长白山区青楷槭抗弓形虫药效物质基础的研究报告

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论文字数:62522 论文编号:sb2022051215184347469 日期:2022-06-01 来源:硕博论文网

本文是一篇药学论文,本文通过MTT法的体外实验,化合物6'-O-没食子酸红景天苷、槲皮素、灰烷苷A、二氢松果醇、(-)-表没食子儿茶素-3-O-没食子酸酯、6-羟基-7-甲氧基香豆素的值大于spi,表明这些化合物对GES-1细胞的细胞毒性较小。九种测试化合物的IC50(µM)均高于spi值,表明该化合物对弓形虫的毒性更大。其中槲皮素、灰烷苷A和6-羟基-7-甲氧基香豆素的SI优于spi,表明这些化合物可能比螺旋霉素具有更好的抗弓形虫活性。

Chapter 1. Introduction
1.1 Chemical Composition
1.1.1 Flavonoids
Park[6] isolated and identified four flavonoids such as morin-3-O-α-L-lyxoside(1), (-)-catechin (2), 3,7,3′,4′-tetramethyl-quercetin (3),5,3’-dihydroxy-3,7,4’-trimethoxy flavone (4) from the stems of A. tegmentosum. Theexperiment of Park[7] showed that the dry stem of A. tegmentosum contains quercetin(5), morin hydrate (6).
Kwon[8]isolated and identified compounds (-)-epicatechin-3-O-gallate (7),(-)-epicatechin (8) from the stems of A. tegmentosum. Yu[9] isolated and identified3′-hydroxyquercetin (9), (-)-gallocatechin (10) from the stem of A. tegmentosum.Heim[10]isolated it from A. tegmentosum 5,7,4’-trihydroxyflav-one glycosides (11).
Lee[11] isolated many flavonoids from the stem of A. tegmentosum, which werehyperin (12), myricitrin (13), kaempfe-3-rhamnoside (14),querceti-n-3-O-[β-D-xylopyranosyl-(1→2)-β–D-glucopyranoside] (15),6-hydroxy-quercetin-3-O-galactoside (16), (+)-catechin (17), dihydromyricetin (18),(+)-catechin-3-O-(3, 4-dihydroxybenzoyl (19), feniculin (20), avicularin (21) (Fig. 1).
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1.2 Pharmacological action
1.2.1 Hepatoprotective effect
Many experts believe that the activation of hepatic stellate cells caused by liverdamage is an important way of liver fibrosis. Because of that, the medical worldgenerally believes that inducing hepatic stellate cell apoptosis or preventing itsactivation is one of the important ways to combat liver fibrosis. The leaves of A.tegmentosum contain a lot of gallic acid, which can produce negative oxygen ions,hydrogen peroxide, hydroxide and other cytotoxic compounds through auto-oxidation.Moreover, it can selectively act on the hepatic stellate cells, which is the accumulationof hydrogen peroxide in the cells that then kills the hepatic stellate cells with a lowlevel of antioxidant capacity.
Feng[16] used carbon tetrachloride and bile duct ligation to establish a mouse liverfibrosis model. The results showed that salidroside can inhibit the expression of miceCol 1, α-SMA, TIMP1, etc. Reduce the production of ECM and then the deposition ofcollagen in the liver.
Seo[11]used A. tegmentosum's EtOH extract (2 mg/kg) to treat carbontetrachloride (CCl4)-induced acute liver injury in rats, and used triglycerides,cholesterol, AST and ALT as liver injury indicators . After treatment of rats, it wasfound that the triglyceride, cholesterol, AST and ALT levels of rats treated with A.tegmentosum EtOH extract were lower than those of rats that did not consume theextract, which proved that the A. tegmentosum EtOH extract treated rats have aprotective effect on the liver and can prevent liver damage.
On the basis of establishing experimental models of liver fibrosis, Feng foundthat salidroside extracted from A. tegmentosum has significantly inhibited the processof liver fibrosis in mice, significantly reduced the liver fibrosis in mice, and protectedthe liver fibrosis in mice to a certain degree.
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Chapter 2. Experimental contents
2.1 Extraction and isolation
2.1.1 Materials and method

药学论文怎么写
药学论文怎么写

2.1.2 Plant material
The stem bark of A. tegmentosum was collected in Changbai Mountain area, JilinProvince, China, in August 2018. Bark mostly about 10 years A. tegmentosum bark,bark gray to dark gray, smooth, with the crack and the resulting bark naturally dried,pulverized to 1-2cm, spare. The materials were identified by Professor Ren-BoAn(Yanbian University College of Pharmacy). A voucher specimen (YB-QKQ-1708) hasbeen deposited in Yanbian University College of Pharmacy.
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2.2 Determination of the content of salidroside and tyrosol in theEtOH extract of A. tegmentosum
2.2.1 Preparation of standard solution
Accurately weigh 15.0 mg of the standard product, add 15 mL of MeOH todissolve, prepare standard stock solutions of salidroside and tyrosol with aconcentration of 1 mg/mL, respectively, prepare standard salidroside and tyrosol bygradually diluting them Methanol solution 1.0 mg/mL, 0.5 mg/mL, 0.25 mg/mL,0.125 mg/mL, 0.0625 mg/mL, 0.03125 mg/mL. Seal and store frozen.
2.2.2 Preparation of test solution
Accurately weigh 9 parts of 15.0 g A. tegmentosum, use orthogonal experiment,use the content of salidroside and tyrosol in the test product as the evaluation standard,use the EtOH reflux extraction method, and use different extraction conditions tocompare 9 parts of A. tegmentosum was extracted, concentrated under reducedpressure, sealed and stored frozen, and their respective extraction rates werecalculated (Table 2).

药学论文参考
药学论文参考

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Chapter 3. Results..................................33
3.1 Structure Identification........................... 33
3.1.1 The structure identification of compound 1...........................34
3.1.2 The structure identification of compound 2...........................35
Chapter 4. Conclusions..............................80
Chapter 3. Results

3.1 Structure Identification
3.1.1 The structure identification of compound 1

药学论文参考
药学论文参考

Physical description: white needles crystals;Melting point: 89-92℃;Molecular weight: 138.16;TOF-MS m/z 119.15 [M-H-H2O]+1H-NMR (300 MHz, DMSO-d6) δH: 9.08 (1H, s, -OH), 6.96 (2H, d, J = 7.7 Hz, H-2,6), 6.63 (2H, d, J = 8.2 Hz, H-3, 5), 4.52 (1H, t, J = 5.2 Hz, -OH), 3.49 (2H, m, H-8),2.56 (2H, t, J = 7.3 Hz, H-7);13C-NMR (75 MHz, DMSO-d6) δC: 155.9 (C-4), 130.1 (C-2, 6), 129.9 (C-1), 115.4(C-3, 5), 63.1 (C-8), 38.7 (C-7). The spectroscopic date was comparable with thosereported in the literature[29], the chemical structure of compound 1 was determined astyrosol (Table 4, Fig. 9), its molecular formula was C8H10O2.
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Chapter 4. Conclusions
1. The stems of air-dried A. tegmentosum were extracted with 90% EtOH,followed by extraction with PE, CH2Cl2 and EtOAc. In this study, the EtOAc extractwas isolated and purified, and a total of 18 compounds were obtained. The results areas follows:tyrosol (1), salidroside (2), β-D-galactopyranoside (3), dihydroconiferylalcohol (4), methyl (p-hydroxyphenyl)acetate (5), 4-hydroxybenzoic acid (6), gallicacid (7), ethyl gallate (8), 4-hydroxyphenylacetic acid (9), 2-(4-hydroxyphenyl)ethyl3,4,5-trihydroxybenzoate (10), 6-hydroxy-7-methoxycoumarin (11), fraxetin (12),asculetine (13), scopoletin (14), (+)-gallocatechin (15), quercetin (16),1,4-benzenedicarboxylic acid (17), 2-ethylhexyl phthalate (18). The isolatedcompounds include 10 phenol compounds, 4 coumarin compounds, 2 flavonoids, and2 benzoic acid lipids. Compounds 3, 4, 5, 6, 8, 9, 10, 11, 12, 13, 17, 18 are compoundsisolated from this plant for the first time. Compounds 3, 4, 5, 17, 18 are the firstcompounds isolated from this Acer.
2. The content of salidroside and tyrosol in the EtOH extract and water extractof A. tegmentosum was determined by high performance liquid chromatography. Theresults showed that the content of salidroside in the EtOH extract of A. tegmentosumwas 3.1227 mg/g. The content of tyrosol in the EtOH extract of A. tegmentosum was1.0900 mg/g. The content of salidroside in the water extract of A. tegmentosum was2.14744 mg/g, and the content of tyrosol in the water extract of A. tegmentosum was0.5458 mg/g.
reference(omitted)


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