Background:Serum-and glucocorticoid-induced kinase 1(SGK1)is a member of the serine/threonine kinase family,playing a crucial role in regulating ion channel function,hormone secretion,cellular growth,survival mechanis...Background:Serum-and glucocorticoid-induced kinase 1(SGK1)is a member of the serine/threonine kinase family,playing a crucial role in regulating ion channel function,hormone secretion,cellular growth,survival mechanisms,and neuronal activity.SGK1 is implicated in kidney diseases,hypertension,and metabolic syndromes,influencing salt intake,renal growth,and renal potassium(K+)excretion duringmineralocorticoid overdose.Although SGK1’s renal functions have been explored,comprehensive identification of SGK1-related genes and signaling cascades remains limited.Objectives:This research sought to explore the cellularmechanisms and signaling pathways influenced by SGK1 in rat kidney cells.Methods:NRK-52E cells,derived from rat kidneys,were exposed to the SGK1 inhibitor GSK 650394(GSK)at doses of 50,100,and 200μM.A comparative transcriptomic approach was employed,utilizing mRNA sequencing(RNA-seq)and differential gene expression(DEG)analysis.To investigate key signaling pathways and molecular mechanisms,analyses including Gene Ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG),andGene Set Enrichment Analysis(GSEA)were carried out.Results:In total,13,987 geneswere detected,with differential expression observed across treatment groups.Specifically,749 DEGs(401 upregulated,348 downregulated)were identified in the 50μM group,2267 DEGs(897 upregulated,1370 downregulated)in the 100μM group,and 5631 DEGs(2547 upregulated,3084 downregulated)in the 200μM group.A total of 176 DEGs were consistently present across all treatment groups(adjusted p-value<0.05;∣log2 fold change∣>1).Enrichment analysis revealed that these genes play a significant role in pathways potentially associatedwith SGK1 regulation.Conclusion:ThroughRNA-seq andDEG analysis,potential target genes and signaling pathways influenced by SGK1 in rat kidney cells were identified,providing a basis for future investigations into SGK1-associated molecular mechanisms and physiological roles in the kidney.展开更多
Background:Chronic prostatitis/chronic pelvic pain syndrome(CP/CPPS)is a frequently encountered disorder characterized by voiding symptoms and pelvic or perineal pain.Proinflammatory T helper 17(Th17)cells are essenti...Background:Chronic prostatitis/chronic pelvic pain syndrome(CP/CPPS)is a frequently encountered disorder characterized by voiding symptoms and pelvic or perineal pain.Proinflammatory T helper 17(Th17)cells are essential for triggering the development of CP/CPPS.High-salt diet(HSD)consumption has been found to cause an accumulation of sodium chloride in peripheral organs,inducing autoimmune responses via the Th17 cell axis.It is currently unknown whether HSD affects the etiology and course of CP/CPPS.Methods:Patients diagnosed with CP/CPPS were evaluated with the National Institutes of Health Chronic Prostatitis Symptom Index scoring system,and the correlation between the symptoms of CP/CPPS with HSD was analyzed.The experimental autoimmune prostatitis(EAP)mouse was established and the mice were fed either a normal-salt diet(NSD)or HSD for 6 weeks to investigate the impact of HSD on CP/CPPS.Then,16S ribosomal RNA sequencing and untargeted metabolomics were introduced to detect the differences in the gut microflora composition and metabolite profiles between NSD-fed and HSD-fed mice,followed by fecal microbiota transplantation,5-hydroxyindole acetic acid(5-HIAA)supplementation,aryl hydrocarbon receptor(AHR)inhibition,and in vitro Th17 differentiation experiments,which were performed to explore the mechanisms underlying HSD-aggravated CP/CPPS.Finally,chromatin immunoprecipitation assay and quantitative polymerase chain reaction were conducted to validate whether AHR can serve as a transcription factor by interacting with the serum and glucocorticoid-regulated kinase 1(Sgk1)promoter in CD4^(+)T cells.Results:Increased salt consumption had a positive correlation with symptom scores of CP/CPPS patients,which was validated by feeding EAP mice with HSD,and HSD worsened the prostate inflammation and tactile allodynia in EAP mice through promoting the differentiation of CD4^(+)T cells to Th17 cells.HSD exacerbated EAP by significantly reducing the relative abundance of beneficial gut microflora,such as Lactobacillaceae,and gut microbiota metabolite 5-HIAA,which is related to tryptophan metabolism.The prostate inflammation,tactile allodynia,and proportion of Th17 cells in mice that received fecal suspensions from the EAP^(+)HSD group were significantly more severe or higher than those in mice that received fecal suspensions from the EAP^(+)NSD group.However,5-HIAA supplementation ameliorated the symptoms of EAP caused by HSD through inhibiting the differentiation of CD4^(+)T cells to Th17 cells,while AHR inhibition abrogated the protective effects of 5-HIAA supplementation on EAP mice fed a HSD through promoting the differentiation of CD4^(+)T cells to Th17 cells.Mechanistically,it has been revealed that the SGK1/forkhead box protein O1(FOXO1)pathway was significantly activated during cytokine-induced Th17 cell differentiation,and AHR has been shown to inhibit SGK1 transcription by interacting with the Sgk1 promoter in CD4^(+)T cells to inhibit FOXO1 phosphorylation,consequently restoring the equilibrium of Th17 cell differentiation.Conclusions:Our findings indicated that high salt intake represented a risk factor for the development of CP/CPPS as it promoted the differentiation of CD4^(+)T cells to Th17 cells through the 5-HIAA/AHR/SGK1/FOXO1 axis,which might be a potential therapeutic target for CP/CPPS.展开更多
目的:探讨百事乐胶囊对慢性应激抑郁模型大鼠学习记忆的保护作用。方法:将SD大鼠随机分为6组,即空白对照组、抑郁模型组、氟西汀组及百事乐胶囊高、中、低(2.88,1.44,0.72 g·kg-1)剂量组,采用慢性温和不可预见性应激加孤养的方法...目的:探讨百事乐胶囊对慢性应激抑郁模型大鼠学习记忆的保护作用。方法:将SD大鼠随机分为6组,即空白对照组、抑郁模型组、氟西汀组及百事乐胶囊高、中、低(2.88,1.44,0.72 g·kg-1)剂量组,采用慢性温和不可预见性应激加孤养的方法建立抑郁模型,造模同时灌胃给药,连续21 d Morris水迷宫测定对大鼠行为学的影响,放射免疫法观察大鼠血浆CORT含量的变化,Western-blotting观察大鼠海马GR,p-GR,SGK1蛋白表达的变化。结果:与空白对照组比较,模型组大鼠逃避潜伏期(EL)、目标象限潜伏时间显著增长(P<0.01);穿越目标象限的次数显著降低(P<0.05);CORT含量显著增高(P<0.01),海马GR、p-GR的表达下调(P<0.01),SGK1的表达增加(P<0.01)。与模型组比较,百事乐胶囊高剂量能缩短模型大鼠的EL(P<0.05),并能缩短目标象限潜伏时间(P<0.05),增加目标象限的穿越次数(P<0.05);同时高、中剂量组大鼠血浆CORT的含量显著下降(P<0.05或P<0.01);高、中剂量组海马GR、p-GR的表达显著增加(P<0.05或P<0.01),高剂量组SGK1的表达显著降低(P<0.05)。结论:百事乐胶囊可能通过GR/p-GR/SGK1信号级联增加应激抑郁模型大鼠海马学习记忆的功能,进而实现抗抑郁的作用。展开更多
目的:观察熟地有效成分甘露三糖对高浓度皮质酮诱导海马神经元损伤及学习记忆相关信号转导分子表达的影响。方法:用24小时新生大鼠原代培养海马神经细胞。第八天用药处理,将细胞分为5组:1:正常对照组;2:1×10-4M高浓度皮质酮模型组;...目的:观察熟地有效成分甘露三糖对高浓度皮质酮诱导海马神经元损伤及学习记忆相关信号转导分子表达的影响。方法:用24小时新生大鼠原代培养海马神经细胞。第八天用药处理,将细胞分为5组:1:正常对照组;2:1×10-4M高浓度皮质酮模型组;3:10-4 M CORT+3nM RU38486拮抗剂组;4:10-4 M CORT+0.1g/L甘露三糖组;5:10-4 M CORT+40uM多奈哌齐阳性对照组。24h后,通过SYTO13-PI双荧光染色观察各组细胞的形态及存活情况,MTT法测定细胞活性,Western Blot检测学习记忆信号转导通路相关分子GCR、BDNF、SGK蛋白表达。结果:与正常组相比,模型组细胞死亡较多,细胞活力下降,GCR、BDNF、SGK蛋白表达均显著性降低;如上改变又均被皮质酮受体拮抗剂RU38486逆转;甘露三糖(0.1g/L)同阳性对照多奈派齐(40μM)均明显减少死细胞数量,提高细胞活力,提高模型细胞GCR、BDNF、SGK蛋白表达。结论:熟地有效成分甘露三糖保护大鼠海马神经细胞免遭高浓度皮质酮的损伤,通过调节学习记忆信号转导途径中的重要蛋白GCR、BDNF、SGK表达从而改善高浓度皮质酮致学习记忆功能退化。展开更多
目的探讨MAL2(myelin and lymphocyte protein 2)在肺癌中的功能及作用机制。方法分析TCGA数据中MAL2在肺癌组织中的表达情况,在H1299和A549细胞中用克隆形成实验检测MAL2基因敲减后对肺癌细胞增殖的影响,探讨MAL2在肺腺癌细胞增殖中的...目的探讨MAL2(myelin and lymphocyte protein 2)在肺癌中的功能及作用机制。方法分析TCGA数据中MAL2在肺癌组织中的表达情况,在H1299和A549细胞中用克隆形成实验检测MAL2基因敲减后对肺癌细胞增殖的影响,探讨MAL2在肺腺癌细胞增殖中的作用,并用CCK8等细胞增殖、转移、凋亡实验进一步验证其功能;将MAL2干扰后,采用Western blot实验检测下游通路基因表达情况以分析MAL2基因可能的作用机制。结果MAL2基因在肺癌组织与癌旁组织相比表达显著升高(P<0.05),且MAL2敲减后抑制细胞增殖和转移、促进细胞凋亡(P<0.05);MAL2可能通过调控SGK1通路发挥作用。结论MAL2基因在肺癌的恶性进展中起重要的作用,该作用通过调控SGK1机制进行,MAL2基因是肺癌的一个新的潜在的分子诊断指标及治疗靶点。展开更多
基金supported by the Kaohsiung Veterans General Hospital(KSVGH-114-047 and KSVGH-114-102)the Zuoying Armed Forces GeneralHospital(KAFGH-ZY_A_111020 and KAFGH-ZY-A-112012).
文摘Background:Serum-and glucocorticoid-induced kinase 1(SGK1)is a member of the serine/threonine kinase family,playing a crucial role in regulating ion channel function,hormone secretion,cellular growth,survival mechanisms,and neuronal activity.SGK1 is implicated in kidney diseases,hypertension,and metabolic syndromes,influencing salt intake,renal growth,and renal potassium(K+)excretion duringmineralocorticoid overdose.Although SGK1’s renal functions have been explored,comprehensive identification of SGK1-related genes and signaling cascades remains limited.Objectives:This research sought to explore the cellularmechanisms and signaling pathways influenced by SGK1 in rat kidney cells.Methods:NRK-52E cells,derived from rat kidneys,were exposed to the SGK1 inhibitor GSK 650394(GSK)at doses of 50,100,and 200μM.A comparative transcriptomic approach was employed,utilizing mRNA sequencing(RNA-seq)and differential gene expression(DEG)analysis.To investigate key signaling pathways and molecular mechanisms,analyses including Gene Ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG),andGene Set Enrichment Analysis(GSEA)were carried out.Results:In total,13,987 geneswere detected,with differential expression observed across treatment groups.Specifically,749 DEGs(401 upregulated,348 downregulated)were identified in the 50μM group,2267 DEGs(897 upregulated,1370 downregulated)in the 100μM group,and 5631 DEGs(2547 upregulated,3084 downregulated)in the 200μM group.A total of 176 DEGs were consistently present across all treatment groups(adjusted p-value<0.05;∣log2 fold change∣>1).Enrichment analysis revealed that these genes play a significant role in pathways potentially associatedwith SGK1 regulation.Conclusion:ThroughRNA-seq andDEG analysis,potential target genes and signaling pathways influenced by SGK1 in rat kidney cells were identified,providing a basis for future investigations into SGK1-associated molecular mechanisms and physiological roles in the kidney.
基金supported by the National Natural Science Foundation of China(82300872 and 82170787)the Anhui Provincial Natural Science Foundation(2408085Y038)+3 种基金the Supporting Projects for Innovative Leading Talents(T000529)the Distinguished Young Scholar of Anhui Colleges(2021-108-10)the Science Foundation for Outstanding Young Scholar of Anhui Colleges(2022AH020073)the Sanming Project of Medicine in Shenzhen Nanshan(SZSM20210300).
文摘Background:Chronic prostatitis/chronic pelvic pain syndrome(CP/CPPS)is a frequently encountered disorder characterized by voiding symptoms and pelvic or perineal pain.Proinflammatory T helper 17(Th17)cells are essential for triggering the development of CP/CPPS.High-salt diet(HSD)consumption has been found to cause an accumulation of sodium chloride in peripheral organs,inducing autoimmune responses via the Th17 cell axis.It is currently unknown whether HSD affects the etiology and course of CP/CPPS.Methods:Patients diagnosed with CP/CPPS were evaluated with the National Institutes of Health Chronic Prostatitis Symptom Index scoring system,and the correlation between the symptoms of CP/CPPS with HSD was analyzed.The experimental autoimmune prostatitis(EAP)mouse was established and the mice were fed either a normal-salt diet(NSD)or HSD for 6 weeks to investigate the impact of HSD on CP/CPPS.Then,16S ribosomal RNA sequencing and untargeted metabolomics were introduced to detect the differences in the gut microflora composition and metabolite profiles between NSD-fed and HSD-fed mice,followed by fecal microbiota transplantation,5-hydroxyindole acetic acid(5-HIAA)supplementation,aryl hydrocarbon receptor(AHR)inhibition,and in vitro Th17 differentiation experiments,which were performed to explore the mechanisms underlying HSD-aggravated CP/CPPS.Finally,chromatin immunoprecipitation assay and quantitative polymerase chain reaction were conducted to validate whether AHR can serve as a transcription factor by interacting with the serum and glucocorticoid-regulated kinase 1(Sgk1)promoter in CD4^(+)T cells.Results:Increased salt consumption had a positive correlation with symptom scores of CP/CPPS patients,which was validated by feeding EAP mice with HSD,and HSD worsened the prostate inflammation and tactile allodynia in EAP mice through promoting the differentiation of CD4^(+)T cells to Th17 cells.HSD exacerbated EAP by significantly reducing the relative abundance of beneficial gut microflora,such as Lactobacillaceae,and gut microbiota metabolite 5-HIAA,which is related to tryptophan metabolism.The prostate inflammation,tactile allodynia,and proportion of Th17 cells in mice that received fecal suspensions from the EAP^(+)HSD group were significantly more severe or higher than those in mice that received fecal suspensions from the EAP^(+)NSD group.However,5-HIAA supplementation ameliorated the symptoms of EAP caused by HSD through inhibiting the differentiation of CD4^(+)T cells to Th17 cells,while AHR inhibition abrogated the protective effects of 5-HIAA supplementation on EAP mice fed a HSD through promoting the differentiation of CD4^(+)T cells to Th17 cells.Mechanistically,it has been revealed that the SGK1/forkhead box protein O1(FOXO1)pathway was significantly activated during cytokine-induced Th17 cell differentiation,and AHR has been shown to inhibit SGK1 transcription by interacting with the Sgk1 promoter in CD4^(+)T cells to inhibit FOXO1 phosphorylation,consequently restoring the equilibrium of Th17 cell differentiation.Conclusions:Our findings indicated that high salt intake represented a risk factor for the development of CP/CPPS as it promoted the differentiation of CD4^(+)T cells to Th17 cells through the 5-HIAA/AHR/SGK1/FOXO1 axis,which might be a potential therapeutic target for CP/CPPS.
文摘目的:探讨百事乐胶囊对慢性应激抑郁模型大鼠学习记忆的保护作用。方法:将SD大鼠随机分为6组,即空白对照组、抑郁模型组、氟西汀组及百事乐胶囊高、中、低(2.88,1.44,0.72 g·kg-1)剂量组,采用慢性温和不可预见性应激加孤养的方法建立抑郁模型,造模同时灌胃给药,连续21 d Morris水迷宫测定对大鼠行为学的影响,放射免疫法观察大鼠血浆CORT含量的变化,Western-blotting观察大鼠海马GR,p-GR,SGK1蛋白表达的变化。结果:与空白对照组比较,模型组大鼠逃避潜伏期(EL)、目标象限潜伏时间显著增长(P<0.01);穿越目标象限的次数显著降低(P<0.05);CORT含量显著增高(P<0.01),海马GR、p-GR的表达下调(P<0.01),SGK1的表达增加(P<0.01)。与模型组比较,百事乐胶囊高剂量能缩短模型大鼠的EL(P<0.05),并能缩短目标象限潜伏时间(P<0.05),增加目标象限的穿越次数(P<0.05);同时高、中剂量组大鼠血浆CORT的含量显著下降(P<0.05或P<0.01);高、中剂量组海马GR、p-GR的表达显著增加(P<0.05或P<0.01),高剂量组SGK1的表达显著降低(P<0.05)。结论:百事乐胶囊可能通过GR/p-GR/SGK1信号级联增加应激抑郁模型大鼠海马学习记忆的功能,进而实现抗抑郁的作用。
文摘目的:观察熟地有效成分甘露三糖对高浓度皮质酮诱导海马神经元损伤及学习记忆相关信号转导分子表达的影响。方法:用24小时新生大鼠原代培养海马神经细胞。第八天用药处理,将细胞分为5组:1:正常对照组;2:1×10-4M高浓度皮质酮模型组;3:10-4 M CORT+3nM RU38486拮抗剂组;4:10-4 M CORT+0.1g/L甘露三糖组;5:10-4 M CORT+40uM多奈哌齐阳性对照组。24h后,通过SYTO13-PI双荧光染色观察各组细胞的形态及存活情况,MTT法测定细胞活性,Western Blot检测学习记忆信号转导通路相关分子GCR、BDNF、SGK蛋白表达。结果:与正常组相比,模型组细胞死亡较多,细胞活力下降,GCR、BDNF、SGK蛋白表达均显著性降低;如上改变又均被皮质酮受体拮抗剂RU38486逆转;甘露三糖(0.1g/L)同阳性对照多奈派齐(40μM)均明显减少死细胞数量,提高细胞活力,提高模型细胞GCR、BDNF、SGK蛋白表达。结论:熟地有效成分甘露三糖保护大鼠海马神经细胞免遭高浓度皮质酮的损伤,通过调节学习记忆信号转导途径中的重要蛋白GCR、BDNF、SGK表达从而改善高浓度皮质酮致学习记忆功能退化。
文摘目的探讨MAL2(myelin and lymphocyte protein 2)在肺癌中的功能及作用机制。方法分析TCGA数据中MAL2在肺癌组织中的表达情况,在H1299和A549细胞中用克隆形成实验检测MAL2基因敲减后对肺癌细胞增殖的影响,探讨MAL2在肺腺癌细胞增殖中的作用,并用CCK8等细胞增殖、转移、凋亡实验进一步验证其功能;将MAL2干扰后,采用Western blot实验检测下游通路基因表达情况以分析MAL2基因可能的作用机制。结果MAL2基因在肺癌组织与癌旁组织相比表达显著升高(P<0.05),且MAL2敲减后抑制细胞增殖和转移、促进细胞凋亡(P<0.05);MAL2可能通过调控SGK1通路发挥作用。结论MAL2基因在肺癌的恶性进展中起重要的作用,该作用通过调控SGK1机制进行,MAL2基因是肺癌的一个新的潜在的分子诊断指标及治疗靶点。
