Ⅲ~Ⅳ期糖尿病肾病不同中医证型的血清蛋白组学研究

Serum proteomics of different traditional Chinese medicine syndrome types of stage Ⅲ-Ⅳ diabetic nephropathy

目的:探索Ⅲ~Ⅳ期糖尿病肾病(DN)患者血清差异蛋白,同时筛选不同中医证型的血清差异蛋白。方法:收集Ⅲ~Ⅳ期DN患者70例(气阴两虚证19例,脾肾气虚证18例,血瘀证16例,湿热证17例),同时选择健康受试者35例(健康对照组),应用表面加强激光解吸电离-飞行时间-质谱(SELDI-TOF-MS)技术检测各组血清蛋白指纹图谱,并对差异蛋白峰进行对比分析。结果:1) DN组与健康对照组之间共有9个蛋白峰存在显著差异,质荷比(M/Z)分别为2042.57、3291.28、4986.15、5312.69、5564.09、9861.47、10786.53、13392.89、17395.27;2)气阴两虚证与脾肾气虚证之间共有4个蛋白峰存在显著差异,M/Z分别为2994.77、4986.15、7937.25、2758.91;3)气阴两虚证与血瘀证之间共有4个蛋白峰存在显著差异,M/Z分别为4986.15、16982.62、6819.69、9947.36;4)气阴两虚证与湿热证之间共有3个蛋白峰存在显著差异,M/Z分别为4986.15、11741.33、7001.54;5)脾肾气虚证与血瘀证之间共有3个蛋白峰存在显著差异,M/Z分别为3448.22、8063.43、9787.21;6)脾肾气虚证与湿热证之间共有5个蛋白峰存在显著差异,M/Z分别为2144.34、3992.01、8871.35、10568.32、14643.47;7)血瘀证与湿热证之间共有6个蛋白峰存在显著差异,M/Z分别为4233.15、5771.32、5987.18、8496.76、6651.25、13551.94。结论:Ⅲ~Ⅳ期DN组与健康对照组血清蛋白表达存在差异,该差异或可作为相关临床诊断的标志物,对探索DN的发病机制具有重要的参考价值;在不同中医证型之间能够找到差异血清蛋白,可能反映"证"的实质内涵,对今后建立诊断决策模型具有积极意义。

Objective: To investigate the differentially expressed proteins in the serum of patients with stage Ⅲ-Ⅳ diabetic nephropathy(DN) or those with different syndrome types.Methods: A total of 70 patients with stage Ⅲ-Ⅳ DN were enrolled as DN group,among whom 19 had deficiency of both Qi and Yin,18 had spleen-kidney Qi deficiency,16 had blood stasis syndrome,and 17 had damp-heat syndrome,and 35 healthy individuals were enrolled as healthy control group.Surface-enhanced laser desorption/ionization-time of flight-mass spectrometry was used to obtain the fingerprints of serum proteins,and the peaks of differentially expressed proteins were analyzed.Results: There were significant differences in 9 protein peaks between the DN group and the healthy control group,with a mass-to-charge ratio(M/Z) of 2042.57,3291.28,4986.15,5312.69,5564.09,9861.47,10786.53,13392.89,and 17395.27,respectively.There were significant differences in 4 protein peaks between the patients with deficiency of both Qi and Yin and those with spleen-kidney Qi deficiency,with an M/Z of 2994.77,4986.15,7937.25,and 2758.91,respectively.There were significant differences in 4 protein peaks between the patients with deficiency of both Qi and Yin and those with blood stasis syndrome,with an M/Z of 4986.15,16982.62,6819.69,and 9947.36,respectively.There were significant differences in 3 protein peaks between the patients with deficiency of both Qi and Yin and those with damp-heat syndrome,with an M/Z of 4986.15,11741.33,and7001.54,respectively.There were significant differences in 3 protein peaks between the patients with spleen-kidney Qi deficiency and those with blood stasis syndrome,with an M/Z of 3448.22,8063.43,and 9787.21,respectively.There were significant differences in 5 protein peaks between the patients with spleen-kidney Qi deficiency and those with damp-heat syndrome,with an M/Z of 2144.34,3992.01,8871.35,10568.32,and 14643.47,respectively.There were significant differences in 6 protein peaks between the patients with blood stasis syndrome and those with damp-heat syndrome,with an M/Z of 4233.15,5771.32,5987.18,8496.76,6651.25,and 13551.94,respectively.Conclusion: There are significant differences in the expression of serum proteins between patients with stage Ⅲ-Ⅳ DN and healthy individuals,which can be used as markers for clinical diagnosis and have an important value in exploring the pathogenesis of DN.Differentially expressed proteins are observed in patients with different syndrome types and may reflect the essence of the syndrome,which has great significance in establishing the decision-making models for diagnosis.