OBJECTIVE:To determine the effect of Wenyang Huazhuo Fang(WHF),a Traditional Chinese Medicine decoction,on renal function in a rat model of doxorubicin-induced nephropathy,and to elucidate the underlying mechanism.MET...OBJECTIVE:To determine the effect of Wenyang Huazhuo Fang(WHF),a Traditional Chinese Medicine decoction,on renal function in a rat model of doxorubicin-induced nephropathy,and to elucidate the underlying mechanism.METHODS:Sprague-Dawley rats were randomly divided into six groups:control,doxorubicin-nephropathy,and prednisone-treated(6.45 mg·kg^-1·d^-1)doxorubicin nephropathy groups,as well as high-(7.26 g·kg^-1·d^-1),medium-(2.42 g·kg-1·d-),and low-dose(0.81 g·kg^-1·d^-1)WHF-treated doxorubicin-nephropathy groups.The nephropathy rat model was established by two tail vein injections of doxorubicin,followed by prednisone or WHF treatment for 8 weeks.Body weights were monitored and urinary protein was measured every 2 weeks.After the end of the treatment period,the rats were euthanized.Serum biochemical indicators were determined and renal morphological alterations were assessed using histological staining.The expression of transient receptor potential cation channel subfamily C member 6(TRPC6),stromal interaction molecule 1(STIM1),and calcium release-activated calcium channel protein 1(Orai1)was detected using western blotting,and their mRNA levels were examined using quantitative real-time reverse transcription-polymerase chain reaction.RESULTS:WHF treatment was found to significantly ameliorate weight loss,proteinuria,hypoalbuminemia,and dyslipidemia in doxorubicin-nephropathy rats.The protein and mRNA levels of TRPC6,STIM1,and Orai1 were partially,but significantly suppressed by prednisone or WHF treatment.CONCLUSION:Treatment with WHF significantly ameliorates renal injury in a rat model of doxorubicin-induced nephropathy,which could be at least partially related to repression of the TRPC6 pathway.展开更多
目的:研究乙型肝炎病毒x基因(hepatitis B virus x gene,HBx)蛋白调节细胞内钙离子可能分子机制,揭示乙型肝炎病毒(hepatitis B virus,HBV)诱导肝癌的可能途径.方法:培养HEK293细胞,取第2代HEK293细胞转染pcDNA-HBx质粒,培养12、24和48 ...目的:研究乙型肝炎病毒x基因(hepatitis B virus x gene,HBx)蛋白调节细胞内钙离子可能分子机制,揭示乙型肝炎病毒(hepatitis B virus,HBV)诱导肝癌的可能途径.方法:培养HEK293细胞,取第2代HEK293细胞转染pcDNA-HBx质粒,培养12、24和48 h后,细胞活力细胞毒性检测(cell counting kit-8,CCK-8)观察转染后细胞生长情况,Western b l o t检测H B x蛋白的表达情况,当共同转染HBx基因、Orai1基因或STIM1基因后co-IP实验、免疫荧光检测观察细胞内蛋白结合情况.结果:转染pcDNA-HBx质粒后HBx蛋白可以在HEK293细胞高表达,转染后的24 h后细胞增殖加快(P<0.05),co-IP实验及免疫荧光检测结果均显示,HBx蛋白在细胞内可以与Orai1蛋白结合.结论:HBx蛋白通过与细胞膜钙离子通道Orai1结合,来升高细胞内钙离子浓度,从而影响细胞增殖等活性.展开更多
基金the Key Research and Development Projects of Shaanxi Province(Study on the Effect of Wenyanghuazhuo Method on Autophagy-related Proteins in Adriamycin Nephropathy Rat Model,No.2020SF-340)a Grant from the Sci-tech Project of Shaanxi Province(Study on the Expression Regulation of STIM1/Orail and TRPC-related Channel Proteins in Renal Tissues of Adriamycin Nephropathy Rats by Aconiti Lateralis Radix and Wenyanghuazhuofang,No.2016SF-068)。
文摘OBJECTIVE:To determine the effect of Wenyang Huazhuo Fang(WHF),a Traditional Chinese Medicine decoction,on renal function in a rat model of doxorubicin-induced nephropathy,and to elucidate the underlying mechanism.METHODS:Sprague-Dawley rats were randomly divided into six groups:control,doxorubicin-nephropathy,and prednisone-treated(6.45 mg·kg^-1·d^-1)doxorubicin nephropathy groups,as well as high-(7.26 g·kg^-1·d^-1),medium-(2.42 g·kg-1·d-),and low-dose(0.81 g·kg^-1·d^-1)WHF-treated doxorubicin-nephropathy groups.The nephropathy rat model was established by two tail vein injections of doxorubicin,followed by prednisone or WHF treatment for 8 weeks.Body weights were monitored and urinary protein was measured every 2 weeks.After the end of the treatment period,the rats were euthanized.Serum biochemical indicators were determined and renal morphological alterations were assessed using histological staining.The expression of transient receptor potential cation channel subfamily C member 6(TRPC6),stromal interaction molecule 1(STIM1),and calcium release-activated calcium channel protein 1(Orai1)was detected using western blotting,and their mRNA levels were examined using quantitative real-time reverse transcription-polymerase chain reaction.RESULTS:WHF treatment was found to significantly ameliorate weight loss,proteinuria,hypoalbuminemia,and dyslipidemia in doxorubicin-nephropathy rats.The protein and mRNA levels of TRPC6,STIM1,and Orai1 were partially,but significantly suppressed by prednisone or WHF treatment.CONCLUSION:Treatment with WHF significantly ameliorates renal injury in a rat model of doxorubicin-induced nephropathy,which could be at least partially related to repression of the TRPC6 pathway.
文摘目的:研究乙型肝炎病毒x基因(hepatitis B virus x gene,HBx)蛋白调节细胞内钙离子可能分子机制,揭示乙型肝炎病毒(hepatitis B virus,HBV)诱导肝癌的可能途径.方法:培养HEK293细胞,取第2代HEK293细胞转染pcDNA-HBx质粒,培养12、24和48 h后,细胞活力细胞毒性检测(cell counting kit-8,CCK-8)观察转染后细胞生长情况,Western b l o t检测H B x蛋白的表达情况,当共同转染HBx基因、Orai1基因或STIM1基因后co-IP实验、免疫荧光检测观察细胞内蛋白结合情况.结果:转染pcDNA-HBx质粒后HBx蛋白可以在HEK293细胞高表达,转染后的24 h后细胞增殖加快(P<0.05),co-IP实验及免疫荧光检测结果均显示,HBx蛋白在细胞内可以与Orai1蛋白结合.结论:HBx蛋白通过与细胞膜钙离子通道Orai1结合,来升高细胞内钙离子浓度,从而影响细胞增殖等活性.
