Influence of dopamine receptor 5 activation on endotoxin induced acute renal inflammation

Wu Lifei; Li Yulin; Kan Xiaoyu; Sun Jie; Wang Luya; Du Jie

doi:10.3760/cma.j.issn.1673-4777.2016.03.019

Abstract：Objective To investigate the effect of dopamine receptor 5 (D5R) agonist fenoldopam on endotoxin induced acute renal inflammation.Methods The protein levels of D1R-D5R in bone marrow mononuclear macrophages in C57BL/6J mouse were detected by Western blotting,the expression level of D5R in bone marrow mononuclear macrophages then was detected by immunofluorescence.Totally 21 C57BL/6J mice were randomly divided into saline group,saline + LPS group,fenoldopam + LPS group (n =7 in each group).The mice were intraperitoneally injected with 0.9％ saline or fenoldopam for 30 minutes,then were intraperitoneally injected with lipopolysaccharide or 0.9％ saline for 6 hours in corresponding group.The mRNA level of D5R,tumor necrosis factor (TNF)-α and interferon (INF)-γ in renal were measured by using real-time fluorescent quantitative polymease chain reaction (qRT-PCR);the mononuclear macrophages infiltration in renal was observed by immunohistochemical staining;the levels of TNF-α,INF-γ in serum were detected.After obtaining bone marrow cells of mice,the bone marrow mononuclear macrophages were induced by macrophage colony-stimulating factors (M-CSF);then the macrophages were pretreated with PBS or fenoldopam for 5 minutes,followed by lipopolysaccharide for 12 hours.The level of cyclic adenosine monophosphate in macrophages was measured by using enzyme linked immunosorbent assay,the mRNA levels of TNF-α and INF-γin mononuclear macrophages were detected by using qRT-PCR.Results In myelomonocyte,D5R was mainly expressed.The mRNA level of D5R in renal in saline + LPS group was significantly higher than that in saline group [(110.5 ± 3.7) vs (1.2 ± 1.0)] (P ＜ 0.001);the mRNA levels of TNF-α and INF-γin renal in fenoldopam + LPS group were significantly lower than those in saline + LPS group [(0.089 ±0.007) vs (0.231 ±0.019),(0.363 ±0.102) vs (0.920 ±0.007)] (P ＜0.01);the infiltration of monocyte in renal in fenoldopam + LPS group was significantly improved compared with saline + LPS group;the serum levels of TNF-α and INF-γin fenoldopam + LPS group were significantly lower than those in saline+LPSgroup [(22±5) vs (236±20),(18±6) vs (76±10)] (P＜0.01 orP＜0.05).In vitro culture,fenoldopam promoted the production of cyclic adenosine monophosphate [(8.726 ± 0.909) nmol/L vs (0.348 ±0.024) nmol/L] (P ＜0.01),and also supressed the production of TNF-α and INF-γinduced by LPS in myelomonocyte [(0.669 ± 0.010) vs (1.362 ± 0.199),(0.896 ± 0.071) vs (2.854 ± 0.270)] (P ＜ 0.05).Conclusions D5R is expressed in bone marrow mononuclear macrophages;D5R agonist fenoldopam can inhibite endotoxin induced acute renal inflammation.