背景:在中国蛇出没的山区,虎耳草[L.]Meeb被广泛用作毒蛇咬伤的即时补救措施。然而,对蛇毒链球菌在蛇咬伤治疗中疗效的科学认识仍然有限,需要进一步研究。
目的:本研究的目的是评估虎耳草酚类提取物(SSPE)对蛇毒(DAV)的抑制作用,并探讨蛇毒蛇毒作为抗蛇毒血清开发有价值的候选药物的潜力。
方法:我们采用了先前优化的提取条件来获得SSPE。利用不同模型进行体外实验,以评估提取的酚类化合物对DAV的抑制作用,特异性靶向磷脂酶A2(PLA2),蛋白水解,纤维蛋白溶解,和透明质酸酶。此外,进行体内研究以评估提取的化合物对DAV诱导的出血和水肿活性的抑制潜力。为了阐明酚类提取物的化学成分,进行超高效液相色谱-质谱(UPLC-MS)分析。
结果:我们的体外抑制研究表明,蛇床子能够抑制PLA2酶的活性,蛋白水解酶,透明质酸酶和纤溶蛋白。SSPE抑制PLA2酶的中位有效剂量(ED50)值,蛋白水解酶和透明质酸酶活性为0.115mg/mL,0.026mg/mL和0.238mg/mL,分别。SSPE的DAV诱导的出血和水肿作用也在体内被成功抑制。高SSPE浓度能完全抑制出血和水肿。值得注意的是,小鼠没有遭受高SSPE浓度的伤害。成分分析表明,SSPE中含有的酚类物质为没食子酸,原儿茶酸,绿原酸,芦丁,山奈酚-3-O-α-L-鼠李糖苷,山奈酚-3-O-β-D-吡喃葡萄糖苷,槲皮素和山奈酚。
结论:本研究提供了对蛇毒蛇咬伤作为紧急治疗的抑制功效的科学验证,为未来针对蛇咬伤的药物开发策略提供理论基础。
BACKGROUND: In the snake-infested mountainous regions of
China, Saxifraga stolonifera [L.] Meeb is widely utilized as an immediate remedy for venomous snake bites. However, the scientific understanding of S. stolonifera\'s efficacy in snakebite treatment remains limited and requires further investigation.
OBJECTIVE: The aim of this study was to assess the inhibitory effects of Saxifraga stolonifera phenolic extracts (SSPE) on Deinagkistrodon acutus venom (DAV) and explore the potential of S. stolonifera as a valuable candidate for antivenom development.
METHODS: We employed our previously optimized extraction conditions to obtain SSPE. In vitro experiments utilizing diverse models were conducted to assess the inhibitory effects of the extracted phenolic compounds on DAV, specifically targeting phospholipase A2 (PLA2), proteolytic, fibrinolytic, and hyaluronidase enzymes. Furthermore, in vivo investigations were conducted to evaluate the inhibitory potential of the extracted compounds against DAV-induced hemorrhagic and edematogenic activity. To elucidate the chemical composition of the phenolic extracts, Ultra Performance Liquid Chromatography-mass spectrometry (UPLC-MS) analysis was performed.
RESULTS: Our in vitro inhibition study showed that S. stolonifera was able to inhibit the activities of PLA2 enzyme, proteolytic enzyme, hyaluronidase and fibrinogenolytic. The median effective dose (ED50) values of SSPE for inhibiting PLA2 enzyme, proteolytic enzyme and hyaluronidase activities were 0.115 mg/mL, 0.026 mg/mL and 0.238 mg/mL, respectively. The DAV-induced hemorrhagic and edematogenic effects of the SSPE were also successfully inhibited in vivo, and the high SSPE concentration was able to completely inhibit the hemorrhage and edema. It is noteworthy that the mice suffered no harm from the high SSPE concentration. The composition analysis showed that the phenolic substances contained in SSPE are gallic acid, protocatechuic acid, chlorogenic acid, rutin, kaempferol-3-O-ɑ-L-rhamnoside, kaempferol-3-O-β-D-glucopyranoside, quercetin and kaempferol.
CONCLUSIONS: This study provides scientific validation of the inhibitory efficacy of S. stolonifera as an emergency treatment for venomous snake bites, offering a theoretical foundation for future drug development strategies targeting snakebite.