BACKGROUND Diabetic nephropathy(DN)is the most frequent chronic microvascular consequence of diabetes,and podocyte injury and malfunction are closely related to the development of DN.Studies have shown that corilagin(...BACKGROUND Diabetic nephropathy(DN)is the most frequent chronic microvascular consequence of diabetes,and podocyte injury and malfunction are closely related to the development of DN.Studies have shown that corilagin(Cor)has hepatoprotective,anti-inflammatory,antibacterial,antioxidant,anti-hypertensive,antidiabetic,and anti-tumor activities.AIM To explore the protective effect of Cor against podocyte injury in DN mice and the underlying mechanisms.METHODS Streptozotocin and a high-fat diet were combined to generate DN mice models,which were then divided into either a Cor group or a DN group(n=8 in each group).Mice in the Cor group were intraperitoneally injected with Cor(30 mg/kg/d)for 12 wk,and mice in the DN group were treated with saline.Biochemical analysis was used to measure the blood lipid profiles.Hematoxylin and eosin staining was used to detect pathological changes in kidney tissue.Immunohistochemistry and Western blotting were used to assess the protein expression of nephrin and podocin.Mouse podocyte cells(MPC5)were cultured and treated with glucose(5 mmol/L),Cor(50μM),high glucose(HG)(30 mmol/L),and HG(30 mmol/L)plus Cor(50μM).Real-time quantitative PCR and Western blotting RESULTS Compared with the control group,the DN mice models had increased fasting blood glucose,glycosylated hemoglobin,triglycerides,and total cholesterol,decreased nephrin and podocin expression,increased apoptosis rate,elevated inflammatory cytokines,and enhanced oxidative stress.All of the conditions mentioned above were alleviated after intervention with Cor.In addition,Cor therapy improved SIRT1 and AMPK expression(P<0.001),inhibited reactive oxygen species and oxidative stress,and elevated autophagy in HG-induced podocytes(P<0.01).CONCLUSION Cor alleviates podocyte injury by regulating autophagy via the SIRT1-AMPK pathway,thereby exerting its protective impact on renal function in DN mice.展开更多
Objective:To explore the regulatory mechanism of transient receptor potential melastatin-7(TRPM7)in high glucose-induced renal tubular epithelial cell injury.Methods:The expression of TRPM7 in the serum of diabetic ne...Objective:To explore the regulatory mechanism of transient receptor potential melastatin-7(TRPM7)in high glucose-induced renal tubular epithelial cell injury.Methods:The expression of TRPM7 in the serum of diabetic nephropathy patients and high glucose-induced HK-2 cells was detected by RT-qPCR.Then,the TRPM7 interference vector was constructed,and the downstream high mobility group box 1(HMGB1)/Toll-like receptor 4(TLR4)signaling pathway proteins were detected.Next,in addition to interference with TRPM7 expression,overexpression of HMGB1 in high glucose-induced HK-2 cells was performed.Cell activity,apoptosis,oxidative stress levels,and inflammation levels were determined by CCK8,TUNEL,Western blotting,immunofluorescence and related kits.Results:TRPM7 expression was upregulated in the serum of diabetic nephropathy patients and high glucose-induced HK-2 cells.Interference with TRPM7 reduced cell damage,epithelial-mesenchymal transition,oxidative stress,and inflammatory response in high glucose-induced HK-2 cells via inhibiting the HMGB1/TLR4 signaling pathway.However,the effects induced by TRPM7 silencing were abrogated by HMGB1 overexpression.Conclusions:Decreased TRPM7 alleviates high glucose-induced renal tubular epithelial cell injury by inhibiting the HMGB1/TLR4 signaling pathway.Further animal experiments and clinical trials are warranted to verify its effect.展开更多
目的:探讨葛根素能否通过调控miR-30e-5p的表达减轻高糖所致的足细胞损伤。方法:高糖诱导小鼠肾足细胞MPC5建立足细胞损伤模型,使用不同剂量的葛根素处理细胞;将miR-NC、miR-30e-5p mimics、anti-miR-NC、anti-miR-30e-5p转染至MPC5细...目的:探讨葛根素能否通过调控miR-30e-5p的表达减轻高糖所致的足细胞损伤。方法:高糖诱导小鼠肾足细胞MPC5建立足细胞损伤模型,使用不同剂量的葛根素处理细胞;将miR-NC、miR-30e-5p mimics、anti-miR-NC、anti-miR-30e-5p转染至MPC5细胞后采用高糖诱导建立足细胞损伤模型,并给予不同剂量的葛根素处理细胞。采用流式细胞术检测细胞凋亡率;采用RT-qPCR法检测miR-30e-5p的表达量;Western Blot法检测半胱氨酰天冬氨酸特异性蛋白酶3(cysteinyl aspartate-specific protease-3,caspase-3)、B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、B淋巴细胞瘤-2相关蛋白(Bcl-2-associated X protein,Bax)、p62、微管相关蛋白轻链3Ⅱ(microtubule-associated protein l light chain 3,LC3-Ⅱ)、LC3-Ⅰ蛋白表达水平并计算LC3-Ⅱ/LC3-Ⅰ的比值。结果:高糖处理小鼠肾足细胞MPC5后,可明显提高MPC5细胞的凋亡率及caspase-3、Bax、p62蛋白表达水平(P<0.05),显著降低Bcl-2蛋白及miR-30e-5p的表达水平和LC3-Ⅱ/LC3-Ⅰ的比值(P<0.05),而葛根素可显著降低细胞凋亡率及caspase-3、Bax、p62的蛋白表达水平(P<0.05),显著升高Bcl-2蛋白、miR-30e-5p的表达水平及LC3-Ⅱ/LC3-Ⅰ的比值(P<0.05);与miR-NC组比较,miR-30e-5p组细胞凋亡率显著降低(P<0.05),caspase-3、Bax、p62蛋白表达水平显著降低(P<0.05),Bcl-2蛋白水平、LC3-Ⅱ/LC3-Ⅰ的比值显著升高(P<0.05);与高剂量葛根素+anti-miR-NC组比较,高剂量葛根素+anti-miR-30e-5p组细胞凋亡率显著升高(P<0.05),caspase-3、Bax、p62蛋白表达水平显著升高(P<0.05),Bcl-2蛋白表达水平显著降低(P<0.05),LC3-Ⅱ/LC3-Ⅰ的比值显著降低(P<0.05)。结论:葛根素可通过上调miR-30e-5p的表达抑制高糖诱导的足细胞凋亡,其作用机制可能与诱导细胞自噬有关。展开更多
基金Supported by Shanghai Pudong New Area Leading Talents Training Program Project,No.PWR12020-02Shanghai Pudong New Area Excellent Young Medical Talents Training Program Project,No.PWRq2023-40Shanghai Pudong New Area Health and Family Planning Scientific Research Project,No.PW2022A-91.
文摘BACKGROUND Diabetic nephropathy(DN)is the most frequent chronic microvascular consequence of diabetes,and podocyte injury and malfunction are closely related to the development of DN.Studies have shown that corilagin(Cor)has hepatoprotective,anti-inflammatory,antibacterial,antioxidant,anti-hypertensive,antidiabetic,and anti-tumor activities.AIM To explore the protective effect of Cor against podocyte injury in DN mice and the underlying mechanisms.METHODS Streptozotocin and a high-fat diet were combined to generate DN mice models,which were then divided into either a Cor group or a DN group(n=8 in each group).Mice in the Cor group were intraperitoneally injected with Cor(30 mg/kg/d)for 12 wk,and mice in the DN group were treated with saline.Biochemical analysis was used to measure the blood lipid profiles.Hematoxylin and eosin staining was used to detect pathological changes in kidney tissue.Immunohistochemistry and Western blotting were used to assess the protein expression of nephrin and podocin.Mouse podocyte cells(MPC5)were cultured and treated with glucose(5 mmol/L),Cor(50μM),high glucose(HG)(30 mmol/L),and HG(30 mmol/L)plus Cor(50μM).Real-time quantitative PCR and Western blotting RESULTS Compared with the control group,the DN mice models had increased fasting blood glucose,glycosylated hemoglobin,triglycerides,and total cholesterol,decreased nephrin and podocin expression,increased apoptosis rate,elevated inflammatory cytokines,and enhanced oxidative stress.All of the conditions mentioned above were alleviated after intervention with Cor.In addition,Cor therapy improved SIRT1 and AMPK expression(P<0.001),inhibited reactive oxygen species and oxidative stress,and elevated autophagy in HG-induced podocytes(P<0.01).CONCLUSION Cor alleviates podocyte injury by regulating autophagy via the SIRT1-AMPK pathway,thereby exerting its protective impact on renal function in DN mice.
文摘Objective:To explore the regulatory mechanism of transient receptor potential melastatin-7(TRPM7)in high glucose-induced renal tubular epithelial cell injury.Methods:The expression of TRPM7 in the serum of diabetic nephropathy patients and high glucose-induced HK-2 cells was detected by RT-qPCR.Then,the TRPM7 interference vector was constructed,and the downstream high mobility group box 1(HMGB1)/Toll-like receptor 4(TLR4)signaling pathway proteins were detected.Next,in addition to interference with TRPM7 expression,overexpression of HMGB1 in high glucose-induced HK-2 cells was performed.Cell activity,apoptosis,oxidative stress levels,and inflammation levels were determined by CCK8,TUNEL,Western blotting,immunofluorescence and related kits.Results:TRPM7 expression was upregulated in the serum of diabetic nephropathy patients and high glucose-induced HK-2 cells.Interference with TRPM7 reduced cell damage,epithelial-mesenchymal transition,oxidative stress,and inflammatory response in high glucose-induced HK-2 cells via inhibiting the HMGB1/TLR4 signaling pathway.However,the effects induced by TRPM7 silencing were abrogated by HMGB1 overexpression.Conclusions:Decreased TRPM7 alleviates high glucose-induced renal tubular epithelial cell injury by inhibiting the HMGB1/TLR4 signaling pathway.Further animal experiments and clinical trials are warranted to verify its effect.
文摘目的:探讨葛根素能否通过调控miR-30e-5p的表达减轻高糖所致的足细胞损伤。方法:高糖诱导小鼠肾足细胞MPC5建立足细胞损伤模型,使用不同剂量的葛根素处理细胞;将miR-NC、miR-30e-5p mimics、anti-miR-NC、anti-miR-30e-5p转染至MPC5细胞后采用高糖诱导建立足细胞损伤模型,并给予不同剂量的葛根素处理细胞。采用流式细胞术检测细胞凋亡率;采用RT-qPCR法检测miR-30e-5p的表达量;Western Blot法检测半胱氨酰天冬氨酸特异性蛋白酶3(cysteinyl aspartate-specific protease-3,caspase-3)、B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、B淋巴细胞瘤-2相关蛋白(Bcl-2-associated X protein,Bax)、p62、微管相关蛋白轻链3Ⅱ(microtubule-associated protein l light chain 3,LC3-Ⅱ)、LC3-Ⅰ蛋白表达水平并计算LC3-Ⅱ/LC3-Ⅰ的比值。结果:高糖处理小鼠肾足细胞MPC5后,可明显提高MPC5细胞的凋亡率及caspase-3、Bax、p62蛋白表达水平(P<0.05),显著降低Bcl-2蛋白及miR-30e-5p的表达水平和LC3-Ⅱ/LC3-Ⅰ的比值(P<0.05),而葛根素可显著降低细胞凋亡率及caspase-3、Bax、p62的蛋白表达水平(P<0.05),显著升高Bcl-2蛋白、miR-30e-5p的表达水平及LC3-Ⅱ/LC3-Ⅰ的比值(P<0.05);与miR-NC组比较,miR-30e-5p组细胞凋亡率显著降低(P<0.05),caspase-3、Bax、p62蛋白表达水平显著降低(P<0.05),Bcl-2蛋白水平、LC3-Ⅱ/LC3-Ⅰ的比值显著升高(P<0.05);与高剂量葛根素+anti-miR-NC组比较,高剂量葛根素+anti-miR-30e-5p组细胞凋亡率显著升高(P<0.05),caspase-3、Bax、p62蛋白表达水平显著升高(P<0.05),Bcl-2蛋白表达水平显著降低(P<0.05),LC3-Ⅱ/LC3-Ⅰ的比值显著降低(P<0.05)。结论:葛根素可通过上调miR-30e-5p的表达抑制高糖诱导的足细胞凋亡,其作用机制可能与诱导细胞自噬有关。
