAIR and Serine/arginine splicing factor 1 (SRSF1), SRSF1 and MLX interacting protein like (MLXIPL) were confirmed by RNA-pull down and RIP assay. HOTAIR was highly expressed in free fatty acids (FFA)-treated HepG2 cells. HOTAIR knockdown alleviated FFA-induced inflammation of HepG2 cells. Then further analysis showed that HOTAIR and SRSF1 had a mutual binding relationship, and HOTAIR maintained MLXIPL mRNA stability via recruiting SRSF1 in HepG2 cells. Moreover, the inhibitory effect of HOTAIR knockdown on FFA-induced inflammation in HepG2 cells was reversed by MLXIPL overexpression. HOTAIR accelerates inflammation of FFA-induced HepG2 cells by recruiting SRSF1 to stabilize MLXIPL mRNA, which will help to find new effective strategies for NAFLD therapy.
The online version contains supplementary material available at 10.1007/s10616-023-00614-x.
据报道，长链非编码RNA HOTAIR与肝脏代谢疾病有关。然而，HOTAIR对非酒精性脂肪性肝病（NAFLD）炎症的影响及其潜在机制尚未见报道。分别采用qRT-PCR和蛋白质免疫印迹法检测基因和蛋白质表达。通过酶联免疫吸附测定法评估炎性细胞因子水平。采用MTT法检测HepG2细胞活力。分别使用检测试剂盒和油红O染色法测定甘油三酯水平和脂质蓄积情况。通过RNA下拉实验和RNA免疫沉淀实验证实了HOTAIR与丝氨酸/精氨酸剪接因子1（SRSF1）、SRSF1与类MLX相互作用蛋白（MLXIPL）之间的直接结合关系。HOTAIR在游离脂肪酸（FFA）处理的HepG2细胞中高表达。敲低HOTAIR可减轻FFA诱导的HepG2细胞炎症。进一步分析表明，HOTAIR与SRSF1存在相互结合关系，并且在HepG2细胞中HOTAIR通过招募SRSF1维持MLXIPL mRNA的稳定性。此外，MLXIPL过表达可逆转敲低HOTAIR对FFA诱导的HepG2细胞炎症的抑制作用。HOTAIR通过招募SRSF1稳定MLXIPL mRNA来加速FFA诱导的HepG2细胞炎症，这将有助于找到治疗NAFLD的新有效策略。
在线版本包含可在10.1007/s10616-023-00614-x获取的补充材料。
