Their fruits have anti-inflammatory, anti-viral, anti-pyretic, anti-liver injury, and other effects [15,16,17,18]
Their fruits have anti-inflammatory, anti-viral, anti-pyretic, anti-liver injury, and other effects [15,16,17,18]. lipase. leaves, affinity screening, pancreatic lipase inhibitor, obesity 1. Introduction Obesity has become one of the greatest threats to human health in this century. It is recognized as the largest global chronic disease by WHO [1,2]. At least 3,400,000 adults die each year as a result of being overweight or obese [3,4]. Because pancreatic lipase (PL) can decompose 50C70% of fat, developing its inhibitor as an anti-obesity drug is preferred. Orlistat is a type of long-term PL inhibitor found and modified from and is currently one of the main drugs for treating obesity. However, this drug produces several side effects including fatty diarrhea, stool urgency, fecal incontinence, allergies, and liver function damage [5,6,7]. AM 103 In recent years, developing safer and more effective PL inhibitors from natural compounds has attracted more and more attention [8,9,10,11], and some new technologies are emerging [12,13]. We also used immobilized enzyme technology to screen PL inhibitors from . (Thunb.) Vahl is widely distributed in China, Japan, and Korea. Their fruits have anti-inflammatory, anti-viral, anti-pyretic, anti-liver injury, and other effects [15,16,17,18]. Their leaves are often used for tea. In this study, in order to find out whether tea, like black tea , oolong tea , and green tea , also possess an inhibitory effect on PL, its subfractions were prepared, and their inhibitive ability on PL were detected; then, the highest inhibiting subfraction was screened by self-made immobilized PL. 2. Results and Discussion 2.1. Inhibition of the Subfractions of F. suspensa Leaves on PL As a positive drug, the inhibition rate of 5 g/mL orlistat reached 58%, and its success proved the feasibility of the enzyme activity assay. The inhibitory effects of crude extracts and various extract subfractions of leaves on PL at 1500 g/mL are shown in Table 1. The table shows that the inhibition rate of the remaining parts on PL was the highest and reached 81.48%. Thus, the remaining precipitation of leaves was selected as the screening object of immobilized enzyme. Table 1 Inhibitory ratio of subfractions of leaves on pancreatic lipase (= 3). leaves (A: 270 nm; B: 340 nm), solution of screened out compounds (C: 270 nm; D: 340 nm), and mixed standard substances (E: 270 nm); 7 standard substances were completely detected at 270 nm, so 340 nm was not done) (1: Rutin; 2: Phillyrin; 4: Chlorogenic acid; 5: Caffeic acid; 8: Forsythiaside A; 9: Hesperidin; 11: Phillygenin). AM 103 Among the ten compounds, Compounds 1, 2, 4, 5, 8 and 9 were rutin, phillyrin, chlorogenic acid, caffeic AM 103 acid, forsythiaside A, and hesperidin respectively by compared with the mixed standard substances (Figure 1E). The undetermined compounds were separated and identified by HPLC-MS/MS; Compounds 6, 7, and 10 were identified as an isomer of forsythiaside A, arctigenin, and kaempferol-3-leaves are shown in Figure 3. Open in a separate window Open in a separate window AM 103 Open in a separate window Figure 2 MS2 spectra (A) and fragmentation schemes (B) of Compounds 6, 7, and 10 of leaves. (6: Isomer of forsythiaside A; 7: Arctigenin; 10: Kaempferol-3-leaves: Rutin (1); Phillyrin (2); Chlorogenic acid (4); Caffeic acid (5); Arctigenin (7); Forsythiaside A (8); Hesperidin (9); Kaempferol-3-leaves. leaves on pancreatic lipase. Table 3 Nine pancreatic lipase ligands screened out from leaves (= 3). leaves (remaining precipitate, Rutin (1), Phillyrin (2), Chlorogenic acid (4), Caffeic acid (5), Arctigenin (7), Forsythiaside A (8), Hesperidin (9), Kaempferol-3-before, was screened out for the first time, and its IC50 notably reached 2.9 0.5 mol/L; moreover, phillyrin, as a promotor of PL was also found. The isomer of forsythiaside A and compound 6 need to be isolated and identified. 3. Materials and Methods 3.1. Materials The following materials were acquired and used: Orlistat (Chongqing Pharscin Pharmaceutical Group Co., Ltd., Chongqing, China); CD213a2 carboxyl-terminated magnetic beads (10 mg/mL, 1 m) (Allrun,.