ABSTRACT
AIM: To investigate the injury of emodin (EMO) in reduce of glomerular mesangial cells (MCs) in lupus nephritis by targeting forkhead protein K2 (FOXK2) through miR-96-5p. METHODS: The contents of 24 h urine protein, serum urea nitrogen (BUN) and serum creatinine (Scr) in MRL / faslpr mice (lupus nephritis group) and MRL / MPJ mice (control group) were detected. MCs were separated, purified and divided into: MCs group (MCs without any treatment), L-EMO group (MCs treated with 10 μmol/L Emodin), M-EMO group (MCs treated with 25 μmol / L Emodin), H-EMO group (MCs treated with 50 μmol / L Emodin), H-EMO + miR-96-5p-NC group (MCs treated with 50 μmol / L Emodin and transfected with miR-96-5p-NC), and H-EMO + miR-96-5p-minic group (MCs treated with 50 μmol/ L Emodin and transfected with miR-96-5p-minic). Double luciferase report experiment was used to verify the targeting relationship between miR-96-5p and FOXK2. The real-time quantitative fluorescent polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-96-5p. Western blot was used to detect the expression of FOXK2 and apoptosis related proteins. The enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of inflammatory factors in MCs. cell count kit 8 (CCK-8) was used to determine the activity of MCs. Annexin-V FITC/PI double staining was used to detect apoptosis of MCs. RESULTS: Compared with the control group, 24 h urinary protein content, serum BUN and Scr levels in the lupus nephritis group were significantly increased (P< 0.05). Compared with the MCs group, the miR-96-5p expression, interleukin1β (IL-1β), interleukin6 (IL-6), tumor necrosis factor-α (TNF-α), A450 value and B-lymphoblastoma-2 (Bcl-2) protein in the L-EMO group, M-EMO group and H-EMO group were significantly decreased (P<0.05), the FOXK2 level, cell apoptosis rate, Bcl-2 related X gene (Bax), aspartate specific cysteine proteinase-3 (cleaved Caspase-3) protein levels were significantly increased, respectively (P<0.05), the effect of Emodin was dose-dependent. Compared with the H-EMO group and H-EMO+miR-96-5p-NC group, H-EMO+miR-96-5p-minic group obviously increased the miR-96-5p expression, inflammatory factor levels, A450 value and Bcl-2 protein level (P<0.05), and obviously decreased FOXK2 level and cell apoptosis rate (P< 0.05). CONCLUSION: EMO can reduce the injury of lupus nephritis MCs by down-regulating miR-96-5p and then up-regulating FOXK2.
ABSTRACT
Objective To investigate the effects of rosmarinic acid on the kidney function,urinary protein,serum MDA and SOD in rats with the 5/6 nephrectomy.Methods We established the rat model with chronic renal dysfunction and randomized them into normal control group,model control group and rosmarinic acid treatment group.There were 6 rats in each group.The blood creatinine level was recorded 2 weeks after operation.We detected the effects of rosmarinic acid on urinary protein,urine creatinine,blood creatinine,serum MDA and SOD in rats with the 5/6 nephretomy which was established by Ponseau S method.The treatment group received rosmarinic acid as an intervention.Results ( 1 ) Compared with control group,the creatinine and The creatinine clearance rate ( Ccr ) of rosmarinic acid treatment group increased ( F =38.912,P ＜ 0.01 ;F =19.968,P ＜ 0.05 ) after treatment,; (2) Rosmarinic acid improved the 24 h urine protein in rats with renal dysfunction significantly(F =16.288 P ＜0.01); (3) Rosmarinic acid had no effect on serum MDA and SOD in rats.Conclusion Rosmarinic acid can decrease urine protein and blood creatinine,increase the Ccr and alleviate kidney failure,but has no effects on MDA and SOD in rat model.