Context: Broussonetia papyrifera (L.) Vent. (Moraceae), a traditional Chinese medicinal herb, has been extensively applied for many years to treat various diseases. Its fruits (Fructus Broussonetiae) have been commonly used as an important tonic for the treatment of age-related disorders with long history; recent research has proved that it contains 32% to 35% fixed oils. The fixed oil is composed mainly of unsaturated fatty acids, including linoleic acid, methyl palmitate, oleic acid and linoleic acid ester. Objective: To investigate the chemistry of the fatty oil from Fructus Broussonetiae (FOFB) and its effects on plasma lipids. Methods: The chemical composition of FOFB was examined and identified by GC-MS. Thirty male Wistar rats fed diet containing FOFB and cholesterol were studied for 28 days. The effect of dietary FOFB on plasma lipids and adipose tissue was tested. Results: Twelve compounds of FOFB were examined and identified, the major components of fatty oil, 8,11-octadecadienoic acid (83.75%), palmitic acid (10.22%), octadecadienoic acid (2.97%) and 9-octadecenoic acid (1.69%) were found. FOFB significantly exhibited the activities of decreasing the rat adipose tissue weight, triacylglycerol, total cholesterol, and low-density lipoprotein cholesterol (LDL-C) concentrations while the rat body weight remained unchanged. Discussion: FOFB contained a large amount of PUFA which had the effect on reducing plasma lipids and adipose.
Unsaturated fatty acid (USFA) is a kind of body fat acid, and the most essential body fatty acids are USFA. USFA, according to the different numbers of the double bond, is divided into monounsaturated fatty acid (MUFA) and polyunsaturated fatty acids (PUFA). In the dietary fats, MUFA includes oleic acid, and PUFA includes linoleic acid, linolenic acid, arachidonic acid, etc. Linoleic acid and linolenic acid cannot be synthesized by human body; they must be supplied by the diet. If the dietary unsaturated fatty acid is insufficient, it is easy to cause the blood low-density lipoprotein and LDL-cholesterol increasing, which can cause atherosclerosis and cardiovascular diseases.
Broussonetia papyrifera (L.) Vent. (Moraceae) is a deciduous plant widely distributed all over China. Fruits of the plant are commonly named Fructus Broussonetiae (FB). In traditional Chinese medicine, FB is used to nourish yin and tonify the blood, replenish liver and kidney, improve eyesight and promote urination, and can be used for treating soreness and weakness of waist and knees, fatigue, severe fever and dizziness [
B. papyrifera fruits were collected from Quanzhou, Fujian Province, China in autumn, 2011. The specimens were identified by Prof. HUANG BK (Second Military Medical University). Voucher specimens (voucher No. 20110915) were deposited at School of Biomedical Sciences, Huaqiao University of China. The fresh fruits were washed, dried and the grey white membranous persistent calyx and impurities were removed. FB ( 2.5 kg ) was obtained and stored in a vacuum dryer for further use.
Samples of FOFB were extruded from FB using an extruder (D-1683, Henan, China) at speeds of 70 - 80 rpm. The obtained orange fatty oil samples were analyzed by Gas chromatography-mass spectrometry (GC-MS) (HP 5890/HP 5973 instrument), which was fitted with capillary glass column DB-wax (30 m × 0.25 mm × 0.25 μm). The column was temperature programmed as follow: 50˚C (2 min), 50˚C - 70˚C at 4˚C/min then 70˚C - 300˚C at 10˚C/min with helium as the carrier gas. Injector temperature was 250˚C. Mass spectra were obtained by scanning from m/z 35 to 500 with a 0.7 s scan time.
The individual oil components were separated and identified by comparison of their retention indices and mass spectra with the NIST 98 and Wiley Mass Spectral Libraries. The relative amounts of individual components were expressed as percentages of the peak area relative to the total peak area.
Experiments in animals were approved by the Laboratory Animal Care Committee of our institution and were conducted in compliance with our institution’s ethical guidelines for animal research.
Thirty male Wistar rats (5-week-old) were housed individually in metallic cages in an animal room kept at 25˚C ± 1˚C and 60% ± 5% relative humidity with a 12:12-h light-dark cycle (lighting from 8: 00 a .m. to 8:00 p.m.). The rats were fed with an adaptation diet for 1 week, and then divided randomly into three groups (each group 10 rats) for designed diets: 1) 10% lard (control) group; 2) 5% FOFB group (5% FOFB + 5% lard); 3) 10% FOFB group (10% FOFB). The recipes of the three experimental diets are shown in
At the end of the feeding period (4 weeks), the rats were deprived of food for 12 h, and then blood samples (with sodium citrate 0.5 mg/ml as anticoagulant) were collected from fossa orbitalis under chloraldurat anesthesia. After the blood sampling, the animals were killed immediately by dislocating cervical, the heart,liver and adipose tissue were isolated, weighted and immediately frozen and stored at −80˚C until used for analysis. Relative heart (liver, adipose tissue) weight was expressed as heart (liver, adipose tissue) weight (g) ×100/final body weight (g).
Plasma samples were obtained by centrifugation (1388 g for 10 min at 10˚C). Plasma total cholesterol (CHO) and triacylglycerol (TG) were determined by CHOD-PAP and GPO-PAP methods using CHO and TG testing kits, respectively. High-density lipoprotein (HDL-C) and Low-density lipoprotein (LDL-C) in plasma were separated by ultracentrifugation (194,000 g for 3 h at 10˚C), and determined by direct method using HDL-C and LDL-C kits. All kits were purchased from Beijing jiuqiang Bio-Tech Com., China. All biochemical indicators were measured using a conventional automated analyzer (Beckman Au680, USA).
Total cholesterol in the liver was assayed from the total lipids extracted by the Folch method [
Each result was given as the mean ± standard deviations (S.D.). Statistical differences among groups were calculated using one-way ANOVA (SAS Institute, Cary, NC), and group means were considered to be significantly different at p < 0.05 as determined by Duncan’s new multiple-range test.
GC and MS are powerful tools for chemical analysis, especially when used together. All of the components in a sample can be separated by GC analysis, and at the same time GC analysis provides a representative spectral output. The size of the peak is proportional to the quantity of the corresponding substances in the specimen analyzed. MS identifies substances by electrically charging the specimen molecules, accelerating them through a magnetic field, breaking the molecules into charged fragments and detecting the different charges. MS analysis is highly reliable if the instrument is of sufficient resolution and the technician’s interpretation of the results is competent.
The chemical components of FOFB were examined by GC-MS thereby 12 components were identified (
Fatty acid compositions of the three experimental diets were determined by GC (
No. | Component | Formula | Molecule weight | Relative content |
---|---|---|---|---|
1 | Tetradecanoic acid | C14H28O2 | 228 | 0.2 |
2 | Palmitic acid | C16H32O2 | 256 | 10.22 |
3 | 8,11-Octadecadienoic acid | C18H32O2 | 280 | 83.75 |
4 | 9-Octadecenoic acid | C18H34O2 | 282 | 1.69 |
5 | Octadecanoic acid | C18H36O2 | 284 | 2.97 |
6 | 9,12-octadecadienoic acid | C18H32O2 | 280 | 0.37 |
7 | 11,14-Eicosadienoic acid | C20H36O2 | 308 | 0.3 |
8 | 11-Eicosenoic acid | C20H38O2 | 310 | 0.21 |
9 | Eicosenoic acid | C20H40O2 | 312 | 0.1 |
10 | Docosanoic acid | C22H44O2 | 340 | 0.08 |
11 | δ-Tocopherol | C27H46O2 | 402 | 0.08 |
12 | Fucosterol | C29H48O | 412 | 0.03 |
Fatty acid (carbons (unsaturated carbons)) | Control | 5% FOFB | 10% FOFB |
---|---|---|---|
14 (0) | 2.21 | 1.15 | 0.20 |
16 (0) | 25.24 | 17.81 | 10.22 |
16 (1) | 4.35 | 2.14 | trace |
18 (0) | 12.99 | 8.04 | 2.97 |
18 (1) | 41.48 | 21.58 | 1.69 |
18 (2) | 13.26 | 48.53 | 84.05 |
SFA | 40.44 | 27.00 | 13.39 |
PUFA | 13.26 | 48.53 | 84.05 |
MUFA | 45.83 | 23.72 | 1.69 |
P/S | 0.33 | 1.80 | 6.28 |
(P + M)/S | 1.46 | 2.68 | 6.40 |
P/M | 0.29 | 2.05 | 49.73 |
M/S | 1.13 | 0.88 | 0.13 |
SFA: saturated fatty acid. PUFA: polyunsaturated fatty acid. MUFA: monounsaturated fatty acid.
Group | Control | 5% FOFB | 10% FOFB |
---|---|---|---|
Initial body weight (g) | 176.13 ± 4.59 | 176.15 ± 4.21 | 176.14 ± 4.29 |
Final body weight (g) | 222.15 ± 19.48 | 219.40 ± 9.34 | 221.00 ± 6.88 |
Food intake (g/day) | 16.96 ± 2.13 | 17.26 ± 1.98 | 17.32 ± 2.15 |
Heart weight (g) | 4.721± 0.66 | 4.78 ± 0.69 | 4.76 ± 0.78 |
Relative heart weight | 2.12 ± 0.71 | 2.18 ± 0.91 | 2.15 ± 0.79 |
Liver weight (g) | 7.06 ± 0.52 | 7.08 ± 1.23 | 7.04 ± 0.91 |
Relative liver weight | 3.73 ± 0.13 | 3.70 ± 0.26 | 3.68 ± 0.49 |
Epididymal fat pad | 3.38 ± 0.20 | 2.98 ± 0.96* | 2.97 ± 0.34* |
Abdominal adipose tissue | 0.72 ± 0.28 | 0.63 ± 0.14 | 0.59 ± 0.15* |
Adipose tissue weight (g) | 4.10 ± 0.41 | 3.61 ± 0.42 | 3.56 ± 0.51* |
Relative adipose weight | 2.17 ± 0.37 | 1.92 ± 0.24 | 1.89 ± 0 .39* |
Each value is expressed as mean ± S.D. for ten rats per dietary group. Values in the same row with asterisk are significantly different. * vs. control group P < 0.05.
This result is in line with previous reports. Diet-induced obesity was related with renal lipid accumulation in mice [
To evaluate the effects of FOFB on the regulation of plasma lipids in rats, the total cholesterol, triacylglycerol, HDL-cholesterol and LDL-cholesterol were measured. Our data showed (
Both of the FOFB groups presented lower level of liver cholesterol (
In conclusion, FOFB contained a large amount of PUFA about 32% to 35% [
Group | Control | 5% FOFB | 10% FOFB |
---|---|---|---|
Total cholesterol (mg/dl) | 326.16 ± 58.78 | 287.59 ± 77.60* | 298.42 ± 56.05* |
Triacylglycerol (mg/dl) | 49.37 ± 8.53 | 42.51 ± 9.18* | 43.24 ± 10.70* |
HDL-cholesterol (mg/dl) | 16.38 ± 2.50 | 16.00 ± 2.67 | 16.51 ± 2.23 |
LDL-cholesterol (mg/dl) | 8.83 ± 1.318 | 7.76 ± 1.99* | 7.59 ± 2.32* |
Each value is expressed as mean ± S.D. for ten rats per dietary group. Values in the same row with asterisk are significantly different. * vs control group P < 0.05.
Group | Control | 5% FOFB | 10% FOFB |
---|---|---|---|
Cholesterol | |||
(mg/liver) | 756.23 ± 67.20 | 669.19 ± 88.11* | 613.88 ± 51.61* |
(mg/g liver) | 107.24 ± 7.53 | 94.50 ± 8.44* | 87.28 ± 10.70* |
Each value is expressed as mean ± S.D. for ten rats per dietary group. Values in the same row with asterisk are significantly different. * vs control group P < 0.05.
each component of FOFB to provide data for development of an antihyperlipidemic agent.
This work was supported by the Quanzhou Science & Technology Fund (2007Z12).
Su-Qiu Pang,Guo-Quan Wang,Xiao-Qin Jin,Ai-Jing Sun,Jun-Sheng Lin,Yong Diao, (2016) Chemical Composition of the Fatty Oil from Fructus Broussonetiae and Its Effects on Rat Plasma Lipids and Adipose Tissue. American Journal of Plant Sciences,07,446-452. doi: 10.4236/ajps.2016.73038