Downregulation of Serum PTEN Expression in Mercury-Exposed Population and PI3K/AKT Pathway-Induced Inflammation

Objective This study investigated the impact of occupational mercury(Hg) exposure on human gene transcription and expression, and its potential biological mechanisms.Methods Differentially expressed genes related to Hg exposure were identified and validated using gene expression microarray analysis and extended validation. Hg-exposed cell models and PTEN lowexpression models were established in vitro using 293T cells. PTEN gene expression was assessed using qRT-PCR, and Western blotting was used to measure PTEN, AKT, and PI3K protein levels. IL-6 expression was determined by ELISA.Results Combined findings from gene expression microarray analysis, bioinformatics, and population expansion validation indicated significant downregulation of the PTEN gene in the high-concentration Hg exposure group. In the Hg-exposed cell model(25 and 10 μmol/L), a significant decrease in PTEN expression was observed, accompanied by a significant increase in PI3K, AKT, and IL-6 expression.Similarly, a low-expression cell model demonstrated that PTEN gene knockdown led to a significant decrease in PTEN protein expression and a substantial increase in PI3K, AKT, and IL-6 levels.Conclusion This is the first study to report that Hg exposure downregulates the PTEN gene, activates the PI3K/AKT regulatory pathway, and increases the expression of inflammatory factors, ultimately resulting in kidney inflammation.

Hypoglycemic mechanism of Tegillarca granosa polysaccharides on type 2 diabetic mice by altering gut microbiota and regulating the PI3K-akt signaling pathwaye

Type 2 diabetes mellitus(T2DM)is a complex metabolic disease threatening human health.We investigated the effects of Tegillarca granosa polysaccharide(TGP)and determined its potential mechanisms in a mouse model of T2DM established through a high-fat diet and streptozotocin.TGP(5.1×10^(3) Da)was composed of mannose,glucosamine,rhamnose,glucuronic acid,galactosamine,glucose,galactose,xylose,and fucose.It could significantly alleviate weight loss,reduce fasting blood glucose levels,reverse dyslipidemia,reduce liver damage from oxidative stress,and improve insulin sensitivity.RT-PCR and Western blotting indicated that TGP could activate the phosphatidylinositol-3-kinase/protein kinase B signaling pathway to regulate disorders in glucolipid metabolism and improve insulin resistance.TGP increased the abundance of Allobaculum,Akkermansia,and Bifidobacterium,restored the microbiota abundance in the intestinal tracts of mice with T2DM,and promoted short-chain fatty acid production.This study provides new insights into the antidiabetic effects of TGP and highlights its potential as a natural hypoglycemic nutraceutical.

Spi1 regulates the microglial/macrophage inflammatory response via the PI3K/AKT/mTOR signaling pathway after intracerebral hemorrhage

Preclinical and clinical studies have shown that microglia and macrophages participate in a multiphasic brain damage repair process following intracerebral hemorrhage.The E26 transformation-specific sequence-related transcription factor Spi1 regulates microglial/macrophage commitment and maturation.However,the effect of Spi1 on intracerebral hemorrhage remains unclear.In this study,we found that Spi1 may regulate recovery from the neuroinflammation and neurofunctional damage caused by intracerebral hemorrhage by modulating the microglial/macrophage transcriptome.We showed that high Spi1expression in microglia/macrophages after intracerebral hemorrhage is associated with the activation of many pathways that promote phagocytosis,glycolysis,and autophagy,as well as debris clearance and sustained remyelination.Notably,microglia with higher levels of Soil expression were chara cterized by activation of pathways associated with a variety of hemorrhage-related cellular processes,such as complement activation,angiogenesis,and coagulation.In conclusion,our results suggest that Spi1 plays a vital role in the microglial/macrophage inflammatory response following intracerebral hemorrhage.This new insight into the regulation of Spi1 and its target genes may advance our understanding of neuroinflammation in intracerebral hemorrhage and provide therapeutic targets for patients with intracerebral hemorrhage.

基于PTEN/PI3K/Akt信号通路探讨糖通饮对糖尿病肾病大鼠足细胞损伤的保护作用

目的探讨糖通饮对糖尿病肾病(DN)大鼠足细胞损伤的保护作用。方法高脂高糖饮食联合链脲佐菌素(STZ)多次注射建立DN大鼠模型,并随机分为模型组、厄贝沙坦组(13.5mg/kg)和糖通饮低、中、高剂量组(930、1860、3720mg/kg),另设置正常组,各组灌胃给予相应剂量药物,每天1次,连续8周。检测各组大鼠空腹血糖(FBG)、24h尿蛋白(24-hUP)、血肌酐(Scr)、血尿素氮(BUN)水平,HE染色观察各组大鼠肾组织病理形态学改变,透射电子显微镜观察肾足细胞超微结构变化,RT-qPCR法检测肾组织PTEN、PI3K、Akt、nephrin、podocin、CD2APmRNA表达,Westernblot法检测肾组织PTEN、PI3K、Akt、p-Akt、nephrin、podocin、CD2AP蛋白表达。结果与模型组比较,糖通饮各剂量组FBG、24-hUP、Scr、BUN水平降低(P<0.05,P<0.01),肾组织病理形态和足细胞超微结构得到改善,PTEN、nephrin、podocin、CD2APmRNA和蛋白表达升高(P<0.05,P<0.01),p-Akt蛋白表达降低(P<0.05,P<0.01);糖通饮中、高剂量组PI3K、AktmRNA和蛋白表达降低(P<0.05,P<0.01)。结论糖通饮对DN大鼠肾组织足细胞有保护作用,其机制可能与调节PTEN/PI3K/Akt信号通路有关。

基于PTEN与JAK对PI3K/Akt信号通路的影响探讨加味小蓟饮子治疗急性放射性膀胱炎的作用机制

目的:探讨加味小蓟饮子治疗急性放射性膀胱炎的作用机制。方法:50只ICR小鼠随机分为空白组12只和造模组38只。采用X射线辐照仪造模,造模后随机取2只小鼠处死鉴定模型是否成功。将剩余造模组小鼠随机分为模型组、小蓟饮子组和加味小蓟饮子组各12只。空白组、模型组、小蓟饮子组、加味小蓟饮子组分别予0.9%氯化钠、0.9%氯化钠、小蓟饮子药液,加味小蓟饮子药液灌胃处理,每日1次,连续7 d。末次灌胃后24 h处死取材,酶联免疫吸附实验(ELISA)法检测血清超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)、总抗氧化能力(T-AOC)含量。Western Blot法检测小鼠膀胱丝氨酸/苏氨酸激酶(AKT)、E钙粘着蛋白(E-Cadherin)、内皮素-1(ET-1)、JAK激酶(JAK)、核转录因子红系2相关因子2(Nrf2)、磷脂酰肌醇3-激酶(PI3K)、人第10号染色体缺失的磷酸酶(PTEN)、Smad蛋白2(Smad2)、Smad蛋白3(Smad3)、转化生长因子β1(TGF-β1)、尿斑蛋白3(Upk3)、血管内皮生长因子(VEGF)蛋白相对表达水平;实时荧光免疫定量-聚合酶链式反应法(RT-PCR)法检测小鼠膀胱微RNA-21(MicroRNA-21,miR-21)、PTEN、JAK、PI3K、AKT、Nrf2信使核糖核酸(mRNA)相对表达水平。结果:相较模型组,加味小蓟饮子组小鼠体内SOD、GSH-Px、T-AOC、AKT、E-Cadherin、JAK、Nrf2、PI3K、Upk3含量升高(P<0.05),MDA、ET-1、PTEN、Smad2、Smad3、TGF-β1、VEGF含量下降(P<0.05);与模型组和小蓟饮子组比较,加味小蓟饮子miR-21 mRNA表达升高(P<0.05)。结论:加味小蓟饮子组可能通过上调miR-21表达,同调PTEN、JAK蛋白以激活下游PI3K/AKT/Nrf2信号通路治疗急性放射性膀胱炎。

Melatonin improves synapse development by PI3K/Akt signaling in a mouse model of autism spectrum disorder

Autism spectrum disorders are a group of neurodevelopmental disorders involving more than 1100 genes,including Ctnnd2 as a candidate gene.Ctnnd2knockout mice,serving as an animal model of autis m,have been demonstrated to exhibit decreased density of dendritic spines.The role of melatonin,as a neuro hormone capable of effectively alleviating social interaction deficits and regulating the development of dendritic spines,in Ctnnd2 deletion-induced nerve injury remains unclea r.In the present study,we discove red that the deletion of exon 2 of the Ctnnd2 gene was linked to social interaction deficits,spine loss,impaired inhibitory neurons,and suppressed phosphatidylinositol-3-kinase(PI3K)/protein kinase B(Akt) signal pathway in the prefrontal cortex.Our findings demonstrated that the long-term oral administration of melatonin for 28 days effectively alleviated the aforementioned abnormalities in Ctnnd2 gene-knockout mice.Furthermore,the administration of melatonin in the prefro ntal cortex was found to improve synaptic function and activate the PI3K/Akt signal pathway in this region.The pharmacological blockade of the PI3K/Akt signal pathway with a PI3K/Akt inhibitor,wo rtmannin,and melatonin receptor antagonists,luzindole and 4-phenyl-2-propionamidotetralin,prevented the melatonin-induced enhancement of GABAergic synaptic function.These findings suggest that melatonin treatment can ameliorate GABAe rgic synaptic function by activating the PI3K/Akt signal pathway,which may contribute to the improvement of dendritic spine abnormalities in autism spectrum disorders.

黄芩清热除痹胶囊通过PTEN/PI3K/AKT信号通路改善痛风性关节炎大鼠的炎症反应及尿酸、脂质代谢失衡

目的探究黄芩清热除痹胶囊(HQC)对痛风性关节炎(GA)大鼠炎症反应及尿酸、脂质代谢的影响,并探讨其可能机制。方法随机将60只SD大鼠分为6组:正常组,模型组,HQC低、中、高剂量组,秋水仙碱组(n=10);以单钠尿酸盐注射右踝关节腔进行GA大鼠造模,HQC低、中、高剂量组及秋水仙碱组予相应剂量药物灌胃,模型组予等量生理盐水,连续7 d。造模后4、8、24、48、72 h检测各组大鼠足趾肿胀度;HE染色法进行滑膜组织学分析;ELISA检测血清中白细胞介素(IL)-10、IL-18、肿瘤坏死因子(TNF)-α、转化生长因子(TGF)-β1、脂联素(APN)、瘦素(LP)、抵抗素(RS)、内脂素(VF)的表达,检测滑膜中APN、LP、RS、VF的表达;生化法检测血清中高密度脂蛋白胆固醇(HDL-C)、甘油三酯(TG)、总胆固醇(TC)、血尿酸(BUA)的水平;RT-qPCR检测滑膜中磷酸酶与紧张素同源物(PTEN)、磷脂酰肌醇-3-激酶(PI3K)、蛋白激酶B(AKT)mRNA的表达;Western blotting检测PTEN、PI3K、p-PI3K、AKT、p-AKT蛋白表达。结果与正常组比较,模型组足趾肿胀度升高(P<0.05),且在48 h达到高峰;HE染色显示大量炎性细胞浸润和滑膜组织增生;TNF-α、TGF-β1、IL-18、TC、TG、LP、RS、VF、BUA、p-PI3K、p-AKT的表达增加(P<0.05),IL-10、APN、HDL-C、PTEN的表达降低(P<0.05)。与模型组比较,HQC和秋水仙碱可降低TNF-α、TGF-β1、IL-18、TC、TG、LP、RS、VF、BUA、p-PI3K、p-AKT的表达(P<0.05),升高IL-10、APN、HDL-C、PTEN的表达(P<0.05),减轻滑膜组织病理损伤,改善足趾肿胀度。结论HQC可改善GA大鼠炎症反应及尿酸、脂质代谢失衡,可能与抑制PTEN/PI3K/AKT信号通路有关。

阿魏酸通过调节PTEN/PI3K/AKT信号通路抑制急性T淋巴细胞白血病进展

目的探究阿魏酸是否可通过调控PTEN/PI3K/AKT信号通路在体内外抑制急性T淋巴细胞白血病进展。方法将急性T淋巴细胞白血病Jurkat细胞分为对照组、阿魏酸处理组和LY294002处理组进行体外实验,对照组正常培养;阿魏酸处理组分别给予不同浓度(1.25、2.5、5、10、20、40、80、160µmol/L)阿魏酸,采用CCK-8法检测细胞增殖能力,筛选实验浓度;LY294002处理组给予50µmol/L PI3K/AKT抑制剂LY294002,采用克隆形成实验、流式细胞术、Transwell实验检测细胞增殖、凋亡、侵袭情况,采用Western blot检测核蛋白Ki67、增殖细胞核抗原(PCNA)、cleaved caspase-3、cleaved caspase-9、E-cadherin、N-cadherin、Vimentin、PTEN、p-PI3K、PI3K、p-AKT和AKT蛋白相对表达量。使用30只雄性BALB/c裸鼠建立移植瘤裸鼠模型,平均分为正常组和阿魏酸处理组进行体内实验,正常组接种Jurkat细胞后以生理盐水灌胃,阿魏酸处理组接种Jurkat细胞后以75 mg/kg阿魏酸灌胃,比较移植瘤质量和体积变化,并检测肿瘤组织中Ki67、cleaved caspase-3/caspase-3、E-cadherin、N-cadherin、PTEN、p-PI3K、PI3K、p-AKT和AKT水平。结果体外实验中,与对照组比较,5、10、20µmol/L阿魏酸处理组和LY294002处理组细胞克隆形成率、细胞侵袭数、Ki67、PCNA、N-cadherin、Vimentin、p-PI3K/PI3K、p-AKT/AKT明显降低/减少(P<0.05),细胞凋亡率、cleaved caspase-3/caspase-3、cleaved caspase-9/caspase-9、E-cadherin、PTEN明显升高(P<0.05)。体内实验中,与正常组比较,阿魏酸处理组裸鼠肿瘤质量减轻,肿瘤体积减小,肿瘤组织中Ki67、N-cadherin、p-PI3K/PI3K、p-AKT/AKT明显降低,cleaved caspase-3/caspase-3、E-cadherin、PTEN明显升高,差异均有统计学意义(P<0.05)。结论阿魏酸在体内外均可抑制急性T淋巴细胞白血病Jurkat细胞的增殖及侵袭,并诱导细胞凋亡,其作用机制可能与调控PTEN/PI3K/AKT信号通路有关。

Alleviatory effect of isoquercetin on benign prostatic hyperplasia via IGF-1/PI3K/Akt/mTOR pathway

We evaluated the effect of isoquercetin(quercetin-O-3-glucoside-quercetin,IQ)as a functional component of Abeliophyllum disistichum Nakai ethanol extract(ADLE)on prostate cell proliferation and apoptosis and its effects on the IGF-1/PI3K/Akt/mTOR pathway in benign prostatic hyperplasia(BPH).Metabolites in ADLE were analyzed using UHPLC-qTOF-MS and HPLC.IQ was orally administered(1 or 10 mg/kg)to a testosterone propionate-induced BPH rat model,and its effects on the prostate weight were evaluated.The effect of IQ on androgen receptor(AR)signaling was analyzed in LNCaP cells.Whether IGF-1 and IQ affect the IGF-1/PI3K/Akt/mTOR pathway in BPH-1 cells was also examined.The metabolites in ADLE were identified and quantified,which confirmed that ADLE contained abundant IQ(20.88 mg/g).IQ significantly reduced the prostate size in a concentration-dependent manner in a BPH rat model,and significantly decreased the expression of AR signaling factors in the rat prostate tissue and LNCaP cells in a concentration-dependent manner.IQ also inhibited the PI3K/AKT/mTOR pathway activated by IGF-1 treatment in BPH-1 cells.In BPH-1 cells,IQ led to G0/G1 arrest and suppressed the expression of proliferation factors while inducing apoptosis.Thus,IQ shows potential for use as a pharmaceutical and nutraceutical for BPH.

Liqi Huoxue dripping pill protects against myocardial ischemia-reperfusion injury via the PI3K/Akt/GSK-3β signaling pathway in rats

Background:Liqi Huoxue dripping pill(LQHXDP),a traditional Chinese drug for coronary heart disease,has a protective effect on the heart of rats with myocardial ischemia-reperfusion injury(MIRI)in previous studies;however,its mechanism of action remains unclear.The purpose of this study was to investigate the protective mechanism of LQHXDP on MIRI in rats and its relationship with the PI3K/Akt signaling pathway.Methods:In this study,Sprague-Dawley rats were pre-infused with LQHXDP(175 mg/kg/d)for 10 days.PI3K inhibitor LY294002(0.3 mg/kg)was intravenously injected 15 minutes before ischemia.The rat model of MIRI was established by ligating the left anterior descending coronary artery.Subsequently,cardiac hemodynamics,serum myocardial injury markers,inflammatory factors,myocardial infarct size,antioxidant indexes,myocardial histopathology,and phosphorylation levels of key proteins of PI3K/Akt signaling pathway were assessed in rats.Results:LQHXDP was found to improve cardiac hemodynamic indexes,reduce serum creatine kinase MB isoenzyme activity and cardiac troponin and heart-type fatty acid binding protein levels,lower serum interleukin-1 beta,interleukin-6 and tumour necrosis factorαlevels,reduce the myocardial infarct size and enhance the antioxidant capacity of myocardial tissue in MIRI rats.Pathological analysis revealed that LQHXDP attenuated the extent of myocardial injury and protected mitochondria from damage in MIRI rats.Immunoblot analysis revealed that LQHXDP increased the expression levels of p-Akt and p-GSK-3βin MIRI rat cardiomyocytes.PI3K inhibitor LY294002 could impair these effects of LQHXDP.Conclusion:LQHXDP attenuated myocardial injury,attenuated oxidative stress injury and reduced inflammatory response in MIRI rats,and its protective effects were mediated by activating of PI3K/Akt/GSK-3βsignaling pathway.

Zuo Gui Wan Promotes Osteogenesis via PI3K/AKT Signaling Pathway:Network Pharmacology Analysis and Experimental Validation

Objective Osteogenesis is vitally important for bone defect repair,and Zuo Gui Wan(ZGW)is a classic prescription in traditional Chinese medicine(TCM)for strengthening bones.However,the specific mechanism by which ZGW regulates osteogenesis is still unclear.The current study is based on a network pharmacology analysis to explore the potential mechanism of ZGW in promoting osteogenesis.Methods A network pharmacology analysis followed by experimental validation was applied to explore the potential mechanisms of ZGW in promoting the osteogenesis of bone marrow mesenchymal stem cells(BMSCs).Results In total,487 no-repeat targets corresponding to the bioactive components of ZGW were screened,and 175 target genes in the intersection of ZGW and osteogenesis were obtained.And 28 core target genes were then obtained from a PPI network analysis.A GO functional enrichment analysis showed that the relevant biological processes mainly involve the cellular response to chemical stress,metal ions,and lipopolysaccharide.Additionally,KEGG pathway enrichment analysis revealed that multiple signaling pathways,including the phosphatidylinositol-3-kinase/protein kinase B(PI3K/AKT)signaling pathway,were associated with ZGW-promoted osteogensis.Further experimental validation showed that ZGW could increase alkaline phosphatase(ALP)activity as well as the mRNA and protein levels of ALP,osteocalcin(OCN),and runt related transcription factor 2(Runx 2).What’s more,Western blot analysis results showed that ZGW significantly increased the protein levels of p-PI3K and p-AKT,and the increases of these protein levels significantly receded after the addition of the PI3K inhibitor LY294002.Finally,the upregulated osteogenic-related indicators were also suppressed by the addition of LY294002.Conclusion ZGW promotes the osteogenesis of BMSCs via PI3K/AKT signaling pathway.

FGF2 promotes the chemotherapy resistance in colon cancer cells through activating PI3K/Akt signaling pathway

Background:To investigate the role of fibroblast growth factor 2(FGF2)in chemotherapy resistance of colon cancer.Methods:An HCT116/5-fluorouracil(5-FU)-resistant cell line was established,and FGF2 levels were detected in a sensitive cell group(HCT116)and a resistant cell group(HCT1116-R)using different methods.Fibroblast growth factor 2 levels in the medium were determined by enzyme-linked immunoassay.The protein expressions of FGF2,fibroblast growth factor receptor 1(FGFR1),and phospho-FGFR1 were assessed by Western blotting,and FGF2 mRNA levels were detected by quantitative real-time polymerase chain reaction.Fibroblast growth factor 2 recombinant protein was added to sensitive cells,and FGFR inhibitor AZD4547 was added to resistant cells,and the cell survival rate was determined using the cell counting kit-8 method and the protein expressions of PI3K(phosphatidylinositol 3 kinase),p-PI3K(phospho-PI3K),Akt(protein kinase B),p-Akt(phospho-Akt),mammalian target of rapamycin(mTOR),p-mTOR(phospho-mTOR),Bad(Bcl-xL/Bcl-2-associated death promoter),NF-κB(nuclear factorκB),GSK-3(glycogen synthase kinase-3),FKHR(forkhead box protein O1),and PTEN(phosphatase and tensin homolog deleted on chromosome ten)were detected by Western blotting.Results:Fibroblast growth factor 2 protein and mRNA expression levels in the HCT116-R group were significantly higher than those in the HCT116 group.Fibroblast growth factor 2 increased the survival rate of HCT116 cells;improved tolerance to 5-FU;upregulated p-PI3K,p-Akt,and p-mTOR;and downregulated Bad.The FGFR inhibitor AZD4547 decreased cell survival rate and tolerance to 5-FU;downregulated p-PI3K,p-Akt,and p-mTOR expression;and upregulated Bad.Conclusions:Fibroblast growth factor 2 promotes chemotherapy tolerance in colon cancer cells by activating the Akt/mTOR and Akt/Bad signaling pathways downstream of PI3K.

XB130 inhibits healing of diabetic skin ulcers through the PI3K/Akt signalling pathway

BACKGROUND Diabetic skin ulcers,a significant global healthcare burden,are mainly caused by the inhibition of cell proliferation and impaired angiogenesis.XB130 is an adaptor protein that regulates cell proliferation and migration.However,the role of XB130 in the development of diabetic skin ulcers remains unclear.AIM To investigate whether XB130 can regulate the inhibition of proliferation and vascular damage induced by high glucose.Additionally,we aim to determine whether XB130 is involved in the healing process of diabetic skin ulcers,along with its molecular mechanisms.METHODS We conducted RNA-sequencing analysis to identify the key genes involved in diabetic skin ulcers.We investigated the effects of XB130 on wound healing using histological analyses.In addition,we used reverse transcription-quantitative polymerase chain reaction,Western blot,terminal deoxynucleotidyl transferasemediated dUTP nick end labeling staining,immunofluorescence,wound healing,and tubule formation experiments to investigate their effects on cellular processes in human umbilical vein endothelial cells(HUVECs)stimulated with high glucose.Finally,we performed functional analysis to elucidate the molecular mechanisms underlying diabetic skin ulcers.RESULTS RNA-sequencing analysis showed that the expression of XB130 was up-regulated in the tissues of diabetic skin ulcers.Knockdown of XB130 promoted the healing of skin wounds in mice,leading to an accelerated wound healing process and shortened wound healing time.At the cellular level,knockdown of XB130 alleviated high glucose-induced inhibition of cell proliferation and angiogenic impairment in HUVECs.Inhibition of the PI3K/Akt pathway removed the proliferative effects and endothelial protection mediated by XB130.CONCLUSION The findings of this study indicated that the expression of XB130 is up-regulated in high glucose-stimulated diabetic skin ulcers and HUVECs.Knockdown of XB130 promotes cell proliferation and angiogenesis via the PI3K/Akt signalling pathway,which accelerates the healing of diabetic skin ulcers.

Simiao Wan alleviates obesity-associated insulin resistance via PKCε/IRS-1/PI3K/Akt signaling pathway based on network pharmacology analysis and experimental validation

Background:The purpose of the study was to investigatethe active ingredients and potential biochemicalmechanisms of Simiao Wan(SMW)in obesity-associated insulin resistance.Methods:An integrated network pharmacology method to screen the active compoundsand candidate targets,construct the protein-protein-interaction network,and ingredients-targets-pathways network was constructed for topological analysis to identify core targets and main ingredients.To find the possible signaling pathways,enrichment analysis was performed.Further,a model of insulin resistance in HL-7702 cells was established to verify the impact of SMW and the regulatory processes.Results:An overall of 63 active components and 151 candidate targets were obtained,in which flavonoids were the main ingredients.Enrichment analysis indicated that the PI3K-Akt signaling pathway was the potential pathway regulated by SMW in obesity-associated insulin resistance treatment.The result showed that SMW could significantly ameliorate insulin sensitivity,increase glucose synthesis and glucose utilization and reduce intracellular lipids accumulation in hepatocytes.Also,SMW inhibited diacylglycerols accumulation-induced PKCεactivity and decreased its translocation to the membrane.Conclusion:SMW ameliorated obesity-associated insulin resistance through PKCε/IRS-1/PI3K/Akt signaling axis in hepatocytes,providing a new strategy for metabolic disease treatment.

The role and research progress of PI3K/AKT signaling pathway in non-traumatic osteonecrosis of the femoral head

Non-traumatic osteonecrosis of the femoral head(NONFH)is one of the most common orthopedic diseases,influenced by multiple signaling pathways and inflammatory factors.The PI3K/AKT signaling pathway is closely related to various biological processes such as apoptosis,autophagy,and metabolism in cells.Increasing evidence suggests that it plays an important role in the development of femoral head necrosis.This paper aims to explore the mechanism of the PI3K/AKT signaling pathway in the pathogenesis of NONFH by analyzing its regulation of lipid metabolism,cell apoptosis and autophagy,and intravascular coagulation.This study provides new insights for the research of NONFH.

过表达CBX7调控PTEN/Akt信号通路干预上皮-间充质转化抑制胃癌细胞迁移、侵袭

目的探讨染色体盒蛋白同源物7(CBX7)对胃癌细胞上皮-间充质转化(EMT)、迁移、侵袭的影响及相关作用机制。方法将CBX7过表达质粒和阴性对照质粒、CBX7 siRNA和阴性对照siRNA转染至体外培养的人胃癌细胞SGC7901。采用qRT-PCR和Western blot法检测细胞中CBX7的mRNA和蛋白表达,CCK-8法检测细胞增殖情况,细胞划痕实验检测细胞迁移情况,Transwell小室检测细胞侵袭情况,Western blot法检测细胞中EMT相关蛋白和磷酸酶张力蛋白同源物基因(PTEN)/蛋白激酶B(Akt)信号通路相关蛋白表达。结果与pcDNA-NC组比较,pcDNA-CBX7组SGC7901细胞存活率、迁移率及侵袭数均显著下降/减少,细胞中E-钙黏蛋白(E-cadherin)、PTEN蛋白表达量均显著升高,N-钙黏蛋白(N-cadherin)和波形蛋白(Vimentin)蛋白表达量、p-Akt/Akt均显著下降,差异有统计学意义(P<0.05)。与si-NC组比较,si-CBX7组SGC7901细胞存活率、迁移率及侵袭数均显著升高/增加,细胞中E-cadherin、PTEN蛋白表达量均显著下降,N-cadherin、Vimentin蛋白表达量和p-Akt/Akt均显著升高,差异有统计学意义(P<0.05)。结论过表达CBX7可通过抑制EMT进程抑制胃癌细胞迁移、侵袭,其作用机制可能与调控PTEN/Akt信号通路有关。

基于PTEN/PI3K/Akt/mTOR信号通路探讨淫羊藿苷联合地塞米松对阿霉素诱导足细胞损伤的保护作用及机制

目的探讨淫羊藿苷联合地塞米松对阿霉素诱导足细胞损伤的保护作用及分子机制。方法体外培养大鼠肾小球足细胞,阿霉素诱导建立足细胞损伤模型,分为:空白组、模型组、淫羊藿苷联合地塞米松组及氯喹组。免疫荧光法测定足细胞特异性骨架蛋白synaptopodin的表达;转染mRFP-GFP-LC3双荧光自噬指示体系,激光共聚焦显微镜观察足细胞内自噬流的强弱;透射电镜观察足细胞内自噬体、自噬溶酶体的变化;Western Blot法检测足细胞自噬相关蛋白LC3-Ⅱ、p62及自噬信号通路蛋白PTEN、p-PI3K、p-Akt、p-mTOR的表达。结果与模型组比,淫羊藿苷联合地塞米松组的synaptopodin的表达升高(P<0.05);足细胞内自噬流水平、自噬体及自噬溶酶体数量均增加(P<0.01);LC3-Ⅱ、PTEN的表达上调(P<0.01),但p62、pPI3K、p-Akt、p-mTOR的表达下调(P<0.01)。与淫羊藿苷联合地塞米松组比,氯喹组synaptopodin的表达降低(P<0.05);足细胞内自噬流的水平降低;自噬体数量增加(P<0.05),自噬溶酶体的数量减少(P<0.01);且LC3-Ⅱ和p62表达同步升高(P<0.05,P<0.01),但PTEN、p-PI3K、p-Akt、p-mTOR的表达无明显变化(P>0.05)。结论淫羊藿苷联合地塞米松可能通过激活PTEN,抑制PI3K/Akt/mTOR信号通路,增强足细胞自噬水平,达到保护足细胞的作用。

miR-21靶向PTEN调控AKT/FoxO1信号通路对胃癌细胞凋亡的机制研究

目的:探究miR-21靶向PTEN调控AKT/FoxO1信号通路对胃癌细胞凋亡的影响。方法:将胃癌SGC-7901细胞分为miR-NC inhibitors组(SGC-7901细胞中转染miR-NC inhibitors质粒)、miR-21 inhibitors组(SGC-7901细胞中转染miR-21 inhibitors质粒)、miR-21 inhibitors+sh-PTEN组(SGC-7901细胞中转染miR-21 inhibitors和sh-PTEN质粒);Control组(不转染任何质粒的SGC-7901细胞)。qRT-PCR检测细胞中miR-21 mRNA表达;采用CCK-8、Transwell小室法和流式细胞仪检测细胞增殖、侵袭能力和凋亡率;蛋白质印迹检测细胞中PTEN、AKT、p-AKT、PI3K、p-PI3K、FoxO1蛋白表达;双荧光素酶报告检测miR-21和PTEN的靶向关系。结果:人正常胃黏膜上皮细胞GES-1(1.00±0.10)相比,胃癌细胞SGC-7901中miR-21(1.89±0.17)表达明显升高(P<0.05)。Control组相比,miR-21 inhibitors组细胞中miR-21表达(0.83±0.10)、增殖率(45.31±4.92)%、侵袭数目(62.34±5.83)个和p-PI3K/PI3K(0.42±0.05)、p-AKT/AKT(0.51±0.05)比值均明显降低,细胞凋亡率(23.48±3.51)%、PTEN(0.98±0.10)和FoxO1(0.76±0.08)蛋白表达明显升高(P<0.0001)。pcDNA-NC组相比,pcDNA-PTEN组细胞增殖率(48.26±5.01)、侵袭细胞数(65.37±6.02)个和细胞中p-PI3K/PI3K(0.46±0.05)、p-AKT/AKT(0.55±0.06)比值均明显降低,细胞凋亡率(25.61±3.27)%及细胞中PTEN(0.91±0.09)和FoxO1(0.70±0.08)蛋白表达升高(P<0.05)。预测发现PTEN的3'UTR端与miR-21有碱基互补结合点位。miR-NC组相比,转染野生型PTEN(PTEN-WT)时miR-21组(0.32±0.03)荧光素酶活性明显降低(P<0.05)。miR-21 inhibitors组相比,miR-21 inhibitors+sh-PTEN组细胞增殖率(90.25±9.14)%和侵袭细胞数(125.69±10.31)个和细胞中p-PI3K/PI3K(0.80±0.08)、p-AKT/AKT(0.76±0.08)比值明显增加,细胞凋亡率(6.24±1.32)和细胞中PTEN(0.30±0.04)、FoxO1(0.38±0.05)蛋白表达降低(P<0.0001)。结论:敲减miR-21可靶向负调控PTEN表达,调控AKT/FoxO1信号通路,抑制胃癌细胞的增殖和侵袭,促进凋亡。

Notch/PTEN/AKT信号通路在跑台运动减轻db/db小鼠肾纤维化中的作用及机制研究

目的:探讨运动对2型糖尿病模型db/db小鼠肾损伤的保护作用及机制。方法:将8周龄的雄性db/db小鼠随机分为4组:db/db组(n=8)、db/db+跑台运动(Exe)组(n=7)、db/db+Exe+Notch抑制剂二苯并氮䓬(DBZ)组(n=7)和db/db+DBZ组(n=6);同月龄雄性m/m小鼠作为阴性对照(Con)组(n=10)。db/db+DBZ组和db/db+Exe+DBZ组小鼠进行DBZ干预(灌胃8周,每周5 d,0.04 mg/kg),db/db+Exe组和db/db+Exe+DBZ组小鼠进行跑台运动(运动8周,每周5 d;db/db+Exe+DBZ组小鼠在灌胃2 h后进行运动)。HE染色和PAS染色评价肾组织形态学变化;Masson染色观察肾组织纤维化程度;试剂盒检测血尿素氮(BUN)和血清肌酐(SCr)水平;Western blot检测肾组织纤维化指标、Notch/PTEN/AKT信号通路相关蛋白和自噬相关蛋白的表达。结果:(1)与Con组相比,db/db组小鼠体重和血糖显著升高(P<0.01),运动干预后显著降低(P<0.05)。(2)与Con组相比,db/db组小鼠BUN和SCr水平显著升高(P<0.01),组织学染色显示肾组织损伤加重,运动干预后BUN和SCr水平均显著降低(P<0.01),肾组织损伤减轻。(3)Western blot结果显示,与db/db组相比,db/db+Exe组I型胶原(Col-I)和α-平滑肌肌动蛋白(α-SMA)表达显著减少(P<0.05),Col-Ⅲ和转化生长因子β_(1)(TGF-β_(1))表达有下降趋势,免疫荧光显示小鼠肾脏中α-SMA、Col-I和纤连蛋白荧光染色阳性信号显著减弱;db/db+DBZ组TGF-β_(1)蛋白表达下降(P<0.01);db/db+Exe+DBZ组Col-I、α-SMA、Col-Ⅲ和TGF-β_(1)蛋白表达有下降趋势,但差异无统计学意义。(4)与db/db组相比,db/db+Exe组和db/db+DBZ组Notch1、Hes1和Hey1表达水平及p-AKT/AKT比值均显著降低(P<0.05),PTEN蛋白表达上调(P<0.05),而db/db+Exe+DBZ组Notch1、Hes1、Hey1和p-AKT/AKT蛋白表达无显著差异,PTEN蛋白显著升高(P<0.01)。(5)与db/db组相比,db/db+Exe组和db/db+DBZ组LC3-Ⅱ/LC3-Ⅰ比值显著升高(P<0.01),p62表达显著减少(P<0.01);db/db+Exe+DBZ组LC3-Ⅱ/LC3-Ⅰ比值升高(P<0.01),p62表达无显著差异。(6)与db/db+DBZ组相比,db/db+Exe组Hey1蛋白下调(P<0.05);与db/db+Exe组相比,db/db+Exe+DBZ组PTEN蛋白上调(P<0.01)。结论:运动减轻db/db小鼠肾功能损伤和肾纤维化,其机制可能与其抑制Notch/PTEN/AKT信号通路而促进细胞自噬有关。

正肝方对二乙基亚硝胺诱导的肝癌前病变模型大鼠肝组织PTEN/PI3K/AKT信号通路的影响

目的观察正肝方对二乙基亚硝胺(Diethylnitrosamine,DEN)诱导的肝癌前病变大鼠的生化指标和PTEN/PI3K/AKT信号通路的影响,探讨其对肝癌前病变大鼠的作用机制。方法将80只大鼠按照随机数字表法分为5组:空白对照组、DEN模型组、正肝方低、中、高剂量组(2 g/kg、4 g/kg、8 g/kg),每组各16只。为建立肝癌前病变大鼠模型,除空白组外,其余各组大鼠连续16周每周1次腹腔注射DEN(50 mg/kg)。同时,从造模第1周开始,空白对照组及模型组灌喂纯水10 ml/kg,正肝方低、中、高剂量组分别灌喂相应剂量的正肝方药液,1次/d,连续灌胃至第16周。禁食不禁水16 h后测量大鼠体质量,麻醉,心脏取血,称量肝脏、胸腺重量。全自动生化法检测肝功能指标,包括丙氨酸氨基转移酶(Alanine aminotransferase,ALT)、碱性磷酸酶(Alkaline phosphatase,ALP)、白蛋白(Albumin,ALB)、γ-谷氨酰转肽酶(γ-glutamyl transpeptidase,γ-GT);HE染色观察肝组织病理改变,免疫组化和蛋白质免疫印迹法(Western blot)检测肝组织中甲胎蛋白(Alpha fetoprotein,AFP)的阳性表达水平,蛋白质免疫印迹法检测肝组织中PTEN/PI3K/AKT信号通路相关蛋白表达水平。结果与正常组比较,DEN模型组大鼠体质量明显降低,肝脏指数增加,胸腺指数下降,肝功能指标除ALB外均明显升高,HE染色可见模型组肝细胞出现不同程度的变性、坏死、排列紊乱、细胞多形性等癌前病变表现,肿瘤组织中AFP、p-PI3K、p-AKT蛋白表达水平明显升高,PTEN蛋白表达水平降低,差异有统计学意义(P<0.01);与DEN模型组比较,正肝方中、高剂量组肝脏指数均有降低,高剂量组较为明显,差异有统计学意义(P<0.05),各组胸腺指数表现出不同程度升高,差异有统计学意义(P<0.05,P<0.001),血清ALT、ALP、GGT水平降低,ALB升高,肿瘤变性坏死区减少,AFP、p-PI3K、p-AKT蛋白表达水平明显降低,差异有统计学意义(P<0.01,P<0.05),PTEN蛋白表达水平提高,差异有统计学意义(P<0.01)。结论正肝方可改善肝功能指标和肝癌前病变状况,提高机体免疫力和PTEN抑癌蛋白的表达水平,降低组织中AFP、p-PI3K、p-AKT等癌症蛋白的表达,其机制可能与抑制PTEN/PI3K/AKT通路的激活,从而抑制癌前病变的发生发展相关。