Interference with miR-210 Alleviated Renal Injury in Septic Rats by Inhibiting JAK-STAT Pathway
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ORIGINAL ARTICLE
Interference with miR-210 Alleviated Renal Injury in Septic Rats by Inhibiting JAK-STAT Pathway Benchao Zhang,1 Ziye Guo,1 Siqi Lai,1 and Hongjian Chen
2,3
Pediatric sepsis has become the leading cause of death in pediatric intensive care units (PICU). The regulation of target genes may be the key to the treatment of pediatric sepsis. The expression of miR-210 in rat serum was detected by RTqPCR. The serum BUN, Scr, and CysC were detected by an automatic biochemical analyzer. The expression of inflammatory factors was detected by ELISA. The apoptosis level of the cells was detected by TUNEL staining. The expression of apoptotic proteins Bcl2, Bax, Cleaved caspase3, caspase3, and JAK/STAT3 pathway-related proteins were detected by western blot. The expression of miR-210 was abnormally elevated in sepsis pups. Interfering with the expression of miR-210 in rats could reduce the degree of renal injury and inhibit the inflammatory response in sepsis pups. In addition, interference with miR-210 could inhibit the apoptosis level of renal tissue cells, and the expression of apoptosis-related proteins was also significantly decreased. During this process, we found that after interfering with the expression of miR-210, the expression of the JAK/STAT pathway was inhibited. Then, pathway agonist SC-39100 can reverse the inhibitory effects of interfering with miR-210 on renal tissue damage, inflammatory response, and apoptosis. Interference with miR-210 alleviated renal injury in septic rats by inhibiting JAK-STAT pathway.
Abstract—
KEY WORDS: miR-210; pediatric sepsis; renal injury; JAK-STAT pathway.
MATERIALS AND METHODS
1
Department of Critical Care Medicine, The People’s Hospital of Longhua, Shenzhen, Shenzhen, 518109, Guangdong Province, China 2 Department of Infection Disease, Children’s Hospital of Nanjing Medical University, No.72, Guangzhou road, Gulou District, Nanjing, Jiangsu Province, 210000, China 3 To whom correspondence should be addressed at Department of Infection Disease, Children’s Hospital of Nanjing Medical University, No.72, Guangzhou road, Gulou District, Nanjing, Jiangsu Province, 210000, China. E-mail: [email protected]
Animals Male Sprague Dawley rats were aged between 17 and 18 days and had an average body weight of 49.4 ± 0.4 g. The model was built after a week of adaptive feeding. The experimental protocol was approved by the commission for animal experimentation of the People’s Hospital of Longhua, Shenzhen.
0360-3997/20/0000-0001/0 # 2020 Springer Science+Business Media, LLC, part of Springer Nature
Zhang, Guo, Lai, and Chen CLP model Preparation
Histological Examination
An animal model of sepsis was established using an internationally recognized cecal ligation and puncture (CLP) sepsis model. The standard steps mainly refer to the report of Rittirsch [1] et al. 24 h before the operation, the rats were injected with the tail vein empty vector or miR-210, and the sepsis model was subsequently induced. After 10% chloral hydrate anesthesia (3 ml/kg, i.p.), the
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