目的探究C1q/肿瘤坏死因子相关蛋白6(C1q/tumor necrosis factor related protein 6,CTRP6)在槲皮素(quercetin,Que)抑制急性心肌梗死(acute myocardial infarction,AMI)后大鼠心肌细胞焦亡中的作用及机制。方法采用冠状动脉左前降支结...目的探究C1q/肿瘤坏死因子相关蛋白6(C1q/tumor necrosis factor related protein 6,CTRP6)在槲皮素(quercetin,Que)抑制急性心肌梗死(acute myocardial infarction,AMI)后大鼠心肌细胞焦亡中的作用及机制。方法采用冠状动脉左前降支结扎法建立AMI大鼠模型。首先,将实验分为假手术组(Sham)、AMI组、槲皮素低剂量组(Que-L,25 mg·kg^(-1))、槲皮素高剂量组(Que-H,100 mg·kg^(-1))、福辛普利钠片组(fosinopril,4 mg·kg^(-1)),每组各10只。各组大鼠按照对应的药物剂量或是生理盐水灌胃,1次/d,连续14 d。多普勒超声检测心功能变化,观察大鼠心肌组织病理学变化,Western blot检测焦亡相关蛋白和CTRP6表达。通过以上实验指标检测筛选得出Que最佳给药剂量。随后,将实验分为Sham、AMI、Que(100 mg·kg^(-1))、Que+si-NC组、Que+si-CTRP6,每组各10只。干预14 d后,检测心肌梗死、心肌损伤指标、细胞焦亡、CTRP6及PI3K/Akt通路蛋白表达。结果与Sham组相比,AMI组大鼠LVEDV、LVESV明显增加,EF、FS明显降低(P<0.05),心肌组织出现明显病理损伤,纤维化程度增加,心肌梗死面积增加,LDH、CK-MB、cTnI水平,TUNEL阳性细胞比率升高,NLRP3、cleaved caspase-1、GSDMD-N、IL-1β、IL-18表达增加,CTRP6表达,p-PI3K/PI3K、p-Akt/Akt比值降低(均P<0.05)。与AMI组相比,Que-L和Que-H大鼠心功能指标以及心肌组织病理损伤程度减轻,心肌梗死面积降低,LDH、CK-MB、cTnI水平,TUNEL阳性率降低( P <0.05),细胞焦亡相关蛋白表达降低,CTRP6及PI3K/Akt通路蛋白表达增加( P <0.05),且呈剂量相关性。与Que组相比,Que+si-CTRP6组大鼠上述指标变化均得到明显逆转。 结论 Que能够抑制心肌细胞焦亡,改善AMI大鼠心肌梗死,其机制与上调CTRP6表达,促进PI3K/Akt信号通路活化相关。展开更多
Potassium-calcium activates channel subfamily N member 3(KCNN3/SK3/KCa2.3)is involved in regulating cellular calcium signaling,muscle contraction and neurotransmitter release.Dysregulation of the KCNN3 channel is asso...Potassium-calcium activates channel subfamily N member 3(KCNN3/SK3/KCa2.3)is involved in regulating cellular calcium signaling,muscle contraction and neurotransmitter release.Dysregulation of the KCNN3 channel is associated with the development of various tumors.We use bioinformatics analysis to identify whether KCNN3 regulates the occurrence and development of stomach adenocarcinoma(STAD)as a prognostic target.By analyzing the Human Protein Atlas(HPA)database and The Cancer Genome Atlas(TCGA)database,we found that the protein and mRNA levels of KCNN3 were dramatically reduced in STAD,and TCGA database showed that KCNN3 significantly correlated with the prognosis and clinical features of STAD.In addition,we found that high expression of KCNN3 in STAD reduced the IC 50 of several drugs in STAD cells,suggesting that high expression of KCNN3 correlated with the drug sensitivity of STAD.To investigate the underlying biological mechanism,we identified a potential KCNN3 interaction factor,tumor necrosis factor receptor superfamily member 7(CD27/TNFRSF7),which is expressed at low levels in STAD.RT-qPCR and Western blotting confirmed that KCNN3 and CD27 positively correlated with each other at protein and mRNA levels,and co-immunoprecipitation and immunofluorescence experiments confirmed that the two proteins interact and colocalize in the cytoplasm.Moreover,we confirmed the inhibitory effect of KCNN3 on the proliferation,migration and invasion of human STAD cells in vitro and in vivo through subcutaneous tumorigenesis and cellular experiments.Furthermore,GO/KEGG enrichment analysis showed that KCNN3 was enriched in signaling pathways regulating the immune response and calcium or metal ion transport.Lastly,we verified through cell co-culture,RT-qPCR and CCK8 assays that high expression of KCNN3 can promote the increase of T cell activating factor and the killing effect of T cells on STAD cells.Therefore,our results suggest that KCNN3 is a potential inhibitory factor affecting the occurrence and progression of STAD.展开更多
文摘Potassium-calcium activates channel subfamily N member 3(KCNN3/SK3/KCa2.3)is involved in regulating cellular calcium signaling,muscle contraction and neurotransmitter release.Dysregulation of the KCNN3 channel is associated with the development of various tumors.We use bioinformatics analysis to identify whether KCNN3 regulates the occurrence and development of stomach adenocarcinoma(STAD)as a prognostic target.By analyzing the Human Protein Atlas(HPA)database and The Cancer Genome Atlas(TCGA)database,we found that the protein and mRNA levels of KCNN3 were dramatically reduced in STAD,and TCGA database showed that KCNN3 significantly correlated with the prognosis and clinical features of STAD.In addition,we found that high expression of KCNN3 in STAD reduced the IC 50 of several drugs in STAD cells,suggesting that high expression of KCNN3 correlated with the drug sensitivity of STAD.To investigate the underlying biological mechanism,we identified a potential KCNN3 interaction factor,tumor necrosis factor receptor superfamily member 7(CD27/TNFRSF7),which is expressed at low levels in STAD.RT-qPCR and Western blotting confirmed that KCNN3 and CD27 positively correlated with each other at protein and mRNA levels,and co-immunoprecipitation and immunofluorescence experiments confirmed that the two proteins interact and colocalize in the cytoplasm.Moreover,we confirmed the inhibitory effect of KCNN3 on the proliferation,migration and invasion of human STAD cells in vitro and in vivo through subcutaneous tumorigenesis and cellular experiments.Furthermore,GO/KEGG enrichment analysis showed that KCNN3 was enriched in signaling pathways regulating the immune response and calcium or metal ion transport.Lastly,we verified through cell co-culture,RT-qPCR and CCK8 assays that high expression of KCNN3 can promote the increase of T cell activating factor and the killing effect of T cells on STAD cells.Therefore,our results suggest that KCNN3 is a potential inhibitory factor affecting the occurrence and progression of STAD.
