Study On Resources And Quality Of Marchantiae Convoiutae Herba In Guangxi

Marchantia convoluta Gao et Chang,a medicinal herb commonly used in Guangxi,is usually used for external treatment of fractures,scalds,tinea and unhealed sores and for internal treatment of icterepatitis,with good efficacy.Previous studies showed that flavones in the plant are effective in physiological activity and can be used as the raw material.Meanwhile,it was found that great differences existed in the quality of Marchantiae Convoiutae Herba from different areas,especially in the content of total flavones.On the other hand,Marchantiae Convoiutae Herba populations are wild and the wild resources are lacking.Therefore,it is urgent to protected and identify producing areas with high quality wild resources and carry out their domestication.For the purpose,this paper carried out the basic research of Marchantiae Convoiutae Herba of development of resources in Guangxi.1.Investigation into resource distribution of Marchantiae Convoiutae Herba in Guangxi;2.Observation and evaluation of the liver protection effect of total flavones from Marchantiae Convoiutae Herba on rats with CCl₄-induced acute liver injury as well as study of the relevant mechanism;3.Design and development of SSR primers of Marchantiae Convoiutae Herba in Guangxi using the SAM method to provide the basis for molecular pharmacognosy identification;4.The method for genomic DNA extraction of Marchantiae Convoiutae Herba and the ISSR-PCR reaction system was optimized:5.The safety limit of heavy metals was detected by atomic absorption spectrophotometry and fluorescence analysis.The safety limit of pesticide residues was detected by gas chromatography;6 The fingerprints of Marchantiae Convoiutae Herba in Guangxi were established by high performance liquid chromatography.Our results are as follows:1 Resource investigation demonstrated that Marchantiae Convoiutae Herba populations were widely distributed in Guangxi,and 11 producing areas were rich in resources.2 By observing and evaluating the liver protection effect of total flavones from Marchantiae Convoiutae Herba in Guangxi on rats with CCl₄-induced acute liver injury,we found that:2.1 Rats in the total flavones group had increased SOD activity and decreased MDA level in the serum compared with the model control group,but no significant differences existed in the GSH-Px activity of both groups,suggesting that the total flavones may protect the liver through inhibition of lipid peroxidation.2.2 Rats in the total flavones group had lower TNF-?,IL-1?and IL-6 levels in the hepatic tissue relative to the model control group,suggesting that the total flavones may protect the liver by inhibiting inflammation.2.3 As compared with the model control group,serous ALT and AST levels dropped and there was less liver damage in the pathological section in the total flavones group,indicating that total flavones from Marchantiae Convoiutae Herba in Guangxi can protect the liver of rats with acute liver damage.The above results show that total flavones from Marchantiae Convoiutae Herba in Guangxi are effective in protecting the liver of rats with acute liver damage caused by CCl₄,and the relevant mechanism may be in relation to the inhibition of lipid peroxidation and inflammation.3 The SAM method was employed to design Marchantiae Convoiutae Herba in Guangxi SSR primers:The modified CTAB method was used for DNA extraction;DNA was subject to enzyme digestion,ligation,inhibitory PCR,preamplification;the expected fragments were obtained through amplification by the SAM method;ligated amplification products were transformed into Escherichia coli;positive clones were obtained by random selection of plagues and used for sequencing,and 86 sequences suitable for SSR primer design were obtained.30 pairs of SSR primers effectively amplified were chosen via PCR,and optimal parameters and key technologies in favor of molecular pharmacognosy identification were learned.Our study filled the gap in respect to domestic research into Marchantiae Convoiutae Herba growing in Guangxi,and established a foundation on which to develop and utilize Marchantiae Convoiutae Herba resources in Guangxi.4 The method for DNA extraction of Marchantiae Convoiutae Herba and the ISSR-PCR reaction system was optimized.The 20?l ISSR-PCR system contained 250?mol/L dNTP mix,1.6mmol/L of Mg²⁺,0.6?mol/L of primers,0.75 U of Taq polymerase,50ng of the template DNA.5 The result of heavy metals and pesticide residue in Marchantiae Convoiutae Herba in Guangxi was that content of cadmium was 0.38mg/kg,and lead was 16.73mg/kg,and iron was 9.49g/kg,and copper was 34.01mg/kg,and mercury was 0.12mg/kg.The total 666??-BHC??-BHC??-BHC??-BHC?were undetected.The total DDT?pp'-DDE,pp'-DDD,op'-DDT,and pp'-DDT?were undetected.PCNB was not detected.The results showed that the heavy metal lead,iron and copper exceeded the standard.It was the first time to study safety evaluation of heavy metals and pesticide residue of Marchantiae Convoiutae Herba in Guangxi.6 The HPLC conditions of the eight batch of Marchantiae Convoiutae Herba in different regions of Guangxi were as follows:ZORBAX SB-C18 column?4.6mm×250mm,5?m?,and the mobile phase was acetonitrile?B?-0.1%phosphoric acid aqueous solution?D?,and Gradient elution were:5%?5%B?0-5 min?,5%?100%B?5⁶0min?,100%?100%B?60⁸0min?.flow rate was 1.0 mL·min^-1,and a wavelength of 230 nm,and a column temperature of 25°C and an injection volume of 20?L.The relative retention time of the common peaks was basically the same,the chromatographic peaks were well separated,and the relative peak area of the common peaks of different batches of crude drugs was different,and the similarity range was 0.943⁰.774.It was the first time to provide HPLC fingerprint of Marchantiae Convoiutae Herba in Guangxi.