Proteomics study of Mongolian medicine warm acupuncture in the treatment of p-chlorophenylalanine-induced insomnia in rats

Figures & data

Figure 1 The entire experimental process. Experimental design for the quantitative proteomic analysis, the experiment is divided into three groups (control group, insomnia model group, and the warm acupuncture treated insomnia group). Protein extraction, trypsin digestion, and labeled with iTRAQ regents. The labeled peptides were separated by SCX chromatography and fractions were analyzed by reversed-phase LC-MS/MS, all data were analyzed by bioinformatics tools from different aspects (GO, KEGG).

Table 1 Quantitative expression analysis of proteins.

Group	Total differentially expressed proteins	up	down
M vs. C	76	45	31
W vs. C	149	101	48
W vs. M	48	16	22

Figure 2 Cluster analysis of differential level proteins between M vs. C. (Colors indicate the differential protein levels, which increase successively from green to red. Increased levels of proteins are indicated in red, and decreased levels are marked in green)

Figure 3 Cluster analysis of differential level proteins between W vs. C. (Colors indicate the differential protein levels, which increase successively from green to red. Increased levels of proteins are indicated in red, and decreased levels are marked in green)

Figure 4 Cluster analysis of differential level proteins between W vs. M. (Colors indicate the differential protein levels, which increase successively from green to red. Increased levels of proteins are indicated in red, and decreased levels are marked in green)

Figure 5 GO analyses of protein functions in M vs. C. The GO functional annotations of 76 differentially expressed proteins in M vs. C. The 76 differentially expressed proteins were classified from three aspects: biological processes, molecular functions, and cellular components.

Figure 6 GO analyses of protein functions in W vs. C. The GO functional annotations of 149 differentially expressed proteins in W vs. C. The 149 differentially expressed proteins were classified from three aspects: biological processes, molecular functions, and cellular components.

Figure 7 GO analyses of protein functions in W vs. M. The GO functional annotations of 48 differentially expressed proteins in W vs. M. The 48 differentially expressed proteins were classified from three aspects: biological processes, molecular functions, and cellular components.

Figure 8 KEGG analyses of protein functions in M vs. C. KEGG database pathway annotation was performed on 76 differentially expressed proteins in the insomnia model group with the control group.

Figure 9 KEGG analyses of protein functions in W vs. C. KEGG database pathway annotation was performed on 149 differentially expressed proteins in the warm acupuncture-treated insomnia group with the control group.

Figure 10 KEGG analyses of protein functions in W vs. M. KEGG database pathway annotation was performed on 48 differentially expressed proteins in the warm acupuncture-treated insomnia group with the insomnia model group.

Table 2 Differentially expressed proteins between W/M vs. M/C groups.

M VS. C	W vs. M
Pro-MCH GN = Pmch up expression	Pro-MCH GN = Pmch down expression
Carbonic anhydrase 3 GN = Ca3up expression	Carbonic anhydrase 3 GN = Ca3down expression
CST complex subunit STN1 GN = Obfc1 up expression	CST complex subunit STN1 GN = Obfc1 down expression
Protein Frmd4b GN = Frmd4bup expression	Protein Frmd4b GN = Frmd4bdown expression
Phosphatidylinositol 3,4,5-trisphosphate 5-phosphatase 1 GN = Inpp5ddown expression	Phosphatidylinositol 3,4,5-trisphosphate 5-phosphatase 1 GN = Inpp5dup expression
Serum deprivation-response protein GN = Sdprdown expression	Serum deprivation-response protein GN = Sdprup expression
Acetoacetyl-CoA synthetase GN = Aacs down expression	Acetoacetyl-CoA synthetase GN = Aacs up expression

Table 3 Differentially expressed proteins between W/M vs. W/C groups.

W VS. C	W VS. M
Protein Frmd4b GN = Frmd4bup expression	Protein Frmd4b GN = Frmd4bup expression
N-ethylmaleimide sensitive factor NSF GN = Nsfup expression	N-ethylmaleimide sensitive factor NSF GN = Nsfup expression
Hydroxyacid oxidase 2 GN = Hao2up expression	Hydroxyacid oxidase 2 GN = Hao2up expression
calcium-independent phospholipase A2 GN = Pla2g6up expression	calcium-independent phospholipase A2 GN = Pla2g6up expression
Protein Dda1 GN = Dda1up expression	Protein Dda1 GN = Dda1up expression
Peptidylprolyl isomerase domain and WD repeat containing 1 GN = Ppwd1up expression	Peptidylprolyl isomerase domain and WD repeat containing 1 GN = Ppwd1up expression
Anamorsin GN = Ciapin1up expression	Anamorsin GN = Ciapin1up expression
Keratin, type II cytoskeletal 5 GN = Krt5up expression	Keratin, type II cytoskeletal 5 GN = Krt5up expression
Phospholemman GN = Fxyd1up expression	Phospholemman GN = Fxyd1up expression
Phosducin-like protein 3 GN = Pdcl3up expression	Phosducin-like protein 3 GN = Pdcl3up expression
Potassium/sodium hyperpolarization-activated cyclic nucleotide-gated channel 4 GN = Hcn4up expression	Potassium/sodium hyperpolarization-activated cyclic nucleotide-gated channel 4 GN = Hcn4up expression
Small G protein signaling modulator 3 GN = Sgsm3down expression	Small G protein signaling modulator 3 GN = Sgsm3down expression
ERC protein 2s GN = Erc2down expression	ERC protein 2s GN = Erc2down expression
Phytanoyl-CoA dioxygenase domain-containing protein 1 GN = Phyhd1 down expression	Phytanoyl-CoA dioxygenase domain-containing protein 1 GN = Phyhd1 down expression
Protein Enpp4 GN = Enpp4down expression	Protein Enpp4 GN = Enpp4down expression

Data availability statement

The data that support the findings of this study are openly available in ‘Zenodo’ at https://doi.org/10.5281/zenodo.8268067

Ethics approval and consent to participate

The animal experiments were approved by the Institutional Animal Care and Use Committee of Inner Mongolia Medical University (YKD2018096).