Cellular and Molecular Neurobiology

, Volume 38, Issue 5, pp 1123–1135 | Cite as

Proteomic Analysis of the Antidepressant Effects of Shen–Zhi–Ling in Depressed Patients: Identification of Proteins Associated with Platelet Activation and Lipid Metabolism

  • Chao Chen
  • Yuan Hu
  • Xian-zhe Dong
  • Xiao-jiang Zhou
  • Li-hua Mu
  • Ping LiuEmail author
Original Research


Shen–Zhi–Ling (SZL) is a Chinese medicine formulated from a Kai–Xin–San decoction that is commonly used to treat depression caused by dual deficiencies in the heart and spleen. However, the underlying mechanisms remain unclear. We investigated biological changes in depression patients (DPs) exhibiting antidepressant responses to SZL treatment using proteomic techniques. We performed label-free quantitative proteomic analysis and liquid chromatography–tandem mass spectrometry to discover and examine altered proteins involved in depression and antidepressant treatment. Serum samples were collected from DPs, DPs who underwent 8 weeks of SZL treatment and healthy controls (HCs). The proteins that differed among the three groups were further validated by Western blot analysis. By performing multivariate analyses, we identified 12 potential serum biomarkers that were differentially expressed among the HC, DP, and SZL groups. We then confirmed the significant changes in alpha-1-antitrypsin, von Willebrand factors, apolipoprotein C-III, and alpha-2-macroglobulin among the three groups by performing Western blot analysis, which supported the proteomic results. Profiling the proteomic changes in DPs treated with SZL could improve our understanding of the pathways involved in SZL responses, such as alterations in platelet activation, inflammatory regulation, and lipid metabolism. Future studies involving larger patient cohorts are necessary to draw more definitive conclusions.


Proteomics Depression Antidepressant effect Shen–Zhi–Ling Kai–Xin–San Platelet activation 



This study was supported by grants from the National Natural Science Foundation (Grant Nos. 81673877, 81373996, and 81173430).

Author Contributions

PL and CC conceived and designed the experiments. CC and XD performed the experiments and wrote the manuscript. CC and PL analyzed the data. YH, XZ, and LM contributed materials. All the authors read and approved the final manuscript.

Compliance with Ethical Standards

Ethical Approval

All the experimental protocols involving human subjects were approved by the Ethics Committee of the Fourth Military Medical University (Xi’an, China).

Conflicts of interest

The authors declare no conflicts of interest.


  1. Amadio P, Colombo GI, Tarantino E, Gianellini S, Ieraci A, Brioschi M, Banfi C, Werba JP, Parolari A, Lee FS, Tremoli E, Barbieri SS (2017) BDNFVal66met polymorphism: a potential bridge between depression and thrombosis. Eur Heart J 38:1426–1435. PubMedCrossRefGoogle Scholar
  2. Armstrong PB (2001) The contribution of proteinase inhibitors to immune defense. Trends Immunol 22:47–52. CrossRefPubMedGoogle Scholar
  3. Bahn S, Chan MK (2015) What can we learn about depression from gene expression in peripheral tissues? Biol Psychiatry 77:207–209. CrossRefPubMedGoogle Scholar
  4. Bao ZX, Zhao GP, Sun W, Chen BJ (2011) Clinical curative effects of kaixin powder on depression with mild or moderate degree. Chin Arch Tradit Chin Med 28:987–988Google Scholar
  5. Bot M, Chan MK, Jansen R, Lamers F, Vogelzangs N, Steiner J, Leweke FM, Rothermundt M, Cooper J, Bahn S, Penninx BW (2015) Serum proteomic profiling of major depressive disorder. Transl Psychiatry 5:e599. CrossRefPubMedPubMedCentralGoogle Scholar
  6. Butler L, Pilkington K (2013) Chinese herbal medicine and depression: the research evidence. Evid Based Complement Alternat Med 2013:739716. CrossRefPubMedPubMedCentralGoogle Scholar
  7. Canan A, Halıcioğlu SS, Gürel S (2012) Mean platelet volume and D-dimer in patients with suspected deep venous thrombosis. J Thromb Thrombolysis 34:283–287. CrossRefPubMedGoogle Scholar
  8. Dang H, Sun L, Liu X, Peng B, Wang Q, Jia W, Chen Y, Pan A, Xiao P (2009) Preventive action of Kai Xin San aqueous extract on depressive-like symptoms and cognition deficit induced by chronic mild stress. Exp Biol Med 234:785–793. CrossRefGoogle Scholar
  9. Dimopoulos N, Piperi C, Salonicioti A, Mitsonis C, Liappas I, Lea RW, Kalofoutis A (2006) Elevation of plasma concentration of adhesion molecules in late-life depression. Int J Geriatr Psychiatry 21:965–971. CrossRefPubMedGoogle Scholar
  10. Domenici E, Willé DR, Tozzi F, Prokopenko I, Miller S, McKeown A, Brittain C, Rujescu D, Giegling I, Turck CW, Holsboer F, Bullmore ET, Middleton L, Merlo-Pich E, Alexander RC, Muglia P (2010) Plasma protein biomarkers for depression and schizophrenia by multi analyte profiling of case-control collections. PLoS ONE 5:e9166. CrossRefPubMedPubMedCentralGoogle Scholar
  11. Dong XZ, Li ZL, Zheng XL, Mu LH, Zhang GQ, Liu P (2013) A representative prescription for emotional disease, Ding-Zhi-Xiao-Wan restores 5-HT system deficit through interfering the synthesis and transshipment in chronic mild stress-induced depressive rats. J Ethnopharmacol 150:1053–1061. CrossRefPubMedGoogle Scholar
  12. Feige JJ, Negoescu A, Keramidas M, Souchelnitskiy S, Chambaz EM (1996) Alpha 2-macroglobulin: a binding protein for transforming growth factor-beta and various cytokines. Horm Res 45:227–232. CrossRefPubMedGoogle Scholar
  13. Feng DD, Tang T, Lin XP, Yang ZY, Yang S, Xia ZA, Wang Y, Zheng P, Wang Y, Zhang CH (2016) Nine traditional Chinese herbal formulas for the treatment of depression: an ethnopharmacology, phytochemistry, and pharmacology review. Neuropsychiatr Dis Treat 12:2387–2402. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Geiser F, Meier C, Wegener I, Imbierowicz K, Conrad R, Liedtke R, Oldenburg J, Harbrecht U (2008) Association between anxiety and factors of coagulation and fibrinolysis. Psychother Psychosom 77:377–383. CrossRefPubMedGoogle Scholar
  15. Hsu MC, Creedy D, Moyle W, Venturato L, Tsay SL, Ouyang WC (2008) Use of complementary and alternative medicine among adult patients for depression in Taiwan. J Affect Disord 111:360–365. CrossRefPubMedGoogle Scholar
  16. Hu Y, Liu P, Dai-Hong G, Rahman K, Wang D-X, Chen M-L, Xie T-T (2008) Behavioral and biochemical effects of Kaixin-San, a traditional Chinese medicinal empirical formula. Drug Dev Res 69:267–271. CrossRefGoogle Scholar
  17. Hu Y, Cao Y, Liu M, Liu P, Cui H, Dai-Hong G (2013) Behavioral and biochemical effects of a formulation of the traditional Chinese medicine, Kai-Xin-San, in fatigued rats. Exp Ther Med 6:973–976. CrossRefPubMedPubMedCentralGoogle Scholar
  18. Hu Y, Zhou XJ, Liu P, Dong XZ, Mu LH, Chen YB, Liu MY, Yu BY (2014) Antidepressant and neuroprotective effect of the Chinese herb kaixinsan against lentiviral shRNA knockdown brain-derived neurotrophic factor-induced injury in vitro and in vivo. Neuropsychobiology 69:129–139. CrossRefPubMedGoogle Scholar
  19. Huang TL (2005) Serum lipid profiles in major depression with clinical subtypes, suicide attempts and episodes. J Affect Disord 86:75–79. CrossRefPubMedGoogle Scholar
  20. Huang TL, Wu SC, Chiang YS, Chen JF (2003) Correlation between serum lipid, lipoprotein concentrations and anxious state, depressive state or major depressive disorder. Psychiatry Res 118:147–153. CrossRefPubMedGoogle Scholar
  21. Huang TL, Sung ML, Chen TY (2014) 2D-DIGE proteome analysis on the platelet proteins of patients with major depression. Proteome Sci 12:1. CrossRefPubMedPubMedCentralGoogle Scholar
  22. Ji Q, Zhu F, Liu X, Li Q, Su SB (2015) Recent advance in applications of proteomics technologies on traditional Chinese medicine research. Evid Based Complement Alternat Med 2015:983139. PubMedPubMedCentralCrossRefGoogle Scholar
  23. Kanehisa M, Goto S, Kawashima S, Okuno Y, Hattori M (2004) The KEGG resource for deciphering the genome. Nucleic Acids Res 32:D277–D280. CrossRefPubMedPubMedCentralGoogle Scholar
  24. Karege F, Bondolfi G, Gervasoni N, Schwald M, Aubry JM, Bertschy G (2005) Low brain-derived neurotrophic factor (BDNF) levels in serum of depressed patients probably results from lowered platelet BDNF release unrelated to platelet reactivity. Biol Psychiatry 57:1068–1072. CrossRefPubMedGoogle Scholar
  25. Kasper S, Fuger J, Möller HJ (1992) Comparative efficacy of antidepressants. Drugs 43:11–22. CrossRefPubMedGoogle Scholar
  26. Kessler RC, Soukup J, Davis RB, Foster DF, Wilkey SA, Van Rompay MI, Eisenberg DM (2001) The use of complementary and alternative therapies to treat anxiety and depression in the United States. Am J Psychiatry 158:289–294. CrossRefPubMedGoogle Scholar
  27. Khetarpal SA, Qamar A, Millar JS, Rader DJ (2016) Targeting apoC-III to reduce coronary disease risk. Curr Atheroscler Rep 18:54. CrossRefPubMedGoogle Scholar
  28. Kou MJ, Chen JX (2012) Integrated traditional and Western medicine for treatment of depression based on syndrome differentiation: a meta-analysis of randomized controlled trials based on the Hamilton depression scale. J Tradit Chin Med 32:1–5. CrossRefPubMedGoogle Scholar
  29. LaMarre J, Wollenberg GK, Gonias SL, Hayes MA (1991) Cytokine binding and clearance properties of proteinase-activated alpha 2-macroglobulins. Lab Investig 65:3–14PubMedGoogle Scholar
  30. Lang UE, Borgwardt S (2013) Molecular mechanisms of depression: perspectives on new treatment strategies. Cell Physiol Biochem 31:761–777. CrossRefPubMedGoogle Scholar
  31. Lee BH, Kim YK (2009) Reduced platelet BDNF level in patients with major depression. Prog Neuropsychopharmacol Biol Psychiatry 33:849–853. CrossRefPubMedGoogle Scholar
  32. Lee J, Kim SH, Choi DS, Lee JS, Kim DK, Go G, Park SM, Kim SH, Shin JH, Chang CL, Gho YS (2015) Proteomic analysis of extracellular vesicles derived from Mycobacterium tuberculosis. Proteomics 15:3331–3337. CrossRefPubMedGoogle Scholar
  33. Lee MY, Kim EY, Kim SH, Cho KC, Ha K, Kim KP, Ahn YM (2016) Discovery of serum protein biomarkers in drug-free patients with major depressive disorder. Prog Neuropsychopharmacol Biol Psychiatry 69:60–68. CrossRefPubMedGoogle Scholar
  34. Li S, Li X, Wang Y, Yang J, Chen Z, Shan S (2014) Global secretome characterization of A549 human alveolar epithelial carcinoma cells during Mycoplasma pneumoniae infection. BMC Microbiol 14:27. CrossRefPubMedPubMedCentralGoogle Scholar
  35. Liu J, Fang Y, Yang L, Qin X, Du G, Gao X (2015) A qualitative, and quantitative determination and pharmacokinetic study of four polyacetylenes from Radix Bupleuri by UPLC-PDA-MS. J Pharm Biomed Anal 111:257–265. CrossRefPubMedGoogle Scholar
  36. Lopez-Vilchez I, Diaz-Ricart M, Navarro V, Torramade S, Zamorano-Leon J, Lopez-Farre A, Galan AM, Gasto C, Escolar G (2016) Endothelial damage in major depression patients is modulated by SSRI treatment, as demonstrated by circulating biomarkers and an in vitro cell model. Transl Psychiatry 6:e886. CrossRefPubMedPubMedCentralGoogle Scholar
  37. Maes M (1993) A review on the acute phase response in major depression. Rev Neurosci 4:407–416. CrossRefPubMedGoogle Scholar
  38. Maes M, Scharpe S, Bosmans E, Vandewoude M, Suy E, Uyttenbroeck W, Cooreman W, Vandervorst C, Raus J (1992) Disturbances in acute phase plasma proteins during melancholia: additional evidence for the presence of an inflammatory process during that illness. Prog Neuropsychopharmacol Biol Psychiatry 16:501–515. CrossRefPubMedGoogle Scholar
  39. Maes M, Nowak G, Caso JR, Leza JC, Song C, Kubera M, Klein H, Galecki P, Noto C, Glaab E, Balling R, Berk M (2016) Toward omics-based, systems biomedicine, and path and drug discovery methodologies for depression-inflammation research. Mol Neurobiol 53:2927–2935. CrossRefPubMedGoogle Scholar
  40. McCaffery JM, Duan QL, Frasure-Smith N, Barhdadi A, Lespérance F, Théroux P, Rouleau GA, Dubé MP (2009) Genetic predictors of depressive symptoms in cardiac patients. Am J Med Genet B Neuropsychiatr Genet 150B:381–388. CrossRefPubMedPubMedCentralGoogle Scholar
  41. Monteggia LM, Malenka RC, Deisseroth K (2014) Depression: the best way forward. Nature 515:200–201. CrossRefPubMedGoogle Scholar
  42. Mu LH, Huang ZX, Liu P, Hu Y, Gao Y (2011) Acute and subchronic oral toxicity assessment of the herbal formula Kai-Xin-San. J Ethnopharmacol 138:351–357. CrossRefPubMedGoogle Scholar
  43. Nemeroff CB, Musselman DL (2000) Are platelets the link between depression and ischemic heart disease? Am Heart J 140:57–62. CrossRefPubMedGoogle Scholar
  44. Nguyen VA, Carey LM, Giummarra L, Faou P, Cooke I, Howells DW, Tse T, Macaulay SL, Ma H, Davis SM, Donnan GA, Crewther SG (2016) A pathway proteomic profile of ischemic stroke survivors reveals innate immune dysfunction in association with mild symptoms of depression—a pilot study. Front Neurol 7:85. CrossRefPubMedPubMedCentralGoogle Scholar
  45. Ormonde do Carmo MBO, Mendes-Ribeiro AC, Matsuura C, Pinto VL, Mury WV, Pinto NO, Moss MB, Ferraz MR, Brunini TMC (2015) Major depression induces oxidative stress and platelet hyperaggregability. J Psychiatr Res 61:19–24. CrossRefPubMedGoogle Scholar
  46. Papakostas GI, Shelton RC, Kinrys G, Henry ME, Bakow BR, Lipkin SH, Pi B, Thurmond L, Bilello JA (2013) Assessment of a multi-assay, serum-based biological diagnostic test for major depressive disorder: a pilot and replication study. Mol Psychiatry 18:332–339. CrossRefPubMedGoogle Scholar
  47. Qin X, Li P, Han M, Liu Z, Liu J (2010) Systematic review of randomize controlled trials of Xiaoyao powder in treatment of depression. J Trad Chin Med 51:500–505Google Scholar
  48. Qin F, Wu XA, Tang Y, Huang Q, Zhang ZJ, Yuan JH (2011) Meta-analysis of randomized controlled trials to assess the effectiveness and safety of Free and Easy Wanderer Plus, a polyherbal preparation for depressive disorders. J Psychiatr Res 45:1518–1524. CrossRefPubMedGoogle Scholar
  49. Qiong W, Yong-Liang Z, Ying-Hui L, Shan-Guang C, Jiang-Hui G, Yi-Xi C, Ning J, Xin-Min L (2016) The memory enhancement effect of Kai Xin San on cognitive deficit induced by simulated weightlessness in rats. J Ethnopharmacol 187:9–16. CrossRefPubMedGoogle Scholar
  50. Qureshi NA, Al-Bedah AM (2013) Mood disorders and complementary and alternative medicine: a literature review. Neuropsychiatr Dis Treat 9:639–658. CrossRefPubMedPubMedCentralGoogle Scholar
  51. Roetker NS, Yonker JA, Lee C, Chang V, Basson JJ, Roan CL, Hauser TS, Hauser RM, Atwood CS (2012) Multigene interactions and the prediction of depression in the Wisconsin Longitudinal Study. BMJ Open 2:e000944. CrossRefPubMedPubMedCentralGoogle Scholar
  52. Sarris J, Panossian A, Schweitzer I, Stough C, Scholey A (2011) Herbal medicine for depression, anxiety and insomnia: a review of psychopharmacology and clinical evidence. Eur Neuropsychopharmacol 21:841–860. CrossRefPubMedGoogle Scholar
  53. Scharinger C, Rabl U, Kasess CH, Meyer BM, Hofmaier T, Diers K, Bartova L, Pail G, Huf W, Uzelac Z, Hartinger B, Kalcher K, Perkmann T, Haslacher H, Meyer-Lindenberg A, Kasper S, Freissmuth M, Windischberger C, Willeit M, Lanzenberger R, Esterbauer H, Brocke B, Moser E, Sitte HH, Pezawas L (2014) Platelet serotonin transporter function predicts default-mode network activity. PLoS ONE 9:e92543. CrossRefPubMedPubMedCentralGoogle Scholar
  54. Schnyer RN, Flaws B (1998) Curing depression naturally with Chinese medicine. Blue Poppy Press, BoulderGoogle Scholar
  55. Serretti A, Mandelli L, Lattuada E, Smeraldi E (2004) Depressive syndrome in major psychoses: a study on 1351 subjects. Psychiatry Res 127:85–99. CrossRefPubMedGoogle Scholar
  56. Simon GE, Savarino J, Operskalski B, Wang PS (2006) Suicide risk during antidepressant treatment. Am J Psychiatry 163:41–47. CrossRefPubMedGoogle Scholar
  57. Valkanova V, Ebmeier KP, Allan CL (2013) CRP, IL-6 and depression: a systematic review and meta-analysis of longitudinal studies. J Affect Disord 150:736–744. CrossRefPubMedGoogle Scholar
  58. van Sloten TT, Schram MT, Adriaanse MC, Dekker JM, Nijpels G, Teerlink T, Scheffer PG, Pouwer F, Schalkwijk CG, Stehouwer CD, Henry RM (2014) Endothelial dysfunction is associated with a greater depressive symptom score in a general elderly population: the Hoorn Study. Psychol Med 44:1403–1416. CrossRefPubMedGoogle Scholar
  59. von Känel R (2004) Platelet hyperactivity in clinical depression and the beneficial effect of antidepressant drug treatment: how strong is the evidence? Acta Psychiatr Scand 110:163–177. CrossRefGoogle Scholar
  60. Wang Y, Li M, Dai X, Fan G (1997) Internal Chinese Medicine. Shanghai Scientific and Technological Documentation Publishing House, ShanghaiGoogle Scholar
  61. Wang Y, Fan R, Huang X (2012) Meta-analysis of the clinical effectiveness of traditional Chinese medicine formula Chaihu-Shugan-San in depression. J Ethnopharmacol 141:571–577. CrossRefPubMedGoogle Scholar
  62. World Health Organization, Western Pacific Region (2007) WHO international standard terminologies on traditional medicine in the western pacific region. World Health Organization, ManilaGoogle Scholar
  63. Wu R, Zhu D, Xia Y, Wang H, Tao W, Xue W, Xia B, Ren L, Zhou X, Li G, Chen G (2015) A role of Yueju in fast-onset antidepressant action on major depressive disorder and serum BDNF expression: a randomly double-blind, fluoxetine-adjunct, placebo-controlled, pilot clinical study. Neuropsychiatr Dis Treat 11:2013–2021. PubMedPubMedCentralCrossRefGoogle Scholar
  64. Xu HB, Zhang RF, Luo D, Zhou Y, Wang Y, Fang L, Li WJ, Mu J, Zhang L, Zhang Y, Xie P (2012) Comparative proteomic analysis of plasma from major depressive patients: identification of proteins associated with lipid metabolism and immunoregulation. Int J Neuropsychopharmacol 15:1413–1425. CrossRefPubMedGoogle Scholar
  65. Yang Y, Chen J, Liu C, Fang L, Liu Z, Guo J, Cheng K, Zhou C, Zhan Y, Melgiri ND, Zhang L, Zhong J, Chen J, Rao C, Xie P (2016) The extrinsic coagulation pathway: a biomarker for suicidal behavior in major depressive disorder. Sci Rep 6:32882. CrossRefPubMedPubMedCentralGoogle Scholar
  66. Yeung WF, Chung KF, Ng KY, Yu YM, Ziea ET, Ng BF (2014) A meta-analysis of the efficacy and safety of traditional Chinese medicine formula Ganmai Dazao decoction for depression. J Ethnopharmacol 153:309–317. CrossRefPubMedGoogle Scholar
  67. Yeung WF, Chung KF, Ng KY, Yu YM, Zhang SP, Ng BF, Ziea ET (2015) Prescription of Chinese herbal medicine in pattern-based traditional Chinese medicine treatment for depression: a systematic review. Evid Based Complement Alternat Med 2015:160189. CrossRefPubMedPubMedCentralGoogle Scholar
  68. Zhan Y, Yang YT, You HM, Cao D, Liu CY, Zhou CJ, Wang ZY, Bai SJ, Mu J, Wu B, Zhan QL, Xie P (2014) Plasma-based proteomics reveals lipid metabolic and immunoregulatory dysregulation in post-stroke depression. Eur Psychiatry 29:307–315. CrossRefPubMedGoogle Scholar
  69. Zhang Y, Han M, Liu Z, Wang J, He Q, Liu J (2012) Chinese herbal formula Xiao Yao San for treatment of depression: a systematic review of randomized controlled trials. Evid Based Complement Alternat Med 2012:931636. PubMedCrossRefGoogle Scholar
  70. Zhao H, Wan X, Chen JX (2009) A mini review of traditional Chinese medicine for the treatment of depression in China. Am J Chin Med 37:207–213. CrossRefPubMedGoogle Scholar
  71. Zheng P, Gao HC, Li Q, Shao WH, Zhang ML, Cheng K, Yang DY, Fan SH, Chen L, Fang L, Xie P (2012) Plasma metabonomics as a novel diagnostic approach for major depressive disorder. J Proteome Res 11:1741–1748. CrossRefPubMedGoogle Scholar
  72. Zhou XJ, Liu M, Yan JJ, Cao Y, Liu P (2012) Antidepressant-like effect of the extracted of Kai Xin San, a traditional Chinese herbal prescription, is explained by modulation of the central monoaminergic neurotransmitter system in mouse. J Ethnopharmacol 139:422–428. CrossRefPubMedGoogle Scholar
  73. Zhu KY, Xu SL, Choi RC, Yan AL, Dong TT, Tsim KW (2013) Kai-Xin-San, a Chinese herbal decoction containing ginseng radix et rhizoma, polygalae radix, acori tatarinowii rhizoma, and poria, stimulates the expression and secretion of neurotrophic factors in cultured astrocytes. Evid Based Complement Alternat Med 2013:731385. PubMedPubMedCentralCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Chao Chen
    • 1
  • Yuan Hu
    • 1
  • Xian-zhe Dong
    • 1
  • Xiao-jiang Zhou
    • 1
  • Li-hua Mu
    • 1
  • Ping Liu
    • 1
    Email author
  1. 1.Department of Clinical PharmacologyGeneral Hospital of Chinese People’s Liberation ArmyBeijingPeople’s Republic of China

Personalised recommendations