Studies On Chemical Constituents And Bioactivities Of Four Medicinal Plants

The dissertation dealt with the chemistry and pharmacological effect of three characteristic uighur medicinal material in Xinjiang(Orobanche coerulescens Steph, Nymphaea candida Presl and Juniperus Sabina L) and one folk medical plant from Yurman in China(Ligularia atroviolacea).The thesis is divided into five main parts.Partâ… Study on chemical constituents and bioactivities of Orobanche coerulescensOrobanche coerulescens Steph(local name:caocongrong,dugencao),is biennial or perennial parasitic herbs,warm in property,sweet in flavour,wide spread in downsand,steppe and Gobi desert.This pant is parasitized at the root of composite artemisia plant,and produced in northwest and northeast of China.O.coerulescens have efficacies of kidney-strengthing and strong gluten,and are used in treatment of kidney deficiency,neurosis and hepatitis.There is about 100 species in genus Orobanche in the world,most of which are distributed over temperate zone.There are 23 species and 3 varieties Orobanche plants in China,most of which are distributed over north,northwest and northeast.The compounds of Orobanche were reported at phenylethanoid glycosides such as acteoside and isoacteoside,and acteoside has significantly protective againstα-galactose induced hepatic injury,which is better than Biphenyl dicarboxylate(DDB).Based on relative literatures,this part has included five resesearch aspects as below.1) Fifteen compounds were isolated and purified by extraction,column chromatography,and their structures were determined on the basis of spectral analysis: acteoside(â… -1),isoacteoside(â… -2),crenatoside(â… -3),cistanoside F(â… -4),sinapoyl-4-O-β-D-glucoside(â… -5),adenosine(â… -6),β-siterol(â… -7),oleanic acid(â… -8), succinic acid(â… -9),caffeic acid(â… -10),protocatechuic aldehyde(â… -11),p-hydro xybenzyl alcohol(â… -12),β-daucosterol(â… -13),D-galacitol(â… -14),D-mannitol(â… -15).â… -4～15 were obtained from this plant for the first time,andâ… -6,7,9,and 13 were isolated from Orobanche genus for the first time.2) The scavenging test of DPPH showed that most compounds have comparative antioxidant activity as L-ascorbic acid and part of them show better activity such as the O.coerulescens extract and phenylethanoid glycosides.Acteoside showed potent free radical scavenging effects with a median inhibition concentration of 25.6μg/ml.3) The anti-HBV activities of acteoside,isoacteoside and crenatoside were measured,and all of them showed suppressive activity on the expression of HBsAg and HBeAg in the HepG2.2.15 cell line.Realtime quantitative PCR analysis showed that the studied compounds have inhibition on the HBV DNA too.4) Hepatoprotective effects were tested using Bacille Calmette-Guerin vaccine (BCG) combined lipopolysaccharides(LPS) induced immunologic injury in primary cultured rat hepatocytes models in vitro.Acteoside exhibited significant hepatoprotective activity by reducing BCG vaccine combined lipopolysaccharides (LPS) induced change in bio-chemical parameters that was evident by enzymatic examination.5) The hepatoprotective effects of acteoside were evaluated in Bacille-Calmette-Guerin and lipopolysaccharide induced immunological liver injury(ILI) in mice in vivo.Acteoside(50,150,300 mg/kg) was administered via gavage daily for 12 days. After following the 12 days treatment,various bio-chemical parameters were evaluated,such as liver index(liver weight/body weight),liver homogenate levels of aspartate aminotransferase(AST),alanine aminotransferase(ALT),hepatic nitric oxide(NO),malondialdehyde(MDA) content,superoxide dismutase(SOD) activity, production of tumor necrosis factor-γ(TNF-γ) and interleukin-2,4,10(IL-2,4,10) by peritoneal macrophages,and histopathologic changes of the liver.Acteoside(50,150, 300 mg/kg) effectively reduced the BCG plus LPS-induced elevated liver index,liver homogenate AST and ALT levels,hepatic NO and MDA content,and restored hepatic SOD activity in ILI mice.Acteoside group and DDB group were much better than the model group on histopathologic changes.Acteosdie treatment also alleviated diminished splenocyte proliferation induced by BCG plus LPS and repressed abnormally high levels of TNF-γand IL-2 from peritoneal macrophages.The histopathological analysis suggested that acteoside reduced the degree of liver injury of ILI mice.The expression of Bax was obviously raised in tissue of liver while Bcl-2 was evidently decreased in serum of model mice.Acteoside group and DDB group were all higher than the model group on the expression of Bcl-2(P<0.01),while lower on the expression of Bax(P<0.01).These results suggest that acteoside has a protective and therapeutic effect on ILI mice,which might be associated with its antioxidant properties,ability to reduce NO production immunoregulatory function and regulating cell apoposis etc. Partâ…¡Study on the chemical constituents of Nymphaea candidaNymphaea candida Presl is a herbaceous hydrophyte native to the southern Xinjiang in China,and the flowers of N.candida has been used as a folk medicine for head pains,common cold,cough,hepatitis and hypertension.It has been reported that the extract of N.candida flowers shows antioxidative,anti-inflammatory and hepatoprotective actions.Twenty compounds were isolated and purified by extraction, column chromatography,and their structures were determined on the basis of spectral analysis:gallic acid(â…¡-1),gallic acid methyl ester(â…¡-2),rutin(â…¡-3),quercetin (â…¡-4),kaempferol(â…¡-5),quercetin 3-methyl ether(â…¡-6),tricin 7-methyl ether(â…¡-7), astragalin(â…¡-8),quercetin 3-methyl ether 3'-O-xyloside(â…¡-9),quercetin 3'-O-xyloside(â…¡-10),isoquercitrin(â…¡-11),kaempferol-3-O-rutinoside(â…¡-12), p-digalloyl acid(â…¡-13),m-digalloyl acid(â…¡-14),stigmasterol(â…¡-15),docosanoic acid,2,3-dihydroxypropyl ester(â…¡-16),tetracosanoic acid,2,3-dihydroxypropyl ester (â…¡-17),hexadecanol(â…¡-18),n-docosane(â…¡-19).These compounds were obtained from this plant for the first time,and conpoundsâ…¡-6～8 andâ…¡-11～19 were isolated from Nymphaea genus for the first time.The scavenging capacity of the extracts of N.candidas to DPPH.is equal to Vc. Moreover,Reducing power,O₂^-,·OH and H₂O₂ radicals scavenging activities of acetic ether extracts of N.candida(NCE) were determined,and NCE showed potently antioxidant capacity.N.candida exhibited significant hepatoprotective activity by reducing BCG vaccine combined lipopolysaccharides(LPS) induced change in bio-chemical parameters that was evident by enzymatic examination.Hepatoprotective effects were also inhibited by NCE in dose-dependent manners.NCE showed suppressive activity on the expression of HBsAg and HBV e antigen(HBeAg) in the HepG2.2.15 cell line. There is hypoglycemic effect by DPP-â…£high throughput screening for the extracts of N.candidas.Partâ…¢Study on the chemical constituents of Juniperus SabinaJuniperus Sabina is groveling shrub plant from Cupressaceaed,and distribut in Xinjiang,Qinghai and Gansu region,China.S.vulgaris has effect of dreving wind and sedation,promoting blood circulation to stop pain,and treat rheumatic arthritis, difficult urination etc.Twenty compounds were isolated and purified by extraction and different kinds of column chromatography,and their structures were determined on the basis of spectral analysis:α-Cedrol(â…¢-1),coumasabin(â…¢-2),isocupressic acid(â…¢-3),β-sterol(â…¢-4),podophyllotoxin(â…¢-5),cupressuflavone(â…¢-6), cupressuflavone 4′,4′″-Di-Me ether(â…¢-7),apigenin(â…¢-8),amentoflavone(â…¢-9), podocarpusflavone A(â…¢-10),quercetin-3-O-(6″-O-acetyl)-β-D- glucopyranoside (â…¢-11),isoscutellarein 7-O-β-D-xylopyranoside(â…¢-12),catechin(â…¢-13),myricetin 3-O-β-D-glucoside(â…¢-14),qucetin(â…¢-15),rutin(â…¢-16),skimm(â…¢-17), undulatoside A(â…¢-18),hypolaetin 7-O-β-xylopyranoside(â…¢-19),qucetin-3-O-α-L-rhaoside(â…¢-20),syfingin(â…¢-21).The test results of DPPH·scavenging capacity showed that compoundsâ…¢-13 andâ…¢-15 showed antioxidative activity close with that of positive control,L-ascorbic acid,and the flavonoids glycosides(â…¢-12,â…¢-14 andâ…¢-16) are more active than the biflavone(â…¢-6,â…¢-7,â…¢-9 andâ…¢-10).It also showed that the extract of J.Sabina has weak hypoglycemic effect by DPP-â…£high throughput screening.Partâ…£Study on the chemical constituents of Ligularia atroviolaceaLigularia species(Asteraceae) is a perennial herb from Compositae,and mainly distributed in damp,shaded areas of western China,especially the provinces of Gansu, Sichuan and Yunnan.There are more twenty species which used to treat desease in folk.They have the efficacy of antiphlogistic,cough relieving,expelling phlegm and activating blood etc.The present study focus on the chemical constituents of the extracted ethyl acetate parts of L.atroviolacea from Lijiang in Yunnan on the basic of the previous research.Four compounds were isolated and purified by extraction and different kinds of column chromatography,and their structures were determined on the basis of spectral analysis:Furanoeremophil-3en-15,6-olide(â…£-1),1α-Chloro-6β-isobutyroxy-9-oxo-10β-hydroxy-furanoeremophilane(â…£-2),8α-[Eremophil-3′,7′(11′)-dien-12′,8′α;5′,6′α-diolide]-eremophil-dien-12,8α;15,6α-3,7(11)-diolide(â…£-3), Ligulatrovine A(â…£-4).Partâ…¤The development of phenylethanoid glycosides...