Jun Shen1, Li Liu2, Facai Zhang3, Jiang Gu3, Guanghui Pan1. 1. Department of Organ Transplantation, The Affiliated Hospital, Guizhou Medical University, Guiyang, Guizhou, China. 2. Department of Urology, The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan, Guangdong, China. 3. Department of Urology, The Affiliated Hospital, Guizhou Medical University, Guiyang, Guizhou, China.
Abstract
OBJECTIVE: To explore the possible mechanism of lncRNA TapSAKI in urine derived sepsis-induced kidney injury. MATERIALS AND METHODS: In vivo urine-derived sepsis (US) rat model and in vitro LPS-induced HK-2 cells were established, and TapSAKI, miR-22, PTEN, TLR4 and p-p65 expressions were detected by qRT-PCR and western blot. RNA precipitation and RNA pull-down were performed to confirm the interaction between TapSAKI and miR-22. RESULTS: TapSAKI was up-regulated, miR-22 was down-regulated, PTEN, TLR4 and p-p65 expressions, and inflammatory factors TNF-α and IL-6 levels were up-regulated in kidney tissue of US rats and LPS-induced HK-2 cells. In addition, TapSAKI interacted with miR-22, and negatively modulate miR-22 expression. We also observed TapSAKI promoted PTEN expression, TLR4/NF-κB pathway related proteins TLR4 and p-p65, and apoptosis protein cleaved-caspase-3 through negatively regulating miR-22. Further experiments proved TapSAKI/miR-22/TLR4/NF-κB pathway could promote HK-2 cell apoptosis. Finally, in vivo experiments showed TapSAKI knockdown negatively regulated miR-22 and positively regulate PTEN, decreased renal function indicators blood urea nitrogen and serum creatinine, and reduced TNF-α and IL-6. CONCLUSION: TapSAKI was elevated in urine derived sepsis-induced kidney injury, and promoted HK-2 cell apoptosis and inflammatory response through miR-22/PTEN/TLR4/NF-κB pathway.
OBJECTIVE: To explore the possible mechanism of lncRNA TapSAKI in urine derived sepsis-induced kidney injury. MATERIALS AND METHODS: In vivo urine-derived sepsis (US) rat model and in vitro LPS-induced HK-2 cells were established, and TapSAKI, miR-22, PTEN, TLR4 and p-p65 expressions were detected by qRT-PCR and western blot. RNA precipitation and RNA pull-down were performed to confirm the interaction between TapSAKI and miR-22. RESULTS: TapSAKI was up-regulated, miR-22 was down-regulated, PTEN, TLR4 and p-p65 expressions, and inflammatory factors TNF-α and IL-6 levels were up-regulated in kidney tissue of US rats and LPS-induced HK-2 cells. In addition, TapSAKI interacted with miR-22, and negatively modulate miR-22 expression. We also observed TapSAKI promoted PTEN expression, TLR4/NF-κB pathway related proteins TLR4 and p-p65, and apoptosis protein cleaved-caspase-3 through negatively regulating miR-22. Further experiments proved TapSAKI/miR-22/TLR4/NF-κB pathway could promote HK-2 cell apoptosis. Finally, in vivo experiments showed TapSAKI knockdown negatively regulated miR-22 and positively regulate PTEN, decreased renal function indicators blood ureanitrogen and serum creatinine, and reduced TNF-α and IL-6. CONCLUSION: TapSAKI was elevated in urine derived sepsis-induced kidney injury, and promoted HK-2 cell apoptosis and inflammatory response through miR-22/PTEN/TLR4/NF-κB pathway.
Authors: Seyed MohammadReza Hashemian; Mohammad Hossein Pourhanifeh; Sara Fadaei; Ali Akbar Velayati; Hamed Mirzaei; Michael R Hamblin Journal: Mol Ther Nucleic Acids Date: 2020-05-15 Impact factor: 8.886