RESUMO
Objective:To evaluate the role of microRNA-10a (miRNA-10a) in renal ischemia-reperfusion (I/R) injury in mice and the relationship with transforming growth factor beta1 (TGF-β 1)/Smad2 signaling pathway. Methods:Twenty-four SPF healthy male adult C57BL/6 mice, aged 8-10 weeks, weighing 20-25 g, were divided into 4 groups ( n=6 each) by the random number table method: sham operation group (S group), renal I/R group (IR group), renal I/R plus miRNA-10a antagonist group (I group), and renal I/R+ miRNA-10a agonist group (M group). The mouse model of renal I/R was developed by clamping the left renal pedicle for 45 min followed by reperfusion in anesthetized animals.miRNA-10a antagonist and agonist 20 nmol were injected via the tail vein once every 24 h for 3 consecutive days starting from 72 h before surgery in group M and group I, respectively, while the equal volume of normal saline was given instead in S and IR groups.Blood samples were collected from the orbital vein at 24 h of reperfusion to determine the concentrations of the serum creatinine (Scr) and blood urea nitrogen (BUN). Then the mice were sacrificed, and the kidney tissues were taken for determination of the malondialdehyde (MDA) content and superoxide dismutase (SOD) activity, contents of interleukin-1 beta (IL-β) and tumor necrosis factor-alpha (TNF-α) (by enzyme-linked immunosorbent assay), and expression of TGF-β 1 and phosphorylated Smad2 (p-Smad2) (by Western blot) and for microscopic examination of the pathological changes.The damage to the renal tubules was scored. Results:Compared with group S, the concentrations of Scr and BUN, contents of MDA, IL-1β and TNF-α in renal tissues and renal tubular damage score were significantly increased in IR, I and M groups, and the activity of SOD was significantly decreased, and the expression of TGF-β 1 and p-Smad2 was up-regulated in IR and M groups ( P<0.05). Compared with group IR, the concentrations of Scr and BUN, contents of MDA, IL-1β and TNF-α in renal tissues and renal tubular damage score were significantly decreased, the activity of SOD was increased, and the expression of TGF-β 1 and p-Smad2 was down-regulated in group I, and the concentrations of Scr and BUN, contents of MDA, IL-1β and TNF-α in renal tissues and renal tubular damage score were significantly increased, the activity of SOD was decreased, and the expression of TGF-β 1 and p-Smad2 was up-regulated in group M ( P<0.05). Compared with group I, the concentrations of Scr and BUN, contents of MDA, IL-1β and TNF-α in renal tissues and renal tubular damage score were significantly increased, the activity of SOD was decreased, and the expression of TGF-β 1 and p-Smad2 was up-regulated in group M ( P<0.05). Conclusions:miRNA-10a is involved in the process of renal I/R injury and is related to activation of TGF-β 1/Smad2 signaling pathway in mice.
RESUMO
Objective:To evaluate the role of ecto-5′-nucleotidase (CD73) in endogenous protective mechanism of hepatic ischemia-reperfusion (I/R) injury in mice and the relationship with transforming growth factor beta1 (TGF-β 1)/Smad3 signaling pathway. Methods:Twenty-four SPF healthy male C57BL/6 mice, aged 8-10 weeks, weighing 20-23 g, were divided into 3 groups ( n=8 each) by the random number table method: sham operation group (S group), hepatic I/R group (IR group) and hepatic I/R plus CD73 specific inhibitor group (APCP group). The hepatic hilum was only exposed but not occluded in group S. The hepatic portal was occluded for 30 min followed by reperfusion to develop the model of hepatic I/R in anesthetized animals in group IR.CD73-specific inhibitor APCP 40 mg/kg was intraperitoneally injected, and 10 min later hepatic I/R was performed.Orbital venous blood samples were collected at 6 h of reperfusion for determination of serum alanine transaminase (ALT) and aspartate transaminase (AST) concentrations.Then the mice were sacrificed, and liver tissues were obtained for determination of the expression of CD73, TGF-β 1 and phosphorylated Smad3 (p-Smad3) (by Western blot), contents of interleukin-1β (IL-1β) and tumor necrosis factor-alpha (TNF-α) (by enzyme-linked immunosorbent assay), and content of malondialdehyde (MDA) and activity of superoxide dismutase (SOD) (with a visible spectrophotometer) and for microscopic examination of the pathological changes of liver tissues (with a light microscope). Results:Compared with group S, the concentrations of AST and ALT in serum and contents of IL-1β, TNF-α and MDA in liver tissues were significantly increased, the activity of SOD was decreased, and the expression of CD73, TGF-β 1 and p-Smad3 was up-regulated in IR and APCP groups ( P<0.05). Compared with group IR, the concentrations of AST and ALT in serum and contents of IL-1β, TNF-α and MDA in liver tissues were significantly increased, the activity of SOD was decreased, and the expression of CD73, TGF-β 1 and p-Smad3 in liver tissues was down-regulated in group APCP ( P<0.05). The pathological changes of liver tissues were accentuated in group APCP as compared with group IR. Conclusions:CD73 is involved in the process of endogenous protective mechanism of hepatic I/R injury in mice, which may be related to the regulation of TGF-β 1/Smad3 signaling pathway.
RESUMO
Objective:To evaluate the effect of sufentanil on activation of Schwann cells after peripheral nerve injury in mice.Methods:Eighty healthy pathogen-free male Balb/c mice, aged 6-8 weeks, weighing 18-22 g, were divided into 4 groups ( n=20 each) using a random number table method: peripheral nerve injury group (group PNI), high dose sufentanil group (group H), medium dose sufentanil group (group M) and low dose sufentanil group (group L). The model of unilateral sciatic nerve transaction was established in ketamine-anesthetized mice.Immediately after establishment of the model, sufentanil 10, 5 and 2.5 μg/kg was injected intraperitoneally once a day for 3 consecutive days in H, M and L groups, respectively, while the equal volume of normal saline was given instead in group PNI.Sciatic function index (SFI) was calculated at 4, 8 and 12 weeks after establishment of the model.At 2, 4, 8 and 12 weeks, 5 mice in each group were sacrificed, and segments of the injuried ipsilateral sciatic nerve were removed for examination of the ultrastructure of the sciatic nerve (with a transmission electron microscope) and for detection of the expression of glial fibrillary acidic protein (GFAP) of sciatic nerve (by immunohistochemistry). Results:Compared with group PNI, SFI was significantly increased, and the expression of GFAP was up-regluated at each time point after establishment of the model in H and M groups ( P<0.05) and no significant change was found in SFI and GFAP expression after establishment of the model in group L ( P>0.05). Compared with group L, SFI was significantly increased, and GFAP expression was up-regluated in H and M groups ( P<0.05). There was no significant difference in SFI and GFAP expression between group H and group M ( P>0.05). The thickness of myelin lamellae was dense, and the proliferation of Schwann cells was not marked in H and M groups.The thickness of myelin lamellae was thin, and the proliferation of Schwann cells was marked in L and MO groups. Conclusion:The mechanism by which sufentanil improves repair after peripheral nerve injury may be related to promoting activation of Schwann cells in mice.