The treatment of wastewaters is crucial to maintain the ecological status of receiving waters,and thereby guarantee the protection of aquatic life and human health.Wastewater quality evaluation is conventionally based...The treatment of wastewaters is crucial to maintain the ecological status of receiving waters,and thereby guarantee the protection of aquatic life and human health.Wastewater quality evaluation is conventionally based on physicochemical parameters,but increasing attention has been paid to integrate physicochemical and biological data.Nevertheless,the regulatory use of fish in biological testing methods has been subject to various ethical and cost concerns,and in vitro cell-based assays have thus become an important topic of interest.Hence,the present study intends:(a) to evaluate the efficiency of two different sample pre-concentration techniques (lyophilisation and solid phase extraction) to assess the toxicity of municipal effluents on rat cardiomyoblast H9c2(2-1) cells,and (b) maximizing the use of the effluent sample collected,to estimate the environmental condition of the receiving environment.The gathered results demonstrate that the H9c2(2-1) sulforhodamine B-based assay is an appropriate in vitro method to assess biological effluent toxicity,and the best results were attained by lyophilising the sample as pre-treatment.Due to its response,the H9c2(2-1) cell line might be a possible alternative in vitro model for fish lethal testing to assess the toxicity of municipal effluents.The physicochemical status of the sample suggests a high potential for eutrophication,and iron exceeded the permissible level for wastewater discharge,possibly due to the addition of ferric chloride for wastewater treatment.In general,the levels of carbamazepine and sulfamethoxazole are higher than those reported for other countries,and both surpassed the aquatic protective values for long-term exposure.展开更多
Background Advances in the understanding of cardiovascular pathogenesis have highlighted that inflammation plays a central role in athemsclemtic coronary heart disease.Therefore,exploring pharmacologically based anti-...Background Advances in the understanding of cardiovascular pathogenesis have highlighted that inflammation plays a central role in athemsclemtic coronary heart disease.Therefore,exploring pharmacologically based anti-inflammatory treatments to be used in cardiovascular therapeutics is worthwhile to promote the discovery of novel ways of treating cardiovascular disorders.Methods The myocardial cell line H9c2（2-1） was exposed to lipopolysacchadde （LPS） in culture and resulted in a cellular pro-inflammation status,miR-21 microRNA levels were detected using quantitative real-time polymerase chain reaction （Q-RT-PCR）.The influence of Iovastatin on miR-21 under normal and pro-inflammatory conditions was tested after being added to the cell culture mixture for 24 hours.Conditional gene function of two predicted cardiovascular system relevant downstream targets of miR-21,protein phosphatase 1 regulatory subunit 3A （PPP1R3A） and signal transducer and activator of transcription 3 （STAT3）,were analyzed with immunoblotting.Results Forty-eight hours of LPS treatment significantly increased the miR-21 to 170.71%±34.32% of control levels （P=0.002）.Co-treatment with Iovastatin for 24 hours before harvesting attenuated the up-regulation of miR-21 （P=0.013）.Twenty-four hours of Iovastatin exposure up-regulated PPP1R3A to 143.85%±21.89% of control levels in cardiomyocytes （P=0.023）.Lovastatin up-regulated the phosphorylation level of STAT3 compared to the background LPS pretreatment （P=0.0077）,this effect was significantly （P=0.018） blunted when miR-21 was functionally inhibited.Conclusions miR-21 plays a major role in the regulation of the cellular anti-inflammation effects of Iovastatin.展开更多
基金The Portuguese Foundation for Science and Technology (FCT) supported the present study through a fellowship attributed to Elsa Teresa Rodrigues(SFRH/BPD/116152/2016),which was funded by the Human Potential Operating Program of the European Social Fund,and by the Portuguese budget through the Ministry of Education and Sciencedeveloped within the Fish Free Project(PTDC/AAG-TEC/4966/2014)+3 种基金supported by the FCT through national funds (3599-PPCDT)the co-funding of the European Regional Development Fund (POCI-01-0145-FEDER-016875)supported within the PT2020 Partnership Agreement and COMPETE 2020 by the centre for Functional Ecology Strategic Project (UID/BIA/04004/2013)the Coimbra Chemistry Center Strategic Project (UID/QUI/00313/2013)。
文摘The treatment of wastewaters is crucial to maintain the ecological status of receiving waters,and thereby guarantee the protection of aquatic life and human health.Wastewater quality evaluation is conventionally based on physicochemical parameters,but increasing attention has been paid to integrate physicochemical and biological data.Nevertheless,the regulatory use of fish in biological testing methods has been subject to various ethical and cost concerns,and in vitro cell-based assays have thus become an important topic of interest.Hence,the present study intends:(a) to evaluate the efficiency of two different sample pre-concentration techniques (lyophilisation and solid phase extraction) to assess the toxicity of municipal effluents on rat cardiomyoblast H9c2(2-1) cells,and (b) maximizing the use of the effluent sample collected,to estimate the environmental condition of the receiving environment.The gathered results demonstrate that the H9c2(2-1) sulforhodamine B-based assay is an appropriate in vitro method to assess biological effluent toxicity,and the best results were attained by lyophilising the sample as pre-treatment.Due to its response,the H9c2(2-1) cell line might be a possible alternative in vitro model for fish lethal testing to assess the toxicity of municipal effluents.The physicochemical status of the sample suggests a high potential for eutrophication,and iron exceeded the permissible level for wastewater discharge,possibly due to the addition of ferric chloride for wastewater treatment.In general,the levels of carbamazepine and sulfamethoxazole are higher than those reported for other countries,and both surpassed the aquatic protective values for long-term exposure.
文摘Background Advances in the understanding of cardiovascular pathogenesis have highlighted that inflammation plays a central role in athemsclemtic coronary heart disease.Therefore,exploring pharmacologically based anti-inflammatory treatments to be used in cardiovascular therapeutics is worthwhile to promote the discovery of novel ways of treating cardiovascular disorders.Methods The myocardial cell line H9c2（2-1） was exposed to lipopolysacchadde （LPS） in culture and resulted in a cellular pro-inflammation status,miR-21 microRNA levels were detected using quantitative real-time polymerase chain reaction （Q-RT-PCR）.The influence of Iovastatin on miR-21 under normal and pro-inflammatory conditions was tested after being added to the cell culture mixture for 24 hours.Conditional gene function of two predicted cardiovascular system relevant downstream targets of miR-21,protein phosphatase 1 regulatory subunit 3A （PPP1R3A） and signal transducer and activator of transcription 3 （STAT3）,were analyzed with immunoblotting.Results Forty-eight hours of LPS treatment significantly increased the miR-21 to 170.71%±34.32% of control levels （P=0.002）.Co-treatment with Iovastatin for 24 hours before harvesting attenuated the up-regulation of miR-21 （P=0.013）.Twenty-four hours of Iovastatin exposure up-regulated PPP1R3A to 143.85%±21.89% of control levels in cardiomyocytes （P=0.023）.Lovastatin up-regulated the phosphorylation level of STAT3 compared to the background LPS pretreatment （P=0.0077）,this effect was significantly （P=0.018） blunted when miR-21 was functionally inhibited.Conclusions miR-21 plays a major role in the regulation of the cellular anti-inflammation effects of Iovastatin.
