Qishen decoction suppresses liver fibrosis by downregulating PI3K/Akt/mTOR signal pathway

Objective:To observe the effect of Qishen decoction on PI3K/Akt/mTOR signal pathway in rats with liver fibrosis;Methods:40 SD rats were randomly divided into four groups: the control group, the model group, Qishen decoction group, and Colchicine group. Except the control group, the remaining three groups were used to establish liver fibrosis model by intraperitoneal injection of carbon tetrachloride. At the end of modeling, Qishen decoction and colchicine group were given corresponding drug gavage treatment, rats in the model group and the control group were treated with equal volume distilled water for 8 weeks. At the end of the treatment, the blood and liver tissues of rats in each group were collected, the liver function indexes and hydroxyproline content were detected by ELISA, the pathological morphology of liver tissue was detected by HE and Masson staining. Immunohistochemical method was used to detect α-SMA protein expression and Western blot was used to detect the expression of α-SMA, Col-I, Col-Ⅲ and key proteins in PI3K/Akt/mTOR signaling pathway.Results: Compared with the model group, Qishen decoction significantly reduced the levels of AST, ALT, and TBIL in serum, and reduced Hyp content, inflammatory score, fibrosis score, and collagen staining area in liver tissue, differences were statistically significant (P<0.05). At meanwhile, Qishen decoction significantly reduce the expression level of α-SMA, CoL-I and Col-Ⅲ in liver tissue(P<0.05). In addition, compared with the model group, Qishen decoction significantly down-regulated the expressions of p-PI3K, p-Akt and p-mTOR in liver tissue, difference were statistically significant (P<0.05).Conclusion: Qishen decoction suppresses liver fibrosis and inhibits the deposition of collagen in liver tissue by down-regulating PI3K/Akt/mTOR signal pathway.

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.

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.

Thymoquinone affects hypoxia-inducible factor-1αexpression in pancreatic cancer cells via HSP90 and PI3K/AKT/mTOR pathways

BACKGROUND Pancreatic cancer(PC)is associated with some of the worst prognoses of all major cancers.Thymoquinone(TQ)has a long history in traditional medical practice and is known for its anti-cancer,anti-inflammatory,anti-fibrosis and antioxidant pharmacological activities.Recent studies on hypoxia-inducible factor-1α(HIF-1α)and PC have shown that HIF-1αaffects the occurrence and development of PC in many aspects.In addition,TQ could inhibit the development of renal cancer by decreasing the expression of HIF-1α.Therefore,we speculate whether TQ affects HIF-1αexpression in PC cells and explore the mechanism.AIM To elucidate the effect of TQ in PC cells and the regulatory mechanism of HIF-1αexpression.METHODS Cell counting kit-8 assay,Transwell assay and flow cytometry were performed to detect the effects of TQ on the proliferative activity,migration and invasion ability and apoptosis of PANC-1 cells and normal pancreatic duct epithelial(hTERTHPNE)cells.Quantitative real-time polymerase chain reaction and western blot assay were performed to detect the expression of HIF-1αmRNA and protein in PC cells.The effects of TQ on the HIF-1αprotein initial expression pathway and ubiquitination degradation in PANC-1 cells were examined by western blot assay and co-immunoprecipitation.RESULTS TQ significantly inhibited proliferative activity,migration,and invasion ability and promoted apoptosis of PANC-1 cells;however,no significant effects on hTERT-HPNE cells were observed.TQ significantly reduced the mRNA and protein expression levels of HIF-1αin PANC-1,AsPC-1,and BxPC-3 cells.TQ significantly inhibited the expression of the HIF-1αinitial expression pathway(PI3K/AKT/mTOR)related proteins,and promoted the ubiquitination degradation of the HIF-1αprotein in PANC-1 cells.TQ had no effect on the hydroxylation and von Hippel Lindau protein mediated ubiquitination degradation of the HIF-1αprotein but affected the stability of the HIF-1αprotein by inhibiting the interaction between HIF-1αand HSP90,thus promoting its ubiquitination degradation.CONCLUSION The regulatory mechanism of TQ on HIF-1αprotein expression in PC cells was mainly to promote the ubiquitination degradation of the HIF-1αprotein by inhibiting the interaction between HIF-1αand HSP90;Secondly,TQ reduced the initial expression of HIF-1αprotein by inhibiting the PI3K/AKT/mTOR pathway.

益智仁-乌药药对调控PI3K/Akt/mTOR通路介导细胞自噬保护肾小球足细胞的作用机制研究

目的研究益智仁-乌药药对通过调控PI3K/Akt/mTOR信号通路促进足细胞自噬治疗糖尿病肾病(Diabetic Nephropathy,DN)的作用。方法60只造模成功的C57BL/KSJ-db/db(以下简称db/db)小鼠随机分为模型组、二甲双胍组、缬沙坦组、益智仁-乌药药对(低、中、高剂量)组,每组10只;另取10只C57BL/KSJ-db/m(以下简称db/m)小鼠为正常组,正常组和模型组给予生理盐水,治疗组小鼠分别给予相应药物,给药8周后检测小鼠肾脏病理学改变,足细胞自噬体数量、结构及相关蛋白表达。结果与模型组相比,益智仁-乌药药对组可显著减轻糖尿病肾病小鼠肾小球基底膜增厚情况,增加足细胞自噬体数量,显著升高自噬相关蛋白表达(P<0.05),降低PI3K/Akt/mTOR信号通路相关蛋白的表达(P<0.05)。其中益智仁-乌药药对高剂量组各指标改善优于益智仁-乌药低、中剂量组。结论益智仁-乌药药对通过抑制PI3K/Akt/mTOR信号通路激活,提高足细胞自噬水平,减轻足细胞损伤,发挥治疗糖尿病肾病的作用。

异莲心碱通过PI3K/Akt/mTOR信号通路影响结肠癌SW480细胞增殖、凋亡和自噬

目的:探讨异莲心碱(Iso)通过PI3K/Akt/mTOR信号通路对结肠癌SW480细胞增殖、凋亡和自噬的影响。方法:用10、20和40μmol/L的Iso处理结肠癌SW480细胞,CCK-8法、流式细胞术和WB法分别检测Iso对细胞增殖活力、凋亡和自噬相关蛋白LC3Ⅰ、LC3Ⅱ、p62表达的影响。然后,用20μmol/L的Iso和25μmol/L的PI3K激活剂740 Y-P分别处理SW480细胞,将细胞分为对照组、740 Y-P组、Iso组和Iso+740 Y-P组,流式细胞术、WB法检测Iso和740 Y-P对各组细胞凋亡及细胞中LC3Ⅰ、LC3Ⅱ、p62、PI3K、p-PI3K、mTOR和p-mTOR蛋白表达的影响。结果:10、20和40μmol/L的Iso处理后,SW480细胞增殖活力均显著下降(均P<0.05),细胞凋亡率均显著升高(均P<0.05),LC3Ⅱ/LC3Ⅰ表达均显著上调(均P<0.05),p26蛋白表达显著下调(P<0.05)。Iso和740 Y-P处理后,与对照组相比,740 Y-P组细胞凋亡率、LC3Ⅱ/LC3Ⅰ表达均显著下降(均P<0.05),p26、p-PI3K/PI3K和p-mTOR/mTOR表达均显著升高(均P<0.05);Iso组细胞凋亡率、LC3Ⅱ/LC3Ⅰ表达升高(均P<0.05),p26、p-PI3K/PI3K和p-mTOR/mTOR表达均显著下降(均P<0.05);与740 Y-P组相比,Iso+740 Y-P组细胞凋亡率、LC3Ⅱ/LC3Ⅰ表达升高(P<0.05),p26、p-PI3K/PI3K和p-mTOR/mTOR表达均显著下降(均P<0.05);与Iso组相比,Iso+740 Y-P组细胞凋亡率、LC3Ⅱ/LC3Ⅰ表达下降(均P<0.05),p26、p-PI3K/PI3K和p-mTOR/mTOR表达均显著升高(均P<0.05)。结论:Iso通过抑制PI3K/Akt/mTOR信号通路抑制结肠癌SW480细胞增殖并诱导细胞凋亡和自噬。

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.

栀子苷调节PI3K/AKT/mTOR信号通路在动脉粥样硬化形成过程中对Th17/Treg功能的影响

目的:观察栀子苷对载脂蛋白E缺乏(ApoE^(-/-))小鼠Th17/调节性T(Treg)细胞失衡的影响及其作用机制。方法:将50只纯合子ApoE^(-/-)雌性小鼠随机分为对照组、模型组和栀子苷低剂量组、栀子苷中剂量组、栀子苷高剂量组。对照组小鼠喂养普通饲料,模型组和栀子苷组小鼠喂养高脂饲料。从第8周开始,栀子苷各剂量组每日灌胃栀子苷(25、50、100 mg/kg),连续8周。试验结束时,采用油红O染色评估主动脉及其根部动脉粥样硬化(AS)病变面积比。采用定量逆转录聚合酶链式反应(RT-PCR)分析主动脉组织肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6、IL-17A和IL-10 mRNA表达;采用流式细胞仪分析脾脏中Th17和Treg细胞百分比;蛋白免疫印迹法(Western Blot)检测主动脉组织磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路相关蛋白表达。结果:油红O染色病变显示,栀子苷中剂量组、栀子苷高剂量组病变百分比低于模型组(P<0.05)。与对照组比较,模型组主动脉TNF-α、IL-6和IL-17A mRNA表达水平升高(P<0.05);栀子苷各剂量组主动脉TNF-α、IL-6和IL-17A mRNA表达水平降低(P<0.05)。与对照组比较,模型组主动脉抗炎细胞因子IL-10 mRNA表达水平降低(P<0.05);栀子苷各剂量组主动脉抗炎细胞因子IL-10 mRNA表达水平升高(P<0.05)。与对照组比较,模型组小鼠脾脏中Th17细胞百分比升高,Treg细胞百分比降低(P<0.05)。栀子苷处理恢复了AS小鼠Th17和Treg细胞的平衡。栀子苷抑制PI3K的表达及AKT和mTOR的磷酸化,MHY1485(mTOR活化剂)减弱了栀子苷对T细胞分化的影响。结论:栀子苷抗AS作用机制可能与抑制PI3K/AKT/mTOR信号引起的Treg细胞增多和Th17细胞减少有关。

ROR2 promotes invasion and chemoresistance of triple-negative breast cancer cells by activating PI3K/AKT/mTOR signaling

Objective:This study aimed to investigate the role of receptor tyrosine kinase-like orphan receptor 2(ROR2)in triple-negative breast cancer(TNBC).Methods:ROR2 expression in primary TNBC and metastatic TNBC tissues was analyzed by immunohistochemical staining and PCR.ROR2 expression in TNBC cell lines was detected by PCR and Western blot analysis.The migration,invasion and chemosensitivity of TNBC cells with overexpression or knockdown of ROR2 were examined.Results:ROR2 expression was high in metastatic TNBC tissues.ROR2 knockdown suppressed the migration,invasion and chemoresistance of TNBC cells.ROR2 overexpression in MDA-MB-435 cells promoted the migration,invasion,and chemoresistance.Moreover,ROR2 knockdown in HC1599 and MDA-MB-435 adriamycin-resistant cells enhanced chemosensitivity to adriamycin.ROR2 could activate PI3K/AKT/mTOR signaling in TNBC cells.Conclusion:ROR2 is upregulated and promotes metastatic phenotypes of TNBC by activating PI3K/AKT/mTOR signaling.

MicroRNA (let-7b-5p)-targeted DARS2 regulates lung adenocarcinoma growth by PI3K/AKT signaling pathway

Background:The aberrant intraellular expression of a mitochondrial aspartyl tRNA synthetase 2(DARS2)has been reported in human cancers.Nevertheless its critical role and detailed mechanism in lung adenocarcinoma(LUAD)remain unexplored.Methods:Initially,The Cancer Genome Atlas(TCGA)based Gene Expression Profiling Interactive Analysis(GEPIA)database (http:/gepia.cancer-pku.cn/)was used to analyze the prognostic relevance of DARS2 expression in LUAD.Further,cell counting kit(CCK)8,immunostaining,and transwell invasion assays in LUAD cell lines in vitro,as well as DARS2 silence on LUAD by tumorigenicity experiments in wivo in nude mice,were performed.Besides,we analyzed the expression levels of p-PI3K(phosphorylated Phosphotylinosital3 kinase),PI3K,AKT(Protein Kinase B),p-AKT(phosphorylated Protein Kinase B),PCNA(proliferating cell nudear antigen),cleaved-caspase 3,E cadherin,and N-cadherin proteins using the Westem blot analysis.Results:LUAD tissues showed higher DARS2 expression compared to normal tissues.Upregulation of DARS2 could be related to Tumor-Node-Metastasis(TNM)stage,high lymph node metastasis,and inferior prognosis.DARS2 silence decreased the proliferation,migration,and invasion abilities of LUAD cells.In addition,the DARS2 downregulation decreased the PCNA and N-cadherin expression and increased cleaved:caspase 3 and E cadherin expressions in LUAD cells,coupled with the inactivation of the PI3K/AKT signaling pathway.Moreover,DARS2 silence impaired the tumonigenicity of LUAD in vivo.Interestingly,let:7b-5p could recognize DARS2 through a complementary sequence.Mechanistically,the increased let 7b 5p expression attenuated the promo oncogenic action of DARS2 during LUAD progression,which were inversely correlated to each other in the LUAD tssues Conclusion:In summary,let 7b-5p,downregulated DARS2 expression,regulating the progression of LUAD cells by the PI3K/AKT signaling pathway.

调补心肾方通过活化PI3K/Akt/mTOR通路促进阿尔茨海默病5xFAD转基因小鼠突触可塑性相关蛋白的合成

目的探讨调补心肾方(党参、制首乌、枸杞子、黄芪等)对阿尔茨海默病5xFAD转基因小鼠突触可塑性的影响及机制。方法取5月龄雄性野生型(WT)小鼠和5xFAD转基因小鼠各18只,分别随机分为对照组(0.9%NaCl)、调补心肾方组(颗粒剂,4.18 g·kg^(-1))、安理申组(盐酸多奈哌齐,0.625 mg·kg^(-1)),每组6只。按照上述分组灌胃给药,每日1次,共60 d。采用透射电镜观察小鼠海马组织超微结构;Western Blot法检测小鼠皮层组织中Synaptophsin、PSD-95、p-NMDAR1、NMDAR1、p-CaMKⅡa、CaMKⅡa、PI3K、p-Akt、Akt、p-mTOR、mTOR蛋白表达水平。结果与WT对照组比较,5xFAD对照组小鼠海马CA1区突触的超微结构不规则,线粒体萎缩、减少,线粒体嵴断裂、消失,突触膜弯曲不规则,突触囊泡减少,突触后致密物(PDS)变薄甚至断裂;皮层组织中Synaptophsin、PSD-95、p-NMDAR1/NMDAR1、p-CaMKⅡa/CaMKⅡa、PI3K、p-Akt/Akt、p-mTOR/mTOR蛋白表达均明显下调(P<0.05)。与5xFAD对照组相比较,5xFAD调补心肾方组小鼠海马CA1区突触的超微结构有明显变化,线粒体数量增加,突触囊泡增多,突触膜完整,突触后致密物有增厚现象;皮层组织中Synaptophsin、PSD-95、p-NMDAR1/NMDAR1、p-CaMKⅡa/CaMKⅡa、PI3K、p-Akt/Akt、p-mTOR/mTOR蛋白表达明显上调(P<0.05)。结论调补心肾方可能通过活化PI3K/Akt/mTOR通路,促进突触可塑性相关蛋白合成,进而改善AD的认知功能障碍。

Acalypha australis L.extract inhibits B16 melanoma cell metastasis through PI3K/AKT signaling pathway

Background:Melanoma is a deadly skin tumor resulting from the malignant transformation of melanocytes.It is highly malignant and invasive,with the highest mortality rate among skin cancers.Acalypha australis L.(AAL),a plant with dual medicinal and culinary purposes,is commonly regarded as an edible wild vegetable in southern China.Additionally,AAL has a long history of medicinal use in China,often employed for its hemostatic,anti-diarrheal,and anti-inflammatory properties.Modern pharmacology has demonstrated that AAL possesses functions such as weight loss,antimicrobial activity,antiviral effects,and treatment for ulcerative colitis.However,there is currently no research available regarding its effectiveness and mechanisms of action on melanoma.Methods:In this investigation,we used methyl thiazolyl tetrazolium assay to detect cell viability,transwell assay to detect cell migration and invasion ability,and Western blot assay to detect relevant signaling pathways.Results:The present study reveals that 2 mg/mL AAL effectively suppresses the metastasis of B16 cells,while simultaneously triggering the expression of key apoptosis-related proteins,including Bcl-2,Bax,and cleaved caspased 3.Subsequent investigations demonstrate that AAL exerts this inhibitory effect via the PI3K/AKT signal transduction pathway,as evidenced by the observed deficits in Ras,AKT,p-AKT,and PI3K expression levels.Conclusion:These findings indicated that AAL could be a valuable therapeutic option for reducing the metastatic potential of B16 melanoma cells.

茯苓酸调控PI3K/AKT/mTOR通路介导葡萄糖代谢促进结直肠癌细胞凋亡的机制研究

目的 研究茯苓酸对结直肠癌细胞葡萄糖代谢及细胞凋亡的调控作用。方法 通过细胞活性测定(Cell Counting Kit-8,CCK8)、克隆形成等实验检测茯苓酸对结直肠癌细胞增殖与细胞活性的影响;采用Annexin V-FITC/PI双染流式细胞凋亡实验检测茯苓酸对结直肠癌细胞凋亡的影响;使用细胞氧气消耗率和细胞外酸化率测定仪器及细胞葡萄糖代谢水平测定试剂盒检测茯苓酸对结直肠癌细胞葡萄糖代谢的影响;通过蛋白免疫印迹(Western blotting, WB)实验验证凋亡、糖酵解、磷脂酰肌醇3-激酶(Phosphatidylinositol3-kinase, PI3K)/丝氨酸、苏氨酸蛋白激酶(AKT Serine/Threonine Kinase 1,AKT)/雷帕霉素靶蛋白(Mammalian Target of Rapamycin, mTOR)通路相关蛋白表达情况。结果 茯苓酸能够有效抑制结直肠癌细胞增殖与细胞活性,并能够通过抑制其糖酵解水平促进其细胞凋亡,同时显著抑制PI3K/AKT/mTOR通路相关蛋白表达水平。结论 茯苓酸可通过调控PI3K/AKT/mTOR通路介导葡萄糖代谢促进CRC细胞凋亡,可作为未来结直肠癌代谢靶向治疗的潜在药物。

布比卡因通过PI3K/AKT/mTOR通路对膀胱癌细胞凋亡和铁死亡的影响

目的 探讨布比卡因对膀胱癌细胞凋亡和铁死亡的影响及其机制。方法 常规培养人膀胱癌细胞(T24和5637)和人尿路上皮细胞(SV-HUC-1),MTT实验检测布比卡因细胞毒性,筛选合适处理浓度,将T24和5637细胞分为对照组、0.25、0.5和1 mmol/L布比卡因组、布比卡因+铁死亡激动剂(Erastin)组、布比卡因+铁死亡抑制剂(Fer-1)组细胞和布比卡因+PI3K激动剂(740Y-P)组。采用流式细胞术检测细胞凋亡率,相应试剂盒检测Fe^(2+)、丙二醛(MDA)、谷胱甘肽(GSH)和活性氧(ROS)水平,Western blot检测Bcl-2、Bax、细胞色素C、xCT、GPX4和磷脂酰肌醇3/蛋白激酶B/哺乳动物雷帕霉素靶蛋白(PI3K/AKT/mTOR)相关蛋白表达。结果 0.25、0.5、1、2、4、8、16 mmol/L布比卡因均可明显抑制T24和5637细胞的活性(P<0.001),选择仅对癌细胞有毒性的0.25、0.5和1 mmol/L布比卡因进行后续研究。与对照组比较,随着布比卡因浓度增加,T24和5637细胞凋亡率、Fe^(2+)、ROS和MDA水平逐渐升高,GSH水平逐渐降低,差异有统计学意义(P<0.001);与1 mmol/L布比卡因组比较,布比卡因+Erastin组Fe^(2+)水平升高,而布比卡因+Fer-1组降低,差异有统计学意义(P<0.05)。Western blot结果显示,与对照组比较,布比卡因组细胞色素C蛋白表达增高,Bax/Bcl-2、xCT、GPX4表达,以及PI3K p85α/PI3K、p-AKT(Thr308)/AKT、p-AKT(Ser473)/AKT和p-mTOR(Ser2448)/mTOR比值降低,差异有统计学意义(P<0.001)。验证实验结果显示,与对照组比较,布比卡因+740Y-P组细胞活力和GSH水平升高,Fe^(2+)水平和细胞色素C蛋白表达降低(P<0.001)。结论 布比卡因可通过抑制PI3K/AKT/mTOR信号通路激活诱导膀胱癌细胞凋亡和铁死亡。

大黄素对精神分裂症大鼠海马组织PI3K/Akt/mTOR信号通路的影响

目的:探讨大黄素调节磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路对精神分裂症(SP)大鼠神经损伤及认知功能的影响。方法:通过注射地卓西平(MK-801)建立SP大鼠模型,并将正常大鼠为对照组,SP大鼠随机分为SP组、不同剂量(20、40、60mg/kg)大黄素组、60mg/kg大黄素+LY294002组,刻板行为及共济失调评分标准评价大鼠行为学功能;Morris水迷宫实验评估大鼠认知功能;试剂盒检测脑组织乙酰胆碱酯酶(AchE)、乙酰胆碱(Ach)活性;HE检测海马组织病理学变化;qRT-PCR检测脑组织中脑源性神经营养因子(BDNF)mRNA、钙依赖性蛋白激酶(CaMKⅡ)mRNA表达;Western blot检测PI3K/Akt/mTOR通路相关蛋白表达。结果:与对照组相比,SP组穿越平台次数、Ach活性、BDNF mRNA、CaMKⅡmRNA表达、p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR表达显著降低,刻板行为及共济失调评分、平均逃避潜伏期、AchE活性显著增加(P<0.05);但不同剂量大黄素干预后,穿越平台次数、Ach活性、BDNF mRNA、CaMKⅡmRNA表达、p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR表达显著增加,刻板行为及共济失调评分、平均逃避潜伏期、AchE活性显著降低,组间有差异(P<0.05);60mg/kg大黄素联合LY294002较60mg/kg大黄素组穿越平台次数、Ach活性、BDNF mRNA、CaMKⅡmRNA表达、p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR表达显著降低,刻板行为及共济失调评分、平均逃避潜伏期、AchE活性显著增加(P<0.05)。结论:大黄素通过调节PI3K/Akt/mTOR信号通路改善SP大鼠认知能力,发挥神经保护功能。

PI3K/AKT/mTOR信号通路在膀胱癌中的作用

目的:分析磷脂酰肌醇3激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白(PI3K/AKT/mTOR)信号通路在膀胱癌中的作用。方法:选取60只SD健康雄性大鼠,采用随机数字法分为对照组、模型组和干预组,每组各20只。模型组和干预组大鼠以致癌物BBN灌胃8周建立膀胱癌模型,建模成功后,干预组大鼠注射PI3K抑制剂,连续干预6周。观察三组大鼠膀胱组织病理学变化,比较各组大鼠血清炎性因子指标、氧化应激指标、PI3K/AKT/mTOR信号通路蛋白表达和CK-19、CYFRA21-1水平的变化。结果:与对照组相比,模型组和干预组大鼠血清转化生长因子-β1(TGF-β1)、白细胞介素-1β(IL-1β)、C反应蛋白(CRP)、丙二醛、一氧化氮、PI3K、AKT、mTOR蛋白表达量、CK-19、CYFRA21-1水平升高,总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)水平降低(P<0.05)。与模型组相比,干预组血清TGF-β1、IL-1β、CRP、丙二醛、一氧化氮、PI3K、AKT、mTOR蛋白表达量、CK-19、CYFRA21-1水平均降低,T-AOC、SOD水平升高(P<0.05)。结论:PI3K抑制剂通过PI3K/AKT/mTOR信号通路遏制膀胱癌的发展。

不同强度运动抑制糖尿病大鼠肾脏PI3K/AKT/mTOR信号通路改善自噬的比较

背景:2型糖尿病损害肾功能。研究表明运动干预可以保护肾脏;鸢尾素可以通过抑制磷脂酰肌醇3-激酶/蛋白激酶B/雷帕霉素靶蛋白信号通路恢复自噬,保护糖尿病肾病患者的肾功能。目的:探讨运动能否通过抑制肾脏磷脂酰肌醇3-激酶/蛋白激酶B/雷帕霉素靶蛋白信号通路过度激活来恢复自噬,改善肾损伤,以及分析不同方式运动产生影响的差异。方法:将6周龄的SD大鼠随机分为空白对照组(正常大鼠)和糖尿病组,其中糖尿病组大鼠经过高脂高糖喂养加腹腔注射低剂量1%链脲佐菌素(30 mg/kg)建立2型糖尿病模型。造模成功后再将糖尿病组大鼠随机分成糖尿病模型组、中强度持续运动组和高强度间歇运动组。两个运动组大鼠分别进行8周不同强度运动干预。取材后采用葡萄糖氧化酶法检测大鼠空腹血糖,使用试剂盒检测糖化血红蛋白水平,Elisa法检测血清胰岛素浓度,计算胰岛素抵抗指数,RT-PCR检测肾组织磷脂酰肌醇3-激酶、蛋白激酶B、雷帕霉素靶蛋白、Beclin-1、podocin、nephrin的基因表达量,Western Blot检测肾组织雷帕霉素靶蛋白及自噬标记蛋白LC3-1、LC3-2、Beclin-1的蛋白表达量。结果与结论:①2型糖尿病大鼠空腹血糖和糖化血红蛋白水平极显著性升高,胰岛素抵抗水平显著上升,胰岛素水平显著下降;两种运动均能使2型糖尿病大鼠空腹血糖和糖化血红蛋白水平极显著下降,胰岛素抵抗水平显著下降,胰岛素水平显著上升;与中强度持续运动组相比,高强度间歇运动组胰岛素水平显著上升。②2型糖尿病大鼠podocin、nephrin基因表达量显著降低;两种不同形式运动均能显著提高其表达;与高强度间歇运动组相比,中等强度持续性运动组足细胞相关蛋白基因表达有进一步上升趋势,但无显著性差异。③2型糖尿病大鼠肾组织磷脂酰肌醇3-激酶、蛋白激酶B、mTORC1的mRNA及蛋白的表达量显著增加,自噬标志蛋白Beclin-1、LC3-2表达量以及LC3-2/LC3-1显著降低;两种不同形式运动均能使肾组织磷脂酰肌醇3-激酶、蛋白激酶B、mTORC1的mRNA及雷帕霉素靶蛋白蛋白的表达量显著降低,自噬标志蛋白Beclin-1、LC3-2以及LC3-2/LC3-1显著升高;与中等强度持续性运动组相比,高强度间歇运动的磷脂酰肌醇3-激酶、蛋白激酶B、mTORC1的mRNA及雷帕霉素靶蛋白的蛋白表达量有进一步下降的趋势,Beclin-1、LC3-2以及LC3-2/LC3-1有进一步升高的趋势,但仅Beclin-1有显著性差异。④结果说明2型糖尿病肾脏足细胞损伤,自噬受到抑制,与磷脂酰肌醇3-激酶/蛋白激酶B/mTORC1信号通路被异常激活密切相关。高强度间歇运动和中等强度持续性运动可以保护糖尿病肾脏,减少足细胞损伤,促进自噬恢复,这可能与运动抑制磷脂酰肌醇3-激酶/蛋白激酶B/雷帕霉素靶蛋白信号通路过度激活有关。与中等强度持续性运动相比,高强度间歇运动恢复自噬的效果呈更优趋势,但足细胞蛋白表达稍有下降。

布托啡诺调节PI3K/AKT/mTOR信号通路对膀胱癌细胞恶性生物学行为的影响

目的:探讨布托啡诺调节磷脂酰肌醇3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白(PI3K/AKT/mTOR)信号通路对膀胱癌细胞恶性生物学行为的影响。方法:体外培养膀胱癌细胞T24;将细胞分为对照组、布托啡诺低剂量组(1 ng/mL)、布托啡诺中剂量组(10 ng/mL)、布托啡诺高剂量组(100 ng/mL)、激活剂组(100 ng/mL布托啡诺+PI3K激动剂Recilisib 10μmol/L)、抑制剂组(100 ng/mL布托啡诺+PI3K抑制剂LY294002 50μmol/L)。CCK-8实验检测细胞增殖;流式细胞术检测细胞凋亡;Transwell检测细胞迁移和侵袭;Western blot检测细胞PI3K/AKT/mTOR信号通路相关蛋白表达;并构建异种移植肿瘤模型,称量肿瘤质量,计算肿瘤体积。结果:与对照组比较,布托啡诺低剂量、布托啡诺中剂量、布托啡诺高剂量组T24细胞OD_(450)值和迁移、侵袭细胞数目及p-PI3K、p-AKT、p-mTOR蛋白表达显著降低,细胞凋亡率显著升高,且呈剂量依赖性(P<0.05);激活剂减弱了布托啡诺抑制T24细胞增殖、迁移和侵袭,促进细胞凋亡的作用,抑制剂加强了布托啡诺抑制T24细胞增殖、迁移和侵袭,促进细胞凋亡的作用。与裸鼠对照组相比,裸鼠布托啡诺低剂量、裸鼠布托啡诺中剂量、裸鼠布托啡诺高剂量组肿瘤体积和质量显著减小,且呈现剂量依赖性(P<0.05);与裸鼠布托啡诺高剂量组比较,裸鼠激活剂组肿瘤体积、肿瘤质量显著增大(P<0.05),裸鼠抑制剂组肿瘤体积、肿瘤质量显著减小(P<0.05)。结论:布托啡诺通过抑制PI3K/AKT/mTOR信号通路抑制膀胱癌细胞增殖、迁移和侵袭。

基于PI3K/AKT/mTOR通路研究脑清通颗粒对脑缺血再灌注大鼠的保护作用

目的:探讨脑清通颗粒对脑缺血再灌注大鼠的保护作用及机制。方法:60只大鼠随机分为假手术对照组、模型对照组、尼莫地平组、脑清通颗粒低、中、高剂量组,每组10只。除假手术对照组外,其余各组采用高脂饲料喂养、复合应激刺激和线栓法制备肝热痰瘀证脑缺血再灌注大鼠模型。从第6周起,假手术对照组和模型对照组用等体积生理盐水灌胃,其余各给药组用相应药物灌胃,1次/d,连续给药7d。观察大鼠体征表现;采用神经功能评分、TTC染色及脑梗死体积测定评价脑缺血再灌注模型;尼氏染色观察神经细胞变化;生化法检测SOD和MDA含量、ELISA法检测TNF-α和IL-6含量;免疫组化检测PI3K、AKT和mTOR的蛋白表达;Western blot法检测p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR的蛋白表达。结果:模型对照组大鼠体征表现符合临床肝热痰瘀证基本特征,神经功能评分、脑梗死体积显著增加(P<0.01),大鼠血清SOD含量明显降低(P<0.01),MDA、TNF-α和IL-6含量明显升高(P<0.01),脑组织PI3K、AKT和mTOR的蛋白表达均明显降低(P<0.01或P<0.05),p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR的蛋白表达均明显降低(P<0.01);脑清通颗粒1.3g/kg、2.6g/kg、5.2g/kg组可改善大鼠体征表现,降低神经功能评分和脑梗死体积(P<0.01或P<0.05),升高血清SOD含量(P<0.01),降低MDA、TNF-α和IL-6含量(P<0.01),升高脑组织PI3K、AKT和mTOR的蛋白表达(P<0.01),升高p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR的蛋白表达(P<0.01)。结论:脑清通颗粒可能是通过激活PI3K/AKT/mTOR信号通路达到对脑缺血再灌注损伤的脑保护作用。

山奈酚通过激活PI3K/AKT/mTOR信号通路改善视网膜氧化应激损伤

目的探讨山奈酚减轻视网膜氧化应激损伤的作用机制。方法H_(2)O_(2)处理ARPE-19细胞,构建体外视网膜氧化应激损伤模型,CCK-8评估细胞活力,流式细胞仪检测细胞凋亡。25只雄性SD大鼠随机分为5组:对照组、模型组及山奈酚低剂量(12.5 mg/kg)组、中剂量(25 mg/kg)组和高剂量(50 mg/kg)组;对照组动物正常饲养,模型组和治疗组动物给予急性视网膜缺血/再灌注损伤处理,构建视网膜氧化应激损伤模型。检测超氧化物歧化酶(SOD)和丙二醛(MDA)的细胞内活性,H-E染色观察视网膜结构完整性,酶联免疫吸附试验(ELISA)检测炎性细胞因子的表达,Western blot评价凋亡自噬的表达和磷脂酰肌醇3激酶(PI3K)/丝氨酸/苏氨酸激酶(AKT)/雷帕霉素机制靶蛋白(mTOR)信号通路的影响。结果体外细胞实验表明,与对照组相比,山奈酚能显著提高ARPE-19氧化应激损伤细胞的活力和减少细胞凋亡(P<0.05)。体内实验表明,与模型组相比,山奈酚能减轻视网膜氧化应激损伤;显著降低MDA表达和升高SOD表达(P<0.05);显著降低血管内皮生长因子(VEGF)和γ-干扰素(IFN-γ)的分泌,增加白细胞介素-10(IL-10)的分泌(P<0.05);同时能显著增强B细胞淋巴瘤/白血病-2(BCL-2)、微管相关蛋白1A/1B-轻链3(LC3)、PI3K、磷酸化mTOR(p-mTOR)和磷酸化AKT(p-AKT)蛋白表达,抑制Bcl-2同源结构域蛋白1抗体(Beclin1)表达(P<0.05)。结论山奈酚通过激活PI3K/AKT/mTOR信号通路减轻视网膜氧化应激损伤。