■与阻塞性睡眠呼吸暂停低通气综合征(OSAHS)相关的慢性肾脏病(CKD)主要由慢性间歇性缺氧(CIH)引起的肾损伤引起。本研究旨在探讨长链非编码RNA(lncRNA)生长停滞特异性转录本5(GAS5)与重组腺嘌呤磷酸核糖基转移酶(APRT)在CIH诱导的肾损伤中的相互作用。
■建立大鼠间歇性缺氧模型,从肾组织中提取总RNA,使用高通量测序技术进行转录组测序。建立CIH大鼠模型,注射sh-GAS5或OE-APRT质粒,测定血清尿素氮(BUN)和肌酐酰胺水解酶,并检测氧化应激相关因子的表达。苏木精和伊红(H&E)和Masson三色染色用于形态学观察,通过TUNEL染色确定细胞凋亡。GAS5,TATA盒结合蛋白相关因子1(TAF1)之间的相互作用,并对APRT进行了预测和验证。转染HK-2细胞后,GAS5、TAF1、APRT、Bax,Bcl-2,凋亡相关因子,纤维化相关因子(胶原蛋白I和IV),和自噬相关蛋白(LC3-Ⅱ,LC3-Ⅰ,p62和Beclin-1)通过RT-qPCR和蛋白质印迹法测量。
■测序结果显示,CIH组TAF1显著升高,APRT显著降低。RNA显著参与CI介导的肾损伤的生物学过程。注射GAS5抑制或APRT过表达质粒的CIH大鼠显示GAS5降低和APRT表达升高,同时抑制血清BUN和肌酐酰胺水解酶水平。同时,GAS5抑制或APRT过表达减弱细胞凋亡和纤维化,抑制氧化应激,并促进CIH诱导的肾小管上皮细胞的自噬。RNA下拉测定和RIP验证了GAS5和TAF1的结合和相互作用。芯片免疫沉淀(ChIP)鉴定了APRT启动子的TAF1调控。GAS5和TAF1负调控APRT表达。
■lncRNAGAS5可以结合TAF1抑制APRT转录,从而增强CIH诱导的大鼠肾损伤。
UNASSIGNED: Chronic kidney disease (CKD) related to obstructive sleep apnea-hypopnea syndrome (OSAHS) mainly results from chronic intermittent hypoxia (CIH)-induced renal injury. This study aimed to explore the interaction between the long noncoding RNA (lncRNA) growth arrest-specific transcript 5 (GAS5) and recombinant adenine phosphoribosyltransferase (APRT) in CIH-induced renal injury.
UNASSIGNED: A rat intermittent hypoxia model was constructed, total RNA was extracted from kidney tissue, and transcriptome sequencing was performed using high-throughput sequencing technology. CIH rat models were established and injected with sh-GAS5 or OE-APRT plasmid, the serum levels of blood urea nitrogen (BUN) and creatinine amidohydrolase were measured, and the expression of oxidative stress-related factors was detected. Hematoxylin and eosin (H&E) and Masson\'s trichrome staining were used for morphological observations, and cell apoptosis was determined by TUNEL staining. Interactions between GAS5, TATA-box binding protein-associated factor 1 (
TAF1), and APRT were predicted and verified. After transfection of HK-2 cells, the expression of GAS5, TAF1, APRT, Bax, Bcl-2, apoptosis-related factors, fibrosis-related factors (collagen I and Ⅳ), and autophagy-related proteins (LC3-Ⅱ, LC3-Ⅰ, p62, and Beclin-1) was measured by RT-qPCR and western blotting.
UNASSIGNED: Sequencing results revealed that
TAF1 was significantly increased and APRT was significantly decreased in the CIH group. RNA was significantly involved in the biological process of kidney injury mediated by CIH. CIH rats injected with GAS5 suppression or APRT overexpression plasmids showed decreased GAS5 and elevated APRT expression, along with suppressed serum levels of BUN and creatinine amidohydrolase. Meanwhile, GAS5 suppression or APRT overexpression attenuated apoptosis and fibrosis, suppressed oxidative stress, and promoted autophagy in CIH-induced renal tubular epithelial cells. The RNA pull-down assay and RIP verified the binding and interaction of GAS5 and
TAF1. Chip immunoprecipitation (ChIP) identified
TAF1 regulation of the APRT promoter. GAS5 and
TAF1 negatively regulated APRT expression.
UNASSIGNED: The lncRNA GAS5 can bind TAF1 to suppress APRT transcription, thereby enhancing CIH-induced renal injury in rats.