Study On The Anti-Agkistrodon Acutus Venom Activities Of Aqueous Extracts Of 24 Species Of Chinese Herbal Medicines And On Isolation And Identification Of The Effective Anti-snake Venom Components From Aqueous Extracts Of Pothos Chinensis

Snakebite have been defined by WHO as a neglected tropical disease that occurs in tropical or subtropical poor rural area.Snakebite envenoming usually causes bodily harm,economic burden,and psychological panic sequelae to snakebitten victims.A common treatment for snakebite first aid is the use of anti-snake venom serum prepared from animals?such as horses?immunized with snake venom to treat patients.However,in some areas?mainly in underdeveloped or developing regions?,herbal medicines are still used to treat snakebite due to various factors,especially in China,the use of Chinese herbal medicine to treat snakebitten victims has achieved satisfactory results,and has relieved the snakebite patients suffering.However,the scientific experimental evidence for the treatment of snakebite by Chinese herbal medicines is not fully sufficient.For this reason,the study used 24species of Chinese herbal medicines commonly used by the doctors in China to treat snakebitten patients.The experiments for anti-snake venom activities were conducted to preliminary reveal the efficacy of these Chinese herbal medicines.Via using ultrasonic extraction,24 species of Chinese herbal medicine aqueous extracts were obtained.The agarose plate method was used to examine the inhibitory effects of the 24 Chinese herbal medicines'extracts against the activities of the main functional components of the A.acutus venom,including PLA₂,hyaluronidase and protease.The experimental results showed that there were Anti-snake venom activities of all the 24 species of Chinese herbal medicines.But the efficacy was different among them.Only P.chinensis has the highest anti-snake venom activities.In order to identify the effective components of anti-snake venom activity in the aqueous extract of P.chinensis,we used macroporous resin to chromatograph,eluting with 25%,50%and 75%ethanol,respectively,then collecting the eluent,and detecting their anti-snake venom activities of PLA₂ and protease in snake venom.The experimental results showed that the eluent which was eluted with 75%ethanol has the highest anti-snake venom activity among them.Then,the eluate was isolated and purified by using HPLC and obtain three peaks P₁,P₂,and P₃.The anti-snake venom activity of PLA₂ and protease were detected in these three peaks.The experimental results showed that P₂ peak had the highest anti-snake venom activity.To further identify the effective substances in the P₂ peak,we used GC-MS to detect and analyze it.The preliminary experimental results showed that the main effective component of the P₂ peak was pyrogallol,which showed the obvious anti-A.acutus venom activities of PLA₂ and protease.In conclusion,this study preliminary revealed that 24 species of Chinese herbal medicines have certain activities against A.acutus venom.Among them,P.chinensis had the highest activity against the A.acutus venom,and one of the effective components was identified as pyrogallol from P.chinensis.These preliminary experimental results may provide,to a certain extent,the scientific experimental evidence for the treatment of snakebitten victims by 24 species of Chinese herbal medicines commonly used by the doctors who treated snakebitten victims in China.However,there are still many effective components that require further identification studies,and how these effective components interact with the main toxic components of snake venom at the molecular level.