BACKGROUND Injury to the glomerular filtration barrier causes diabetic kidney disease(DKD),and glomerular endothelial-mesenchymal transition damages the filtration barrier of glomerular endothelial cells.Shenfushu gra...BACKGROUND Injury to the glomerular filtration barrier causes diabetic kidney disease(DKD),and glomerular endothelial-mesenchymal transition damages the filtration barrier of glomerular endothelial cells.Shenfushu granules(SFSGs)can treat chronic renal failure;however,their role and mechanism in DKD remain unclear.AIM To investigate the role of SFSGs in delaying DKD progression and their underlying mechanism in a streptozotocin-induced DKD mouse model.METHODS The microalbumin content in the urine and the blood glucose,creatinine,and blood urea nitrogen levels in the serum were measured.The expression and distribution ofα-smooth muscle actin(α-SMA),heparan sulfate(HS)and cluster of differentiation(CD)31 were observed through immunofluorescence or immunohistochemistry.Western blotting was conducted to measure the expression of CD31,α-SMA,PIK3R1,protein kinase B(AKT),phospho-PIK3R1,phospho-AKT,and heparanase-1.Network pharmacology was conducted to screen and identify the core components and targets of SFSGs.Molecular docking and dynamic simulations were performed to evaluate the binding ability of the core components of SFSGs to their core targets.RESULTS Compared with those in the model group,the 24-hour microalbuminuria(188.2±20.1 and 140.4±24.7 vs 323.2±44.4),serum creatinine(79.4±2.6 and 68.7±6.0 vs 110.2±4.8),blood urea nitrogen(14.4±1.1 and 13.1±0.5 vs 19.5±1.1),and renal index(20.3±1.0 and 19.6±0.8 vs 25.3±1.7)were significantly lower in the SFSGs(2.08 and 4.16 g/kg/day extract)-treated DKD mice.SFSGs inhibited the down regulation of CD31 and the upregulation ofα-SMA in the glomerular endothelial cells of DKD mice.Additionally,SFSGs suppressed the decrease in glycocalyx thickness and the expression of its component HS.Network pharmacology revealed that PIK3R1 was the core target of SFSGs.SFSGs markedly downregulate the expression of phospho-PIK3R1,phospho-AKT,and heparanase-1.However,the PIK3R1 agonist abolished the regulatory effect of SFSGs on the expression of CD31,α-SMA,and heparanase-1.CONCLUSION Collectively,these results suggest that SFSGs can significantly delay DKD progression and inhibit injury to the glycocalyx and the endothelial-mesenchymal transition of glomerular endothelial cells.This mechanism is related to PIK3R1/AKT/heparanase-1 signaling pathway regulation.展开更多
基金Supported by the Changsha Science and Technology Major Project,No.kh2205035the Hunan Traditional Chinese Medicine Science and Technology Project,No.2021061the Natural Science Foundation of Hunan Province,No.2025JJ80080。
文摘BACKGROUND Injury to the glomerular filtration barrier causes diabetic kidney disease(DKD),and glomerular endothelial-mesenchymal transition damages the filtration barrier of glomerular endothelial cells.Shenfushu granules(SFSGs)can treat chronic renal failure;however,their role and mechanism in DKD remain unclear.AIM To investigate the role of SFSGs in delaying DKD progression and their underlying mechanism in a streptozotocin-induced DKD mouse model.METHODS The microalbumin content in the urine and the blood glucose,creatinine,and blood urea nitrogen levels in the serum were measured.The expression and distribution ofα-smooth muscle actin(α-SMA),heparan sulfate(HS)and cluster of differentiation(CD)31 were observed through immunofluorescence or immunohistochemistry.Western blotting was conducted to measure the expression of CD31,α-SMA,PIK3R1,protein kinase B(AKT),phospho-PIK3R1,phospho-AKT,and heparanase-1.Network pharmacology was conducted to screen and identify the core components and targets of SFSGs.Molecular docking and dynamic simulations were performed to evaluate the binding ability of the core components of SFSGs to their core targets.RESULTS Compared with those in the model group,the 24-hour microalbuminuria(188.2±20.1 and 140.4±24.7 vs 323.2±44.4),serum creatinine(79.4±2.6 and 68.7±6.0 vs 110.2±4.8),blood urea nitrogen(14.4±1.1 and 13.1±0.5 vs 19.5±1.1),and renal index(20.3±1.0 and 19.6±0.8 vs 25.3±1.7)were significantly lower in the SFSGs(2.08 and 4.16 g/kg/day extract)-treated DKD mice.SFSGs inhibited the down regulation of CD31 and the upregulation ofα-SMA in the glomerular endothelial cells of DKD mice.Additionally,SFSGs suppressed the decrease in glycocalyx thickness and the expression of its component HS.Network pharmacology revealed that PIK3R1 was the core target of SFSGs.SFSGs markedly downregulate the expression of phospho-PIK3R1,phospho-AKT,and heparanase-1.However,the PIK3R1 agonist abolished the regulatory effect of SFSGs on the expression of CD31,α-SMA,and heparanase-1.CONCLUSION Collectively,these results suggest that SFSGs can significantly delay DKD progression and inhibit injury to the glycocalyx and the endothelial-mesenchymal transition of glomerular endothelial cells.This mechanism is related to PIK3R1/AKT/heparanase-1 signaling pathway regulation.
文摘目的:探讨磷脂酰肌醇-3激酶调节亚基1(phosphoinositide-3-kinase regulatory subunit 1,PIK3R1)及其编码蛋白p85α在宫颈鳞癌中的表达及临床价值。方法:选取38例宫颈鳞癌组织和39例良性宫颈组织,采用HE染色观察鳞癌宫颈和良性宫颈的形态,免疫组织化学法检测p85α蛋白的表达;分析宫颈鳞癌组织中p85α的表达与各临床参数的相关性;选取19例宫颈鳞癌组织和19例良性宫颈组织,采用实时荧光定量PCR(qRT-PCR)技术检测PIK3R1和人第10号染色体缺失的磷酸酶及张力蛋白同源的基因(gene of phosphatase and tension homology deleted on chromsome ten,PTEN)的表达,并分析二者的相关性。结果:与良性宫颈组织相比,p85α蛋白在宫颈鳞癌组织中表达明显降低(P<0.01);病理分期越高,p85α的表达越低(P<0.01),有远处转移的肿瘤组织中p85α的表达较无远处转移的肿瘤组织表达低(P<0.05)。与良性宫颈组织相比,PIK3R1和PTEN在宫颈鳞癌组织中的表达均明显下调(P<0.01)。相关分析表明,PIK3R1和PTEN在宫颈鳞癌中表达呈中度相关(r=0.567,P=0.011)。结论:PIK3R1及编码蛋白p85α在宫颈鳞癌中表达均下调,二者在宫颈鳞癌中可能作为抑癌基因发挥关键作用。
