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1、1Comparison of Polyphenol Contents, Antioxidant, and Anti-inflammatory Activities of Wild and Cultivated Lactuca indica 汇报人汇报人: :罗虹建罗虹建 2Literature source3ContentsIntroduction 12 Experimental section Conclusions234Results and discussion41. Introduction Dietary antioxidants protect the human body fro

2、m damage by free radicals, which have been associated with many degenerative（退行性）diseases. Fruits and vegetables are good sources of natural antioxidants, such as vitamins, carotenoids, flavonoids,and phenolics that prevent free radical damage. Phenolic compounds, such as flavonoids,phenolic acids,

3、and tannins play important roles in the prevention of degenerative diseases, such as, cancer, arteriosclerosis（动脉粥（动脉粥样硬化）样硬化）, and those associated with inflammation.51. Introduction Lactuca indica belongs to the family Compositeae（菊科）. It is an edible medicinal plant and is widely distributed in A

4、sian countries, including South Korea. This indigenous herb isused as folk medicine due to its anti-inflammatory and anti-bacterial activities, and is used traditionally to treat intestinal disorders. Various compounds, including terpenoids, sterols, and flavonoids, and the methanol extract of the L

5、. indica has been shown to exhibit anti-diabetic, antioxidant, anti-chol esterolemic, and hepato protective activities.2. Experimental sectionSample Extraction :The ground CL and WL samples 80% methanol -1:20 (w:v). The mixtures were shaken at 90 rpm in a shaking water bath at 60 for 5 h, and centri

6、fuged at 12,000 at -4 for 20 min. repeated three times using the residue obtained from the filtration process. Extracts were then pooled and condensed using a rotary evaporator under reduced pressure at 60 . Finally, extracts were frozen at -42 and concentrated using a freeze-dryer.72. Experimental

7、section2.1 The total polyphenols content (TPC)gallic acid as the standard phenol 1 mL of extract+0.2 mL of Folin Ciocalteau dark for 3 min at room T+0.5 mL Na 2 CO 3 (10%) and 1 mL of distilled water.UV-VISl l=725nm=725nmAbsorbance valuesdark for 1h at room Tmixtures82. Experimental section2.2Total

8、flavonoids content (TFC) Quercetin（槲皮素）（槲皮素） as the standard phenol0.5 mL of each extract+0.1 mL of 10% aluminum nitrate（硝酸铝）+0.1 mL of 1 M potassium acetate（醋酸钾）（醋酸钾）+3 mL of 80% ethanolUV-VISl l=415nm=415nmAbsorbance valuesdark for 40min at room Tmixtures92. Experimental section2.3 HPLC Analysis o

9、f Phenolics50 mg dried sample +10 mL of 80% methanol (passed through a 0.45m membrane filter.) UV detector (Waters 2489) l=l=280 nm. V=0.8 mLmin -1 . RP-18 column (4.4 mm 250 mm, 5 m;). Timesolvent A(9% acetic acid )solvent B(100% methanol )0-5min9555-28831728-40831740-606931102. Experimental sectio

10、n2.4 Evaluation of Antioxidant Activities （ IC 50 values）113. Results and discussion IC50 IC50半抑制浓度（或称半抑制率）是指半抑制浓度（或称半抑制率）是指“反应反应”被被抑制一半时抑制剂的浓度，这里的反应可以是酶催化反抑制一半时抑制剂的浓度，这里的反应可以是酶催化反应、抗原抗体反应等。应、抗原抗体反应等。 在凋亡方面，可以理解为一定浓度的某种药物诱导在凋亡方面，可以理解为一定浓度的某种药物诱导肿瘤细胞凋亡肿瘤细胞凋亡50%50%，该浓度称为，该浓度称为50%50%抑制浓度，即凋亡细抑制浓度，即凋亡细胞

11、与全部细胞数之比等于胞与全部细胞数之比等于50%50%时所对应的浓度。时所对应的浓度。 IC50 IC50值可以用来衡量药物诱导凋亡的能力，值可以用来衡量药物诱导凋亡的能力，该数值该数值越低，诱导能力越强，越低，诱导能力越强，当然也可以反向说明某种细胞对当然也可以反向说明某种细胞对药物的耐受程度。药物的耐受程度。 122. Experimental section2.5 Measurement of Anti-inflammatory Activity133. Results and discussionTable 1. Total polyphenols and flavonoids con

12、tents of wild and cultivated L. indica.WL CLLeaf RootCultivated plants were considered qualitatively inferior to specimens grown in the wild for medicinal and aromatic plants. This was attributed to the impacts of stress factors, such as drought, salinity, temperature, and soil characteristics, whic

13、h might increase secondary metabolite synthesis in wild specimens . 143. Results and discussionTable 2. Phenolic contents of wild and cultivated L. indica.咖啡酸、咖啡酸、 绿原酸、绿原酸、 、阿魏酸、阿魏酸、 柚皮苷、柚皮苷、15163. Results and discussionWang et al. (2003)identify 6 major phenolics (protocatechuic acid, methyl -hydro

14、xybenzoate, caffeic acid, 3,5-dicaffeoylquinic acid, luteolin 7-O- -glucopyranoside, and quercetin 3-O- -glucopyranoside) in L. indica extract.（对羟苯甲酸、原儿茶酸、咖啡酸、异绿原酸、毛地黄黄酮、木犀草素（对羟苯甲酸、原儿茶酸、咖啡酸、异绿原酸、毛地黄黄酮、木犀草素-7-O-吡喃葡萄糖苷）吡喃葡萄糖苷）The majority of naturally occurring phenolics retain antioxidative and anti-

15、 inflammatory properties that appear to promote chemoprotective(抗癌抗癌) activities. Gallic acid isolated from Caesalpinia decapetala wood（云实木）（云实木） was found to have significant in vitro free radical scavenging activity in ABTS and DPPH assays (Bhadoriya et al., 2012). Thus, it appears that gallic aci

16、d significantly contributes to the antioxidant activity of L. indica, especially in leaf extracts.173. Results and discussionTable 3. IC50 values of L. indica extracts as determined by DPPH radical scavenging ability, ABTS radical scavenging ability, and carotene bleaching activity.183. Results and

17、discussionIn the present study, -carotene bleaching activity was found to exhibit a pattern similar to that of radical scavenging activity. The CL root and leaf extracts had significantly higher activities than corresponding WL extracts The antioxidant power of phenolic compounds is related to struc

18、tural factors, such as the number and positions of hydroxyl（羟基） and/or methoxyl（甲氧基） groups. Therefore, the results of the current study imply that the high reducing power of WL is due to relatively high polyphenol content. Previous studies have also demonstrated that wild plants have higher antioxi

19、dant activities than cultivated plants. 193. Results and discussion返回返回203. Results and discussion返回返回213. Results and discussionWe first tested cell viabilities in the presence of test extracts to exclude possible cytotoxic effects on LPS- stimulated RAW 264.7 macrophages and results are shown in F

20、igs. 1 and 2. The viabilities of RAW 264.7 macrophages was over 80% when treated with CL or WL extracts at concentrations of 100-750 gmL -1 . These results demonstrate that L. indica extracts were not cytotoxic.22 Conclusions4234. conclusionsIn conclusion, the results of the present study indicate that L. indica grown in the wild has significantly higher antioxidant and anti-inflammatory acti