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To establish and characterize two chronic stress-related dry-eye models using chronic unpredictable mild restraint (CUMR) and chronic unpredictable rhythm disturbance (CURD), respectively, and to compare their phenotypic presentations, metabolomic profiles, and correlations with traditional Chinese medicine syndrome patterns.
Healthy male specific-pathogen-free rats were divided into control, CUMR, and CURD groups. Rats in the CUMR group underwent single-cage housing combined with behavioral restraint daily, while CURD rats received random daily rhythm interventions. After 6 weeks of modeling, dry-eye rats associated with Yu syndrome were assessed for general conditions, behavioral performance, and dry-eye parameters. Histopathological alterations in the conjunctiva, cornea, and lacrimal gland were examined by hematoxylin-eosin staining. Serum concentrations of 5-hydroxytryptamine (5-HT) and cortisol, and cornea levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), were quantified by enzyme-linked immunosorbent assay. Serum metabolomic profiling was conducted using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry.
(1) There were significant differences in appearance and behavior between the CUMR and CURD groups. (2) Compared with the control group, cornea fluorescein staining(CFS) was significantly increased in the CUMR and CURD groups, while tear flow and tear-film breakup time (BUT) were significantly decreased, especially in the CUMR group. (3) There was no obvious inflammatory cell infiltration in the cornea in the CUMR or CURD group. Compared with the control group, there were significantly fewer conjunctival goblet cells in the CUMR and CURD groups, and the area of lacrimal gland vacuolation was significantly increased. (4) Serum 5-HT was significantly decreased and cortisol was significantly increased in the CUMR group compared with the CURD group, but there were no significant differences in cornea tissue TNF-α and IL-1β levels. (5) Serum metabolomic analysis revealed 235 differential metabolites in the CUMR and CURD groups, demonstrating a significant separation trend.
Distinct stress-modeling paradigms differentially impact host metabolism, culminating in divergent TCM syndrome manifestations and dry-eye phenotypes. The CUMR model exemplifies a dry-eye phenotype consistent with "Liver Depression and Spleen Deficiency," potentially mediated by perturbations in tryptophan metabolism, while the CURD model aligns with "Stagnation Heat in the Liver Channel," likely arising from disruptions in the tricarboxylic acid cycle.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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