| Artemisia hedinii Ostenf.et Pauls.is a plant of the genus Artemisia L.Its Tibetan name is " Sangzi Nabu ".The traditional medicinal efficacy of A.hedinii is to clear heat and cool the blood,reduce yellowing and reduce inflammation,which has been used by Tibetan doctors since ancient times to treat "Chiba"disease,acute jaundice hepatitis and cholecystitis.(Ministerial Drug Standards(Tibetan Medicine Volume One)"(1995 edition),"Common Tibetan Medicine Chronicle","Chinese Tibetan Medicine Chronicle","Gansu Chinese Herbal Medicine Resources Chronicle","Chinese Materia Medica")and other related monographs record that its root is the dried above-ground parts of A.hedinii.In previous studies,sesquiterpenoids were found to be the main structural type of its chemical composition,and the plant and its components were often reported as anti-inflammatory and antibacterial potential drugs.At the same time,in the"Ministerial Drug Standards(Tibetan Medicine Volume One)"(1995 edition),there are only regulations on character,powder and physical and chemical color identification,and the quality standards of A.hedinii medicinal materials are extremely imperfect.In order to verify the traditional medicinal efficacy of Tibetan medicine A.hedinii and to investigate the material basis of its efficacy against intrahepatic cholestatic liver injury,this paper uses α-naphthylisothiocyanate(ANIT)as a moulding agent to construct a mouse model of intrahepatic cholestasis,to compare the efficacy of the total extract of A.hedinii and its polar parts,to screen the effective parts,and the chemical components of the effective parts were systematically separated.On the basis of this study,10 batches of A.hedinii medicinal materials were studied for quality standard.The specific research content is as follows.1.Pharmacodynamic evaluation and screening of effective parts of Artemisia hedinii Ostenf.et Pauls.against intrahepatic cholestatic liver injury.In this experiment,ANIT(60 mg/kg)was used to construct a mouse model of acute intrahepatic cholestatic liver.13 groups of 10 mice were divided into normal group,model group and positive drug group,and high and low dose groups were set for the total extract of A.hedinii,petroleum ether,ethyl acetate,n-butanol and water.According to the clinical dosage of A.hedinii and the results of the preliminary experiments,the doses of mice in the high and low dose groups were set at 2.4 and 0.8 g/kg of raw drug respectively.Blood and liver were collected after 7 days of gavage administration,and the relevant indexes in the serum and liver homogenate of the mice were measured to observe the morphological changes and histopathological sections of the liver.The results showed that the high/low dose groups of A.hedinii total extract,ethyl acetate site and petroleum ether site could significantly reduce the levels of ALT,AST,ALP,TBIL,DBIL and TBA in mice serum,and increase the levels of SOD and GSH in mice liver tissue,as well as reduce the activity of MDA,while the other sites had no significant effect.The morphological changes and histopathological findings in the livers of the mice at these three sites further suggest that they alleviate the ANIT-induced liver damage in mice.Therefore,the ethyl acetate and petroleum ether sites were identified as effective sites against intrahepatic cholestatic liver.2.Study on the chemical composition of the active parts of Artemisia hedinii Ostenf.et Pauls.against intrahepatic cholestatic liver injury.A total of 23 compounds were isolated and identified from the ethyl acetate and petroleum ether sites,including 9 flavonoids,5 sesquiterpenes,4phenylpropanoids,2 polyacetylenes,1 triterpene,1 lignan and 1 phenolic acid.These compounds are: brickellin(CH1)、 apigenin(CH2)、 3α,16α-dihydroxy-4,4,9,14-tetramethyl-3,5-di-O-caffeoyl-muco-quinic acid(CH6)、 2′,5,5′-trihydroxy-3,4′,6,7-tetramethoxyflavone-2′-O-β-D-glucoside(CH8)、rel-(2α,3β)-7-O-Methylcedrusin(CH9)、(E)-Tonghaosu(CH10)、(Z)-Tonghaosu(CH11)、(9β,10α)-19-norpregn-5-ene-11,20-dione(CH13)、Artetourneforin B(CH18)、9-Oxo-6α,7α(H)-eudesm-4,11(13)-dien-12,6-olide(CH19)、 luteolin 5-O-β-Dglucoside(CH23)、luteolin 7-O-β-D-glucoside(CH25)、11,12-dihydroxyeudesm-4-en-3-one(CH33)、 Hispiduloside(CH41)、 5,5′-trihydroxy-3,4′,7-tetramethoxyflavone-2′-O-β-D-glucoside(CH43)、 methyl 4,5-di-O-caffeoylquinate(CH50)、ethyl 3,3′,4,4′-tetrahydroxy-δ-truxinate(CH51)3,5-di-O-caffeoylmuco-quinic acid methyl ester(CH53)、 1,3-O-di-E-caffeoyl glycerol(CH55)、luteolin(CH60)、(7R,9S,10S)-9-acetoxyeudesm-3,5,11(13)-trien-12-oic acid(CH62)、9β-Aceroxy4,5-dehydro-4(15)-dihydrocosric acid(5)(CH63)、quercetin-3-O-β-D-glucoside(CH69).Among them,CH1,CH8,CH9,CH13,CH23,CH33,CH43 and CH51 are the first division of Artemisia L,and the other 15 compounds are the first division of this plant.3.Study on the quality standard of Artemisia hedinii Ostenf.et Pauls.Ten batches of A.hedinii herbs from the Tibetan region were studied for quality standards according to the four General Principles of the 2020 edition to provide a methodological reference for the establishment of quality standards for the herbs.The powder characteristics of the herbs were characterized by microscopic identification;A thin-layer identification method was established using(E)-Tonghaosu(CH10)and(Z)-Tonghaosu(CH11)as controls;The moisture,total ash and alcohol-soluble extract contents of 10 batches of A.hedinii herbs were measured,and the moisture content ranged from 8.26% to 9.39%,the total ash from 8.98% to 21.67% and the alcohol-soluble extract from 12.30% to 29.75%.The content of chlorogenic acid in 10 batches of A.hedinii herbs ranged from 1.90mg/g to 13.45 mg/g,and the content of isochlorogenic acid A ranged from 1.31mg/g to 8.66 mg/g.In summary,this paper was the first to evaluate the protective effect of the Tibetan medicine A.hedinii on intrahepatic cholestatic liver injury using an animal model,and the effective parts were screened for ethyl acetate and petroleum ether.A total of 23 compounds were isolated and identified from the active parts,and their main chemical components were flavonoids,sesquiterpenoids and phenylpropanoids.On the basis of the previous pharmacological and chemical studies,the quality standards of 10 batches of A.hedinii herbs from Tibet were further investigated and a draft quality standard for A.hedinii was developed.The results of this study provide basic data for the clarification of the material basis of the efficacy of the Tibetan medicine A.hedinii against intrahepatic cholestatic liver injury and the quality control of A.hedinii herbs,and lay the foundation for the further development and utilization of A.hedinii resources. |
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