基于TMT技术的益气法干预压力负荷心衰大鼠心肌线粒体蛋白质组学研究

目的：通过TMT技术分析益气法对压力负荷心衰大鼠心肌细胞线粒体蛋白质组学差异,探讨观察益气中药对心衰的干预作用机制。方法：运用腹主动脉结扎法建立压力负荷心衰大鼠模型,使用卡托普利、黄芪党参及中西药联合连续灌胃给药8周,通过TMT技术对压力负荷心衰大鼠心肌细胞线粒体进行蛋白质组质谱检测。结果：共鉴定到889个蛋白,对生物意义重要的蛋白进行生物学分析。心衰模型组的能量代谢相关蛋白水平明显上调,涉及ATP合成及β氧化等功能,卡托普利可抑制模型组相关蛋白的上调。而益气法则不仅涵盖这些生物功能,还参与了抑制心室重构及增强心肌收缩力等功能过程,中西医结合组则可更好地提高能量代谢、心肌收缩等相关蛋白水平,并涉及了中药组及西药组均未涉及的蛋白酶体降解功能,同时降低了卡托普利对β氧化的强化作用。结论：益气法联合ACEI类药物可更好地优化心衰能量代谢障碍,并且还有可能通过抑制心室重构和蛋白酶体降解体系等途径改善心衰患者预后。

基于TMT技术分析益气法干预心衰大鼠的机制

目的利用串联质谱标签(TMT)技术分析益气法对心梗后心衰大鼠心肌线粒体蛋白质差异,探讨益气法对心梗后心衰的干预机制。方法运用左冠前降支结扎法建立心梗后心衰大鼠模型,分别予黄芪党参、卡托普利及中西药联合干预8周,利用TMT技术对各组大鼠进行线粒体蛋白质组学分析。结果共鉴定到633个蛋白,分析有重要生物意义的蛋白。结果显示,模型组抗心肌细胞凋亡相关蛋白水平上调,西药组保护心功能相关蛋白上调,细胞凋亡相关蛋白下调。中药组改善心肌能量代谢的蛋白上调,促心肌细胞凋亡蛋白下调,中西医结合组较其余各组可更好地抑制心肌细胞凋亡相关蛋白的表达。结论益气法联合ACEI类药物,可更好地优化心肌梗死后心衰的心肌能量代谢,减少心肌细胞凋亡。

Effect of acupuncture on hippocampal mitochondrial proteome expression in SAMP8 mouse model with Alzheimer disease

Objective：To observe the effect of acupuncture on the expression of mitochondrial proteome in hippocampus of senescence-accelerated mouse prone g （SAMPg） mice models with Alzheimer disease （AD）,and to explore the possible protective mechanism of acupuncture on mitochondria.Methods：Sixty 6-month-old male SAMP8 mice were randomly divided into an acupuncture at acupoint group,an acupuncture at non-acupoint group and a model group,20 mice in each group.The 20 male senescence-accelerated mouse/resistance 1 （SAMR1） mice of the same age were used as a normal control group.Shenshu （BL 23）,Baihui （GV 20）,Xuehai （SP 10） and Geshu （BL 17） were selected for acupuncture intervention in acupuncture at acupoint group.After an 8-week intervention,mitochondrial tissues were extracted from the hippocampus.Differentially expressed proteins were identified by subcellular organelle proteomics.Western blot was used to verify the expressions of some related proteins in hippocampal mitochondria.Results：Compared with the model group,there were 13 differentially expressed protein spots in the acupuncture at acupoint group,of which,9 were up-regulated,including neurofilament light polypeptide （NFL）,actin （cytoplasmic 1,database ID：ACTB）,tubulin beta-2A chain （TBB2A）,tropomodulin-2 （TMOD2）,pyruvate dehydrogenase E1 component subunit beta （PDHE1-β）,NADH-ubiquinone oxidoreductase 75 kDa subunit （database ID：NDUS1）,heat shock cognate 71 kDa protein （HSC71）,pyruvate dehydrogenase E1 component subunit alpha （PDHE1-α） and ATP synthase beta subunit （ATP-β）;4 were down-regulated,including glial fibrillary acidic protein （GFAP）,pyruvate dehydrogenase phosphatase 1 （PDP1）,mitochondrial-processing peptidase subunit alpha （MMP-α） and adenosine kinase （ADK）.According to the information provided in the protein database,most of the differentially expressed proteins involve the regulation of mitochondrial function and structure.The expression levels of NFL and TBB2A in the normal control group and the acupuncture at acupoint group were significantly higher than those in the acupuncture at non-acupoint group （P〈0.05）.ATP-β and NDUS1 expression levels were significantly higher in the acupuncture at acupoint group than those in the acupuncture at non-acupoint group （P〈0.05）;there was no significant difference between the acupuncture at non-acupoint group and the model group （P〉0.05）.Conclusion：Acupuncture may achieve the potential therapeutic effect on AD by regulating the structure and functional proteins of hippocampal mitochondria.