CN116159085A - Application of bear gall heart-saving preparation in preparation of medicine for treating hyperlipidemia-related diseases - Google Patents
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Abstract
The application provides an application of bear gall heart-saving preparation in preparing a medicament for treating hyperlipidemia-related diseases, and relates to the field of medicament application. Hyperlipidemia-associated diseases include hyperlipidemia, fatty liver, cognitive dysfunction, cardiovascular disease, and cardiac insufficiency.
Description
Technical Field
The application relates to the field of pharmaceutical application, in particular to application of a bear gall heart-saving preparation in preparation of a medicament for treating hyperlipidemia-related diseases.
Background
Hyperlipidemia (HLP) is a major risk factor for the development of cardiovascular and cerebrovascular diseases and fatty liver. It was found that total cholesterol (total cholesterol, TC), low density lipoprotein cholesterol (LDL-C) are the main cause of hyperlipidemia, while when the levels of TC, triglyceride (TG), low density lipoprotein cholesterol (low density lipoprotein cholesterol, LDL-C) in blood are elevated, lipids deposit on the vessel wall in large amounts, cause vascular endothelial cell injury, induce expression of various adhesion factors, promote foam cell formation, and cause cardiovascular disease; on the other hand, the liver uptake of free fatty acids increases, leading to accumulation of TG transport disorders in the liver to form fatty liver.
Currently, statins and fibrates are common drugs for the clinical treatment of hyperlipidemia. The statin drugs can obviously reduce TC, have good blood lipid reducing effect, but have side effects such as hepatotoxicity, rhabdomyolysis and the like; fibrates can lower TG and raise high density lipoprotein cholesterol (high density lipoprotein cholesterol, HDL-C), but have side effects such as hepatorenal toxicity. Therefore, development of high-efficiency low-toxicity hypolipidemic drugs is necessary.
In recent years, the traditional Chinese medicine has obvious advantages and characteristics for treating the hyperlipidemia, and is mainly characterized by multiple ways, multiple targets and less side effects. The traditional Chinese medicine mainly reduces blood fat in terms of inhibiting lipid absorption, affecting bile acid circulation, inhibiting lipid synthesis, resisting peroxidation, improving HDL and the like.
A preparation of fel Ursi heart-saving preparation is fel Ursi heart-saving pill, which is prepared from fel Ursi powder, bufonis venenum, borneolum Syntheticum, artificial Moschus, ginseng radix, margarita, artificial bezoar, fel Sus Domestica powder, cornu Bubali concentrated powder, has effects of tonifying heart, invigorating qi, and inducing resuscitation with aromatics, and is mainly used for treating chest obstruction due to heart qi deficiency, manifested by chest distress, heart pain, short breath, and palpitation.
Disclosure of Invention
The application aims to provide an application of bear gall heart-saving preparation in preparing a medicament for treating hyperlipidemia-related diseases.
The technical problem of the application is solved by adopting the following technical scheme.
On one hand, the embodiment of the application provides application of the bear gall heart-saving preparation in preparing medicines for treating hyperlipidemia-related diseases.
In some embodiments of the present application, the bear gall heart-saving preparation is a pharmaceutical preparation prepared from bear gall powder, venenum bufonis, borneol, artificial musk, ginseng, pearl, artificial bezoar, pig gall powder, buffalo horn concentrated powder and a proper amount of pharmaceutically acceptable carrier, and can be an oral liquid or solid preparation.
The bear gall heart-saving preparation comprises pills, capsules, tablets and the like. The pill is taken as a sample in the content of the invention, and the protection scope is not limited.
In some embodiments of the present application, the hyperlipidemia-associated diseases described above include hyperlipidemia, fatty liver, cognitive dysfunction, cardiovascular disease, and cardiac insufficiency.
In some embodiments of the present application, the cognitive dysfunction includes senile dementia.
In some embodiments of the present application, the hyperlipidemia includes hypercholesterolemia, hypertriglyceridemia, mixed hyperlipidemia, and low high density lipoprotein cholesterol.
Compared with the prior art, the application has at least the following advantages or beneficial effects:
(1) In the application, the experimental result shows that the bear gall heart-saving pill can reduce the content of TC, TG, LDL-C in serum of a rat with hyperlipidemia and increase the content of HDL-C.
(2) In the application, the bear gall heart-saving pill can improve the steatosis of the liver from two indexes of glutamic-oxaloacetic transaminase (glutamic oxaloacetic transaminase, AST) and glutamic-pyruvic transaminase (glutamic pyruvic transaminase, ALT) and HE staining result analysis of the liver.
(3) In the application, the vascular endothelial injury, intimal thickening and smooth muscle cell arrangement disorder of the model group rats are proved by experimental results to be capable of reducing the vascular endothelial injury by improving the blood lipid level of the rat with hyperlipidemia.
(4) In the application, experimental results show that the bear gall heart-saving pill can reduce cholesterol synthesis and metabolism by reducing the content of cholesterol ester transfer protein (cholesterol ester transfer protein, CETP) of rats with hyperlipidemia, and the activity of 3-hydroxy-3-mevalonate monoacyl-CoA reductase (recombiant 3-hydroxy-3-methylglutaryl coenzyme A reductase, HMGCR) and the activity of acyl transferase (acyl carbonyl a-cholesterol acyltransferase, ACAT) of rats in liver tissues. The bear gall heart-saving pill can lower the expression of HMGCR to inhibit the biosynthesis of cholesterol, regulate the levels of ACAT and CETP to increase the metabolism and transportation of lipid, thereby improving the blood lipid level and playing a certain role in treating hyperlipidemia.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings that are needed in the embodiments will be briefly described below, it being understood that the following drawings only illustrate some embodiments of the present application and therefore should not be considered limiting the scope, and that other related drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is the effect of XP on liver histopathology in hyperlipidemic rats (HE staining, ×400);
FIG. 2 shows the pathological changes of the aorta of the hyperlipidemic rats (HE staining);
FIG. 3 is a water maze test result;
FIG. 4 is a Y maze test result;
FIG. 5 is a graph showing the results of a new object discrimination experiment;
FIG. 6 shows the effect of XP on heart histopathology in hyperlipidemic rats (HE staining).
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present application more clear, the technical solutions in the embodiments of the present application will be clearly and completely described below. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
It should be noted that, in the case of no conflict, the embodiments and features in the embodiments may be combined with each other. The present application will be described in detail with reference to specific examples.
The features and capabilities of the present application are described in further detail below in connection with the examples.
Example 1
Experimental animals: SPF-grade SD male rats, 200+ -20 g, purchased from ChengdoenWeibull biotechnology Co., ltd. [ license number: SCXK (Yu) 2018-0003, which is conventionally raised in SPF-class animal houses, at a constant temperature (24+ -2) deg.C and a relative humidity (60+ -5)%. During the experiment, 25g of maintenance feed or high-fat feed (Beijing Bo Aigang Biotechnology Co., ltd.) was administered per day to each rat, and drinking water was free.
Reagents and instrumentation: bear gall heart-rescuing pill (XP, shenyang Qinggong pharmaceutical Co., ltd.); simvastatin (moesadong, hangzhou pharmaceutical limited); TC, TG, LDL-C, HDL-C, AST, ALT, nitric Oxide (NO) were purchased from Nanjing's institute of biological engineering; endothelin (ET-1), CETP, ACAT, HMGCR are all available from the biotechnology company, inc. Of Rui, spring; multifunctional microplate reader Varioskan TM LUX (Thermo Fisher); forward fluorescence microscope (Nikon).
Modeling and grouping: SD male rats were randomly divided into a Normal control group (Normal control group) and a model control group according to body weight after being fed with Normal feed for 3 days. Normal control group was given normal diet, model control group was given continuously 6 weeks high fat diet. After the 6-week molding was completed, the high-fat rats successfully molded were randomly divided into a Model control group (Model control group), a Simvastatin group (Simvastatin group), and an XP dose group (XP group), each group being 12.
Wherein, the configuration of simvastatin liquid medicine (4 mg/kg) comprises: each tablet of simvastatin is 20mg, 1 tablet of simvastatin tablet is taken, 50ml of 0.5% CMCNA is added to dissolve the simvastatin tablet, and 0.4mg/ml simvastatin liquid medicine is prepared;
the XP liquid medicine (15.75 mg/kg) is configured to comprise: the preparation method comprises grinding fel Ursi heart-saving pill into powder with mortar, adding 0.5% CMCNA, and preparing into 1.575mg/ml fel Ursi heart-saving pill liquid medicine.
During the administration period, each administration group was given the corresponding drug (1 ml/100 g) by gavage, and the model control group and the normal control group were given the same volume of physiological saline 1 time a day for 9 consecutive weeks, and the administration group and the model group were continuously fed with the high-fat feed. Rats were sacrificed 9 weeks after dosing and the samples were stored in a-80 ℃ refrigerator for testing.
The experimental method comprises the following steps: during the experiment, rat body weight was recorded weekly. After the end of the administration, the anesthetized rat, the abdominal aorta, blood collection, serum separation, and determination of TC, TG, LDL-C, HDL-C, NO, ET-1, CETP content and AST, ALT, HMGCR activity were performed according to the kit instructions; taking liver tissues to refer to a kit instruction book for determining TC and TG contents and ACAT activities; after the aorta, heart and liver were fixed in 4% paraformaldehyde for 24 hours, each was dehydrated with 70%, 85%, 95% and 100% ethanol solutions sequentially for 1 hour, embedded in paraffin, cut into sections with a thickness of 4 μm, and then HE stained.
The statistical treatment method comprises the following steps: all data were statistically analyzed using SPSS 26.0 software and the results were obtained using mean.+ -. Standard deviation
The comparison among multiple groups adopts single-factor analysis of variance, P<0.05 is statistically significant.
Experimental results:
(1) Influence of bear gall heart-saving pill on hyperlipidemia rat blood fat
The experimental results are shown in table 1:
TABLE 1 influence of XP on blood lipid levels in hyperlipidemic rats [ ]
n＝12)
In comparison with the control group of the model, * P<0.05， ** P<0.01
the results in Table 1 show that the model control group showed significantly higher TC, TG, LDL-C content and significantly lower HDL-C content (P < 0.01) than the normal control group, indicating that the high fat diet feeding method can successfully replicate the hyperlipidemic model of rats. Compared with the model control group, the TC, TG, LDL-C content of the XP group and the Simvastatin group is reduced, the HDL-C content is increased (P <0.01 and P < 0.05) after 9 weeks of administration, and the effect of reducing blood fat of the XP group is the most obvious and similar to that of the Simvastatin group.
(2) Influence of bear gall heart-saving pill on lipid content and pathology in liver tissue of hyperlipidemic rat
The experimental results are shown in table 2:
TABLE 2 influence of XP on lipid content in liver tissue of hyperlipidemic rats
n＝12)
| Group of | Dose/(mg/kg) | TC/(mmol/g prot) | TG/(mmol/g prot) |
| Normal control | / | 0.045±0.014** | 0.127±0.035** |
| Model control | / | 0.142±0.042 | 0.434±0.142 |
| |
4 | 0.077±0.026* | 0.264±0.062** |
| XP | 15.75 | 0.084±0.036* | 0.258±0.106* |
In comparison with the control group of the model, * P<0.05， ** P<0.01
from the analysis in table 2, the TG and TC contents in the rat liver tissue of the model control group were significantly increased (P < 0.01) compared to the normal control group; the XP group and the Simvastatin group reduce the TG and TC contents (P is less than 0.05) in liver tissues, which suggests that the XP can reduce the pathological changes of fatty liver of rats with hyperlipidemia to a certain extent.
And from the liver HE staining of FIG. 1, the liver cells of the rats in the normal control group are complete in structure and have no formation of fat vacuoles; the liver cells of the rats in the model control group are distributed in disorder, and a large amount of fatty vacuoles can be seen in the cells, which indicates the formation of fatty liver. Compared with the model control group, the pathological change of liver tissue of the XP group is obviously improved, the liver cells are distributed regularly, and the large vesicular lipid drops are reduced and are similar to those of the Simvastatin group. Taken together, XP is shown to improve lipid deposition in the liver of hyperlipidemic rats.
(3) Influence of bear gall heart-saving pill on NO and ET-1 in serum of rat with hyperlipidemia and aortic morphology
The experimental results are shown in table 3 and fig. 2:
TABLE 3 XP concentration of NO, ET-1 in serum of hyperlipidemic rats
n＝12)
In comparison with the control group of the model, * P<0.05， ** P<0.01
ET-1 and No are important vasodilator and vasoconstrictor substances, respectively, which interact to maintain normal function of blood vessels. When hyperlipidemia occurs, NO and ET-1 metabolism in the vascular endothelium is abnormal, resulting in impaired vascular endothelial function.
From the analysis in Table 3, compared with the normal control group, the serum ET-1 content of the rats in the model control group is obviously increased (P < 0.01), and the NO content is obviously reduced (P < 0.01); compared with a model control group, the XP dosage group can obviously reduce the content of serum ET-1 and increase the content of NO in serum, and is similar to the SIM group. The results show that the XP group can reduce vascular endothelial injury by improving the blood lipid level of the hyperlipidemic rats.
The results in fig. 2 show that the normal control rats had intact aortic intima and smooth muscle cells were ordered and no foam cells were seen; the rat aortic intima of the model control group is damaged, the intima media is thickened, smooth muscle cells are arranged in disorder, and lipid vacuoles at the media are obviously increased; the rat aortic intima of XP group and simvastatin group has more complete structure, less fat vacuoles and more orderly arrangement of middle smooth muscle; the XP group is suggested to have a certain protection effect on vascular endothelial structure injury of the rat with hyperlipidemia.
(4) Influence of bear gall heart-saving pill on AST and ALT content and liver tissue structure in serum of rat with hyperlipidemia
The experimental results are shown in table 4:
TABLE 4 influence of XP on liver function in hyperlipidemic rats
n＝12)
| Group of | Dose/(mg/kg) | ALT/(U/L) | AST/(U/L) |
| Normal control | / | 6.4±1.0** | 10.5±1.8** |
| Model control | / | 13.1±2.6 | 17.4±3.8 |
| |
4 | 9.2±1.9** | 11.9±1.7** |
| XP | 15.75 | 9.2±1.9** | 11.9±2.4** |
In comparison with the control group of the model, * P<0.05， ** P<0.01
from the analysis in table 4, AST and ALT activities in the serum of rats of the model control group are obviously increased (P < 0.01) compared with the normal control group; compared with the model control group, the XP group significantly reduces AST and ALT activities, and is similar to the Simvastatin group.
(5) Influence of bear gall heart-saving pill on CETP and HMGCR in serum of rat with hyperlipidemia
The experimental results are shown in table 5:
TABLE 5 influence of XP on the active content of HMGCR and ACAT in hyperlipidemic rats
n＝12)
| Group of | Dose/(mg/kg) | HMGCR/(U/L) | CETP/(μg/L) |
| Normal control | / | 87.1±42.7 ** | 1510.7±133.5 ** |
| Model control | / | 264.8±109.2 | 2641.5±465.9 |
| |
4 | 128.2±77.8 * | 1869.7±387.5 ** |
| XP | 15.75 | 134.1±70.3 * | 2004.1±283.3 * |
In comparison with the control group of the model, * P<0.05， ** P<0.01
HMGCR is the rate-limiting enzyme for cholesterol synthesis by the liver, HMGCR expression is up-regulated and cholesterol synthesis in the body is increased, currently clinical statins are lipid-lowering by inhibiting HMGCR activity, ACAT is the only enzyme in cells that synthesizes Cholesterol Ester (CE), cholesterol ester production and Very Low Density Lipoprotein (VLDL) secretion can be reduced by lowering ACAT, and CETP promotes transport of CE and TG between HDL-C and LDL-C and VLDL in the blood, and lipid metabolism balance can be maintained by regulating CETP expression levels.
Table 5 shows that the serum CETP and HMGCR levels in the rats of the model control group are increased (P < 0.01) compared with the normal control group; the serum CETP and HMGCR levels were reduced in the XP and simvastatin groups of rats compared to the model group.
(6) Influence of bear gall heart-saving pill on ACAT in liver tissue of hyperlipidemic rat
The experimental results are shown in table 6:
TABLE 6 influence of XP on the active content of HMGCR and ACAT in hyperlipidemic rats
n＝12)
| Group of | Dose/(mg/kg) | ACAT/(U/L) |
| Normal control | / | 5.5±1.1 ** |
| Model control | / | 13.4±1.2 |
| |
4 | 8.1±0.8 ** |
| XP | 15.75 | 9.1±1.5 ** |
In comparison with the control group of the model, * P<0.05， ** P<0.01
table 6 the results show that the ACAT activity in liver tissue of rats is increased in the model control group compared to the normal control group (P < 0.01); the XP group had a significant decrease compared to the model group.
(5) And (6) the result shows that the bear gall heart-saving pill can reduce cholesterol synthesis and accelerate metabolism by reducing the content of CETP in serum, HMGCR activity and ACAT activity in liver tissues of a rat with hyperlipidemia, thereby improving blood lipid level and playing a certain role in treating hyperlipidemia.
(7) Influence of bear gall heart-rescuing pill on cognitive dysfunction of hyperlipoidemia rat
At the end of the experiment, we used water maze, Y maze, new object discrimination experiments to evaluate the learning and memory ability of rats.
As shown in fig. 3, the analysis of the results of the water maze experiment shows that, in the hidden platform experiment, as the training time of the rats increases, the latency period for three groups of rats to find the platform decreases, and the latency period of each group of rats is shortest on the fifth day, which indicates that all three groups of rats obtain a certain learning and memory capacity in training.
The latency time for model control rats to find the hidden station three days after training was significantly prolonged compared to the normal control (P <0.01, P < 0.05); the XP dose group found a significant reduction in the time to hidden station latency on the last day of training (P < 0.05) compared to model control rats. In the probe experiment, the number of times of crossing the platform by the normal control group rat and the percentage of time of the platform quadrant are respectively 3.2+/-1.0 times and 31.08+/-6.1 percent, the number of times of crossing the platform by the model control group rat is reduced to 2.0+/-0.9 times, meanwhile, the percentage of time of the platform quadrant is reduced to 24.58+/-5.18 percent, and compared with the normal control group, the two groups of data have obvious differences (P < 0.01), so that the memory of the model control group rat on the position of the safety platform is reduced; XP dose group rats recovered to a different extent from the memory of the safety platform position (P < 0.05), thus demonstrating that XP can improve the learning and memory capacity of hyperlipidemic rats; this is further verified by a typical path diagram of the probe experiment.
The Y maze experiment result is shown in figure 4, and the alternation rate of the rats in the model control group is higher than that of the rats in the normal control group (P < 0.01), which indicates that the rats in the model control group have short-time working memory decline; whereas the short-term working memory of the XP dose group rats was restored (P < 0.05).
As shown in fig. 5, compared with the normal control group rats, the new object distinguishing experiment result shows that the distinguishing index of the model control group rats is obviously reduced (P < 0.01), which indicates that the learning and memory ability of the model control group rats is weakened; XP dose group rats had a significant difference in discrimination index rise (P < 0.05) compared to model control group rats.
In conclusion, the learning and memory ability of the hyperlipidemic rats established by the 9-week hyperlipidemic diet feeding method is reduced, and the XP can improve the cognitive dysfunction of the hyperlipidemic rats.
(8) Influence of bear gall heart-rescuing pill on heart pathology of hyperlipoidemia rat
FIG. 6 shows that the heart is complete in myocardial tissue structure of the normal control group rats, and myocardial cells are orderly arranged without inflammatory infiltration as can be seen from the HE staining result; the model control group has the advantages that the myocardial cells of the rat are arranged in disorder, a large number of myocardial cells are hypertrophic, deformed and the like, and a small amount of inflammatory cells infiltrate; the administration groups can relieve the heart pathological changes of the rats with hyperlipidemia to different degrees, wherein the XP dose group has the most obvious pathological changes, the myocardial cells are arranged slightly orderly, a small number of myocardial cells are hypertrophic, denatured and the like, and the cell nuclei are uniformly distributed.
In conclusion, XP can have certain protective action to the heart function of hyperlipidemic rats.
The embodiments described above are some, but not all, of the embodiments of the present application. The detailed description of the embodiments of the present application is not intended to limit the scope of the application, as claimed, but is merely representative of selected embodiments of the application. All other embodiments, which can be made by one of ordinary skill in the art based on the embodiments herein without making any inventive effort, are intended to be within the scope of the present application.
Claims (5)
1. The application of bear gall heart-saving preparation in preparing medicament for treating hyperlipidemia-related diseases.
2. The use according to claim 1, wherein the bear gall heart-saving formulation comprises an oral liquid or solid formulation, such as: pills, pellets, capsules and tablets.
3. The use according to claim 1, wherein the hyperlipidemia-related diseases comprise hyperlipidemia, fatty liver, cognitive dysfunction, cardiovascular disease and cardiac insufficiency.
4. The use according to claim 3, wherein the cognitive dysfunction comprises senile dementia.
5. The use according to claim 3, wherein the hyperlipidemia comprises hypercholesterolemia, hypertriglyceridemia, mixed hyperlipidemia and low high density lipoprotein cholesterol.
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| CN108670979A (en) * | 2018-06-21 | 2018-10-19 | 齐光伟 | Adjust-blood lipid norcholesterol prevents the bear gall powder of cardiovascular and cerebrovascular diseases and atherosclerosis |
| CN115089655A (en) * | 2022-05-12 | 2022-09-23 | 北京宜生堂医药科技研究有限公司 | Traditional Chinese medicine composition for treating Alzheimer's disease, extract, preparation method and application |
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| CN108670979A (en) * | 2018-06-21 | 2018-10-19 | 齐光伟 | Adjust-blood lipid norcholesterol prevents the bear gall powder of cardiovascular and cerebrovascular diseases and atherosclerosis |
| CN115089655A (en) * | 2022-05-12 | 2022-09-23 | 北京宜生堂医药科技研究有限公司 | Traditional Chinese medicine composition for treating Alzheimer's disease, extract, preparation method and application |
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| 周鸿缘 等: "牛磺熊去氧胆酸的药理作用及应用研究进展", 动物医学进展, vol. 43, no. 8, pages 103 * |
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