A gastrodin-EGCG combination and its application in treating and improving lipid metabolism disorder

Abstract

The application discloses a gastrodin-EGCG composition and application thereof in treating and improving lipid metabolism disorder, wherein the composition takes gastrodin and epigallocatechin gallate as main active ingredients, and the gastrodin and the epigallocatechin gallate are combined in a certain proportion. In a HepG2 cell lipid deposition model induced by mixed fatty acids, the gastrodin-EGCG composition can reduce intracellular lipid droplet deposition and triglyceride content, and the improvement effect thereof is better than that of the gastrodin or the EGCG alone, and the composition has a significant synergistic effect. The composition can be used for preparing a medicine or a health food for improving liver cell lipid deposition, regulating lipid metabolism disorder or assisting in improving abnormal states related to lipid accumulation.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to a composition with gastrodin and epigallocatechin gallate as the main active ingredients, and the application of the composition in treating and improving lipid metabolism. Background Technology

[0002] Lipid metabolism disorders refer to abnormalities in the body's lipid absorption, synthesis, breakdown, transport, and utilization processes. They are often manifested as abnormal levels of triglycerides, cholesterol, and low-density lipoprotein cholesterol, as well as excessive lipid deposition in tissues such as the liver. Long-term lipid metabolism disorders can further contribute to the development of obesity, fatty liver, hyperlipidemia, insulin resistance, and cardiovascular metabolic diseases. Therefore, finding safe, effective, and long-term interventional lipid metabolism regulatory components is of great significance.

[0003] While single-dose combinations can improve the efficacy of lipid metabolism disorders with increasing dosage, higher doses often lead to more side effects. Therefore, developing drugs with synergistic effects for treating and improving lipid metabolism disorders is an urgent technical challenge. Summary of the Invention

[0004] The purpose of this invention is to address the shortcomings of the prior art by providing a gastrodin-EGCG composition, the composition comprising (A) gastrodin and / or gastrodin derivatives; and (B) epigallocatechin gallate and / or epigallocatechin gallate derivatives.

[0005] Furthermore, the mass ratio of gastrodin to epigallocatechin gallate is 2-3:1.

[0006] Preferably, the mass ratio of gastrodin to epigallocatechin gallate is 2.5:1.

[0007] Furthermore, the gastrodin is derived from one or more of the following: gastrodin extract, gastrodin purified product, chemically synthesized gastrodin, or commercially available gastrodin standard.

[0008] Furthermore, the EGCG is derived from one or more of the following: tea extract, purified tea polyphenols, chemically synthesized EGCG, or commercially available EGCG standards.

[0009] Furthermore, gastrodin derivatives include salts, esters, acids, ketones, cocrystals, chelates, complexes, glycosides, hydrates, etc., formed from gastrodin.

[0010] Furthermore, EGCG includes salts, esters, acids, ketones, cocrystals, chelates, complexes, glycosides, hydrates, etc., formed from EGCG, as well as its hydrolysis products.

[0011] Furthermore, the composition may be supplemented with pharmaceutically, food-grade, or health food-grade excipients to prepare tablets, capsules, granules, powders, oral liquids, lyophilized powders, pills, suspensions, beverages, or solid beverages.

[0012] A drug for treating lipid metabolism disorders, comprising (A) gastrodin and / or gastrodin derivatives; and (B) epigallocatechin gallate and / or epigallocatechin gallate derivatives; optionally, pharmaceutically acceptable excipients. The amount of gastrodin and / or gastrodin derivatives is 25-100 mg, and the amount of epigallocatechin gallate is 10-40 mg.

[0013] Furthermore, the amount of gastrodin and / or gastrodin derivatives is 50 mg, and the amount of epigallocatechin gallate is 20 mg.

[0014] The present invention also provides the use of the gastrodin-EGCG composition in the preparation of products for treating and / or improving hepatocyte lipid deposition.

[0015] Furthermore, the hepatocyte lipid deposition is a mixed fatty acid-induced lipid deposition in HepG2 cells.

[0016] Furthermore, the treatment and / or improvement of hepatocyte lipid deposition includes reducing intracellular lipid droplet deposition, reducing intracellular triglyceride and cholesterol levels.

[0017] The present invention also provides the use of the gastrodin-EGCG composition in the preparation of products for treating and / or improving lipid metabolism disorders.

[0018] Compared with the prior art, the present invention has the following beneficial effects: The gastrodin-EGCG composition provided by the present invention has gastrodin and EGCG as the main active ingredients, with clear components and wide sources, and has a good foundation for development and application. A lipid deposition model was established by inducing HepG2 cells with mixed fatty acids. The results showed that the combined use of gastrodin and EGCG can improve lipid deposition in hepatocytes. Compared with the use of gastrodin or EGCG alone, the gastrodin-EGCG composition has a more significant improvement effect on lipid droplet deposition and increased intracellular TG content in HepG2 cells induced by mixed fatty acids, suggesting that the combination of the two has good synergistic or enhanced potential. Attached Figure Description

[0019] Figure 1 Effects of different concentrations of gastrodin on the viability of HepG2 cells Figure 2 Effects of different concentrations of EGCG on the viability of HepG2 cells Figure 3 Effects of combined gastrodin and EGCG on lipid droplet formation in HepG2 cells Figure 4 Effects of combined gastrodin and EGCG on intracellular triglyceride levels in HepG2 cells Figure 5 Effects of different concentrations of EGCG on lipid droplet formation in HepG2 cells Figure 6 Effects of different concentrations of EGCG on intracellular triglyceride levels in HepG2 cells Figure 7 Effects of different concentrations of gastrodin on lipid droplet formation in HepG2 cells Figure 8 Effects of different concentrations of gastrodin on intracellular triglyceride levels in HepG2 cells. Detailed Implementation

[0020] To facilitate understanding by those skilled in the art, the present invention will be further described below with reference to embodiments. The content mentioned in the embodiments is not intended to limit the present invention. Example 1

[0021] A gastrodin-EGCG composition comprising (A) gastrodin and (B) epigallocatechin gallate. The mass ratio of gastrodin to epigallocatechin gallate is 2.5:1. Example 2

[0022] A gastrodin-EGCG composition comprising (A) gastrodin and (B) epigallocatechin gallate. The mass ratio of gastrodin to epigallocatechin gallate is 2:1. Example 3

[0023] A gastrodin-EGCG composition comprising (A) gastrodin and (B) epigallocatechin gallate. The mass ratio of gastrodin to epigallocatechin gallate is 3:1. Example 4

[0024] A gastrodin-EGCG composition comprising (A) gastrodin and (B) epigallocatechin gallate. The gastrodin is present in a concentration of 25 mg, and the epigallocatechin gallate is present in a concentration of 10 mg. Example 5

[0025] A gastrodin-EGCG composition comprising (A) gastrodin and (B) epigallocatechin gallate. The gastrodin is 50 mg and the epigallocatechin gallate is 20 mg. Example 6

[0026] A gastrodin-EGCG composition comprising (A) gastrodin and (B) epigallocatechin gallate. The gastrodin is 100 mg and the epigallocatechin gallate is 40 mg.

[0027] The effects of gastrodin and EGCG on the viability of HepG2 cells were evaluated using the CCK-8 assay. Cells were seeded in 96-well plates, and after adhesion, different concentrations of the test agents were added. Cells were cultured for 24 h, 48 h, or 72 h. After culture, the culture medium was discarded, and basal medium containing 10% CCK-8 reagent was added to each well. Cells were incubated in the dark for 1 h, and absorbance was measured at 450 nm. Cell viability was calculated by normalization to the control group.

[0028] The results are as follows Figure 1 Treatment with gastrodin concentrations over 24-48 hours showed minimal impact on cell viability; no significant cytotoxicity was observed below 1000 μM within 24 hours. Figure 2 Low concentrations of EGCG can enhance cell viability; viability gradually decreases at concentrations ≥100 μM, and toxicity is significant after prolonged treatment at concentrations of 200-400 μM. Therefore, EGCG can be considered to have no significant toxicity below 100 μM.

[0029] HepG2 cells were divided into four groups: a blank control group, a free fatty acid model group, a free fatty acid + 100 μM gastrodin group, a free fatty acid + 25 μM EGCG group, and a free fatty acid + 100 μM gastrodin + 25 μM EGCG group. Except for the blank control group, each group was treated with 0.30 mM free fatty acid and the corresponding test substance simultaneously for 24 h.

[0030] Lipid droplet formation was observed using Oil Red O staining. After culturing in medium containing the modeling agent for 24 h, the culture medium was discarded, and the cells were washed twice with PBS buffer and fixed with 4% paraformaldehyde for 10 min at room temperature. The paraformaldehyde solution was discarded, and the cells were washed twice with PBS. Cells were stained with Oil Red O staining solution at room temperature in the dark for 20 min, the Oil Red O staining solution was discarded, and the cells were washed twice with PBS. The cells were then observed, photographed, and stored under a microscope. The results of Oil Red O staining are shown below. Figure 3 The results showed that, compared with the free fatty acid model group, the number of red lipid droplets in cells was reduced to varying degrees in both the gastrodin and EGCG single-drug groups. The combination of gastrodin and EGCG resulted in fewer red lipid droplets in cells, with significantly lower staining intensity than the free fatty acid model group, and showed a further improvement trend compared to the two single-drug groups.

[0031] The intracellular TG content in each group was measured using a TG detection kit, and the total protein content was determined using the BCA method for correction. The results are shown in Table 1.

[0032] Table 1. Effects of gastrodin combined with EGCG on intracellular TG levels induced by free fatty acids in HepG2 cells. Group Processing conditions TG (mmol / g prot) Compared to the free fatty acid group, it decreased by (%) Blank control group No free fatty acids and test substance added 0.307±0.085 — Free fatty acid model group 0.30 mM free fatty acids 2.548±0.118 — Gastrodin group Free fatty acids + 100 μM gastrodin 1.690±0.073 33.65 EGCG Group Free fatty acids + 25 μM EGCG 1.845±0.105 27.56 Gastrodin + EGCG group Free fatty acids + 100 μM gastrodin + 25 μM EGCG 1.353±0.096 46.88 The TG levels in the single-drug intervention groups were lower than those in the free fatty acid model group. The TG content in the gastrodin and EGCG combination group was 1.353 ± 0.096 mmol / g prot, a decrease of approximately 46.88% compared to the free fatty acid model group. Compared to the EGCG monotherapy group, the TG content in the combination group was further reduced by approximately 26.66%, and compared to the gastrodin monotherapy group, it was further reduced by approximately 19.94%. These results indicate that the combined treatment of 100 μM gastrodin and 25 μM EGCG has a stronger inhibitory effect on free fatty acid-induced TG accumulation.

[0033] Under the concentration conditions of this embodiment, the molar ratio of gastrodin to EGCG is 4:1; the equivalent mass ratio based on free compounds is approximately 2.5:1. This combination can reduce intracellular lipid droplet and TG accumulation while maintaining cell viability.

[0034] HepG2 cells were induced and treated with 50 μM EGCG according to the method described in the above embodiments. After the intervention, Oil Red O staining was used to observe intracellular lipid droplet formation and lipid deposition, and intracellular triglyceride (TG) levels were detected. The results are as follows: Figure 5 and Figure 6 This indicates that, compared with the FFA model group, intervention with 50 μM EGCG alone can reduce intracellular lipid droplet formation and decrease intracellular lipid accumulation levels in HepG2 cells to some extent.

[0035] The effect of 50 μM EGCG alone on improving cellular lipid deposition was weaker than that of the combined intervention group of 100 μM gastrodin and 25 μM MEGCG. In this comparative example, the concentration of EGCG used was twice that of the combined intervention group, but its overall effect of reducing lipid droplet formation and improving intracellular lipid accumulation was still not as good as that of the combined intervention group, thus eliminating the dose effect.

[0036] HepG2 cells were induced and treated with 200 μM gastrodin according to the method described in the above embodiments. After the intervention, Oil Red O staining was used to observe intracellular lipid droplet formation and lipid deposition, and intracellular TG levels were detected. The results are as follows: Figure 7 and Figure 8 This indicates that, compared with the FFA model group, intervention with 200 μM gastrodin alone can reduce intracellular lipid deposition in HepG2 cells and has a certain ameliorative effect on FFA-induced lipid metabolism abnormalities.

[0037] The effect of 200 μM gastrodin alone on improving lipid droplet formation and intracellular lipid accumulation was weaker than that of the combined intervention group of 100 μM gastrodin and 25 μM EGCG. In this comparative example, the concentration of gastrodin used was twice that in the combined intervention group, but its overall effect on reducing lipid deposition was still inferior to that of the combined intervention group. This indicates that the lipid deposition inhibition effect produced by the combined use of gastrodin and EGCG is not a simple additive effect of the two components individually, nor is it caused by an increase in the dosage of either component.

[0038] Table 2 Comparative TG Levels Group Processing conditions TG (mmol / g prot) Compared to the free fatty acid group, it decreased by (%) Comparative Example 1 Free fatty acids + 50 μM EGCG 1.625±0.010 34.74 Comparative Example 2 Free fatty acids + 200 μM gastrodin 1.424±0.034 34.98 It is hypothesized that gastrodin and EGCG may have a synergistic effect in improving lipid metabolism. EGCG mainly activates the AMPK pathway, inhibits SREBP-1c-mediated lipid synthesis, and promotes PPARα / CPT1A-mediated fatty acid oxidation; gastrodin, on the other hand, mainly reduces oxidative stress, lipid peroxidation, and mitochondrial damage by activating the Nrf2 antioxidant pathway. The protective effect of gastrodin on cells and mitochondria can enhance the regulatory effect of EGCG on lipid synthesis and breakdown. Together, they reduce lipid production, promote lipid consumption, and inhibit lipid damage, thereby synergistically alleviating cellular lipid accumulation and improving lipid metabolism disorders.

[0039] The specific embodiments of the present invention have been described above. It should be noted that the present invention is not limited to the specific embodiments described above, and those skilled in the art can make various modifications or variations within the scope of the claims, which do not affect the essence of the present invention.

Claims

1. A gastrodin-EGCG composition, characterized in that, The composition includes gastrodin and epigallocatechin gallate.

2. The gastrodin-EGCG composition according to claim 1, characterized in that, The mass ratio of gastrodin to epigallocatechin gallate is 2.0~3.0:

1.

3. The gastrodin-EGCG composition according to any one of claims 1–2, characterized in that, The composition also includes pharmaceutically, food-grade, or health food-grade excipients.

4. A medicament for treating lipid metabolism disorders, comprising (A) gastrodin and / or gastrodin derivatives; and (B) epigallocatechin gallate and / or epigallocatechin gallate derivatives; optionally, pharmaceutically acceptable excipients; wherein the gastrodin and / or gastrodin derivatives are in the amount of 25-100 mg, and the epigallocatechin gallate is in the amount of 10-40 mg.

5. The drug according to claim 4, characterized in that, Gastrodin and / or gastrodin derivatives are 50 mg, and epigallocatechin gallate is 20 mg.

6. The use of the gastrodin-EGCG composition according to any one of claims 1–5 in the preparation of products for treating and / or improving lipid metabolism.

7. The application according to claim 6, characterized in that, The improvement of hepatocyte lipid deposition includes reducing intracellular lipid droplet deposition, reducing intracellular triglyceride and cholesterol levels, or one or more of these.

8. The use of the gastrodin-EGCG composition according to any one of claims 1–7 in the preparation of products for treating and / or improving lipid metabolism disorders, wherein the products are pharmaceuticals, health foods or dietary supplements.

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