The role of miRNA-181 targeting phosphatase and tensin homologue deleted on chromosome ten in the regulation of phosphatidylinositol-3-kinase/Akt signaling pathway in renal injury of hyperuricemia rats / 中华风湿病学杂志

ABSTRACT

Objective:To investigate the role of miRNA-181 targeting phosphatase and tensin homologue deleted on chromosome ten (PTEN) in regulating phosphatidylinositol-3-kinase/Akt signaling pathway (PI3K/Akt) signaling pathway in renal injury of hyperuricemia rats.Methods:Forty male Wistar rats were randomly divided into control group, model group, negative control group and miRNA-181 inhibition group. Their serum uric acid, creatinine and urea nitrogen were tested. HE staining was used to observe the renal histopathological changes in each group. The expression of miRNA-181, PTEN, PI3K and Akt mRNA in renal tissue of rats in each group was detected by quantitative real time-polymerase chain reaction (qRT-PCR). Western blotting analysis of PTEN, PI3K, Akt and p-Akt protein expression in renal tissue of rats in each group. The targeting relationship between miRNA-181 and PTEN was confirmed by double luciferase reporter gene experiment. One-way analysis of variance (ANOVA) was used for the comparison between multiple groups, with the same variance. LSD- t test was used for further comparison between the two groups. If the variance was not the same, Tamhane's T2 test was used for further comparison between the two groups. Independent sample t-test was used to compare between the two groups. Results:Compared with the control group (135±21) mmol/L; (27.8±2.1) μmol/L; (6.8±0.5) μmol/L, the contents of uric acid [(213±28) mmol/L, (214±23) mmol/L, creatinine (49.2±2.3) μmol/L, (48.6±2.2) μmol/L and urea nitrogen (11.5±2.7) μmol/L; (11.7±2.5) μmol/L] in the model group and the negative control group were significantly increased ( Furic acid=26.739, Fcreatinine=259.055, Furea nitrogen=12.921, all P<0.05); compared with the nega-tive control group, the contents of uric acid (169±21) mmol/L, creatinine (33.7±1.8) μmol/L and urea nitrogen (9.1±1.7) μmol/L in the miRNA-181 inhibition group were decreased (LSD- turic acid=4.356, LSD- tcreatinine=15.773, LSD- turea nitrogen=2.858, all P<0.05). The expression level of miRNA-181 in renal tissue of the model group and the negative control group (1.88±0.16, 1.84±0.18) was significantly higher than that of the control group (0.53±0.08) ( F=193.554, P<0.05), while the expression level of PTEN protein (0.18±0.02, 0.16±0.02) and mRNA (0.48±0.08, 0.44±0.07) were lower than that of the control group (1.27±0.06, 1.27±0.16) ( Fprotein=515.116, FmRNA=141.470, all P<0.05) ); after inhibiting miRNA-181, the expression level of miRNA-181 (1.35±0.58) in renal tissue increased significantly (LSD- t=10.341, P<0.05), and the expression level of PTEN protein (0.84±0.05) and mRNA (0.90±0.08) increased on average (LSD- tprotein=20.471, Tamhane's T2 mRNA=13.881, all P<0.05). The results of double luciferase reporter gene analysis showed that PTEN was the target gene of miRNA-181. Compared with the control group (0.18±0.02, 0.09±0.01, 0.05±0.02, 1.06±0.07, 0.96±0.06), the expression level of PI3K (1.01±0.06, 1.00±0.06), Akt (0.90±0.05, 0.95±0.04), p-Akt protein (0.99±0.07, 0.97±0.05) and the expression level of PI3K (3.63±0.18, 3.68±0.22), Akt mRNA (2.38±0.05, 2.34±0.12) in the renal tissue of the model group and the negative control group were significantly increased ( FPI3K protein=169.979, FAkt protein=393.411, Fp-Akt protein=164.201, FPI3K mRNA=563.944, FAkt mRNA=141.470, all P<0.05); after inhibiting the expression of miRNA-181, the expression level of PI3K (0.69±0.06), Akt (0.42±0.03), p-Akt protein (0.50±0.05) and the expression level of PI3K (2.40±0.09), Akt mRNA (1.40±0.12) in the renal tissue of the rats were decreased (LSD- tPI3K protein=7.432, LSD- tAkt protein=18.291, LSD- tp-Akt protein=9.595, Tamhane's T2 PI3K mRNA=17.070, Tamhane's T2 Akt mRNA=17.357, all P<0.05). Conclusion:Inhibition of miRNA-181 expression can target PTEN to inhibit PI3K / Akt signaling pathway to protect renal injury in hyperuricemia rats.