Page 75 - 《中国药房》2025年3期
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1 000                                                              mitochondrial translation  68(8.65%)
                      921
                                                                   positive regulation of transcription by RNA polymerase Ⅱ  46(5.85%)
                                                                      regulation of transcription by RNA polymerase Ⅱ  45(5.73%)
                               4                                                  translation      38(4.83%)
            750                                                    negative regulation of transcription by RNA polymerase Ⅱ  36(4.58%)
                   667                                上调:254                    signal transduction  31(3.94%)
                                                                                apoptotic process
           差异蛋白数/个  500   上调  -log 10 P  2            无差异:7 763      positive regulation of transcription, DNA-templated  21(2.67%)  biological
                                                      下调:667
                                                                                                   23(2.93%)
                                                                        regulation of transcription, DNA-templated
                                                                                                   21(2.67%)
                                                                              protein phosphorylation
                                                                                                         process
                          下调
                          总数
                                                                                                   21(2.67%)
                                                                                  cell adhesion
                                                                                                   21(2.67%)
                 254                                                     positive regulation of apoptotic process  21(2.67%)
            250                                                      negative regulation of transcription, DNA-templated  19(2.42%)
                               0                                              innate immune response  18(2.29%)
                                   -4     -2            0             2            4  cell differentiation  18(2.29%)
             0                           log 2 FC                           mitochondrial inner membrane  53(6.21%)
                                                                                                   51(5.98%)
                                                                               mitochondrial matrix
              A.差异蛋白筛选结果               B.火山图                                    extracellular region  48(5.63%)
                                                                                 Golgi apparatus
                                                                                                   43(5.04%)
                                                                                extracellular space  41(4.81%)
              图3 GAA组与对照组的差异蛋白分析结果                                                 chromatin       39(4.57%)
                                                                          mitochondrial large ribosomal subunit  38(4.45%)
                                                                        intracellular membrane-bounded organelle  31(3.63%)
                                                                           endoplasmic reticulum membrane  30(3.52%)  cellular
                                       线粒体:290(31.49%)                       protein-containing complex  27(3.17%)  component
                                       细胞核:204(22.15%)                            cell surface     27(3.17%)
                                       细胞外基质:138(14.98%)                 integral component of plasma membrane  121(14.19%)
                                       细胞质:129(14.01%)                          Golgi membrane     96(11.25%)
                                       细胞膜:106(11.51%)                   mitochondrial small ribosomal subunit  23(2.7%)
                                       细胞质核:23(2.5%)                        perinuclear region of cytoplasm  21(2.46%)
                                                                                neuronal cell body
                                       过氧化物酶体:12(1.3%)                         lysosomal membrane  20(2.34%)
                                                                                                   20(2.34%)
                                       内质网:8(0.87%)                                dendrite        19(2.23%)
                                       细胞骨架:4(0.43%)                      mitochondrial intermembrane space  18(2.11%)
                                       高尔基体:2(0.22%)                       structural constituent of ribosome  58(7.95%)
                                       线粒体核:2(0.22%)                              DNA binding      44(6.03%)
                                       内质网线粒体:1(0.11%)        RNA polymerase Ⅱ cis-regulatory region sequence-specific DNA binding  42(5.75%)
                                       溶酶体:1(0.11%)                              zinc ion binding  40(5.48%)
                                       线粒体过氧化物酶体:1(0.11%)      DNA-binding transcription factor activity,RNA polymerase Ⅱ-specific  38(5.21%)
                                                                           protein homodimerization activity  34(4.66%)
                                                              DNA-binding transcription activator activity, RNA polymerase Ⅱ-specific  28(3.84%)  molecular
                                                                               calcium ion binding
                   图4 差异蛋白的亚细胞定位结果                                       DNA-binding transcription factor activity  27(3.7%)  function
                                                                                                   25(3.42%)
                                                                                  GTP binding      24(3.29%)
                                                                                 enzyme binding    23(3.15%)
          3.2.4 差异蛋白的GO分析和KEGG信号通路分析结果                                sequence-specific double-stranded DNA binding  22(3.01%)
                                                                               protein kinase binding  20(2.74%)
                                                                                chromatin binding
              选取GO分析中P值排前50名的条目绘制富集条目                                         magnesium ion binding  20(2.74%)
                                                                                                   19(2.6%)
                                                                              ATP hydrolysis activity  18(2.47%)
          柱形图(见图 5A),由图 5A 可知,生物过程主要集中在
                                                                                      0            5           10         15              20
          线粒体蛋白翻译、RNA 聚合酶Ⅱ对转录的调控、蛋白翻                                                        Percent/%
                                                                                   A. GO分析
          译、凋亡、细胞粘连及分化;细胞组分主要集中在线粒体
                                                                          apoptosis       14(3.09%)
                                                                          lysosome
          膜、基质、染色质、细胞膜;分子功能主要集中在核糖体                                    cellular senescence  11(2.43%)  cellular processes
                                                                                          11(2.43%)
                                                                         motor proteins   10(2.21%)
          的构建、转录因子与 DNA 的结合等。选取排前 50 名的                             PI3K-Akt signaling pathway  22(4.86%)
                                                                 cytokine-cytokine receptor interaction  16(3.53%)
          KEGG 信号通路绘制富集条目柱形图(见图 5B),由图                               MAPK signaling pathway  15(3.31%)
                                                                                          13(2.87%)
                                                                      TNF signaling pathway
                                                                   NF-kappa B signaling pathway  12(2.65%)  environmental information processing
          5B可知,信号通路主要为PI3K/AKT(phosphoinositide 3-                   AMPK signaling pathway  11(2.43%)
                                                                      Ras signaling pathway
                                                                                          10(2.21%)
                                                                          Ribosome        39(8.61%)
          kinase/protein kinase B,磷脂酰肌醇3激酶/蛋白激酶B)、                  aminoacyl-tRNA biosynthesis  12(2.65%)  genetic information processing
                                                              pathways of neurodegeneration-multiple diseases  46(10.15%)
          MAPK(mitogen-activated protein kinase,促分裂原活化              amyotrophic lateral sclerosis  42(9.27%)
                                                                                          41(9.05%)
                                                                       Alzheimer disease
                                                                       huntington disease  40(8.83%)
                                                                         prion disease
          的蛋白质激酶)、TNF(tumor necrosis factor,肿瘤坏死因                       Parkinson disease  37(8.17%)
                                                                                          37(8.17%)
                                                             chemical carcinogenesis -reactive oxygen species  35(7.73%)
          子)、NF-κB(nuclear factor κB,NF-κB)、AMPK(AMP-                diabetic cardiomyopathy  34(7.51%)
                                                                   non-alcoholic fatty liver disease  31(6.84%)
                                                                       pathways in cancer
          activated protein kinase,AMP 活化的蛋白质激酶)、Ras             human T-cell leukemia virus 1 infection  26(5.74%)
                                                                                          21(4.64%)
                                                                      lipid and atherosclerosis  18(3.97%)
          信号通路,主要涉及细胞凋亡、衰老、运动等细胞过程。                                  epstein-Barr virus infection  18(3.97%)  human diseases
                                                                                          17(3.75%)
                                                                       viral carcinogenesis
          3.3 GAA对差异蛋白表达的影响结果                                      herpes simplex virus 1 infection  17(3.75%)
                                                                   human papillomavirus infection
                                                                                          15(3.31%)
                                                                 transcriptional misregulation in cancer  14(3.09%)
              选取 FC 排前 3 名的蛋白 NDRG1、ERRFI1、CXCL3                  fluid shear stress and atherosclerosis  13(2.87%)
                                                                       Salmonella infection
                                                                                          13(2.87%)
                                                                  central carbon metabolism in cancer  13(2.87%)
          进行验证。与正常组比较,模型组细胞中 NDRG1、                              pathogenic Escherichia coli infection  12(2.65%)
                                                                      microRNAs in cancer  12(2.65%)
          CXCL3 蛋白表达水平均显著升高,ERRFI1 蛋白表达水                      Kaposi sarcoma-associated herpesvirus infection  12(2.65%)
                                                                      alcoholic liver disease
                                                                                          12(2.65%)
                                                               chemical carcinogenesis-receptor activation  11(2.43%)
          平显著降低(P<0.05);与模型组比较,阳性对照组和                                    breast cancer    11(2.43%)
                                                                     oxidative phosphorylation  31(6.84%)
          GAA各浓度组细胞中NDRG1、CXCL3蛋白表达水平均                               biosynthesis of cofactors  17(3.75%)  metabolism
                                                                       carbon metabolism
                                                                                          13(2.87%)
                                                                      fatty acid metabolism  10(2.21%)
          显著降低(P<0.05),ERRFI1 蛋白表达水平均显著升高                           biosynthesis of amino acids  10(2.21%)
                                                                                          43(9.49%)
                                                                         thermogenesis
                                                                 retrograde endocannabinoid signaling  22(4.86%)
         (P<0.05)。与阳性对照组和GAA高浓度组比较,GAA                               osteoclast differentiation  13(2.87%)  organismal systems
                                                                 NOD-like receptor signaling pathway  12(2.65%)
                                                                     IL-17 signaling pathway
          中、低浓度组细胞中CXCL3蛋白表达水平均显著升高,                             complement and coagulation cascades  12(2.65%)
                                                                                          10(221%)
          ERRFI1 蛋白表达水平显著降低(P<0.05)。与阳性对                                      0              4              8               12              16
                                                                                    Percent/%
          照组比较,GAA高、中、低浓度组细胞中NDRG1蛋白表                                             B. KEGG分析
          达水平均显著升高(P<0.05)。结果见图6。                            图5 差异蛋白的GO分析和KEGG信号通路分析结果
          中国药房  2025年第36卷第3期                                                 China Pharmacy  2025 Vol. 36  No. 3    · 321 ·
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