BACKGROUND NLRP3-mediated pyroptosis is recognized as an essential modulator of renal disease pathology.Long noncoding RNAs(lncRNAs)are active participators of diabetic nephropathy(DN).X inactive specific transcript(X...BACKGROUND NLRP3-mediated pyroptosis is recognized as an essential modulator of renal disease pathology.Long noncoding RNAs(lncRNAs)are active participators of diabetic nephropathy(DN).X inactive specific transcript(XIST)expression has been reported to be elevated in the serum of DN patients.AIM To evaluate the mechanism of lncRNA XIST in renal tubular epithelial cell(RTEC)pyroptosis in DN.METHODS A DN rat model was established through streptozotocin injection,and XIST was knocked down by tail vein injection of the lentivirus LV sh-XIST.Renal metabolic and biochemical indices were detected,and pathological changes in the renal tissue were assessed.The expression of indicators related to inflammation and pyroptosis was also detected.High glucose(HG)was used to treat HK2 cells,and cell viability and lactate dehydrogenase(LDH)activity were detected after silencing XIST.The subcellular localization and downstream mechanism of XIST were investigated.Finally,a rescue experiment was carried out to verify that XIST regulates NLR family pyrin domain containing 3(NLRP3)/caspase-1-mediated RTEC pyroptosis through the microRNA-15-5p(miR-15b-5p)/Toll-like receptor 4(TLR4)axis.RESULTS XIST was highly expressed in the DN models.XIST silencing improved renal metabolism and biochemical indices and mitigated renal injury.The expression of inflammation and pyroptosis indicators was significantly increased in DN rats and HG-treated HK2 cells;cell viability was decreased and LDH activity was increased after HGtreatment. Silencing XIST inhibited RTEC pyroptosis by inhibiting NLRP3/caspase-1. Mechanistically,XIST sponged miR-15b-5p to regulate TLR4. Silencing XIST inhibited TLR4 by promotingmiR-15b-5p. miR-15b-5p inhibition or TLR4 overexpression averted the inhibitory effect ofsilencing XIST on HG-induced RTEC pyroptosis.CONCLUSIONSilencing XIST inhibits TLR4 by upregulating miR-15b-5p and ultimately inhibits renal injury inDN by inhibiting NLRP3/caspase-1-mediated RTEC pyroptosis.展开更多
Non-alcoholic fatty liver disease(NAFLD)is emerging as a common cause of chronic liver disease in children and adults.NAFLD can progress to steatohepa-titis and potentially even hepatocellular carcinoma.Early identifi...Non-alcoholic fatty liver disease(NAFLD)is emerging as a common cause of chronic liver disease in children and adults.NAFLD can progress to steatohepa-titis and potentially even hepatocellular carcinoma.Early identification of pati-ents at risk for progressive disease is crucial for managing NAFLD.Recent studies have identified long noncoding RNAs(lncRNAs),circular RNAs,and microRNAs as playing important roles in the pathogenesis of NAFLD.These noncoding RNAs are involved in modulating several metabolic pathways such as hepatic glucose and lipid metabolism,oxidative stress,and even carcinogenesis.Elevated levels of lncARSR and lncRNA nuclear-enriched abundant transcript 1 have been found in patients with NAFLD.In addition,lncRNAs such as PRYP4-3 and RP11-128N14.5 can distinguish patients with NAFLD from healthy indi-viduals.Increased MEG3 expression has been observed in both NAFLD and non-alcoholic steatohepatitis,suggesting that it may help predict patients at risk for disease progression.With advances in transcriptomics,we may discover additional targets to help in the identification and prognostication of NAFLD.展开更多
目的观察敲降长链非编码RNA-肺腺癌转移相关转录因子1(lncRNA-MALAT1)调控微小RNA-194-5p(miR-194-5p)/叉头框蛋白A1(FOXA1)通路对脂多糖(LPS)诱导的人肺泡上皮细胞(HPAEpiC)凋亡的影响.方法采用1 mg/L LPS处理HPAEpiC复制脓毒症急性肺...目的观察敲降长链非编码RNA-肺腺癌转移相关转录因子1(lncRNA-MALAT1)调控微小RNA-194-5p(miR-194-5p)/叉头框蛋白A1(FOXA1)通路对脂多糖(LPS)诱导的人肺泡上皮细胞(HPAEpiC)凋亡的影响.方法采用1 mg/L LPS处理HPAEpiC复制脓毒症急性肺损伤(ALI)体外模型.采用荧光定量聚合酶链反应(qPCR)检测HPAEpiC细胞中MALAT1、miR-194-5p的表达水平,构建MALAT1敲降载体、miR-194-5p抑制剂及FOXA1抑制剂转染HPAEpiC,将细胞分为空白对照组、LPS模型组、LPS+生理盐水组、LPS+MALAT1抑制剂组、LPS+MALAT1抑制剂+生理盐水组、LPS+MALAT1抑制剂+miR-194-5p抑制剂组和LPS+FOXA1抑制剂组.采用CCK-8细胞增殖及毒性检测试剂盒、流式细胞术、蛋白质免疫印迹试验(Western Blot)检测MALAT1或其敲降后对LPS诱导的HPAEpiC增殖及凋亡和FOXA1表达的影响;采用双荧光素酶报告基因检测lncRNA-MALAT1、miR-194-5p及FOXA1的靶向调控关系.结果与空白对照组比较,LPS可上调HPAEpiC-MALAT1的表达,促进HPAEpiC凋亡并抑制其增殖〔MALAT1(2^(-ΔΔCt)):0.83±0.09比0.15±0.02,HPAEpiC凋亡率:(21.31±2.31)%比(5.41±0.42)%,24 h HPAEpiC增殖活性(A值):0.42±0.03比0.54±0.02,均P<0.05〕;与LPS+生理盐水组比较,敲降MALAT1可抑制LPS诱导的HPAEpiC凋亡并促进其增殖〔细胞凋亡率:(6.40±0.40)%比(21.38±2.31)%,24 h HPAEpiC增殖活性(A值):0.53±0.03比0.40±0.02,均P<0.05〕,降低MALAT1的表达(2-ΔΔCt:0.20±0.03比1.02±0.09,P<0.05).双荧光素酶报告基因及Western Blot证实,敲降MALAT1通过靶向上调miR-194-5p从而抑制FOXA1在LPS诱导的HPAEpiC中的表达〔miR-194-5p(2^(-ΔΔCt)):5.27±0.15比1.21±0.09,FOXA1蛋白表达(灰度值):0.36±0.05比1.00±0.07,均P<0.05〕,miR-194-5p抑制剂能逆转MALAT1敲降在LPS诱导的HPAEpiC中对FOXA1的作用〔FOXA1蛋白表达(灰度值):2.76±0.20比1.00±0.07,P<0.05〕,抑制FOXA1表达能减轻LPS诱导的HPAEpiC凋亡并促进其增殖〔FOXA1蛋白表达(灰度值):0.28±0.03比1.00±0.03,HPAEpiC凋亡率:(6.78±0.38)%比(19.21±0.70)%,24 h HPAEpiC增殖活性(A值):0.59±0.20比0.41±0.02,均P<0.05〕.结论敲降MALAT1通过靶向上调miR-194-5p抑制FOXA1的表达,进而抑制LPS诱导的HPAEpiC凋亡,为脓毒症ALI的诊断和治疗提供了潜在的分子靶点.展开更多
基金Supported by Natural Science Foundation of Shenzhen University General Hospital (SUGH2020QD011)
文摘BACKGROUND NLRP3-mediated pyroptosis is recognized as an essential modulator of renal disease pathology.Long noncoding RNAs(lncRNAs)are active participators of diabetic nephropathy(DN).X inactive specific transcript(XIST)expression has been reported to be elevated in the serum of DN patients.AIM To evaluate the mechanism of lncRNA XIST in renal tubular epithelial cell(RTEC)pyroptosis in DN.METHODS A DN rat model was established through streptozotocin injection,and XIST was knocked down by tail vein injection of the lentivirus LV sh-XIST.Renal metabolic and biochemical indices were detected,and pathological changes in the renal tissue were assessed.The expression of indicators related to inflammation and pyroptosis was also detected.High glucose(HG)was used to treat HK2 cells,and cell viability and lactate dehydrogenase(LDH)activity were detected after silencing XIST.The subcellular localization and downstream mechanism of XIST were investigated.Finally,a rescue experiment was carried out to verify that XIST regulates NLR family pyrin domain containing 3(NLRP3)/caspase-1-mediated RTEC pyroptosis through the microRNA-15-5p(miR-15b-5p)/Toll-like receptor 4(TLR4)axis.RESULTS XIST was highly expressed in the DN models.XIST silencing improved renal metabolism and biochemical indices and mitigated renal injury.The expression of inflammation and pyroptosis indicators was significantly increased in DN rats and HG-treated HK2 cells;cell viability was decreased and LDH activity was increased after HGtreatment. Silencing XIST inhibited RTEC pyroptosis by inhibiting NLRP3/caspase-1. Mechanistically,XIST sponged miR-15b-5p to regulate TLR4. Silencing XIST inhibited TLR4 by promotingmiR-15b-5p. miR-15b-5p inhibition or TLR4 overexpression averted the inhibitory effect ofsilencing XIST on HG-induced RTEC pyroptosis.CONCLUSIONSilencing XIST inhibits TLR4 by upregulating miR-15b-5p and ultimately inhibits renal injury inDN by inhibiting NLRP3/caspase-1-mediated RTEC pyroptosis.
文摘Non-alcoholic fatty liver disease(NAFLD)is emerging as a common cause of chronic liver disease in children and adults.NAFLD can progress to steatohepa-titis and potentially even hepatocellular carcinoma.Early identification of pati-ents at risk for progressive disease is crucial for managing NAFLD.Recent studies have identified long noncoding RNAs(lncRNAs),circular RNAs,and microRNAs as playing important roles in the pathogenesis of NAFLD.These noncoding RNAs are involved in modulating several metabolic pathways such as hepatic glucose and lipid metabolism,oxidative stress,and even carcinogenesis.Elevated levels of lncARSR and lncRNA nuclear-enriched abundant transcript 1 have been found in patients with NAFLD.In addition,lncRNAs such as PRYP4-3 and RP11-128N14.5 can distinguish patients with NAFLD from healthy indi-viduals.Increased MEG3 expression has been observed in both NAFLD and non-alcoholic steatohepatitis,suggesting that it may help predict patients at risk for disease progression.With advances in transcriptomics,we may discover additional targets to help in the identification and prognostication of NAFLD.
文摘目的观察敲降长链非编码RNA-肺腺癌转移相关转录因子1(lncRNA-MALAT1)调控微小RNA-194-5p(miR-194-5p)/叉头框蛋白A1(FOXA1)通路对脂多糖(LPS)诱导的人肺泡上皮细胞(HPAEpiC)凋亡的影响.方法采用1 mg/L LPS处理HPAEpiC复制脓毒症急性肺损伤(ALI)体外模型.采用荧光定量聚合酶链反应(qPCR)检测HPAEpiC细胞中MALAT1、miR-194-5p的表达水平,构建MALAT1敲降载体、miR-194-5p抑制剂及FOXA1抑制剂转染HPAEpiC,将细胞分为空白对照组、LPS模型组、LPS+生理盐水组、LPS+MALAT1抑制剂组、LPS+MALAT1抑制剂+生理盐水组、LPS+MALAT1抑制剂+miR-194-5p抑制剂组和LPS+FOXA1抑制剂组.采用CCK-8细胞增殖及毒性检测试剂盒、流式细胞术、蛋白质免疫印迹试验(Western Blot)检测MALAT1或其敲降后对LPS诱导的HPAEpiC增殖及凋亡和FOXA1表达的影响;采用双荧光素酶报告基因检测lncRNA-MALAT1、miR-194-5p及FOXA1的靶向调控关系.结果与空白对照组比较,LPS可上调HPAEpiC-MALAT1的表达,促进HPAEpiC凋亡并抑制其增殖〔MALAT1(2^(-ΔΔCt)):0.83±0.09比0.15±0.02,HPAEpiC凋亡率:(21.31±2.31)%比(5.41±0.42)%,24 h HPAEpiC增殖活性(A值):0.42±0.03比0.54±0.02,均P<0.05〕;与LPS+生理盐水组比较,敲降MALAT1可抑制LPS诱导的HPAEpiC凋亡并促进其增殖〔细胞凋亡率:(6.40±0.40)%比(21.38±2.31)%,24 h HPAEpiC增殖活性(A值):0.53±0.03比0.40±0.02,均P<0.05〕,降低MALAT1的表达(2-ΔΔCt:0.20±0.03比1.02±0.09,P<0.05).双荧光素酶报告基因及Western Blot证实,敲降MALAT1通过靶向上调miR-194-5p从而抑制FOXA1在LPS诱导的HPAEpiC中的表达〔miR-194-5p(2^(-ΔΔCt)):5.27±0.15比1.21±0.09,FOXA1蛋白表达(灰度值):0.36±0.05比1.00±0.07,均P<0.05〕,miR-194-5p抑制剂能逆转MALAT1敲降在LPS诱导的HPAEpiC中对FOXA1的作用〔FOXA1蛋白表达(灰度值):2.76±0.20比1.00±0.07,P<0.05〕,抑制FOXA1表达能减轻LPS诱导的HPAEpiC凋亡并促进其增殖〔FOXA1蛋白表达(灰度值):0.28±0.03比1.00±0.03,HPAEpiC凋亡率:(6.78±0.38)%比(19.21±0.70)%,24 h HPAEpiC增殖活性(A值):0.59±0.20比0.41±0.02,均P<0.05〕.结论敲降MALAT1通过靶向上调miR-194-5p抑制FOXA1的表达,进而抑制LPS诱导的HPAEpiC凋亡,为脓毒症ALI的诊断和治疗提供了潜在的分子靶点.
