Optimization Of Extraction And Function Of Buthus Martensii Karsch In Traditional Chinese Medicine

The dry body of Buthus martensii Karsch is used as a traditional Chinese medicine,which is processed by complicated procedures.It has more than 1,000 years of medicinal history in China,and has many pharmacological effects such as anti-epilepsy,anticonvulsant,analgesic,anti-tumor and immune regulation.Due to the limitations of the understanding of medicinal scorpion,the quality control methods of medicinal scorpion have not been updated for several decades.On the contrary,the outdated quality control method of medicinal scorpion has limited the understanding and modernization of medicinal scorpion.This thesis intends to carry out the extraction method of medicinal scorpion,explore the active ingredients and biological functions of medicinal scorpion,and provide reference for establishing a more scientific quality control standard.First,this paper optimizes the extraction method of protein component in medicinal scorpion.This study started from the medicinal scorpion tablets,and divided medicinal scorpion into two parts:the body and the tail.By comprehensively comparing the effects of ultraviolet sterilization,⁶⁰Co-γray irradiation sterilization and high pressure steam sterilization on the sterilization effect and the extraction components of medicinal scorpion,it was found that the ⁶⁰Co-γray irradiation method was effective for the sterilization of medicinal scorpion.The temperature,materail-liquid ratio,solvent and time were extensively investigated.The orthogonal test showed that the temperature was the main factor affecting the extraction of the active constituents of medicinal scorpion.The temperature was determined to be 60℃,the ratio of material to liquid was 1:30,and the concentration of ethanol was 50%,extraction time 6 h was the optimal extraction method for the total active ingredient.In order to avoid the effect of salt in medicinal scorpion on the biological function evaluation,this paper used dialysis method to effectively remove the salt in medicinal scorpion with a cutoff flow of 100 Da dialysis bag.It was also proved irrational that some content detection of medicinal scorpion used as quality control indicators in current pharmacopoeia.Secondly,this paper initially carried out the component analysis and function exploration of medicinal scorpion extract.According to the optimized extraction method,we found that the medicinal scorpion extracts from different sources presented similar UV spectral characteristics,but showed different chromatographic separation characteristics.In vitro cell experiments showed that medicinal scorpion had a certain anti-tumor effect,in which the anti-tumor effect of the body was stronger than that of the tail.Different tumor cells showed different sensitivity to the extract,gastric cancer cell AGS,colon cancer cell HCT116,leukemia cell line HL-60,Jurkat,Raji were more sensitive to the extract of the body than other cells.Electrophysiological experiments showed that medicinal scorpion extract still had strong ion channel modulating activity,and the extract of medicinal scorpion tail could inhibit different potassium channel currents,such as human potassium channels Kv1.2,Kv1.3 and Kv4.1.Their inhibitory concentration IC₅₀ values were 46.2±4.5μg/mL,192.8±22.6μg/mL,and 141.2±34.3μg/mL,respectively.The body had only a weak inhibitory effect on some channels.In addition,the extracts of boiled medicinal scorpion expressed lower effects than those of untreated ones,which indicated that the processed medicinal scorpion might not be suitable for further boiling.In summary,this paper develops a new method for extracting medicinal scorpion,explores the composition of the scorpion and conducts preliminary research on its function,providing a biological basis for the establishment of more scientific and reasonable quality standards.