Yuling Li1, Xingjuan Gao2, Zhihua Wang3, Wei Liu1, Fang Xu1, Yejia Hu1, Yanuo Li1, Lei Shi4. 1. Department of Pathophysiology, Binzhou Medical University, Yantai 264003, Shandong, China. 2. Department of Pediatrics, The Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai 264000, Shandong, China. 3. Department of Gastroenterology, The Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai 264000, Shandong, China. 4. Department of Pathophysiology, Binzhou Medical University, Yantai 264003, Shandong, China. Electronic address: shilei0043@126.com.
Abstract
AIMS: Hepatitis is a kind of liver dysfunction and usually refers to a variety of pathogenic factors. Circular RNA (circRNA) participates in diverse diseases. This study aimed to explore the roles and regulatory mechanisms of the circRNA-4099 in L02 cell line. MAIN METHODS: Cells were induced with H2O2 dilutions and transfected with circRNA-4099 overproduction vectoer and the specific siRNA (si-circRNA-4099), as well as microRNA-706 (miR-706) mimic or the corresponding controls. Cell viability was detected with the CCK-8. The apoptotic rate was determined by the annexin v-FITC/PI with flow cytometer. The expression of circRNA-4099 or miR-706 was quantified depending on qRT-PCR. The reactive oxygen species (ROS) level was examined through ROS assay. The relative proteins were individually determined via western blot. The relationship between the circRNA-4099 and miR-706 was detected through bioinformatics analysis and luciferase reporter assay. KEY FINDINGS: H2O2 induced inhibition of viability and promotion of apoptosis, ROS production and cell fibrosis as well as keap1/Nrf2 and p38MAPK cascades on L02 cell line. circRNA-4099 was stimulated by H2O2. Plentiful circRNA-4099 augmented H2O2-resulted damage by inhibiting miR-706. miR-706 mimic partly abolished the influence of circRNA-4099 on L02 cells. Meanwhile, circRNA-4099 silence exerted opposite effect on these elements above. SIGNIFICANCE: circRNA-4099 aggravated H2O2-induced injury by inhibiting miR-706 through triggering keap1/Nrf2 and p38MAPK in the L02 cells.
AIMS: Hepatitis is a kind of liver dysfunction and usually refers to a variety of pathogenic factors. Circular RNA (circRNA) participates in diverse diseases. This study aimed to explore the roles and regulatory mechanisms of the circRNA-4099 in L02 cell line. MAIN METHODS: Cells were induced with H2O2 dilutions and transfected with circRNA-4099 overproduction vectoer and the specific siRNA (si-circRNA-4099), as well as microRNA-706 (miR-706) mimic or the corresponding controls. Cell viability was detected with the CCK-8. The apoptotic rate was determined by the annexin v-FITC/PI with flow cytometer. The expression of circRNA-4099 or miR-706 was quantified depending on qRT-PCR. The reactive oxygen species (ROS) level was examined through ROS assay. The relative proteins were individually determined via western blot. The relationship between the circRNA-4099 and miR-706 was detected through bioinformatics analysis and luciferase reporter assay. KEY FINDINGS: H2O2 induced inhibition of viability and promotion of apoptosis, ROS production and cell fibrosis as well as keap1/Nrf2 and p38MAPK cascades on L02 cell line. circRNA-4099 was stimulated by H2O2. Plentiful circRNA-4099 augmented H2O2-resulted damage by inhibiting miR-706. miR-706 mimic partly abolished the influence of circRNA-4099 on L02 cells. Meanwhile, circRNA-4099 silence exerted opposite effect on these elements above. SIGNIFICANCE: circRNA-4099 aggravated H2O2-induced injury by inhibiting miR-706 through triggering keap1/Nrf2 and p38MAPK in the L02 cells.