Study On The Folk Application And Anti-tuberculosis Activity Of Hedyotis Scandens Roxb

The global tuberculosis epidemic is still grim.In 2017,there are an estimated 10 million people infected with tuberculosis worldwide,and 1.6 million deaths(including 300,000 HIV-positive tuberculosis patients).The rapid increase of multidrug-resistant Mycobacterium tuberculosis(M.tb)and extensively drug-resistant M.tb worldwide make the prevention and treatment of tuberculosis face tough challenges.Therefore,development of new drug against tuberculosis is urgent needed.Ethnomedicine is a treasure trove of resources accumulated by human beings for a long time.It is the wisdom of all ethnic groups in the struggle against diseases by using traditional medicines in the long-term,and they provide important resources and key clue for modern medical research inadvertently.Based on the potential medicinal value and price advantage on the treatment of tuberculosis by using traditional medicines,it is important to collect,collate and research on traditional Chinese medicine for the treatment of tuberculosis,which will promote the protection of traditional medicine knowledge and enrich the modern medicine.In this paper,several methods like interdisciplinary research method,literature research method,ethnological plant investigation method,ethnological investigation method and phytochemical research method was united to survey the traditional single prescriptions and compound prescriptions used by 15 ethnic groups for the treatment of tuberculosis.Among them,18 ethnic medicines are selected for activity verification and the experimental results show that some ethnic medicines do have antimicrobial activity against Mycobacterium smegmatis,a non-pathogenic and fast growth mycobacterium.Based on the antimicrobial activity,procurability and folk application,Hedyotis scandens Roxb.has been chosen for further research on effective part screening and related chemical composition analysis.Overall,the main research contents of this paper are as follows:(1)Collating anti-tuberculosis Ethnomedicine theory and prescription.The clinical basic theories of some Ethnomedicine(mainly Miao,Yi,Yao,and Yi)and their understanding,classification,diagnosis and treatment of tuberculosis and the common medication characteristics of Ethnomedicine was summarized.At last,1154 prescriptions of Ethnomedicine for tuberculosis treatment was collected,which includes 677 simple prescription and 477 compound prescription from 15 ethnic groups(Miao,Yao,Don,Yi,Tujia,Ji' nuo,Zhuang,Li,De' ang,Zang,Mao' nan,Ge' lao,Shui,Dai,Hui and dominated in Miao and Yi of Guizhou).For these prescriptions,the characteristics of families,forms,medicines,place names,Chinese translations,places of origin,best collection methods and time,medication contraindications were catalogued.(2)Preliminary qualitative verification of anti-tuberculosis activity for some ethnic medicine.To verify the effectiveness of the prescriptions we collected,18 ethnic medicines were screened and their crude alcohol extractions were used for antibacterial activity test against Mycobacterium smegmatis(M.sm)by disk diffusion test.The results showed that some of them,such as Fagopyrum dibotrys(D.Don)Hara?Hedyotis scandens Roxb.?Lysionotus pauciflorus Maxim and Ardisia japonica(Thunb)Blume had relatively obvious antibacterial effects.The extraction of Fagopyrum dibotrys(D.Don)Hara and Hedyotis scandens Roxb exhibit the best bacteriostatic activity: the inhibition zone diameter was greater than 10 mm and 13 mm at the concentration of 200 mg/mL and 500 mg/mL respectively.And these drugs have almost no antibacterial activity against Escherichia coli,Staphylococcus aureus,Bacillus subtilis and Rhodotorula mucilaginosa(only extracts of Fagopyrum dibotrys(D.Don)Hara and Lysionotus pauciflorus Maxim show antimicrobial activity against Staphylococcus aureus;and the extracts of Cynanchum atratum Bunge show antimicrobial activity against Rhodotorula mucilaginosa).these results indicate that these drugs have specific antibacterial activity against mycobacteria,and the folk treatment of tuberculosis has a high degree of specificity.(3)Active site screening of Hedyotis scandens Roxb.Basis on the above results,the crude extracts of Hedyotis scandens Roxb.was furtherly extracted with petroleum ether,dichloromethane and n-butanol for different polar extraction and their antibacterial activity was tested.The results showed that the active components against M.sm were mainly distributed in dichloromethane phase and n-butylate phase.The dichloromethane phase was further separated by thin layer chromatography,column chromatography,semi-preparative high-performance liquid chromatography(HPLC),etc.,and the active site of dichloromethane phase of Hedyotis scandens Roxb.was located in the methanol ratio of 25:1,10:1.5:1 segment.And the active Fragment 13 sample was further separated and the active compounds was mainly concentrated in a dichloromethane: methanol ratio of 15:1.(4)Identification of volatile substances in Hedyotis scandens Roxb.To further understand the volatile components in Hedyotis scandens Roxb.,for the first time,37 volatile components involving aldehydes,ketones,alcohols,esters,and unsaturated alkanes was analyzed and identified by GC-MS.To a certain extent,this research has promoted the process of collecting and collating of anti-tuberculosis ethnic medicines.The activity of some ethnic medicines with anti-tuberculosis activity were preliminarily verified by test against M.sm by disk diffusion test in vitro.Hedyotis scandens Roxb.,whose crude extraction exhibits the best antibacterial activity,was chosen for activity part separation and tracking.This research preliminary confirms that Hedyotis scandens Roxb.is an effective ethnic pharmacological use for the treatment of tuberculosis and laid a foundation for the further screening and isolation of anti-tuberculosis active substances from Hedyotis scandens Roxb.in future.