MiR-372 Participates In The Progression Of Acute Hyperlipidemic Pancreatitis By Regulating Pten

Objective:To prove the high expression of miR-372 in mice with acute hyperlipidemic pancreatitis and explore the relationship between miR-372 and downstream signaling pathway.Methods:Using TargetScan database to predict miR-372 target gene,dual luciferase reporter gene detection was used to detect whether miR-372 could bind to PTEN promoter.20 male C57BL/6 mice were divided into Control group,HLAP group,AD-NC group,and AD-sh-miR-372 treatment group,with 5 mice in each group.Ad-sh-mir-372 and ad-nc were injected through tail vein,fed with high-fat diet for three weeks,and then intraperitoneal injection of cerulein combined with lipopolysaccharide was used to establish mice model of hyperlipidemic acute pancreatitis.After 12 hours of modeling,the mice were killed,the pancreatic tissue and blood were collected for examination.①Biochemical analyzer was used to detect the levels of LDL-C,HDL-C,TG,TG.② The expression of miR-372 in serum was detected by qRT-PCR.③Histopathological examination:hematoxylin eosin staining and periodic acid Schiff staining were used to observe the histopathological situation of pancreas;④molecular biological examination:Western blot was used to quantitatively detect the protein levels of phosphatase and tensin homolog deleted on chromosome ten(PTEN),pAKT,P50,P65 and β-Catenin in pancreas tissues.Enzyme linked immunosorbent assay was used to detect the expression of serum interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α),IL-1β and TGF-β.ANOVA and Bonferroni test were used to analyze the data.Results:(1)The target of PTEN binding on miR-372 was predicted by TargetScan database.Dual luciferase reporter gene detection showed that mir-372 combined with 3’UTR of PTEN and inhibited its transcription.(2)In HLAP model,the expression of miR-372 was increased,the expression of PTEN was decreased,and the expression levels of p-AKT,P65 and β-Catenin were increased.(3)After miR-372 expression was inhibited by lentivirus transfection,the expression of miR-372 was significantly decreased,the infiltration of inflammatory cells was reduced,the levels of TNF-α and TGF-β were decreased,the levels of triglyceride and cholesterol were decreased,the expression of PTEN was increased,and the expression of p-AKT was decreased.Conclusion:In mice with acute hyperlipidemic pancreatitis,inhibition of miR-372 can reduce inflammation and blood lipid,which may be due to the down-regulation of PTEN,p-Akt and TGF-β pathway by miR-372.This study provides a new idea for studying the molecular mechanism of acute hyperlipidemic pancreatitis and its new therapeutic targets,and may become a therapeutic target for HLAP in the future.Objective:The current study aims to investigate the expression of miR-372 in serum of patients with hyperlipidemic acute pancreatitis(HLAP)and its clinical significance.Methods:Patients with AP admitted to our hospital from November 2018 to December 2019 were selected.AP patients with HTG diagnosis were included in group A,and then they were divided into mild(HTG-MAP)and severe(HTG-SAP)subgroups according to AP classification criteria.The rest were non htgap patients and were included in group B.Another 25 patients with hyperlipidemia(Group C)and 30 healthy volunteers(Group D)were selected as the control group.The serum levels of triglyceride and amylase in all patients were examined and Acute Physiology and Chronic Health Evaluation(APACHE)Ⅱ scores were recorded.The level of miR-372 in serum was detected by RT-PCR.The correlation between the level of miR-372 and clinical indicators of HLAP was analyzed,and its role in predicting the severity of acute pancreatitis was evaluated.Results:Our data showed that the level of miR-372 in serum of group A(4.76±2.60)was significantly higher than that of group B(0.98±0.80)(p<0.001),group C(0.85±0.62)(p<0.001)and group D(0.76±0.44)(p<0.001),but there was no significant difference in the expression of miR-372 between groups B,C and D.ROC curve analysis showed that the area under the curve of detection of HLAP by miR-372 was 0.785(0.634-0.935).The expression level of miR-372 in was significantly increased in the severe HLAP group(6.45±2.20)than that in mild HLAP group group(3.08± 1.74),with significant difference(P<0.05).Further study showed that the expression level of miR-372 was correlated with the level of triacylglycerol(r=0.666,P<0.001),but not with amylase(r=0.145,P>0.05).Conclusion:The expression of miR-372 in HLAP was significantly up-regulated,and it increased with the severity of the disease,which may shed light on the clinical diagnosis and severity assessment of HLAP.