Exploring the in vitro anti-inflammatory activity of gross saponins of Tribulus terrestris L. fruit by using liquid chromatography-mass spectrometry-based cell metabolomics approach.

Our previous studies confirmed the efficacy of gross saponins of Tribulus terrestris L. fruit in treating cerebral ischemia. This study aimed to investigate the related mechanisms in vitro. The lipopolysaccharide-induced BV2 cells model was constructed and treated with gross saponins at different concentrations to explore its anti-inflammatory activity. The cell metabolite changes were tracked by liquid chromatography-mass spectrometry (LC-MS)-based metabolomics, and the metabolic biomarkers and related metabolic pathways were analyzed. Molecular biochemistry analysis was further used to verify the relevant inflammatory pathways. The results showed that the saponins reduced nitric oxide release and the secretion of tumor necrosis factor-alpha, interleukin-1ß, and interleukin-6 from lipopolysaccharide-induced BV2 cells. Metabolic perturbations occurred in lipopolysaccharide-treated BV2 cells, which could be reversed by drug treatment via mainly regulating glycerophospholipid metabolism, tryptophan metabolism, purine metabolism pathways, etc. The western blot analysis demonstrated that saponin could suppress the activation of the inflammatory-related signaling pathway. The present study explored the in vitro anti-inflammatory mechanism of gross saponins of Tribulus terrestris L. fruit using an LC-MS-based cell metabolomics approach, which confirms the great potential of LC-MS for drug efficacy evaluation and can be applied in other herbal medicine-related analyses.

GC-MS-Based Metabolomics to Reveal the Protective Effect of Gross Saponins of Tribulus terrestris Fruit against Ischemic Stroke in Rat.

Stroke is one of the most common neurological disorders and seriously threatens human life. Gross saponins of Tribulus terrestris fruit (GSTTF) are used for neuroprotective treatment on convalescents of ischemic stroke. However, the therapeutic effects and mechanisms have not yet well understood, especially from the metabolic perspective. In this study, the protective effect of GSTTF on ischemic stroke in a middle cerebral artery occlusion (MCAO) rat model was investigated by the GC-MS-based metabolomics approach. 2,3,5-triphenyltetrazolium chloride (TTC) staining of brain tissues showed that GSTTF significantly reduced the infarct area after MCAO surgery. Metabolomic profiling showed a series of metabolic perturbation occurs in ischemic stroke compared with sham group. GSTTF can reverse the MCAO-induced serum metabolic deviations by regulating multiple metabolic pathways including fatty acids metabolism, amino acids metabolism, and carbohydrates metabolism. The current study provided a useful approach for understanding the mechanism of MCAO-induced ischemic stroke and a reliable basis for evaluating the efficacy of GSTTF in the treatment of ischemic stroke.

[Ultrastructural changes in rabbits cornea and sclera caused by radio frequency burn].

OBJECTIVE: To assess the influence of radio frequency (RF) burn on the ultrastructure of rabbits cornea, sclera, lens and retina. METHODS: The cornea and sclera of 24 adult New Zealand rabbits were burned by RF (power 15 W) for 10 seconds. The specimens of cornea, sclera, lens and retina were collected and examined under transmission electron microscopy (TEM) at 2 hours, 24 hours, 3 days and 14 days after burning. RESULTS: The TEM showed corneal epithelial cells nuclear chromatin pyknosis, stromal partial colloid fibers rupture, endothelial cells mitochondria turgescence and vacuolization; scleral fibroblastic nuclear chromatin pyknosis, rough endoplasmic reticulum distention, mitochondria turgescence; lens normal; retinal pigment epithelial cells mitochondria turgescence, photoreceptor cells outer segment disarrangement, inner segment mitochondria turgescence, and outer nuclear layer cells' nuclear chromatin pyknosis, 2 hours after burning. Corneal epithelial cells structure was fairly clear, some mitochondria of endothelial cells were turgescent, scleral fibroblastic mitochondria were slightly turgescent, and retinal structure was almost normal, 3 days after burning. There were no significant changes of cornea, sclera and retina in ultrustructure when the burnt groups were compared with the control group, 14 days after burning. CONCLUSION: These findings suggest that RF can cause direct lesion of cornea, sclera and indirect lesion of retina by burning cornea and selera, but the ultrastructural changes will almost recover 14 days after RF burning.