Chip-based serum proteomics approach to reveal the potential protein markers in the sub-acute stroke patients receiving the treatment of Ginkgo Diterpene Lactone Meglumine Injection

J Ethnopharmacol. 2020 Oct 5:260:112964. doi: 10.1016/j.jep.2020.112964. Epub 2020 May 13.

Abstract

Ethnopharmacological relevance: Ginkgo biloba L. is a kind of traditional Chinese medicinal material with a long history. Its main active ingredients, ginkgolides, can be used for the treatment of stroke and other cardio-cerebrovascular diseases. Ginkgo Diterpene Lactone Meglumine Injection (GDLI), a modernized TCM, has attracted much attention because of its neuroprotective and anti-inflammatory properties.

Aim of the study: To uncover the effects of GDLI on ischemic stroke patients, as well as the underlying biomarkers involved in sub-acute stroke.

Materials and methods: We used a state-of-the-art targeted proteomics chip to investigate the association between numerous serum proteins (1101 proteins) and the sub-acute phase post-ischemic stroke. Then, the relative proteins of anti-apoptosis, anticoagulant, and neuroprotection of GDLI were verified in animal models.

Results: Compared with the serum from healthy volunteers, we identified 15 up-regulated proteins and 26 down-regulated proteins (FC ≥ 1.5) involved in inflammatory response, immune response, and nervous system development in the sub-acute ischemic stroke. The pro-inflammatory proteins, such as IL17, MSP-R, G-CSF-R, TLR3, MIP-3β, TNFRSF19, and TNFRSF12, were significantly increased in serum, illustrating that the chronic inflammatory state was evident in the sub-acute stage of ischemic stroke. However, the common pro-inflammatory proteins, such as IL-1β, IL-6, IL-8, TNF-α, IFN-γ, and IL-10, known to be up-regulated in acute stroke, had close or lightly lower levels than healthy humans (FC ≥ 1.5, P > 0.05). And some cytokines (IL3, CCL13, TNFRSF3, IL10 R beta, HLA-A, IL-1 F8/FIL1 eta, TNFRSF8, CCL18) were also markedly down-regulated in the sub-acute phase of stroke. These proteins are highly associated with the onset of stroke-induced immunosuppression and post-stroke infection. Moreover, we noticed that Ginkgo Diterpene Lactone Meglumine Injection (GDLI) treatment for 14 days was helpful to the recovery of patients in the subacute period. After the treatment of GDLI, it was observed that several inflammatory cytokines (i.e. IL-17 and IL-28A), chemokine (i.e. CCL14), and Coagulation Factor III were reduced. Meanwhile, the anti-inflammatory cytokines (IL-10 R alpha, GREMLIN, and Activin C) and neurotrophic factors (Neurturin and IGFBP2) were found to be up-regulated in stroke patients through self-control observation. Finally, we identified the IGFBP2 as a novel marker in the animal models.

Conclusions: In summary, the potential markers in sub-acute stroke patients were highly different from known protein markers in the acute phase of ischemic stroke. The serum protein IGFBP2 could be novel biomarkers for the treatment of GDLI in sub-acute stroke patients. Our present findings provide an innovative insight into the novel treatment of GDLI in ischemic stroke therapy.

Keywords: Antibody chip; Ginkgo diterpene lactone; Protein markers; Sub-acute ischemic stroke.

MeSH terms

  • Aged
  • Animals
  • Anti-Inflammatory Agents / administration & dosage
  • Anti-Inflammatory Agents / isolation & purification
  • Anti-Inflammatory Agents / therapeutic use*
  • Biomarkers / blood
  • Blood Proteins / metabolism*
  • Case-Control Studies
  • Disease Models, Animal
  • Diterpenes / administration & dosage
  • Diterpenes / isolation & purification
  • Diterpenes / therapeutic use*
  • Female
  • Ginkgo biloba* / chemistry
  • Humans
  • Infusions, Intravenous
  • Ischemic Stroke / blood
  • Ischemic Stroke / diagnosis
  • Ischemic Stroke / drug therapy*
  • Male
  • Middle Aged
  • Neuroprotective Agents / administration & dosage
  • Neuroprotective Agents / isolation & purification
  • Neuroprotective Agents / therapeutic use*
  • Plant Extracts / administration & dosage
  • Plant Extracts / isolation & purification
  • Plant Extracts / therapeutic use*
  • Protein Array Analysis*
  • Proteomics*
  • Rats, Sprague-Dawley
  • Signal Transduction
  • Time Factors
  • Treatment Outcome

Substances

  • Anti-Inflammatory Agents
  • Biomarkers
  • Blood Proteins
  • Diterpenes
  • Neuroprotective Agents
  • Plant Extracts