Phosphoproteomic analysis reveals differential associations between liver-spleen disharmony and qi-blood deficiency syndromes in [CFS] 2026 Zhang+

[Andy]

Abstract​

Background​

Chronic fatigue syndrome (CFS) is a debilitating disorder characterized by persistent fatigue that is not alleviated by rest and is often accompanied by multiple somatic symptoms. The etiology of CFS remains poorly understood, and conventional Western medicine offers limited effective targeted therapies. In contrast, Traditional Chinese Medicine (TCM), which utilizes pattern differentiation—particularly the Liver-Spleen Disharmony Pattern (LSDP) and the Qi-Blood Deficiency Pattern (QBDP)—has demonstrated clinical efficacy in managing CFS. However, the molecular mechanisms underpinning TCM pattern classification in CFS remain largely unexplored.

Methods​

A total of 30 participants were enrolled in this study, including 10 CFS patients with LSDP, 10 CFS patients with QBDP, and 10 age- and sex-matched healthy controls (HC). Serum phosphoproteomic profiling was conducted using liquid chromatography-tandem mass spectrometry (LC-MS/MS), which incorporated data-dependent acquisition (DDA) for spectral library construction and data-independent acquisition (DIA) for label-free quantification. Differentially phosphorylated sites (DPSs) and proteins (DPPs) were identified with thresholds of absolute fold change (|FC|) ≥ 1.2 and a Benjamini-Hochberg (BH)-corrected false discovery rate (FDR) < 0.05. Principal component analysis (PCA) was employed to assess global differences in phosphorylation profiles across groups, and functional enrichment analyses were performed to elucidate the biological functions of differential molecules.

Results​

PCA revealed distinct clustering of phosphoproteomic profiles among the three groups, with high consistency across biological replicates (PC1 explained 19.3% of the total variance, and PC2 explained 15.9%). A total of 849 non-redundant DPSs and 586 non-redundant DPPs were identified across the three pairwise comparisons. The HC vs. LSDP comparison yielded the highest number of differential molecules (406 DPSs and 351 DPPs), with a balanced distribution of upregulated and downregulated events. In contrast, the HC vs. QBDP comparison was dominated by phosphorylation upregulation (61.2% of DPSs), while the QBDP vs. LSDP comparison showed a higher proportion of downregulated DPSs (56.7%). Functional enrichment analysis indicated that upregulated DPPs in the HC vs. LSDP comparison were primarily involved in MAPK signaling and cytoskeletal remodeling, while downregulated DPPs were enriched in pathways associated with neurodegenerative diseases and nucleocytoplasmic transport. Notably, we identified a candidate differential phosphorylation site, DENND3 S472 (S472@DENND3_HUMAN), with moderate discriminatory power (raw p = 0.042, BH-corrected FDR < 0.05, AUC = 0.72).

Conclusion​

This exploratory study identified significant differences in serum phosphoproteomic profiles between CFS patients with LSDP and QBDP. The distinct phosphoproteomic signatures observed in LSDP and QBDP provide preliminary molecular evidence supporting TCM pattern differentiation in CFS. These findings enhance the understanding of CFS pathogenesis and lay the groundwork for precision-based TCM diagnosis and individualized therapeutic strategies for CFS.

Open access

 

[Hutan]

LSDP: The diagnosis of LSDP was based on the《Expert Consensus on TCM Diagnosis and Treatment of Liver-Spleen Disharmony Syndrome (2023)》 The main symptoms include chest and hypochondriac distension, emotional depression, poor appetite, and loose stools. Secondary symptoms include frequent sighing, chest or hypochondriac pain, abdominal pain with a tendency toward diarrhea, abdominal distension, borborygmus, or diarrhea that relieves pain. The tongue typically has a white coating, and the pulse is wiry or weak. A diagnosis of LSDP was made if at least two main symptoms and two or more secondary symptoms were present.

QBDP: The diagnosis of QBDP was based on the《Guidelines for Clinical Research of New Chinese Medicines (Trial)》The main symptoms include fatigue, shortness of breath, reluctance to speak, and a pale or sallow complexion. Secondary symptoms include dizziness, spontaneous sweating, poor appetite, and insomnia. The tongue appears pale with a thin white coating,and the pulse is thin and weak. A diagnosis of QBDP was made if at least two main symptoms and two or more secondary symptoms were present.

I thought a different cultural view of ME/CFS could be interesting - I'm quite willing to believe that there are actually two sorts of ME/CFS.

I think there is a problem with the translation there, I don't think 'hypochondriac' means what the authors think it means. I think with 'chest and hypochondriac distension' and 'chest and hypochondriac pain', maybe they were thinking of something like 'below the ribs' when they wrote 'hypochondriac'?

Anyway, I don't recognise ME/CFS in either of those definitions. (Although I did catch myself sighing as I read the first definition, which was a bit funny, but I reckon it was due to disappointment, not due to sighing being a symptom of my disease. (Perhaps sighing and eye-rolling are symptoms of reading too many ME/CFS research papers.)

I'm surprised there isn't a TCM diagnosis that fits better.