目的探讨PIWI蛋白互作RNA-10555(piR-10555)对心肌细胞坏死的调控作用及其机制。方法选用500μmol/L的过氧化氢(H 2 O 2)处理H9c2细胞0、6、12、24 h,应用实时荧光定量PCR(RT-qPCR)技术检测H9c2细胞中piR-10555的表达水平。H9c2细胞分...目的探讨PIWI蛋白互作RNA-10555(piR-10555)对心肌细胞坏死的调控作用及其机制。方法选用500μmol/L的过氧化氢(H 2 O 2)处理H9c2细胞0、6、12、24 h,应用实时荧光定量PCR(RT-qPCR)技术检测H9c2细胞中piR-10555的表达水平。H9c2细胞分别转染piR-10555类似物agomir-piR-10555及转染piR-10555抑制剂antagomir-piR-10555后使用H 2 O 2处理,通过RT-qPCR方法检测H9c2细胞中piR-10555的表达,并使用碘化丙啶(PI)染色方法和乳酸脱氢酶(LDH)测定试剂盒检测细胞的坏死情况。结果在H 2 O 2处理24 h后,H9c2细胞中piR-10555的表达水平显著上调(F=14.51,P<0.05)。转染agomir-piR-10555以后,H9c2细胞中piR-10555表达水平明显升高(F=154.00,P<0.05),H9c2细胞的PI阳性率显著增加(F=32.61,P<0.05),H9c2细胞培养基上清液中的LDH活性明显提高(F=126.50,P<0.05)。转染antagomir-piR-10555后,H9c2细胞中piR-10555表达水平显著降低(F=33.31,P<0.05),PI阳性率显著下降(F=138.70,P<0.05),细胞培养基上清液中的LDH活性明显降低(F=58.60,P<0.05)。结论piR-10555可能参与调控由H 2 O 2诱导的H9c2细胞的坏死。展开更多
Microalgae can be cultivated for producing high-valued products through the production of enzymes to offset the cost of CO_(2) sequestration,providing financial incentives.The viability of algae in the photobioreactor...Microalgae can be cultivated for producing high-valued products through the production of enzymes to offset the cost of CO_(2) sequestration,providing financial incentives.The viability of algae in the photobioreactor needs to be monitored to ensure biologically active live cells.In this study,we explored a simple fluorometry method for differentiation of live and dead algal cells in photobioreactors by fluorescein diacetate(FDA)and propidium iodide(PI)fluorescence staining.FDA stains fluorescent green to the living cells while PI stains the dead cells,allowing the discrimination of live and dead cells.The method was evaluated using two green algae and two strains of cyanobacteria grown in shake flasks and a continuously stirred photobioreactor.The method was found applicable for Chlorella pyrenoidosa and Synechococcus 7002 but was not applicable for the cultures of Scenedesmus dimorphus and Synechococcus elongatus 7942.We conclude that FDA is a good stain for monitoring live algal cells in photobioreactors but its applicability to individual species of algae must be evaluated.展开更多
文摘目的探讨PIWI蛋白互作RNA-10555(piR-10555)对心肌细胞坏死的调控作用及其机制。方法选用500μmol/L的过氧化氢(H 2 O 2)处理H9c2细胞0、6、12、24 h,应用实时荧光定量PCR(RT-qPCR)技术检测H9c2细胞中piR-10555的表达水平。H9c2细胞分别转染piR-10555类似物agomir-piR-10555及转染piR-10555抑制剂antagomir-piR-10555后使用H 2 O 2处理,通过RT-qPCR方法检测H9c2细胞中piR-10555的表达,并使用碘化丙啶(PI)染色方法和乳酸脱氢酶(LDH)测定试剂盒检测细胞的坏死情况。结果在H 2 O 2处理24 h后,H9c2细胞中piR-10555的表达水平显著上调(F=14.51,P<0.05)。转染agomir-piR-10555以后,H9c2细胞中piR-10555表达水平明显升高(F=154.00,P<0.05),H9c2细胞的PI阳性率显著增加(F=32.61,P<0.05),H9c2细胞培养基上清液中的LDH活性明显提高(F=126.50,P<0.05)。转染antagomir-piR-10555后,H9c2细胞中piR-10555表达水平显著降低(F=33.31,P<0.05),PI阳性率显著下降(F=138.70,P<0.05),细胞培养基上清液中的LDH活性明显降低(F=58.60,P<0.05)。结论piR-10555可能参与调控由H 2 O 2诱导的H9c2细胞的坏死。
基金the Natural Sciences and Engineering Research Council(NSERC)of Canada in the form of a strategic grant(STPGP 380768-09)。
文摘Microalgae can be cultivated for producing high-valued products through the production of enzymes to offset the cost of CO_(2) sequestration,providing financial incentives.The viability of algae in the photobioreactor needs to be monitored to ensure biologically active live cells.In this study,we explored a simple fluorometry method for differentiation of live and dead algal cells in photobioreactors by fluorescein diacetate(FDA)and propidium iodide(PI)fluorescence staining.FDA stains fluorescent green to the living cells while PI stains the dead cells,allowing the discrimination of live and dead cells.The method was evaluated using two green algae and two strains of cyanobacteria grown in shake flasks and a continuously stirred photobioreactor.The method was found applicable for Chlorella pyrenoidosa and Synechococcus 7002 but was not applicable for the cultures of Scenedesmus dimorphus and Synechococcus elongatus 7942.We conclude that FDA is a good stain for monitoring live algal cells in photobioreactors but its applicability to individual species of algae must be evaluated.
