Application of dammarane saponin compound in preparation of lipid-lowering product

Abstract

The invention belongs to the technical field of natural products and medicinal chemistry, and particularly relates to application of a dammarane type saponin compound in preparation of a lipid-lowering product, and the dammarane type saponin compound is one or more of gypenoside L, gypenoside LI, damulin A and damulin B. The invention further discloses a preparation method of the dammarane type saponin compound. The invention also provides a composition. The composition is composed of the dammarane type saponin compound and the statins. The application finds that the single dammarane saponin compound can be applied to lipid lowering, and after the dammarane saponin compound is combined with the sitavastatin for use, a synergistic effect can be generated, and the lipid lowering performance of the sitavastatin can be promoted, so that the use of a large amount of sitavastatin medicines can be reduced, and side effects caused by the use of the large amount of sitavastatin medicines can be reduced. The dammarane type saponin compound provided by the invention is extracted from gynostemma pentaphylla, is a natural plant extract, is safe and effective in components, and has a relatively good application prospect in carrying out lipid-lowering products or carrying out synergistic lipid lowering by combining with other medicines.

Description

Technical Field

[0001] The present invention belongs to the technical field of natural products and medicinal chemistry, and more specifically, relates to the application of dammarane saponin compounds in the preparation of lipid-lowering products. Background Art

[0002] Gypenoside L, Gypenoside LI, Damulin A and Damulin B are dammarane-type saponin compounds isolated from Gynostemma pentaphyllum, which are two pairs of isomers (the C-20 position of Gypenoside L is S-configuration, while the C-20 position of Gypenoside LI is R-configuration. The double bond positions of Damulin A and Damulin B are different). Studies have found that heat-treated Gynostemma pentaphyllum total saponins and gypenoside L, gypenoside LI and damulin B can inhibit the proliferation of lung cancer cells and breast cancer cells. Since the application of lipid-lowering drugs such as statins, some side effects have been found in lipid-lowering chemical synthetic drugs while treating diseases, such as rhabdomyolysis, pancreatitis and diabetes; for some patients (such as hereditary hyperlipidemia), there is still no drug available. Therefore, more research on lipid-lowering products that can reduce the use of statins and have good efficacy is needed. At present, no research on the above compounds in lipid-lowering has been retrieved. Summary of the invention

[0003] This application uses the HepG2 cell model to explore the effects of gypenoside L, gypenoside LI, damulin A, and damulin B on lipid-lowering and their structure-activity relationship, providing evidence for further research on natural compounds with lipid-lowering effects.

[0004] This application also studies the above compounds in combination with statins, and significantly enhances the lipid-lowering effect of statins.

[0005] The dammarane saponin compounds provided in the present application have a good lipid-lowering effect when used alone or in combination with statins.

[0006] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0007] Application of a dammarane saponin compound in the preparation of a lipid-lowering product, wherein the dammarane saponin compound is one or more of gypenoside L, gypenoside LI, damulin A, and damulin B.

[0008] The CAS numbers of the compounds are as follows:

[0009] gypenoside L:CAS:94987-09-4;

[0010] gypenoside LI:CAS:94987-10-7;

[0011] damulin A: CAS:1202868-74-3;

[0012] damulin B: CAS:1202868-75-4;

[0013] The above compounds are all known compounds.

[0014] Preferably, the dammarane-type saponin compound reduces TC, TG and fat accumulation.

[0015] The present application also provides a composition consisting of a dammarane-type saponin compound and a statin.

[0016] Preferably, the statin is simvastatin.

[0017] The present application also claims to protect the use of the composition in the preparation of lipid-lowering products.

[0018] Furthermore, the statin is simvastatin; and the dammarane saponin compound is one or more of gypenoside L, gypenoside LI, damulin A, and damulin B.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] The present application found that a single dammarane saponin compound can be used for lipid-lowering, and when used in combination with simtavatin, it can significantly produce a synergistic effect, promote the lipid-lowering performance of simtavatin, and thus reduce the use of a large number of simtavatin drugs. The dammarane saponin compound provided in the present application is extracted from Gynostemma pentaphyllum, which is a natural plant extract with safe and effective ingredients. It has good application prospects in the development of lipid-lowering products or in combination with other drugs for synergistic lipid-lowering. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 The effect of oleic acid (A) 25-OH cholesterol + cholesterol (B) on the survival rate of HepG2 cells. Reagent treatment group vs. normal control group * p≤0.05, ** p≤0.01.

[0022] Figure 2The effects of heat-treated gypenosides HGyp (A), gypenoside L (B), gypenoside LI (C), damulin A (D), and damulin B (E) on the survival rate of HepG2 cells. Drug treatment group vs. normal control group * p≤0.05, ** p≤0.01.

[0023] Figure 3 The effect of oleic acid 500μM and cholesterol 100μM + 25-OH cholesterol 10μM on TC (A) and TG (B) of HepG2 cells. Reagent treatment group vs. normal control group * p≤0.05, ** p≤0.01.

[0024] Figure 4 Cell staining. (A) Control group; (B) Model group; (C) 10μM simvastatin group; (D) 100μM damulin A group; (E) 50μM damulin A group; (F) 50μM damulin B group; (G) 25μM damulin B group; (H) 50μM gypenoside L group; (I) 25μM gypenoside L group; (J) 10μM gypenoside LI group; (K) 5μM gypenoside LI group.

[0025] Figure 5 The effects of Danulin A, damulin B, gypenoside L, and gypenoside LI on protein. Model group vs. Control group # p≤0.05, ## p≤0.01. Gynostemma pentaphyllum saponin groups vs. Model group * p≤0.05, ** p≤0.01. DETAILED DESCRIPTION

[0026] The technical scheme of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention and the figures. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative work are within the scope of protection of the present invention. If the specific conditions are not specified in the embodiments, they are prepared according to the conditions recommended by the normal conditions or the manufacturers. If the manufacturer is not specified in the reagents or instruments used, they are all conventional products that can be purchased commercially.

[0027] Example 1 Effect of Gynostemma pentaphyllum saponins on the survival rate of HepG2 cells

[0028] Cell viability was detected by CCK8, and the inhibitory effects of gypenoside L, gypenoside LI, damulin A, damulin B, modeling agent OA, and cholesterol on HepG2 cells were observed.

[0029] 1 Experimental methods

[0030] 1.1 Drug configuration

[0031] Take 10 mg each of gypenoside L, gypenoside LI, damulin A, damulin B, and simvastatin, add DMSO to prepare a 100 mM solution. Take 5 μL of oleic acid (0.89 mg / μL), add 11 μL of DMSO solution to prepare a 1000 mM oleic acid solution.

[0032] 1.2 Cell viability assay

[0033] The experiment was divided into a blank control group, a model group, and 5, 10, 50, 100, 150, and 200 μM gypenoside L, gypenosideLI, damulin A, and damulin B compound groups. Cells in the logarithmic growth phase were selected, washed with PBS to remove impurities, and after trypsin digestion, centrifuged at 1200 rpm, a certain volume of complete medium was added to mix, the cell suspension was aspirated, and a certain multiple was diluted with PBS for counting. The counted cells were diluted with complete medium according to the experimental design. After mixing, they were added to a 96-well plate, and 200 μL of cell suspension was added to each well. The number of cells in each well was 1×10 4 , and culture in an incubator for 24 hours. When the cells are observed to grow attached to the wall, add the prepared drugs to the 96-well plate at the designed concentration. Continue to culture in the incubator for 24 hours. Take out the 96-well plate, add CCK8 solution to each well, 200 μL per well, and culture at 37°C in the dark for 30 minutes. Measure the absorbance at 560nm with an enzyme reader. Calculate the cell survival rate according to the following formula: Survival rate (%) = (OD 实验组 -OD 空白组 ) / (OD 对照组 -OD 空白组 )×100%.

[0034] 1.3 Cell model experiments

[0035] The experiment set up a blank group and a model group. The cells in the logarithmic growth phase were digested and counted, and plated in a 6-well plate, with 2×10 cells per well. 5According to the cell viability experiment, appropriate oleic acid concentration and cholesterol concentration were selected to incubate the cells for 24 h, and the cell TC and TG contents were determined according to the method in the kit instructions.

[0036] 2 Experimental results

[0037] 2.1 Effects of compounds on cell viability

[0038] High concentrations of oleic acid, cholesterol, and gypenosides have certain cytotoxicity. In this experiment, the CCK8 method was used to detect the effects of different gypenosides at different concentrations on the survival rate of HepG2 cells through a cell viability experiment, and the concentration with the least cytotoxic effect was selected as the subsequent experimental condition.

[0039] As the concentration of the membrane-forming agent OA and 25-OH cholesterol + cholesterol changes, the cell survival rate will be affected, so choosing the appropriate concentration of the membrane-forming agent is conducive to the smooth progress of the experiment. Figure 1 As shown in A, when the OA concentration is within 700 μM, the cell survival rate is above 90%, indicating that it can be used for cell membrane formation below this concentration. Figure 1 As shown in B, when the concentration of 25-OH cholesterol + cholesterol is 16+160 μM, the cell survival rate is still above 90%, indicating that 25-OH cholesterol + cholesterol has little effect on cells.

[0040] As the concentration of gypenosides increased, the cell survival rate gradually decreased, indicating that gypenosides had a certain toxic effect on HepG2 cells. Figure 2 As shown in A, when the HGyp concentration is within 50 μg / mL, the cell survival rate is above 90%, indicating that the toxic effect on cells is relatively small within this concentration range. Therefore, the dosing concentration of HGyp can be set within 50 μg / mL. Figure 2 As shown in B, when the concentration of gypenoside L is below 50 μM, the cell survival rate is above 90%, indicating that the toxic effect on cells is relatively small within this concentration range. Therefore, the administration concentration of gypenoside L can be set within 50 μM. Figure 2 As shown in Figure C, when the concentration of gypenoside LI is below 10 μM, the cell survival rate is above 90%, indicating that the toxic effect on cells is relatively small within this concentration range. Therefore, the administration concentration of gypenoside LI can be set within 10 μM. Figure 2 As shown in D, when the concentration of damulin A is below 200 μM, the cell survival rate is above 90%, indicating that the compound has a relatively small toxic effect on cells. Figure 2As shown in Figure E, when the concentration of damulin B is below 50 μM, the cell survival rate is above 90%, indicating that the toxic effect on cells is relatively small within this concentration range. Therefore, the administration concentration of damulin B can be set within 50 μM.

[0041] 2.2 Cell model construction

[0042] According to the effects of oleic acid and cholesterol on cell viability, this experiment selected oleic acid concentration of 100μM, cholesterol concentration of 100μM, and 25-OH cholesterol concentration of 10μM. The cholesterol and triglyceride content of each group was measured, and the results are as follows Figure 3 As shown, TC and TG of HepG2 cells incubated with oleic acid were significantly increased; TC of HepG2 cells incubated with cholesterol + 25-OH cholesterol was significantly increased and TG was decreased.

[0043] In summary, oleic acid concentrations below 700 μM have little effect on HepG2 cells. Different concentrations of 25-OH cholesterol + cholesterol have little effect on HepG2 cells. The concentrations of HGyp, gypenoside L, gypenoside LI, and damulin B that affect the 90% survival rate of HepG2 cells are 50 μM, 50 μM, 10 μM, and 25 μM, respectively. Among them, damulin A has almost no effect on cell survival rate.

[0044] Example 2: Effect of Gynostemma pentaphyllum saponins on lipid-lowering effects of oleic acid-induced HepG2 cells

[0045] In order to further study the lipid-lowering effect of gypenosides, the oleic acid-induced HepG2 hyperlipidemia cell model was used to explore the lipid-lowering effects of gypenoside L, gypenoside LI, damulin A, damulin B and combined with simvastatin as well as their effects on lipid synthesis.

[0046] 1 Experimental methods

[0047] 1.1 Experimental Grouping

[0048] Blank control group; 0.5mM OA group; 0.5mM OA+10μM simvastatin intervention group; 0.5mM OA+50μMgypenoside L intervention group; 0.5mM OA+25μM gypenoside L intervention group; 0.5mM OA+10μMgypenosideLI intervention group; 0.5mM OA+5μM gypenoside LI intervention group; 0.5mM OA+100μMdamulin A intervention group; 0.5mMOA+50μM damulin A intervention group; 0.5mM OA+50μM damulin B intervention group; 0.5mM OA+25μM damulin B intervention group. Simvastatin + gypenoside L: 10μM + 25μM; simvastatin + gypenoside LI: 10μM + 10μM; simvastatin + damulin A: 10μM + 50μM; simvastatin + damulin B: 10μM + 25μM.

[0049] 1.2 Cell culture

[0050] Select cells in logarithmic growth phase for the experiment. Dilute the counted cell suspension and add it to a 24-well plate. The number of cells in each well is 5×10 4 After 24 hours of culture, when the cells are attached to the wall and in good condition, proceed to the next experiment. Add 500mM oleic acid as a membrane-forming agent and continue to culture for 24 hours. After the high-lipid model cells are formed, add the prepared drugs to the 24-well plate according to the designed concentration. Put it in the incubator and culture it for 24 hours. After 24 hours, take out the 24-well plate, wash it twice with PBS, add 40μL IP lysis solution and lyse it on ice for 40 minutes. Take a certain amount of cell suspension for measurement.

[0051] 1.3 TC and TG content determination

[0052] The sample and working solution were added to a 96-well plate according to the ratio, and the cell TC and TG contents were determined according to the kit instructions.

[0053] 1.4 Protein immunoblotting

[0054] Gel electrophoresis was used to determine the expression of SREBP1, SREBP2, PPARα, PPARδ, PPARγ and HMGCR proteins in cells. The method is as follows: First, extract cell proteins, collect the cells treated with drugs in 24-well plates, and wash them three times with PBS; add 40 μL of cell lysis solution containing 1 mM PMSF to each well and place on ice for 40 minutes, during which time the cells are blown several times with a gun to fully lyse; centrifuge at 12000g, 4℃ for 10 minutes, and collect the supernatant. Secondly, the protein concentration is determined, and the electrophoresis development and other processes are the same as the reference Western blot experimental method.

[0055] 1.5 Statistical analysis

[0056] The experimental data were expressed as mean ± standard deviation (SD) and statistically analyzed using SPSS25.0 software. The experimental data were analyzed using one-way analysis of variance, and p < 0.05 was considered to be statistically different, and p < 0.01 was considered to be statistically significant.

[0057] 2 Experimental results

[0058] 2.1 Effect of Gynostemma pentaphyllum saponins on TC in HepG2 cell model induced by oleic acid

[0059] The experiment selected a concentration with less toxicity to cells and measured the effect of gypenosides on cholesterol in HepG2 cells incubated with oleic acid within this concentration range.

[0060] Table 1 Clearance rate of TC in HepG2 cells by gypenosides alone

[0061]

[0062]

[0063] HFD group vs. ND group # p≤0.05, ## p≤0.01. Gynostemma pentaphyllum saponin groups vs. HFD group * p≤0.05, ** p≤0.01.

[0064] Table 2 Clearance rate of TC in HepG2 cells by Gynostemma pentaphyllum saponin combination group

[0065]

[0066] Gynostemma pentaphyllum saponin group vs. corresponding combined drug group § p≤0.05, §§ p≤0.01.

[0067] Table 1 shows that the total cholesterol content in the model group increased significantly compared with the blank group, with a very significant difference (p≤0.05), indicating that the oleic acid-induced HepG2 cell fat accumulation model was successfully established. Compared with the model group, the Damulin A group had a significant difference, and the clearance rate of TC in the cells reached 64.98% and 67.97%. Compared with the model group, the Damulin B group had a significant difference, and the clearance rate of TC in the cells reached 69.72% and 54.34%. Compared with the model group, the Gypenoside L group had a significant difference, and the clearance rate of TC in the cells reached 57.69% and 72.38%, and within a certain concentration range, with the increase of drug concentration, the clearance rate of TC also gradually increased, indicating that within a certain concentration range, the clearance rate of TC by the compound was positively correlated. Compared with the model group, the Gypenoside LI group had a significant difference, and the clearance rate of TC in the cells reached 57.58% and 61.42%. Among them, gypenoside L 25μM had the strongest effect on reducing cellular TC, with a TC clearance rate of up to 72.38%.

[0068] In summary, according to the above experimental results, it can be clearly found that the four compounds have a good scavenging effect on total cholesterol in HepG2 cells incubated with oleic acid. At a concentration of 25 μM, the clearance rate of cholesterol in cells by gypenoside L is close to that of simvastatin. Damulin B has the lowest toxicity and the clearance rate of total cholesterol is positively correlated with the concentration of monomer saponins. Gypenoside LI is more toxic and its effect on total cholesterol in cells is slightly lower than that of other compounds. In short, gypenosides all affect total cholesterol in cells, and the clearance rate of total cholesterol in cells is related to the concentration of monomers.

[0069] In addition, it can be directly seen from Table 2 that the combined administration group also showed extremely significant differences (p≤0.01) compared with the gypenoside group, i.e., the single-drug group. This shows that after combined administration, the cholesterol-lowering abilities of damulin A, damulin B, gypenoside L, and gypenoside LI were significantly enhanced, and the above substances, when used in combination with simtavatin, could produce significant synergistic effects.

[0070] 2.2 Effect of Gynostemma pentaphyllum saponins on TG in HepG2 cell model induced by oleic acid

[0071] The experiment selected a concentration with less toxicity to cells and determined the effect of gypenosides on triglycerides in HepG2 cells incubated with oleic acid within this concentration range.

[0072] Table 3 Clearance rate of TG in HepG2 cells by Gynostemma pentaphyllum saponin alone

[0073]

[0074] HFD group vs. ND group # p≤0.05, ## p≤0.01. Gynostemma pentaphyllum saponin groups vs. HFD group * p≤0.05, ** p≤0.01.

[0075] Table 4 The clearance rate of TG in HepG2 cells by Gynostemma pentaphyllum saponin combination

[0076]

[0077] Gynostemma pentaphyllum saponin groups (except Damulin A high-dose group) vs. corresponding combined drug groups § p≤0.05, §§ p≤0.01.

[0078] As can be seen from Table 3, compared with the blank group, the content of total triglyceride in the model group increased significantly, with a very significant difference (p ≤ 0.05), indicating that the oleic acid-induced HepG2 cell fat accumulation model was successfully established. Compared with the model group, the Damulin A group had a significant difference, and the clearance rate of TG in the cells reached 18.76% and 37.09%. Compared with the model group, the Damulin B group had a significant difference, and the clearance rate of TG in the cells reached 25.10% and 35.95%. Compared with the model group, the Gypenoside L group had a significant difference, and the clearance rate of TG in the cells reached 32.80% and 42.86%, and within a certain concentration range, with the increase of drug concentration, the clearance rate of TG also gradually increased, indicating that within a certain concentration range, the clearance rate of compounds to TG was positively correlated. Compared with the model group, the Gypenoside LI group had a significant difference, and the clearance rate of TG in the cells reached 24.69% and 10.80%. Among them, gypenoside L 25μM had the strongest effect on reducing cell TG, and the clearance rate of TG was as high as 42.86%.

[0079] In summary, according to the above experimental results, it can be clearly found that the four compounds have a certain scavenging effect on triglycerides in HepG2 cells incubated with oleic acid, and gypenoside L 25μM has the highest clearance rate of triglycerides in cells. Damulin B has the lowest toxicity and the second highest clearance rate of triglycerides. Gypenoside LI is more toxic and has a lower effect on triglycerides in cells than other compounds. In short, gypenosides all affect triglycerides in cells, and the clearance rate of intracellular triglycerides is related to the concentration of monomers.

[0080] In addition, it can be seen from Table 4 that the combined administration group showed extremely significant differences (p≤0.01) compared with the gypenoside group, i.e., the single-drug group. This indicates that after combined administration, the triglyceride-lowering abilities of damulin A, damulin B, gypenoside L, and gypenoside LI were significantly enhanced, and the above substances, when used in combination with simtavatin, could produce significant synergistic effects.

[0081] 2.3 Effect of oleic acid on fat accumulation in HepG2 cells

[0082] The liver is the main organ for lipid metabolism. It is involved in the production, storage, transportation and oxidation of lipids and is crucial to all processes of lipid metabolism. Hepatocytes are the main site for the implementation of liver metabolic functions. Under normal circumstances, there is a small amount of lipid in the cytoplasm of hepatocytes. When the balance between fatty acid uptake and new fat is broken, the body produces a series of stress responses, resulting in a large accumulation of lipids in hepatocytes. Therefore, hepatocytes can be used to explore the effects of drugs on fat accumulation.

[0083] The liver can regulate the lipid level in the blood. It plays a role in lowering lipids by inhibiting the synthesis of lipids in the liver or increasing the oxidation of lipids. In this experiment, the effect of gypenosides on lipid-lowering was observed by Oil Red O staining. Compared with the model group, intracellular lipids decreased after gypenosides intervention, and biochemical indicators also indicated a decrease in triglycerides. Therefore, it can be determined that gypenosides can reduce fat accumulation to varying degrees.

[0084] Through microscopic observation of cell morphology after Oil Red O staining, compared with the blank control group, the model group had a large amount of stained oil, the oils were fused with each other and the number was large, indicating that the fat accumulation model incubated with oleic acid was successful. After the administration of four Gynostemma pentaphyllum saponin compounds, the area of ​​red-stained oil in HepG2 cells became smaller, the number decreased, and the color became lighter than that of the model group. Figure 4 It can be seen that the high-dose and low-dose groups of damulin A, damulin B, gypenoside L, and gypenoside LI all reduced lipid accumulation to varying degrees.

[0085] 2.4 Effects on cellular SREBP2, SREBP1, PPARα, PPARδ, PPARγ and HMGCR

[0086] Hepatic lipid metabolism is a complex process involving a variety of lipid metabolism-related proteins. SREBP plays an important role in lipid production. The mevalonate pathway mediated by 3-hydroxy-3-methyl glutaryl coenzyme A reductase (HMGCR) is a key step in the de novo synthesis of cholesterol in the body. The expression and regulation of HMGCR are very important for maintaining cholesterol homeostasis in the body.

[0087] Western blot was used to detect the expression of SREBP2, SREBP1, PPARα, PPARδ, PPARγ and HMGCR, which are related proteins in lipid metabolism, to explore the effect of gypenosides in reducing lipid accumulation. Figure 5 As shown in the figure, compared with the control group, the SREBP2 protein level in the model group was significantly increased. In contrast, the low-dose damulin A group significantly inhibited the expression of SREBP2 in OA-treated HepG2 cells. Compared with the blank control group, the SREBP1 protein in the model group did not change significantly, but the high- and low-dose damulin A groups, the high- and low-dose damulin B groups, and the high- and low-dose gypenoside L groups significantly inhibited the expression of SREBP1 in HepG2 cells. The PPARA protein in OA-treated HepG2 cells increased, the PPARA protein in the high-dose damuin A and low-dose gypenoside LI groups decreased, and the protein expression in other groups increased. For HMGCR and PPARD proteins, the high- and low-dose damulin B, gypenoside L, and gypenoside LI groups were lower than the model group, and also lower than the blank control group. From the results, it was found that the PPARG protein in each drug-treated group was lower than that in the model group. The above results show that the lipid-lowering effects of the four compounds are different in strength and the proteins they act on are also different.

[0088] Hepatic lipid metabolism is a complex process, which is participated by a variety of lipid metabolism-related proteins. SREBP plays an important role in lipid production. The mevalonate pathway mediated by 3-hydroxy-3-methyl glutaryl coenzyme A reductase (HMGCR) is a key step in the de novo synthesis of cholesterol in the body. The expression and regulation of HMGCR are very important for the maintenance of cholesterol homeostasis in the body. This study showed that damulin A, damulin B, and gypenoside L reduced the expression of SREBP1. The expression of HMGCR protein was significantly reduced after intervention with damulin B, gypenoside L, and gypenoside LI. As for SREBP2, only the expression of the low-dose damulin A group was reduced. This indicates that gypenosides may cause changes in the expression levels of important proteins for lipogenesis in HepG2 cells and inhibit liver fatty acid synthesis. PPARα plays an important regulatory role in the four-step β-oxidation reaction of dehydrogenation, hydration, rehydrogenation, and sulfhydration. The results of this study showed that damulin B had an upward trend compared with the model group, which was consistent with the description in the literature. In the results of this article, compared with the blank group, the expression of PPARα in the AL, BH, BL, LH, LL, and LIH groups increased. This shows that Gynostemma pentaphyllum saponins may cause changes in the expression levels of important proteins for fatty acid oxidation in HepG2 cells and inhibit excessive liver fat accumulation. PPARγ is a lipid-regulating factor that participates in lipid synthesis, lipid storage, and lipogenesis. In this experiment, the expression of PPARγ was basically the same before and after OA treatment, but the expression of PPARγ decreased significantly after treatment with the four compounds. This is consistent with the literature survey. PPARγ is not only involved in lipid regulation of adipocyte differentiation, but also regulates the levels of liver fatty acid transporters and apolipoproteins, and participates in lipid-lowering effects.

[0089] In summary, this application used the oleic acid-induced HepG2 cell model to study the lipid-lowering activity of four compounds, damulin A, damulin B, gypenoside L, and gypenoside LI, and found that the pharmacodynamic characteristics of the four compounds have different focuses. Gypenoside L has a significant effect on lowering cholesterol and triglycerides, followed by damulin A and damulin B; the cholesterol-lowering and triglyceride-lowering effects of gypenoside LI are slightly weaker. From the perspective of targets, damulin B and gypenoside L have the same target. Damulin A and damulin B have quite different targets. The differences in the lipid-lowering activity and targets of the four compounds may be related to the structure of the compounds.

[0090] In addition, after combined use with simtavatin, its lipid-lowering performance was significantly different from that of the single-drug group, and both showed a combined synergistic effect. This further shows that the above substances have a more prominent lipid-lowering effect when used in combination with simtavatin.

[0091] The above embodiments of the present invention are merely examples for clearly illustrating the technical solution of the present invention, and are not intended to limit the specific implementation methods of the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the claims of the present invention shall be included in the protection scope of the claims of the present invention.

Claims

1. The use of dammarane saponin compounds in the preparation of lipid-lowering products, characterized in that: The dammarane saponin compound is one or more of gypenoside L, gypenoside LI, damulin A and damulin B.

2. The use according to claim 1, characterized in that: The dammarane-type saponin compound reduces TC, TG and fat accumulation.

3. A composition, characterized in that The invention is composed of a dammarane saponin compound and a statin; the dammarane saponin compound is one or more of gypenoside L, gypenoside LI, damulin A and damulin B.

4. The composition according to claim 3, characterized in that The statin is simvastatin.

5. Use of the composition according to claim 3 or 4 in the preparation of lipid-lowering products.

6. Use of a dammarane saponin compound in the preparation of a product for enhancing the lipid-lowering effect of statins, characterized in that: The statin is simvastatin; the dammarane saponin compound is one or more of gypenoside L, gypenoside LI, damulin A and damulin B.

7. The use according to claim 6, characterized in that: The dosage form of the product is powder, capsule, tablet, injection or ointment.

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