| 英文摘要 |
Trifolium repens (white clover) is rich in bioactive volatiles with therapeutic potential. This study developed an advanced hybrid ultrasound—infrared—microwave hydro-distillation (UIMHD) method for efficient essential oil (EO) isolation and investigated its active sesquiterpene, pseudoionone, for multi-mechanistic neuroprotection in diabetic neuropathy (DN). The optimized UIMHD system synergistically combined ultrasound cavitation, infrared heating, and microwave hydro-distillation, enhancing yield and reducing time versus conventional extraction. Gas chromatography—mass spectrometry (GC—MS) profiling identified pseudoionone as the dominant constituent. Pharmacological evaluations were performed in alloxan-induced diabetic mice to assess antidiabetic, anti-inflammatory, neuroprotective, and anticonvulsant activities. Cytokine modulation (TNF-a, IL-6, IL-10), oxidative stress markers (CAT, GSH, TBARS), and strychnine-induced seizure assays were conducted to elucidate mechanisms. The UIMHD technique increased T. repens essential oil (TR-EO) yield by 62.5% and shortened isolation time by 50%. Pseudoionone (49.16%) exhibited potent hypoglycemic and insulin-restorative effects, alleviated thermal hyperalgesia and tactile allodynia, and improved antioxidant defenses. Both the TR-EO and pseudoionone significantly down-regulated TNF-a and IL-6 while up-regulating IL-10, indicating anti-inflammatory cytokine balance restoration. Additionally, pseudoionone delayed strychnine-induced tonic seizures, suggesting functional involvement of glycinergic pathways. Strong antibacterial and antibiofilm activities were also observed against multidrug-resistant Shigella dysenteriae. The advanced UIMHD method provides a sustainable platform for isolating bioactive volatiles from T. repens. Pseudoionone emerges as a multi-target natural agent mitigating diabetic neuropathy via TNF-a/IL-6/IL-10 modulation, oxidative- stress restoration, and strychnine-sensitive GlyR potentiation. |
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