Diabetic kidney disease (DKD)is a microvascular complication of type 2 diabetes.The study of DKD mechanisms is the most important target for the prevention of DKD.Renal senescence is one of the important pathogeneses ...Diabetic kidney disease (DKD)is a microvascular complication of type 2 diabetes.The study of DKD mechanisms is the most important target for the prevention of DKD.Renal senescence is one of the important pathogeneses for DKD,but the mechanism of renal and cellular senescence is unclear.Decreased expression of circulating miR-126 is associated with the development of DKD and may be a promising blood-based biomarker for DKD.This study is to probe the effect and mechanism of miR-126 on the aging of human glomerular mesangial cells (HGMCs)induced by high glucose.HGMCs were cultured with Roswell Park Memorial Institute (RPMI-1640)in vitro.The effect of high glucose on morphology of HGMCs was observed 72h after intervention.The cell cycle was examined by flow cytometry.The telomere length was measured by Southern blotting.The expression levels of p53,p21 and Rb proteins in p53-p21-Rb signaling pathway and p-statl,p-stat3 in JAK/STAT signaling pathway were detected by Western blotting respectively.The expression of miR-126 was examined by qRT-PCR.MiR-126 mimics was transfected into HGMCs.The effects of miR-126 mimics transfection on cell morphology,cell cycle,telomere length,p53,p21,Rb,p-stat1 and p-stat3 were observed. The results showed that high glucose not only arrested the cell cycle in G1phase but also shortened the telomere length.High glucose led to high expression of p53,p21,Rb,p-statl and p-stat3 and premature senescence of HGMCs by activating the telomere-p53-p21-Rb and JAK/STAT signaling pathways.Moreover,the miR-126 was decreased in HGMCs induced by high glucose.It was suggested that the transfection of miR-126 mimics could inhibit the telomere-p53-p21-Rb and JAK/STAT signaling pathway activity in vitro and delay the senescence of HGMCs.The results may serve as a new strategy for the treatment of DKD.展开更多
ObjectiveTo explore the regulatory effect of Pien Tze Huang (PZH) on targeting partner of NOB1 (PNO1) and it’s down-stream mediators in colorectal cancer (CRC) cells.MethodsQuantitative polymerase chain reaction was ...ObjectiveTo explore the regulatory effect of Pien Tze Huang (PZH) on targeting partner of NOB1 (PNO1) and it’s down-stream mediators in colorectal cancer (CRC) cells.MethodsQuantitative polymerase chain reaction was performed to determine mRNA levels of PNO1, TP53, and CDKN1A. Western blotting was performed to determine protein levels of PNO1, p53, and p21. HCT-8 cells were transduced with a lentivirus over-expressing PNO1. Colony formation assay was used to detect cell survival in PNO1 overexpression of HCT-8 cells after PZH treatment. Cell-cycle distribution, cell viability and cell apoptosis were performed to identify the effect of PNO1 overexpression on cell proliferation and apoptosis of HCT-8 cells after PZH treatment. Xenograft BALB/c nude mice bearing HCT116 cells transduced with sh-PNO1 or sh-Ctrl lentivirus were evaluated. Western blot assay was performed to detect PNO1, p53, p21 and PCNA expression in tumor sections. Terminal deoxynucleotidyl transferase dUTP nick end labling (TUNEL) assay was used to determine the apoptotic cells in tissues.ResultsPZH treatment decreased cell viability, down-regulated PNO1 expression, and up-regulated p53 and p21 expressions in HCT-8 cells (P<0.05). PNO1 overexpression attenuated the effects of PZH treatment, including the expression of p53 and p21, cell growth, cell viability, cell cycle arrest and cell apoptosis in vitro (P<0.05). PNO1 knockdown eliminated the effects of PZH treatment on tumor growth, inhibiting cell proliferation inhibition and apoptosis induction in vivo (P<0.05). Similarly, PNO1 knockdown attenuated the effects of PZH treatment on the down-regulation of PNO1 and up-regulation of p53 and p21 in vivo (P<0.05).ConclusionThe mechanism by which PZH induces its CRC anti-proliferative effect is at least in part by regulating the expression of PNO1 and its downstream targets p53 and p21.展开更多
目的:探讨五味子水提取液(SWE)对叔丁基过氧化氢诱导的神经元衰老的保护作用并探讨其机制。方法:将原代培养的小鼠皮层神经元培养14 d后,分为正常组、模型组(叔丁基过氧化氢100μmol·L-1孵育6 h)和给药组(衰老处理前加不同浓度SWE)...目的:探讨五味子水提取液(SWE)对叔丁基过氧化氢诱导的神经元衰老的保护作用并探讨其机制。方法:将原代培养的小鼠皮层神经元培养14 d后,分为正常组、模型组(叔丁基过氧化氢100μmol·L-1孵育6 h)和给药组(衰老处理前加不同浓度SWE);MTT法检测各组神经细胞的活力;硫代巴比妥酸检测丙二醛含量;黄嘌呤氧化酶法检测超氧化物歧化酶活性;RT-PCR法检测各组皮层神经元中p53和p21 m RNA的表达。结果:给药组小鼠皮层神经元的活力和SOD活性较模型组显著增强(P<0.01);MDA含量显著降低(P<0.01);p53和p21 m RNA的表达明显升高(P<0.01),以750μg/m L效果最好。结论:SWE对叔丁基过氧化氢诱导的小鼠皮层神经元衰老有明显的神经保护作用,这可能与抑制了p53-p21通路有关。展开更多
目的:探讨五味子水提取液(SWE)对叔丁基过氧化氢(t-BHP)诱导的人神经母细胞瘤SH-SY5Y细胞衰老的保护作用并探讨其机制。方法:将SH-SY5Y细胞分为正常组、模型组(叔丁基过氧化氢100μmol·L-1孵育12 h)和给药组(衰老处理后加不同浓度S...目的:探讨五味子水提取液(SWE)对叔丁基过氧化氢(t-BHP)诱导的人神经母细胞瘤SH-SY5Y细胞衰老的保护作用并探讨其机制。方法:将SH-SY5Y细胞分为正常组、模型组(叔丁基过氧化氢100μmol·L-1孵育12 h)和给药组(衰老处理后加不同浓度SWE);MTT法检测各组神经细胞的活力,检测乳酸脱氢酶释放量,RT-PCR法检测各组SH-SY5Y细胞中p53和p21 m RNA的表达。结果:与正常组比较,用不同浓度的t-BHP处理的SH-SY5Y细胞存活能力显著降低,并得到t-BHP诱导SH-SY5Y细胞衰老的最适条件为100μmol/L孵育12 h。与模型组比较,给予不同浓度的SWE能显著提高t-BHP损伤细胞的存活率,改善受损SH-SY5Y细胞的形态,降低LDH的释放量。给药组SH-SY5Y细胞的活力较模型组显著增强(P<0.01);LDH释放量显著降低(P<0.01);p53和p21 m RNA的表达明显降低(P<0.01),以500μg/m L效果最好。结论:SWE对t-BHP诱导的SH-SY5Y细胞衰老有明显的神经保护作用,这可能与抑制了p53-p21通路有关。展开更多
基金This project was supported by grants from the Key Science and Technology Development Program of Nanjing City of the People's Republic of China (No. YKK15057 and No.YKK16097)and the National Natural Science Foundation of China (No.81473684).
文摘Diabetic kidney disease (DKD)is a microvascular complication of type 2 diabetes.The study of DKD mechanisms is the most important target for the prevention of DKD.Renal senescence is one of the important pathogeneses for DKD,but the mechanism of renal and cellular senescence is unclear.Decreased expression of circulating miR-126 is associated with the development of DKD and may be a promising blood-based biomarker for DKD.This study is to probe the effect and mechanism of miR-126 on the aging of human glomerular mesangial cells (HGMCs)induced by high glucose.HGMCs were cultured with Roswell Park Memorial Institute (RPMI-1640)in vitro.The effect of high glucose on morphology of HGMCs was observed 72h after intervention.The cell cycle was examined by flow cytometry.The telomere length was measured by Southern blotting.The expression levels of p53,p21 and Rb proteins in p53-p21-Rb signaling pathway and p-statl,p-stat3 in JAK/STAT signaling pathway were detected by Western blotting respectively.The expression of miR-126 was examined by qRT-PCR.MiR-126 mimics was transfected into HGMCs.The effects of miR-126 mimics transfection on cell morphology,cell cycle,telomere length,p53,p21,Rb,p-stat1 and p-stat3 were observed. The results showed that high glucose not only arrested the cell cycle in G1phase but also shortened the telomere length.High glucose led to high expression of p53,p21,Rb,p-statl and p-stat3 and premature senescence of HGMCs by activating the telomere-p53-p21-Rb and JAK/STAT signaling pathways.Moreover,the miR-126 was decreased in HGMCs induced by high glucose.It was suggested that the transfection of miR-126 mimics could inhibit the telomere-p53-p21-Rb and JAK/STAT signaling pathway activity in vitro and delay the senescence of HGMCs.The results may serve as a new strategy for the treatment of DKD.
基金Supported by National Natural Science Foundation of China(Nos.81803882,82274188 and 82274148)Natural Science Foundation of Fujian Province(No.2020J06026)。
文摘ObjectiveTo explore the regulatory effect of Pien Tze Huang (PZH) on targeting partner of NOB1 (PNO1) and it’s down-stream mediators in colorectal cancer (CRC) cells.MethodsQuantitative polymerase chain reaction was performed to determine mRNA levels of PNO1, TP53, and CDKN1A. Western blotting was performed to determine protein levels of PNO1, p53, and p21. HCT-8 cells were transduced with a lentivirus over-expressing PNO1. Colony formation assay was used to detect cell survival in PNO1 overexpression of HCT-8 cells after PZH treatment. Cell-cycle distribution, cell viability and cell apoptosis were performed to identify the effect of PNO1 overexpression on cell proliferation and apoptosis of HCT-8 cells after PZH treatment. Xenograft BALB/c nude mice bearing HCT116 cells transduced with sh-PNO1 or sh-Ctrl lentivirus were evaluated. Western blot assay was performed to detect PNO1, p53, p21 and PCNA expression in tumor sections. Terminal deoxynucleotidyl transferase dUTP nick end labling (TUNEL) assay was used to determine the apoptotic cells in tissues.ResultsPZH treatment decreased cell viability, down-regulated PNO1 expression, and up-regulated p53 and p21 expressions in HCT-8 cells (P<0.05). PNO1 overexpression attenuated the effects of PZH treatment, including the expression of p53 and p21, cell growth, cell viability, cell cycle arrest and cell apoptosis in vitro (P<0.05). PNO1 knockdown eliminated the effects of PZH treatment on tumor growth, inhibiting cell proliferation inhibition and apoptosis induction in vivo (P<0.05). Similarly, PNO1 knockdown attenuated the effects of PZH treatment on the down-regulation of PNO1 and up-regulation of p53 and p21 in vivo (P<0.05).ConclusionThe mechanism by which PZH induces its CRC anti-proliferative effect is at least in part by regulating the expression of PNO1 and its downstream targets p53 and p21.
文摘目的:探讨五味子水提取液(SWE)对叔丁基过氧化氢诱导的神经元衰老的保护作用并探讨其机制。方法:将原代培养的小鼠皮层神经元培养14 d后,分为正常组、模型组(叔丁基过氧化氢100μmol·L-1孵育6 h)和给药组(衰老处理前加不同浓度SWE);MTT法检测各组神经细胞的活力;硫代巴比妥酸检测丙二醛含量;黄嘌呤氧化酶法检测超氧化物歧化酶活性;RT-PCR法检测各组皮层神经元中p53和p21 m RNA的表达。结果:给药组小鼠皮层神经元的活力和SOD活性较模型组显著增强(P<0.01);MDA含量显著降低(P<0.01);p53和p21 m RNA的表达明显升高(P<0.01),以750μg/m L效果最好。结论:SWE对叔丁基过氧化氢诱导的小鼠皮层神经元衰老有明显的神经保护作用,这可能与抑制了p53-p21通路有关。
文摘目的:探讨五味子水提取液(SWE)对叔丁基过氧化氢(t-BHP)诱导的人神经母细胞瘤SH-SY5Y细胞衰老的保护作用并探讨其机制。方法:将SH-SY5Y细胞分为正常组、模型组(叔丁基过氧化氢100μmol·L-1孵育12 h)和给药组(衰老处理后加不同浓度SWE);MTT法检测各组神经细胞的活力,检测乳酸脱氢酶释放量,RT-PCR法检测各组SH-SY5Y细胞中p53和p21 m RNA的表达。结果:与正常组比较,用不同浓度的t-BHP处理的SH-SY5Y细胞存活能力显著降低,并得到t-BHP诱导SH-SY5Y细胞衰老的最适条件为100μmol/L孵育12 h。与模型组比较,给予不同浓度的SWE能显著提高t-BHP损伤细胞的存活率,改善受损SH-SY5Y细胞的形态,降低LDH的释放量。给药组SH-SY5Y细胞的活力较模型组显著增强(P<0.01);LDH释放量显著降低(P<0.01);p53和p21 m RNA的表达明显降低(P<0.01),以500μg/m L效果最好。结论:SWE对t-BHP诱导的SH-SY5Y细胞衰老有明显的神经保护作用,这可能与抑制了p53-p21通路有关。
