AU2018101556A4 - A kind of nitraria tangutorum bobr seed extract - Google Patents
A kind of nitraria tangutorum bobr seed extract Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/06—Antihyperlipidemics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
Abstract
Abstract The present invention provides a kind of Nitraria tangutorum Bobr seed extract, which is prepared using the following method: Take Nitraria tangutorum Bobr seeds, after extraction with ethanol water solution as the solvent, concentrate the extracted solution till the solution has no ethanol odor, and then extract with petroleum ether-water system, take the water layer, remove the solvent, and obtain the Nitraria tangutorum Bobr seed extract. As shown in the present invention research, the Nitraria tangutorum Bobr seed extract can reduce the total cholesterol, total triglyceride, LDL-C, and increase HDL-C in the serum of rats with hyperlipidemia, and counteract the abnormal body weight growth of hyperlipidemia rats to a certain extent. At the same time, it is discovered in the pathological sections of liver tissues of rats in different groups that the extract can improve changes in the adipose tissue and cell morphology in fatty livers to different extents. Therefore, the extract provides a novel choice for the clinical prevention or treatment of hyperlipidemia or fatty liver. Meanwhile, the present invention has also discovered a novel active position other than the Nitraria tangutorum Bobr seed oil, providing a new orientation for the exploitation of Nitraria tangutorum Bobr seeds.
Description
A kind of Nitraria tangutorum Bobr seed extract
CLAIM OF PRIORITY
This application claims priority to Chinese Application No.201810245500.X, filed on March 23, 2018, all of which are hereby incorporated by reference.
BACKGROUND OF THE DISCLOSURE
FIELD OF THE DISLOSURE
The present invention relates to a kind of Nitraria tangutorum Bobr seed extract.
DESCRIPTION OF THE RELATED ART
Hyperlipidemia is a major cause for arteriosclerosis and arteriosclerosis-related diseases such as coronary artery cardiac diseases, ischemic encephalopathy and peripheral blood vessel diseases. Although there have been improvement in arteriosclerosis related disease morbidity due to medical treatment, as well as prevention, the situation mentioned above are still covering a relatively large proportion in the morbidity and mortality of senile and middle aged population. Because of the ever-increasing obesity and aging of population, the annual morbidity and the case number will increase in large scale in the future decade. Hyperlipidemia is the increase of total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C) and the reduction of high density lipoprotein cholesterol (HDL-C) caused by lipids metabolism abnormality. In particular, researches indicated that the increase of LDL-C concentration plays a major role in the pathogenesis of arteriosclerosis, and has a strong correlation with higher morbidity of cardiovascular diseases. Reducing the LDL-C level by diet or medicines plays a major role in the reduction in morbidity of cardiovascular and cerebrovascular diseases. Facts indicated that hyperlipidemia is one of the risky factors for arteriosclerosis, and therefore it promotes the R&D of medicines to reduce the cholesterol level. These medicines play their roles mainly by reducing the level of total cholesterol and low density lipoprotein. Therefore, it is vital to R&D novel medicines capable of reducing cardiovascular diseases risks. In particular novel medicines with strong blood lipids level reduction capacity and without side or toxic effects would be most appreciated. Early stage researches indicated that the fruit of Nitraria tangutorum Bobr (Zygophyllaceae family Nitraria Tangutorum Bobr category) has significant inhibition effects for TG and TC in mice and rats with hyperlipidemia and for high LDL-C level in rats induced by high-fat diet, and at the same time it can significantly increase the 0
2018101556 18 Oct 2018 ratio of HDL/TC, HDL/LDL in hyperlipidemia rats, demonstrating that Nitraria Tangutorum Bobr fruit has significant blood lipids level reduction and arteriosclerosis prevention as well as anti-oxidation effects. At the same time, researches also demonstrated that the medium and high doses, namely 5g/kg and 10g/kg, of seed oil 5 of Nitraria Tangutorum Bobr have significant inhibition effects for TC, TG and LDL in serum of rats induced by high-fat diet, and it can significantly increase the ratio of HDL/TC and HDL/LDL in hyperlipidemia rats, demonstrating that Nitraria Tangutorum Bobr seed oil has significant blood lipid reduction effects.
At present, the researches on blood lipids reduction effects of Nitraria Tangutorum
Bobr seeds are limited only in the blood lipids reduction effects of Nitraria Tangutorum Bobr seed oil, and there are no available reports on the blood lipids reduction activity of other parts of the Nitraria Tangutorum Bobr seed.
BRIEF DESCRIPTION OF THE DISCLOSURE
Nitraria Tangutorum Bobr seed oil has blood lipids reduction effects, and is composed of non-polar or low-polar compounds. It can be obtained with carbon dioxide supercritical extraction, or extracted with non-polar or low-polar organic solvents, such as ethanol, because the dissolution property of ethanol is between polar and non-polar solvents, and ethanol can dissolve in water at any ratio, and 20 therefore, it can selectively dissolve aqueous soluble ingredients or some ingredients soluble in non-polar solvents. In general, when the ethanol content is over 90%, it can extract volatile oil, organic acid, resin, and chlorophyll, etc (“Pharmaceutics of TOM”, Zhang Zhaowang, Chinese TCM Press, First Edition 2003, page 99, the last third paragraph). According to the technology content mentioned above, to extract 25 Nitraria Tangutorum Bobr seed oil ingredients with ethanol, use ethanol of a concentration over 90% V/V as the solvent.
Different from the technology mentioned above, the present invention aims to provide a novel extract other than the seed oil. Therefore, the present invention provides a kind of Nitraria Tangutorum Bobr seed extract, which is prepared using the following 30 method:
Take Nitraria tangutorum Bobr seeds, after extraction with ethanol water solution as the solvent, concentrate the extracted solution till the solution has no ethanol odor, and then extract with petroleum ether-water system, take the water layer, remove the solvent, and obtain the Nitraria tangutorum Bobr seed extract.
During the extraction preparation course of the present invention, the ethanol extract must be extracted with petroleum ether, which is a non-polar solvent and an excellent solvent for extracting oil and lipid type ingredients. After extraction with petroleum
2018101556 18 Oct 2018 ether, non-polar or low-polar ingredients should be dissolved in the petroleum ether layer, and the remaining water layer has almost no oil and lipid ingredients. Therefore, the extract is completely different from the chemical ingredients of seed oil.
Wherein, the concentration of the ethanol water solution mentioned is between 50% and 95% V/V, and between 70% and 80% further, and the optimal concentration is 75% V/V.
Wherein, the extraction temperature is between 50°C and 80°C, and between 60°C and 70°C further, and the optimal choice is 65°C.
Furthermore, before extraction with ethanol water solution, the Nitraria tangutorum 10 Bobr seeds must be pulverized.
Wherein, the seeds are in Zygophyllaceae family Nitraria Tangutorum Bobr category.
The present invention also provides the application of the Nitraria Tangutorum Bobr seed extract in the preparation of medicines or health care products for improving hyperlipidemia.
Wherein, the medicines or health care products mentioned reduce TC, TG, and LDLC and improve HDL-C in serum.
The present invention also provides the application of the Nitraria Tangutorum Bobr seed extract in the preparation of medicines or health care products for prevention or treatment of fatty liver.
Wherein, the fatty liver mentioned is nonalcoholic fatty liver.
Wherein, the fatty liver mentioned is simple fatty liver.
As shown in the present invention research, the Nitraria tangutorum Bobr seed extract can reduce the total cholesterol, total triglyceride, LDL-C, and increase HDLC in the serum of rats with hyperlipidemia, and counteract the abnormal body weight 25 growth of hyperlipidemia rats to a certain extent. At the same time, it is discovered in the pathological sections of liver tissues of rats in different groups that the extract can improve changes in the adipose tissue and cell morphology in fatty livers to different extents. Therefore, the extract provides a novel choice for the clinical prevention or treatment of hyperlipidemia or fatty liver.
Meanwhile, the present invention has also discovered a novel active position other than the Nitraria tangutorum Bobr seed oil, providing a new orientation for the exploitation of Nitraria tangutorum Bobr seeds.
Brief Description of Drawings
Figure 1 illustrates the dynamic influence of high-fat diet modeling method on rats' body weights.
Figure 2 is the renal coefficient for rats in different test groups.
Figure 3 is the hepatic coefficient for rats in different test groups.
Figure 4 is the spleen coefficient for rats in different test groups.
Figure 5 is the cardiac coefficient for rats in different test groups.
Figure 6 is total cholesterol content of rats in different test groups.
Figure 7 is total triglyceride content of rats in different test groups.
Figure 8 is HDL-C content of rats in different test groups.
Figure 9 is LDL-C content of rats in different test groups.
Figure 10 is liver tissue pathologic sections of rats in different test groups (200X).
DETAILED DESCRIPTION OF EMBODIMENTS OF THE DISCLOSURE
Embodiment 1. Preparation method for the extract of the present invention
Take 5kg Nitraria tangutorum Bobr seeds, pulverize them with a high-speed universal pulverizer, and extract in an Automatic Decoction Machine (feed liquid ratio: 1:1 Okg/L; duration: two hours; extraction solvent: 75% ethanol; three times; extraction temperature: 65°C). After extracting three times, converge the extraction solution and recover the ethanol on a rotary evaporator until there is basically no ethanol odor, and then add petroleum ether and extract repeatedly for five times. The volume ratio of petroleum ether to ethanol extract is 2:1. Concentrate the water remained after petroleum ether extraction for desiccation, and obtain the extract of the present invention.
Embodiment 2. Pharmacodynamics research on the extract of the present invention
1.1 Instruments and Reagents
DFY-1000C high-speed universal pulverizer, Wenling Linda Machinery Co., Ltd; HYY401 Automatic decoction packaging machine, Tianjing Huayanyuan Machinery Co., Ltd; R1050 large-scale rotary evaporator, Zhengzhou Greatwall Scientific Industrial and Trade Co., Ltd; TF-FD-1 vacuum freeze dryer, Shanghai Tianfeng Industrial Co., Ltd; Table-Top Filtration Centrifuge TF-3-C, Shanghai Shengke Instruments and Equipment Co., Ltd; Tecan Infinite F200/M200 multi-functional Microplate Reader, Tecan Co., Ltd in Switzerland; Olympus Microscope, Olympus Co., Ltd; ALC-110.4 electronic balance (Acculab Co.,Ltd in Germany). Ethanol, petroleum ether, tween 80, xylene, and liquid paraffin are all analytically pure. Common food specifically for rats and mice, Beijing Heli Kechuang Science and Technology Development Co.,Ltd; cholesterol, cholate, Zhengzhou Hongxin Food and Chemical Co., Ltd; propylthiouracil, Shanghai Regal Biological Technology Development Co., Ltd; Hypocol capsule, Beijing Beida Weixin Biotechnology Co.,Ltd;
2018101556 18 Oct 2018
TC test reagent kit, TG test reagent kit, LDL-C test reagent kit, and HDL-C test reagent kit are all provided by Nanjing Jiancheng Bioengineering Institute.
1.2 Sample Preparation
Take 5kg Nitraria tangutorum Bobr seeds, pulverize them with a high-speed universal pulverizer, and extract in an Automatic Decoction Machine (feed liquid ratio: 1:1 Okg/L; duration: two hours; extraction solvent: 75% ethanol; three times). After extracting three times, converge the extraction solution and recover the ethanol on a rotary evaporator until there is basically no ethanol odor, and then add petroleum ether and extract repeatedly for five times. The volume ratio of petroleum ether to ethanol 10 extract is 2:1. Concentrate the water remained after petroleum ether extraction for desiccation, and obtain the extract of the present invention .After extracting three times, converge the extraction solution and recover the ethanol on a rotary evaporator until there is no ethanol odor, and take out a part of the extract for freezing desiccation to obtain the ethanol extraction. For the other part, add 15 petroleum ether and extract repeatedly for five times. The volume ratio of petroleum ether to ethanol extract is 2:1. Concentrate the crude extract from the water part after petroleum ether extraction on a rotary evaporator, and then desiccate by freezing on a vacuum freeze dryer to obtain the water part. The configured concentration of hypocol capsule is 0.02g/ml_, and the concentrations of other groups of samples are 20 low concentration 0.02g/ml_, final concentration 0.06g/ml_, and high concentration 0.18g/ml_, respectively.
1.3 Animal Grouping and Hyperlipidemia Model Establishment
120 Wistar rats, half of which are male and the other half of which are female, purchased from Medical Experiment Center in Lanzhou University, with body weight 25 170±1 Og, after feeding with common food for four days, randomly grouped, with 10 animals in each group, 5 male and 5 female. The groups are respectively blank control group (N), high-fat diet model group (M), positive control group with Hypocol (P), ethanol extract high dosage group (BZH), ethanol extract medium dosage group (BZM), ethanol extract low dosage group(BZL), water part high dosage group 30 (BZWH), water part medium dosage group (BZWM), and water part low dosage group (BZWL). Among them, the blank control group is fed with common food, and other groups are fed with high-fat diet. High-fat diet formula: 15% lard, 1% cholesterol, 0.2% cholate, 12% yolk and 71.8% basic food. Furthermore, in the course of modeling, in order to guarantee the success of modeling, the dosage groups, model 35 group and positive control group are given 1mL/100g of lipid emulsion by gavage together with high-fat diet. Lipid emulsion formula: 15% lard, 2.5% cholesterol, 0.5% cholate, 0.1% propylthiouracil, 1% tween, and the rest is distilled water. The blank
2018101556 18 Oct 2018 control group is given distilled water of the same volume by gavage. The lipid emulsion is added with anti-thyroid drug, namely propylthiouracil, which can inhibit the breaking down of cholesterol, and added with cholate, which can promote the absorption of cholesterol, and therefore, increase TC and TG in the blood, and push 5 the establishment of hyperlipidemia model. Three weeks after feeding with high-fat diet and gavage with lipid emulsion, apply drug therapy for four weeks. The gavage volume for rats are 1ml_/100g. Their grouping situation and dosage volume for each group are shown in Table 1. During rat feeding, observe the rats' status and health conditions daily, change padding and clean the drinking water bottle once every 4 10 days to avoid bacteria from generating, and at the same time measure and record body weights once every two weeks.
1.4 Blood Lipid Determination and Statistics for the Weights of Organs
On the last night of drug therapy, feed the rats with no diet but feed them with water. On the second day, anesthetize rats with 25% urethane with a dose of 1mL/100g, 15 and use blood collection tubes to collect blood from the rats’ aorta. Keep the blood samples still under room temperature for two hours, and centrifuge the samples for 15 minutes with a high speed centrifuge, with the speed set to 5000rpm, and pipette the serum on the upper layer, and use test reagent kits to measure TC, TG, LDL-C, and HDL-C in the serum. After blood collection, separate the hearts, livers, kidneys, 20 and spleens of the rats, rinse them with normal saline, remove the superfluous fluid with filter paper, and weigh the organs. After weighing, on the same position of each liver, cut off one piece of liver tissues, drop them into 10% formalin solution for stabilizing, and change the new formalin solution for soaking on the second day.
Table 1. Rat grouping situation and dosage volume for each group
| group | Test drug | Dosing concentration (mgOni-) | Dosage (8¾) |
| l | N | — | — |
| J | M | — | — |
| 3 | P | 20 | 0.2 |
| 4 | BZH | 20 | 0.2 |
| 5 | BZM | ήθ | D.fi |
| 6 | BZL | ISO | 1.8 |
| 7 | BZWH | 20 | 0.2 |
| Ji | BZWM | fiO | D.fi |
| 9 | BZWL | 1R0 | 1.8 |
The dosage in the present invention is calculated based on the extract weight.
1.5 Histopathological Sections
Take out the liver tissues stabilized in 10% formalin, section the tissues of suitable thickness, and carry out paraffin section. Procedures in details are as follows: (1)
Dehydration: Dehydrate in 70% and 80% ethanol for 40 minutes, dehydrate in 95%l and 95%ll ethanol for 30 minutes, and dehydrate in 100% I and 100%ll absolute ethanol for 20 minutes and 15 minutes, respectively. (2) Hyalinization: Drop the tissues after dehydration into chloroform I and II for hyalinization. (3) Waxing: Place the tissues into melted liquid paraffin, the waxing durations are 40, 30, and 20 minutes, respectively. (4) Embedding: Place the waxed tissue materials into a metal embedding box filled with paraffin liquid (in the middle of the paraffin). When the paraffin surface solidifies, immediately place it into cold water for cooling, and obtain a paraffin block containing a tissue block. (5) Sectioning: Clamp the paraffin block already embedded into the paraffin block pliers of a rotary microtome, keep the paraffin block section in parallel with the slicing blade, and tighten. Section thickness is between 4 and 7 pm. Place the sections gently into a water bath tub (about 50°C) with a brush.(6) Section patching and baking: Separate a paraffin belt (continuous sections) of a certain length into single paraffin sections with a forcep and flatten the sections in warm water. Then, spread the sections onto slides, and dry the slides in a 60°C constant temperature incubator, or in a 37°C incubator in a long period. (7) Section dewaxing and hydration: Dewax dry sections in xylene I and II as well as 95% ethanol I and II for five minutes, respectively, and clear paraffin from sections in flowing water for several minutes. (8) Staining: Place the sections into hematoxylin to stain for five minutes, and clean them with flowing water. Place the sections into 1% HCI for several seconds, and clean them in clear water. Place the sections into ammonium water for about one minute, and clean them with clear water. Finally, place the sections in Eosin solution to stain for two to three minutes, and rinse them in clear water for about 15 minutes. (9) Dehydration, hyalinization and sealing: Dehydrate stained sections in 70% (several seconds), 80% (one minute), as well as 95% I, 95% II, 100% I, and 100% II solutions for five minutes each in sequence. Then, hyalinize the sections in xylene I and II for five minutes, respectively, and drip a drop of resin for sealing. After drying, observe the sections.
1.6 Statistical Analysis
The test results are all expressed in X±S.D, and all the test data applies SPSS16.0 software for single factor analysis of variance, p < 0.05 indicates that there is statistical difference, while p < 0.01 indicates that the difference is extremely significant.
2. Experiment Results
2.1. Basic Health Conditions of Rats During Experiment
During the experiment, the ingestion and imbibing of each group of rats are normal; the rats are lustrous in fur, their feces are normal in color and volume, and their spirit
2018101556 18 Oct 2018 statuses are active and healthy. Sometimes there is no fight, resulting in the death of individual rats during the experiment.
2.2. Analysis of Ratios of Body Weights to Organs Weights of Rats in Different Groups
During the eight weeks feeding of the rats, the body weights of experimented rats in different groups all have significant trends of increase.
Table 2. Influence on the body weights of rats in different experiment groups in the feeding period (g)
| group | 0 week | 2 Week | 4 week | 6 week | 8 week |
| 1 | 179.017.2 | 259.418.9· | 289.1110.2* | 311.518.9** | 322.9114.5** |
| 2 | 183.216.8 | 264.419.1 | 295J±9.S | 324.8112.6 | 344.8112.3 |
| 3 | 180.415.4 | 260.217.9· | 285.718.7·* | 303.9111.8** | 319.9113.5** |
| 4 | 184.213.5 | 261.fi±fi.5 | 286.8*11.5*· | 302.6113.5*· | 315.3*11.8** |
| 5 | 1 | 261.318.3 | 283 0,ή·* | 324.5112.4*· | |
| 6 | 176.516.8 | 263.417.8 | 282.8111.3·· | 298.619.7** | 318.9110.1*· |
| 7 | 184.615.9 | 259.618.4· | 282.515.8·· | 305.7112.1·· | 320.5112.8*· |
| ft | 182.S16.2 | 260.219.0* | 285.819.2·* | 311.5110.5·* | 338.9112.7· |
| 9 | 181.517.3 | 262.517.7 | 286.818.7** | 308.411.1.4** | 331.5111.7** |
Compared with the model group: *: P < 0.05, the difference is significant; **: P < 0.01, the difference is extremely significant.
It can be learnt from Table 2 and Figure 1 that, after the administration of high-fat diet for four weeks and the application of drug therapy for a week on the fourth week at the same time, the body weights of rats in the model group are significantly higher 15 than those in other medicine groups (P < 0.01). After five weeks of related drug therapy, the body weights of rats in the normal group, positive Hypocol group, BZ high, medium and low dosage groups, and BZW high, medium and low dosage group are all significantly lower than those in the model group (P < 0.05 or P < 0.01). As shown in the results mentioned above, all medicine groups have certain trends of 20 resistance for the abnormal body weight growth in rats.
It can be learnt from Table 3 and Figures 2, 3, 4, and 5 that the kidney, spleen and heart coefficients of rats in different experiment groups have no statistical difference (P < 0.05), demonstrating that there is no significant difference in medicine intervention during this stage. However, after the application of drug therapy for five 25 weeks, the liver coefficients in the normal group, positive Hypocol group, and BZ high, medium, and low dosage groups, and BZW high dosage group are significantly lower than the liver coefficient in the model group, indicating a statistically significant difference (P < 0.01); the liver coefficient in the BZW medium dosage group is also significantly different from that in the model group (P < 0.05); the liver coefficient in
2018101556 18 Oct 2018 the BZW low dosage group is not significantly different from that in the model group, without a statistically significant difference.
Table 3. Ratio of heart, liver, spleen, and kidney weights to body weights of rats in different experiment groups
| group | Sample | Size kidney | liver | spleen | heart |
| 1 | 10 | 0.68it*0.022 | 2.42*0.108** | 0.158*0.007 | 0.342*0.0'14 |
| 2 | 9 | 0.675*0.027 | 2.76*0.087 | 0.165*0.005 | 0.326*0.012 |
| 3 | 10 | 0.692*0.025 | 2.59*0.078** | 0.163*0.008 | 0.348*0.015 |
| 4 | 9 | 0.695*0.018 | 2.46*0.065** | 0.161*0.004 | 0.322*0.012 |
| 5 | 10 | 0.704*0.023 | 2.40*0.061** | 0.159*0.005 | 0.303*0.009 |
| 6 | 10 | 0.706*0.020 | 2.60*0.078** | 0.162*0.005 | 0.324*0.011 |
| 7 | 10 | 0.684*0.021 | 2.64*0,12D** | 0.168*0.005 | 0,324*0.014 |
| S | 9 | 0,666*0,014 | 2.71 ±0.094* | 0.166*0,007 | 0.325*0.013 |
| 9 | 10 | 0.683 *0.0] 3 | 2.76*0.088 | 0.165*0.006 | 0.319*0.01 i |
Compared with the model group: *: P < 0.05, the difference is significant; **: P < 0.01, the difference is extremely significant.
2.3. Influences on the Blood Lipid Level of Rats with Hyperlipidemia in Different Medicine Groups
Table 4. Blood lipid level of rats in different experiment groups
| group Sample Size TC | TG | HDL-C | LDL-C | ||
| 1 | 10 | I.SOD *0.072** | 0.374 ±0.015·· | 0.928*0,035·* | 1,43610.054·· |
| 2 | 9 | 3.384 ±0.135 | 0.757 ±0,033 | 0.643*0,024 | 5.46810.198 |
| 3 | 10 | 2.784 *0.124** | 0.367 *0.014·· | 0,799*0.027** | 2.454*0.076*· |
| 4 | 9 | 2.426 *0.102** | 0.490 *0.022·· | 0,822*0.029** | 1,986*0.059·· |
| 5 | 10 | 2.562*0.098** | 0.530*0.024·* | 0.761*0.024** | 2.414*0.065·· |
| 6 | 10 | 2.616*0.121** | 0.625* 0.027·· | 0.714*0.021** | 2.667*0.074·· |
| 7 | 10 | 2.501 ±0.111·* | 0.522 *0.024·* | 0.784*0.021** | 2.234*0.061“ |
| 8 | 9 | 2.662*0.123** | 0.582 *0.026** | 0.722*0.02** | 2.849*0.081·· |
| 9 | 10 | 2.814*0.125** | 0.639*0.03·* | 0.684*0.014* | 3.159*0.102·· |
Compared with the model group: *: P < 0.05, the difference is significant; **: P < 0.01, the difference is extremely significant.
(1). Total Cholesterol (TC)
It can be learnt from Table 4 and Figure 6 that, TC content in the high-fat diet model group is significantly higher than that in the normal control group (P < 0.01), while TC level in the positive medicine group is significantly lower than that in the model group (P < 0.01) but higher than that in the normal control group. TC content in BZ high, medium, and low dosage groups and BZW high, medium, and low dosage groups is 20 significantly lower than that in the high-fat diet model group (P < 0.01), while TC content in BZ high, medium, and low dosage groups and BZW high, medium, and low dosage groups is lower than that in the positive medicine group, demonstrating that its effects to reduce TC level are superior to the effects in the positive medicine group. Furthermore, high dosage groups of all medicines have better effects to reduce TC content in serum of hyperlipidemia rats than medium dosage groups, and even better than low dosage groups. The results mentioned above demonstrate that various medicine groups, namely BZ and BZW groups, have the effects to reduce the TC content in serum of hyperlipidemia rats, with dosage-dependent effects.
(2) . Total Triglyceride (TG)
It can be learnt from Table 4 and Figure 8 that, TG content in the high-fat diet model group is significantly higher than that in the normal control group (P < 0.01), while TG level in the positive medicine group is significantly lower than that in the model group (P < 0.01) but equivalent to that in the normal control group. TG content in BZ high, medium, and low dosage groups and BZW high, medium, and low dosage groups is significantly lower than that in the high-fat diet model group (P < 0.01). Furthermore, high dosage groups of all medicines have better effects to reduce TG content in serum of hyperlipidemia rats than medium dosage groups, and even better than low dosage groups. The results mentioned above demonstrate that various medicine groups, namely BZ and BZW groups, have the significant effects to reduce the TG content in serum of hyperlipidemia rats, with dosage-dependent effects.
(3) High density lipoprotein cholesterol (HDL-C)
It can be learnt from Table 4 and Figure 8 that, HDL-C content in the high-fat diet model group is significantly lower than that in the normal control group (P < 0.01), while HDL-C level in the positive medicine group is significantly higher than that in the model group (P < 0.01). HDL-C content in BZ high, medium, and low dosage groups and BZW high and medium dosage groups is significantly higher than that in the high-fat diet model group (P < 0.01), while the HDL-C level in the BZW low dosage group demonstrates only a significant difference compared with that in the model group (P < 0.05). The HDL-C level in the BZ high dosage group is higher than that in the positive medicine group. Furthermore, high dosage groups of all medicines have better effects to increase HDL-C content in serum of hyperlipidemia rats than medium dosage groups, and even better than low dosage groups. The results mentioned above demonstrate that various medicine groups, namely BZ and BZW groups, have the effects to increase the HDL-C content in serum of hyperlipidemia rats, with dosage-dependent effects.
(4) Low density lipoprotein cholesterol (LDL-C)
It can be learnt from Table 4 and Figure 9 that, LDL-C content in the high-fat diet model group is extremely significantly higher than that in the normal control group (P < 0.01), and the content is more than twice as much as that in the normal group. The
LDL-C level in the positive medicine group is significantly lower than that in the model group (P < 0.01). LDL-C content in BZ high, medium, and low dosage groups and BZW high, medium, and low dosage groups is significantly lower than that in the high-fat diet model group (P < 0.01), while the LDL-C level in BZ high and medium dosage groups and BZW low dosage group is lower than that in the positive medicine group. Furthermore, high dosage groups of all medicines have better effects to reduce LDL-C content in serum of hyperlipidemia rats than medium dosage groups, and even better than low dosage groups. The results mentioned above demonstrate that various medicine groups, namely BZ and BZW groups, have the effects to reduce the LDL-C content in serum of hyperlipidemia rats, with dosage-dependent effects.
2.4. Histopathological Observation of the Liver
Influences on the appearance of in-vitro liver tissues of rats in different groups: Observe liver tissues just removed from rats in various groups by naked eyes, and discover that the liver tissues of rats in the normal group demonstrate fresh bright red, with no liver edema phenomenon in general, smooth in surface, rich in elasticity, soft in texture, and sharp and neat in edges; the liver tissues of rats in the model group have tight envelopes, pale white in color like lard, rough in surface, blunt in edges, relatively harder in texture compared with liver tissues from rats in the normal group, greasy in sense when you touch the cutting section, and larger in size compared with liver tissues from rats in the normal group, and the phenomena mentioned above demonstrate that the self-produced high-fat diet has the trend to induce the formation of fatty liver in rats; the liver tissues of rats in the positive group and BZ high dosage group are close to those in the normal group in appearance; and the abnormal morphology of liver tissues of rats in BZ medium and low dosage groups and BZW high and medium dosage groups is improved to certain extents. The liver tissues of rats in the BZ medium dosage group are gradually changed from pale white to red in color; the liver tissues of rats in the BZ low dosage group and BZW high and medium dosage groups are changed from hard to soft in texture, and from tight to loose in surface envelopes gradually; the liver tissues of rats in the BZW low dosage group still have relatively severe edema, with the worst improvement effects.
It is discovered in pathological sections of livers that: as shown in Figure 10, the hepatic lobules of rats in the normal group are integral in structure, with clearly visible hepatic venous sinusand neat arrangement of liver cells, and the liver cells around the central vein are arranged in radial shape, with rich and even cytoplasm and without lipid droplet cells; in the model group rat livers, there are a large quantity of fat physaliphores of different sizes around the central vein, and the hepatic cells are swelling and demonstrating ballooning degeneration with a certain extent of necrosis of some liver cells, at the same time, the hepatic cell nucleus are squeezed by lipid droplet to one side, and the central vein also demonstrates abnormal dilation, with a trend to develop into nonalcoholic fatty liver; in the positive medicine group, the physaliphores are reduced significantly compared with those in the model group, but the liver tissues have slight watery lesion. Basically, there are no physaliphores in the liver tissues of rats in the BZ high dosage group, and the hepatic edema and ballooning degeneration have been basically recovered, without necrosis liver cells, and the central vein is shrinked and rich in cytoplasm, with the cell nucleus arranged in the center of liver cells; the changes of liver cells in BZ medium and low dosage groups and BZW high and medium dosage groups are mainly demonstrated in the significant reduction of adipocytes and edema cells, rich cytoplasm content, and becoming normal of the hepatic cell structure. However, there are still a large number of swelling cells and adipocytes in the hepatic tissue of rats in the BZW low dosage group.
3. Conclusion
The present research establishes a hyperlipidemia model in rats by feeding of highfat diet and gavage with fatty emulsion at the same time, with the results demonstrating that the TC, TG and LDL-C levels in serum of rats in the model group are significantly higher than those in the normal control group, and at the same time, HDL-C content in the serum of rats in the model group is significantly lower than that in the normal control group, indicating the successful setup of the blood lipid reduction model. A research is carried out on the blood lipid reducing activity of Nitraria tangutorum Bobr seed extract (BZ) and the water part after the extraction of petroleum ether (BZWH) to screen a more effective blood lipid reducing medicine, and the results demonstrate that the high, medium, and low dosage groups of various medicines can significantly reduce TC,TG and LDL-C and improve HDL-C in serum of hyperlipidemia rats, indicating that the medicines both have the effects to reduce blood lipid level in hyperlipidemia rats. Meanwhile, the medicine groups can resist the trend of abnormal growth of body weights of hyperlipidemia rats. The observation results of pathological sections of hepatic tissues also demonstrate that different medicine groups have the effects to improve the hepatic tissue and cell morphology to different extents. Furthermore, the effect to reduce blood lipid is dependent on the medicine dosage to a certain extent.
Claims (10)
- THE CLAIMS DEFINING THE INVENTION ARE AS FOLLOWS:1. A kind of Nitraria tangutorum Bobr seed extract, wherein prepared in the following method:Take Nitraria tangutorum Bobr seeds, after extraction with ethanol water solution as the solvent, concentrate the extracted solution till the solution has no ethanol odor, and then extract with petroleum ether system, take the water layer, remove the solvent, and obtain the Nitraria tangutorum Bobr seed extract.
- 2. The Nitraria tangutorum Bobr seed extract according to claim 1, wherein the ethanol water solution concentration is between 50% and 95% V/V, and between 70% and 80% V/V further, with the optimal choice as 75% V/V.
- 3. The Nitraria tangutorum Bobr seed extract according to claim 1 or 2, wherein the extraction temperature is between 50°C and 80°C, and between 60°C and 70°C further, with the optimal choice as 65°C.
- 4. The Nitraria tangutorum Bobr seed extract according to claim 1, wherein Nitraria tangutorum Bobr seeds are pulverized before extraction with ethanol water solution.
- 5. The Nitraria tangutorum Bobr seed extract according to any one of claims 1 to 4, wherein the Nitraria tangutorum Bobr is in Zygophyllaceae family Nitraria Tangutorum Bobr category.
- 6. Application of the Nitraria tangutorum Bobr seed extract according to any one of claims 1 to 5, wherein the medicines or health care products for improving hyperlipidemia abnormality are prepared.
- 7. The application according to claim 6, wherein the medicines or health care products reduce TC, TG, and LDL-C and improve HDL-C in serum.
- 8. Application of the Nitraria tangutorum Bobr seed extract according to any one of claims 1 to 5, wherein the medicines or health care products for prevention or treatment of fatty liver are prepared.
- 9. The application according to claim 8, wherein the fatty liver mentioned is nonalcoholic fatty liver.
- 10. The application according to claim 8 or 9, wherein the fatty liver mentioned is simple fatty liver.
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