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Objective To investigate the effect of Dapagliflozin on high glucose-induced podocyte proliferation and apoptosis through p38 mitogen-activated protein kinase(p38 MAPK)pathway.Methods Human glomerular podocytes(HGPC)were divided into control(Con)group,low/medium/high D-glucose(Glu 10,Glu 20,Glu 30)group,high glucose(HG)group,low/medium/high concentration Dapagliflozin(HG+Dap 12.5,HG+Dap 25,HG+Dap 50)group,Dapagliflozin(HG+Dap)group,inhibitor(HG+ SB 203580)group,Dapagliflozin + inhibitor(HG+Dap+SB 203580)group and Dapagliflozin + activator(HG+Dap+C16-PAF)group.After 24 hours of intervention,the cell viability,proliferation rate,apoptosis rate and levels of related factors were tested.Results Compared with Con group,IL-1β,TNF-α,apoptosis rate,Caspase-3 mRNA and protein expression,p53,p-p38 MAPK protein expression were increased(P＜0.05),while cell proliferation rate,Cyclin D1 mRNA and protein expression were decreased in HG group(P＜0.05).Compared with HG group,the proliferation rate,Cyclin D1 mRNA and protein expression were increased(P＜0.05),while IL-1β,TNF-α,apoptosis rate,Caspase-3 mRNA and protein expression,p53,p-p38 MAPK protein expression were decreased in the HG+Dap and HG+SB 203580 groups(P＜0.05).Compared with HG+Dap group,cell proliferation rate,Cyclin D1 mRNA and protein expression were increased(P＜0.05),while IL-1β,TNF-α,apoptosis rate,Caspase-3 mRNA and protein expression,p53,p-p38 MAPK protein expression were decreased in HG+Dap+SB 203580 group(P＜0.05).In HG+Dap+C16-PAF group,IL-1β,TNF-α,apoptosis rate,Caspase-3 mRNA and protein expression,p53,p-p38 MAPK protein expression were increased(P＜0.05),while cell proliferation rate,Cyclin D1 mRNA and protein expression were decreased(P＜0.05).Conclusion Dagagliflozin can promote HGPC proliferation and inhibit apoptosis and inflammation in high D-glucose environment,and its mechanism may be related to the inhibition of p38 MAPK pathway signal transduction.
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Objective To evaluate the effect of ambroxol on p38 mitogen-activated protein kinase pathway in mice with sepsis-induced lung injury.Methods Sixty male C57BL/6 mice were equally and randomly divided into 3 groups (n=20 each) using a random number table:sham operation group (group S),sepsis-induced lung injury group (group CLP),and sepsis-induced lung injury + ambroxol group (group AMB).Sespsis was produced by cecal ligation and puncture(CLP).Ambroxol 50 mg/kg preconditioning was injected intraperitoneally for 3 days in group AMB,while the equal volume of normal saline instead was given in S and VILI groups.The arterial blood gas was detected 24 h after CLP.Then the mice were sacrificed and broncho-alveolar lavage fluid (BALF) was collected for determination of the concentrations of total protein,interleukin-1β (IL-1β),tumor necrosis factor-α (TNF-α),IL-6 and intercellular adhesion molecule-1 (ICAM-1).The lung tissues were taken for determination of wet to dry lung weight ratio (W/D ratio),expression of p-p38 MAPK,IL-1β mRNA,TNF-α mRNA,IL-6 mRNA and ICAM-1 mRNA,and for examination of the pathological changes which were scored.Results Compared with group S,partial pressure of oxygen in arterial blood(PaO2) was decreased (P＜0.05),and W/D ratio,lung injury score,concentrations of total protein,IL-1β,TNF-α,IL-6 and ICAM-1 in BALF,and expression of p-p38 MAPK,IL-1β,TNF-α,IL-6 and ICAM-1 mRNA were significantly increased in CLP group (P＜0.05).Compared with group CLP,PaO2 was increased (P＜0.05),W/D ratio,lung injury score,concentrations of total protein,IL-1β,TNF-α,IL-6 and ICAM-1 in BALF,and expression of p-p38 MAPK,IL-1β,TNF-α,IL-6 and ICAM-1 mRNA were decreased in group AMB (P＜0.05).Conclusion Ambroxol can attenuate sepsis-induced lung injury probably through inhibiting p38 mitogen-activated protein kinase pathway in mice.
RESUMO
Objective To investigate the effect of silment information regulator factor related enzymes 1 (SIRT1) on the apoptosis of retinal ganglion cells (RGCs) in rats with diabetic retinopathy and its downstream molecular mechanisms.Methods Together 60 healthy male Sprague-Dawley rats were collected and randomly divided into normal group,diabetic group,SIRTI activator-resveratrol treatment group (treatment group),and diabetic rat model was induced by intraperitoneal injection of streptozotocin at 60 mg · kg-1 in the latter two group rats,while the normal group was injected with sodium citrate buffer at 60 mg · kg-1.Then,after 72 h,rats with blood glucose ＞ 16.7 mmol · L-1 were designated as diabetic rats by blood glucose test.Then each rat in the treatment group was treated with SIRT1 activator-resveratrol at 20 g · kg-1 once a day at the 2nd day after the success of the model,and the normal group and diabetic group were given methylene chloride.Finally,after immunohistochemical staining for retina,TUNEL assay was used to evaluate the apoptosis of RGCs,while the expression of SIRTI,p38 MAPK and Caspase-3 protein was detected by Western blot.Results The apoptotic index of RGCs in the normal group,diabetic group and treatment group was (0.848+0.131)％,(19.038 + 1.327)％,(10.461 + 1.089)％ respectively at 8 weeks,and the difference among the three groups was statistically significant (F =670.497,P =0.000),while the differences between each two groups were also statistically significant (all P =0.000).Furthermore,when compared with the normal group (0.132 ± 0.043),the expression of SIRT1 protein in the diabetic group (0.060 ± 0.028) and the treatment group (0.073 ± 0.026) was significantly decreased,and the overall difference among the three groups was statistically significant (F =1 310.663,P =0.000),while the differences between each two groups were also statistically significant (all P =0.000).The expression levels of p38 MAPK and Caspase-3 were increased in diabetic group (1.121 ± 0.082,0.266 ± 0.005) and treatment group (0.574 ± 0.012,0.190 ±0.060) respectively,and the overall difference and pairwise comparison in the three groups approached statistically significance (all P =0.000,0.000).Conelusion Up-regulation of SIRT1,can inhibit the apoptosis of RGCs,and protect RGCs against apoptosis in rat model of diabetic retinopathy,which may be correlated with the downregulation of p38 MAPK signal pathway.
RESUMO
Objective To investigate the effect of silment information regulator factor related enzymes 1 (SIRT1) on the apoptosis of retinal ganglion cells (RGCs) in rats with diabetic retinopathy and its downstream molecular mechanisms.Methods Together 60 healthy male Sprague-Dawley rats were collected and randomly divided into normal group,diabetic group,SIRTI activator-resveratrol treatment group (treatment group),and diabetic rat model was induced by intraperitoneal injection of streptozotocin at 60 mg · kg-1 in the latter two group rats,while the normal group was injected with sodium citrate buffer at 60 mg · kg-1.Then,after 72 h,rats with blood glucose ＞ 16.7 mmol · L-1 were designated as diabetic rats by blood glucose test.Then each rat in the treatment group was treated with SIRT1 activator-resveratrol at 20 g · kg-1 once a day at the 2nd day after the success of the model,and the normal group and diabetic group were given methylene chloride.Finally,after immunohistochemical staining for retina,TUNEL assay was used to evaluate the apoptosis of RGCs,while the expression of SIRTI,p38 MAPK and Caspase-3 protein was detected by Western blot.Results The apoptotic index of RGCs in the normal group,diabetic group and treatment group was (0.848+0.131)％,(19.038 + 1.327)％,(10.461 + 1.089)％ respectively at 8 weeks,and the difference among the three groups was statistically significant (F =670.497,P =0.000),while the differences between each two groups were also statistically significant (all P =0.000).Furthermore,when compared with the normal group (0.132 ± 0.043),the expression of SIRT1 protein in the diabetic group (0.060 ± 0.028) and the treatment group (0.073 ± 0.026) was significantly decreased,and the overall difference among the three groups was statistically significant (F =1 310.663,P =0.000),while the differences between each two groups were also statistically significant (all P =0.000).The expression levels of p38 MAPK and Caspase-3 were increased in diabetic group (1.121 ± 0.082,0.266 ± 0.005) and treatment group (0.574 ± 0.012,0.190 ±0.060) respectively,and the overall difference and pairwise comparison in the three groups approached statistically significance (all P =0.000,0.000).Conelusion Up-regulation of SIRT1,can inhibit the apoptosis of RGCs,and protect RGCs against apoptosis in rat model of diabetic retinopathy,which may be correlated with the downregulation of p38 MAPK signal pathway.