中医肝阳化风证本质蛋白质组学研究

作者	单位
张雨星	中南大学湘雅医院中西医结合研究所
曾年菊	中南大学湘雅医院中西医结合研究所
熊新贵	中南大学湘雅医院中西医结合研究所
萧梅芳	中南大学湘雅医院中西医结合研究所
区健刚	中南大学湘雅医院中西医结合研究所
梁清华	中南大学湘雅医院中西医结合研究所
范荣	中南大学湘雅医院中西医结合研究所
陈疆	中南大学湘雅医院远程医疗中心

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中文摘要:

目的从蛋白质表达水平初步探讨中医肝阳化风证的本质内涵。方法严格按照中医辨证标准选择脑出血(intracerebral hemorrhage,ICH)肝阳化风证与阴虚风动证、脑梗死(cerebral infarction,CI)肝阳化风证与阴虚风动证、颈椎病(cervical spondylosis,CS)肝阳化风证与肝阳上亢证、帕金森病(Parkinson′s disease,PD)肝阳化风证与血虚生风证患者作对照研究,分离上述各证型组患者及健康人外周血单核细胞(peripheral blood mononuclear cell,PBMC),提取总蛋白后进行双向凝胶电泳(2-DE),PDQuest软件进行分析,采用基质辅助激光解析电离质谱(MALDI-TOF-MS)测定肽质量指纹图,用Mascot查询系统查询SwissProt数据库,比较同病异证、异病同证及同类证患者PBMC差异表达与相同表达的蛋白。结果建立了上述各证型组与健康人PBMC总蛋白的2-DE图谱,通过比较分析并鉴定出同病异证中ICH肝阳化风证与阴虚风动证相同表达蛋白质点1个,差异表达蛋白质点22个;CI肝阳化风证与阴虚风动证相同表达蛋白质点6个,差异表达蛋白质点21个;CS肝阳化风证与肝阳上亢证相同表达蛋白质点3个,差异表达蛋白质点12个;PD肝阳化风证与血虚生风证无相同表达蛋白质点,差异表达蛋白质点12个。异病同证中相同表达蛋白质点13个。四病同类证有1个相同表达的蛋白质点为硫氧环蛋白依赖性过氧化物酶(Thioredoxin-dependent peroxide reductase,TPx)。结论同病异证有不同的本质内涵(有多个不同的蛋白表达),异病同证有共同的物质基础(有共同的蛋白表达),同类证有相同的物质基础(有相同的蛋白表达),提示中医学的证有物质基础,有本质可寻。

英文摘要:

Objective To preliminarily study the essence of wind syndrome caused by Gan-yang hyperactivity(WSGH) in Chinese medicine at the protein expression level.Methods WSGH was strictly differentiated from wind stirring due to yin deficiency syndrome in patients with intracerebral hemorrhage(ICH) and those with cerebral infarction(CI);from Gan-yang hyperactivity syndrome in patients with cervical spondylosis(CS);from wind syndrome induced by blood deficiency in patients with Parkinson’s disease(PD) according to Chinese medicine syndrome typing standard.Control studies were performed.Peripheral blood mononuclear cells(PBMCs) were isolated from all patients of the aforesaid syndromes and healthy subjects.The total proteins were extracted,two-dimensional gel electrophoresis(2-DE) conducted and analyzed by PDQuest software.The peptide mass fingerprint(PMF) was determined using matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS).The SwissProt database was inquired using Mascot reference system.Proteins of different and same expressions in PBMCs of patients suffering from the same disease of different syndromes,different diseases of the same syndrome,and syndromes of the same kind were compared.Results The 2-DE map of PBMCs’ total proteins in the aforesaid syndrome groups and healthy subjects was established.Through comparison,analysis,and appraisement,there was 1 protein dot of the same expression and 22 protein dots of different expressions between ICH patients of WSGH and ICH patients of wind stirring due to yin deficiency syndrome.There were 6 protein dots of the same expression and 21 protein dots of different expressions between CI patients of WSGH and CI patients of wind stirring due to yin deficiency syndrome.There were 3 protein dots of the same expression and 12 protein dots of different expressions between CS patients of WSGH and CS patients of Gan-yang hyperactivity syndrome.There was no protein dot of the same expression and 12 protein dots of different expressions between PD patients of WSGH and PD patients of wind syndrome induced by blood deficiency.There were 13 protein dots of the same expression in different diseases of the same syndrome.There was 1 protein dot(Thioredoxin-dependent peroxide reductase,TPx) of the same expression in the four diseases of the same kind syndrome.Conclusions Different connotations of the essence existed(having multiple different protein expressions) in patients with the same disease of different syndromes.Syndromes of the same kind share the same material bases(having the same protein expression).These suggested that Chinese medicine syndrome has its own material bases and essence findable.

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