目的探讨全反式维甲酸(ATRA)对高糖环境下正常人肾小管上皮细胞(HK-2细胞)炎症因子表达的影响及其可能机制。方法将体外培养的HK-2细胞随机分为7组:空白对照组,高糖组(D-葡萄糖30 mmol·L^(-1)),高渗组(甘露醇24.5 mmol·L^(-1)...目的探讨全反式维甲酸(ATRA)对高糖环境下正常人肾小管上皮细胞(HK-2细胞)炎症因子表达的影响及其可能机制。方法将体外培养的HK-2细胞随机分为7组:空白对照组,高糖组(D-葡萄糖30 mmol·L^(-1)),高渗组(甘露醇24.5 mmol·L^(-1)),ATRA干预组[包括高糖+低浓度ATRA组(ATRA 10-7mol·L^(-1))、高糖+中浓度ATRA组(ATRA 10-6mol·L^(-1))、高糖+高浓度ATRA组(ATRA 10-5mol·L^(-1))],Rho激酶抑制药(Y27632)干预组[高糖+Y27632(Y27632为30μmol·L^(-1))],均干预48 h。逆转录-聚合酶链反应(RT-PCR)法检测细胞Rho A mRNA、ROCK1 mRNA表达水平,酶联免疫吸附法(ELISA)检测HK-2细胞白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)变化。结果 RT-PCR结果显示,空白对照组与高渗组Rho A mRNA、ROCK1 mRNA表达差异无统计学意义(P>0.05);高糖组Rho A mRNA、ROCK1 mRNA表达较空白对照组、高渗组显著升高(P<0.05);ATRA各干预组Rho A mRNA、ROCK1 mRNA表达较高糖组显著减少(P<0.05),且呈现ATRA浓度依赖性;Y27632干预组Rho A mRNA表达与高糖组差异无统计学意义,ROCK1mRNA表达显著减少(P<0.05)。Person直线相关分析显示,高糖组及ATRA干预组Rho A mRNA与ROCK1 mRNA表达呈正相关。ELISA检测结果示空白对照组与高渗组表达IL-6、TNF-α无明显差异(P>0.05);高糖组表达IL-6、TNF-α较空白对照组明显升高(P<0.05);ATRA或Y27632干预后,IL-6、TNF-α表达明显减少(P<0.05)。结论 ATRA可抑制高糖环境下HK-2细胞IL-6、TNF-α表达,其机制可能与抑制Rho A/ROCK信号通路有关。展开更多
The hedgehog signaling cascade is an evolutionarily conserved pathway that regulates multiple aspects of embryonic development and plays a decisive role in tissue homeostasis. As the best studied member of three hedge...The hedgehog signaling cascade is an evolutionarily conserved pathway that regulates multiple aspects of embryonic development and plays a decisive role in tissue homeostasis. As the best studied member of three hedgehog ligands, sonic hedgehog(Shh) is known to be associated with kidney development and tissue repair after various insults. Recent studies uncover an intrinsic link between dysregulated Shh signaling and renal fibrogenesis. In various types of chronic kidney disease(CKD), Shh is upregulated specifically in renal tubular epithelium but targets interstitial fibroblasts, thereby mediating a dynamic epithelialmesenchymal communication(EMC). Tubule-derived Shh acts as a growth factor for interstitial fibroblasts and controls a hierarchy of fibrosis-related genes, which lead to the excessive deposition of extracellular matrix in renal interstitium. In this review, we recapitulate the principle of Shh signaling, its activation and regulation in a variety of kidney diseases. We also discuss the potential mechanisms by which Shh promotes renal fibrosis and assess the efficacy of blocking this signaling in preclinical settings. Continuing these lines of investigations will provide novel opportunities for designing effective therapies to improve CKD prognosis in patients.展开更多
文摘目的探讨全反式维甲酸(ATRA)对高糖环境下正常人肾小管上皮细胞(HK-2细胞)炎症因子表达的影响及其可能机制。方法将体外培养的HK-2细胞随机分为7组:空白对照组,高糖组(D-葡萄糖30 mmol·L^(-1)),高渗组(甘露醇24.5 mmol·L^(-1)),ATRA干预组[包括高糖+低浓度ATRA组(ATRA 10-7mol·L^(-1))、高糖+中浓度ATRA组(ATRA 10-6mol·L^(-1))、高糖+高浓度ATRA组(ATRA 10-5mol·L^(-1))],Rho激酶抑制药(Y27632)干预组[高糖+Y27632(Y27632为30μmol·L^(-1))],均干预48 h。逆转录-聚合酶链反应(RT-PCR)法检测细胞Rho A mRNA、ROCK1 mRNA表达水平,酶联免疫吸附法(ELISA)检测HK-2细胞白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)变化。结果 RT-PCR结果显示,空白对照组与高渗组Rho A mRNA、ROCK1 mRNA表达差异无统计学意义(P>0.05);高糖组Rho A mRNA、ROCK1 mRNA表达较空白对照组、高渗组显著升高(P<0.05);ATRA各干预组Rho A mRNA、ROCK1 mRNA表达较高糖组显著减少(P<0.05),且呈现ATRA浓度依赖性;Y27632干预组Rho A mRNA表达与高糖组差异无统计学意义,ROCK1mRNA表达显著减少(P<0.05)。Person直线相关分析显示,高糖组及ATRA干预组Rho A mRNA与ROCK1 mRNA表达呈正相关。ELISA检测结果示空白对照组与高渗组表达IL-6、TNF-α无明显差异(P>0.05);高糖组表达IL-6、TNF-α较空白对照组明显升高(P<0.05);ATRA或Y27632干预后,IL-6、TNF-α表达明显减少(P<0.05)。结论 ATRA可抑制高糖环境下HK-2细胞IL-6、TNF-α表达,其机制可能与抑制Rho A/ROCK信号通路有关。
基金supported by the National Natural Science Foundation of China(81130011,81370839,81521003)Guangdong Science Foundation(2014A030312014)+2 种基金Guangzhou Projects Grant(15020025)American Heart Association FTF(16990086)National Institutes of Health Grants(DK064005,DK091239,DK106049)
文摘The hedgehog signaling cascade is an evolutionarily conserved pathway that regulates multiple aspects of embryonic development and plays a decisive role in tissue homeostasis. As the best studied member of three hedgehog ligands, sonic hedgehog(Shh) is known to be associated with kidney development and tissue repair after various insults. Recent studies uncover an intrinsic link between dysregulated Shh signaling and renal fibrogenesis. In various types of chronic kidney disease(CKD), Shh is upregulated specifically in renal tubular epithelium but targets interstitial fibroblasts, thereby mediating a dynamic epithelialmesenchymal communication(EMC). Tubule-derived Shh acts as a growth factor for interstitial fibroblasts and controls a hierarchy of fibrosis-related genes, which lead to the excessive deposition of extracellular matrix in renal interstitium. In this review, we recapitulate the principle of Shh signaling, its activation and regulation in a variety of kidney diseases. We also discuss the potential mechanisms by which Shh promotes renal fibrosis and assess the efficacy of blocking this signaling in preclinical settings. Continuing these lines of investigations will provide novel opportunities for designing effective therapies to improve CKD prognosis in patients.
