Application of green lemon pulp alcohol-water extract in preparation of medicine for improving dyslipidemia

Abstract

The invention relates to the technical field of medicines, and particularly provides application of a green lemon pulp alcohol-water extract in preparation of a medicine for improving dyslipidemia. The invention provides a new natural product application scheme for preparing the medicine for treating the dyslipidemia, and has a good medicinal value.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and more specifically, to the use of a lemon pulp alcohol water extract in the preparation of a drug for improving dyslipidemia. Background Art

[0002] Dyslipidemia refers to abnormal levels of lipids such as cholesterol and triglycerides in the blood, and is an important risk factor for cardiovascular diseases such as atherosclerosis and coronary heart disease. At present, the drugs used in the clinical treatment of dyslipidemia are mainly chemical synthetic drugs such as statins. Although they can effectively lower cholesterol levels, long-term use may be accompanied by side effects such as liver damage and muscle pain, and some patients have drug intolerance. At the same time, natural products are gradually gaining attention in the field of blood lipid regulation due to their high safety and few side effects. However, existing research has mostly focused on the extraction of single components, and the comprehensive utilization of active ingredients in plant waste is insufficient.

[0003] Lemons are a common fruit, but the pulp (including the peel, pomace, and seeds) after juicing is often treated as waste, resulting in a waste of resources. Prior research on lemon pulp is limited, and there have been no reports of its extracts being systematically used in medications to improve dyslipidemia. Summary of the Invention

[0004] The present invention provides an application of a lemon fruit cake alcohol water extract in preparing a drug for improving dyslipidemia, which provides a new natural product application scheme for the preparation of drugs for dyslipidemia and has good medicinal value.

[0005] In order to achieve these purposes and other advantages according to the present invention, a use of an alcoholic water extract of lemon pulp in preparing a drug for improving dyslipidemia is provided.

[0006] Preferably, dyslipidemia is manifested as abnormal plasma lipid parameters caused by obesity.

[0007] Preferably, the drug downregulates the levels of inflammatory factors in subjects with dyslipidemia caused by obesity.

[0008] Preferably, the drug downregulates the levels of biomarkers of myocyte damage and metabolic abnormalities in subjects with dyslipidemia caused by obesity.

[0009] Preferably, the kaffir lime dregs alcohol-water extract is prepared by the following steps: adding the dregs after squeezing the juice from fresh kaffir lime fruits to a 95% ethanol aqueous solution at a mass volume ratio of 1 kg / 4-5 L, heating to boiling, reflux extraction for 3-4 hours, filtering to remove the pomace, and volatilizing the extract until there is no alcohol taste, to obtain an extract concentrate, which is the kaffir lime dregs alcohol-water extract.

[0010] Preferably, the medicine is a pharmaceutical composition, which comprises the alcoholic water extract of kaffir lime fruit meal and a pharmaceutically acceptable carrier, and the alcoholic water extract of kaffir lime fruit meal serves as the sole active ingredient of the pharmaceutical composition.

[0011] Preferably, the pharmaceutically acceptable carrier comprises one or more of a filler, a binder, a disintegrant, a lubricant and a flavoring agent.

[0012] Preferably, the drug is prepared into a pharmaceutically acceptable dosage form.

[0013] Preferably, the dosage forms include pills, tablets, powders, capsules, granules, powders, pellets, drops, sprays, injections, suspensions, ointments, gels, and suppositories.

[0014] The present invention has at least the following beneficial effects: The present invention utilizes the active ingredients in the lemon pulp to develop a safe and effective blood lipid regulating drug, which can improve blood lipid disorders, solve the problems of many side effects of existing blood lipid regulating drugs and insufficient utilization of natural products, and provide a safe and effective drug option for clinical practice.

[0015] Other advantages, objectives and features of the present invention will be reflected in part from the following description and will be understood by those skilled in the art through study and practice of the present invention. DETAILED DESCRIPTION

[0016] The present invention is further described in detail below with reference to the accompanying examples so that those skilled in the art can implement the invention with reference to the description.

[0017] It should be understood that terms such as “having”, “including” and “comprising” used herein do not preclude the existence or addition of one or more other elements or combinations thereof.

[0018] It should be noted that the experimental methods described in the following embodiments are conventional methods unless otherwise specified, and the reagents and materials can be obtained from commercial channels unless otherwise specified.

[0019] The application of the alcohol-water extract of lemon pulp in the preparation of drugs for improving dyslipidemia. Dyslipidemia refers to abnormal levels of lipids (such as cholesterol and triglycerides) in the blood, which is an important risk factor for cardiovascular diseases such as atherosclerosis and coronary heart disease. The alcohol-water extract of lemon pulp is used to improve dyslipidemia, which is manifested by improving the levels of indicators such as total cholesterol in the blood, especially the abnormal levels of indicators such as plasma total cholesterol caused by obesity. It can also significantly reduce the levels of abnormally elevated inflammatory factors in subjects with lipid metabolism disorders caused by obesity, and can also significantly reduce the levels of biomarkers of muscle cell damage and metabolic abnormalities in subjects with dyslipidemia caused by obesity. The drug has a targeted intervention effect, effectively alleviating the symptoms of inflammation and muscle soreness caused by hyperlipidemia caused by obesity, and provides a new research direction for natural products.

[0020] Lemon, scientific name Citrus aurantifolia (Christm.) Swingle. Lime pulp refers to the residual pulp (i.e., waste product, including peel, pomace, and seeds) left after fresh fruit is washed and directly squeezed for juice. The aqueous extract refers to the mixture of active ingredients obtained by extraction with ethanol and water as the solvent. Preparation of the aqueous extract involves raw material pretreatment (collection and washing, drying, and pulverization).

[0021] The drug for improving dyslipidemia is a pharmaceutical composition, which includes an alcohol-water extract of kaffir lime fruit meal and a pharmaceutically acceptable carrier. The alcohol-water extract of kaffir lime fruit meal is the only active ingredient of the pharmaceutical composition. The pharmaceutically acceptable carrier is a pharmaceutical excipient that does not affect the efficacy of the active ingredient and meets pharmaceutical requirements. The active ingredient and the pharmaceutical excipient are mixed to form a pharmaceutical preparation.

[0022] The pharmaceutically acceptable carrier includes one or more of a filler (lactose, microcrystalline cellulose, corn starch, mannitol, pregelatinized starch, etc.), a binder (hydroxypropyl methylcellulose, polyvinylpyrrolidone, starch slurry, gelatin, polyvinyl alcohol, etc.), a disintegrant (cross-linked polyvinylpolypyrrolidone, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, dry starch, cross-linked sodium carboxymethyl cellulose, etc.), a lubricant (magnesium stearate, micropowdered silica gel, talc, hydrogenated vegetable oil, polyethylene glycol, etc.), and a flavoring agent (sweetener, aromatic, acidulant, masking agent, etc.).

[0023] The drug is prepared into pharmaceutically acceptable dosage forms, including pills, tablets, powders, capsules, granules, powders, dripping pills, drops, sprays, injections, suspensions, ointments, gels, and suppositories. For example, when preparing pills, a lime fruit dregs alcohol water extract extract is taken, dried and crushed into a fine powder, honey is heated and refined into medium honey, mixed with the extract powder in a pan pill pot, and molded with water or dilute ethanol to form a pill core, and the powder and refined honey are gradually added. The pill core is rolled to a suitable size, dried after molding, and can be coated with cinnabar or film as needed to obtain a smooth-looking pill. When preparing tablets, the lime extract dry paste powder is evenly mixed with a filler and a disintegrant, a binder is added to make a soft material, sieved and granulated, and dried. A lubricant is added after the dried granules are granulated, and the tablets are pressed into tablets using a tablet press after mixing. Sugar coating can be performed as needed, and an isolation layer, a powder coating layer, a sugar coating layer, etc. are wrapped in sequence to form a complete tablet. When preparing the injection, the lemon extract is dissolved in water for injection, the pH is adjusted and then filtered and purified, an isotonicity regulator and an antioxidant are added to the filtrate, the mixture is stirred evenly and then filtered through a fused glass funnel, the liquid medicine is filled into an ampoule and sealed, sterilized, and then leak tested with a methylene blue solution, unqualified products are removed, and a sterile injection is obtained.

[0024] 1 Study on the cholesterol-lowering effect of lemon pulp alcohol water extract 1.1 Preparation of lemon fruit pulp alcohol water extract: Fresh lemon fruits were washed and squeezed directly, and the residual fruit residue (peel, pomace, and seeds) after juicing was heated to boiling with a 95% ethanol-water solution at a mass volume ratio of 1 kg / 4 L. The mixture was refluxed for 3 hours, filtered through 6 layers of gauze to remove the pomace, and the extract was evaporated to dryness until there was no alcohol taste to obtain the extract concentrate.

[0025] 1.2 Materials and Methods 1.2.1 Experimental animals: Female C57BL / 6J mice 1.2.2 Experimental grouping, modeling and drug administration: The mice were divided into the following groups and fed continuously for 16 weeks.

[0026] Normal control group: mice were fed with ordinary feed (D12492B feed), and grew normally without obesity; High-fat modeling group: mice fed with high-fat diet (D12492) became obese; Positive drug group: fed with high-fat diet (D12492), and gavage with atorvastatin calcium tablets at 10 mg / kg body weight per day; Group 1: fed a high-fat diet (D12492) and given 5 mg / kg body weight of lemon extract by gavage daily. Group 2, the alcohol-water extract of lemon pulp, was fed a high-fat diet (D12492) and given lemon extract by gavage at 10 mg / kg body weight daily; Lime pulp alcohol-water extract 3: The mice were fed a high-fat diet (D12492) and given lime extract by gavage at 20 mg / kg body weight every day.

[0027] 1.3 Plasma total cholesterol test: At the end of the experiment, the mice were fasted for 12 hours, blood was collected from the orbit and quickly centrifuged to obtain serum. The total cholesterol (TC) detection kit, high-density lipoprotein cholesterol (HDL-C) detection kit, low-density lipoprotein cholesterol (LDL-C) detection kit, and triglyceride (TG) detection kit were used to measure the levels of TC, HDL-C, LDL-C, and TG in the samples, respectively. The results are shown in Table 1.

[0028] Table 1 As shown in Table 1, the average plasma total cholesterol level in the obese model group was 4.61 mmol / L, significantly higher than the 3.51 mmol / L level in the normal control group, indicating that the modeled mice had clear dyslipidemia, which was directly related to their obesity. The kaffir lime fruit meal alcohol-water extract lowered cholesterol in a dose-dependent manner. All dose groups (2.5-20 mg / kg) reduced the plasma total cholesterol level in obese mice. The average level of the kaffir lime fruit meal alcohol-water extract at 20 mg / kg was 4.09 mmol / L, which was more effective than 10 mg / kg of atorvastatin calcium tablets. The kaffir lime fruit meal alcohol-water extract significantly lowered total cholesterol in obese mice induced by a high-fat diet, particularly targeting elevated plasma cholesterol caused by obesity. Its mechanism of action may involve inhibiting hepatic HMG-CoA reductase, reducing cholesterol synthesis, promoting its conversion to bile acid excretion, upregulating LDL receptors to accelerate clearance, and inhibiting intestinal cholesterol absorption, thereby reducing plasma total cholesterol through multiple pathways and thus systemically regulating lipid metabolism.

[0029] As shown in Table 1, the mean TC level in the obese mice was 4.61 mmol / L, significantly higher than the 3.51 mmol / L level in the control group, consistent with high-fat diet-induced cholesterol metabolism disorders. This confirms that the mice exhibited clear hypercholesterolemia, directly related to their obesity. The aqueous extract of lemon pulp reduced TC in a dose-dependent manner, with all doses (5-20 mg / kg) significantly improving hypercholesterolemia in obese mice. The mean TC level in the 20 mg / kg group was 4.09 mmol / L, significantly superior to the 10 mg / kg group (4.32 mmol / L), suggesting a significant inhibitory effect on obesity-induced TC elevation. This mechanism may involve multiple pathways: inhibiting hepatic HMG-CoA reductase activity, reducing endogenous cholesterol synthesis; promoting the conversion of cholesterol to bile acids for intestinal excretion; and upregulating hepatic membrane LDL receptor expression, accelerating plasma LDL-C clearance. Furthermore, the polyphenolic components in the extract may inhibit intestinal NPC1L1-mediated cholesterol absorption, thereby systemically reducing TC levels.

[0030] As shown in Table 1, the mean HDL-C level in the obese mice was 2.85 mmol / L, significantly lower than the 3.20 mmol / L in the control group. This suggests that a high-fat diet disrupts the balance between HDL-C synthesis and metabolism, leading to a decrease in "good cholesterol" levels. This is consistent with the typical characteristics of high-fat diet-induced low HDL-C and is directly related to obesity. Lime pulp alcohol extract significantly reversed the HDL-C decrease, with doses of 5-20 mg / kg increasing HDL-C in a concentration-dependent manner. The 20 mg / kg group achieved HDL-C levels of 4.50 mmol / L, significantly superior to the 10 mg / kg group (4.03 mmol / L), suggesting a significant effect on obesity-induced HDL-C reduction. The mechanism may involve multiple pathways: enhancing HDL-C synthesis or inhibiting HDL-C oxidative modification or renal clearance, thereby increasing plasma HDL-C levels and function and enhancing reverse cholesterol transport.

[0031] As shown in Table 1, the mean LDL-C level in the obese mice was 0.74 mmol / L, significantly higher than the 0.53 mmol / L level in the normal control group. This indicates that a high-fat diet induces LDL-C metabolic disturbances, leading to the accumulation of large amounts of "bad cholesterol" in the plasma, a high-risk signal for atherosclerosis and directly related to obesity. The water-alcohol extract of lemon pulp reduced LDL-C in a dose-dependent manner, with all doses (5-20 mg / kg) significantly lowering LDL-C in obese mice. Among them, the LDL-C in the 20 mg / kg group dropped to 0.41 mmol / L, which was better than the 10 mg / kg positive drug group (0.45 mmol / L) and lower than the normal control group, suggesting that it has a strong regulatory effect on the abnormal increase of LDL-C caused by obesity. Its mechanism may involve multi-pathway regulation: upregulating liver LDLR gene expression, accelerating LDL-C receptor-mediated endocytosis and degradation, and on the other hand inhibiting liver microsomal triglyceride transfer protein (MTP) activity, reducing VLDL synthesis, and indirectly reducing LDL-C precursors, thereby blocking the generation and accumulation of LDL-C.

[0032] As shown in Table 1, the mean plasma TG level in the obesity model group was 1.36 mmol / L, significantly higher than the 1.08 mmol / L in the normal control group. This reflects the hyperdecomposition of adipose tissue and increased TG synthesis in obesity, leading to lipid metabolism disorders. The improvement of TG by the alcohol-water extract of lemon pulp was dose-dependent, with TG in the 20 mg / kg group decreasing to 1.24 mmol / L, demonstrating its potential to regulate obesity-related hypertriglyceridemia and comparable to that of the positive drug group (1.25 mmol / L). This mechanism may involve multiple pathways of regulation: it may activate lipoprotein lipase (LPL) activity in peripheral tissues such as adipose tissue and skeletal muscle, promoting the breakdown of TG in chylomicrons and VLDL into free fatty acids, accelerating peripheral utilization, and thus reducing plasma TG levels.

[0033] 1.4 Inflammatory factor detection: At the end of the experiment, the mice were fasted for 12 hours, blood was collected from the orbits and quickly centrifuged, and serum was collected. The IL-6, IL-1β, and TNF-α levels were detected according to the instructions of the ELISA kit. The results are shown in Table 2.

[0034] Table 2 Obesity can cause chronic inflammation, which is essentially a systemic, chronic low-grade inflammation induced by various inflammatory factors. The hallmark proinflammatory cytokines of chronic inflammation include abnormal production of IL-6, IL-1β, and TNF-α. As shown in Table 2, levels of proinflammatory cytokines were significantly elevated in the obese mice. Administration of the kaffir lime fruit meal alcohol-water extract at various doses (10-20 mg / kg) significantly suppressed serum levels of IL-6, IL-1β, and TNF-α, suggesting that this may modulate the inflammatory response by suppressing proinflammatory cytokine levels. Serum levels of these proinflammatory cytokines were significantly elevated in the high-fat model mice compared with the normal control group. However, treatment with the kaffir lime fruit meal alcohol-water extract at doses of 10-20 mg / kg significantly suppressed serum IL-6, IL-1β, and TNF-α. This suggests that the extract may modulate the inflammatory response by regulating the proinflammatory cytokine network, inhibiting proinflammatory cytokine levels, and thereby intervening in the inflammatory cascade.

[0035] 1.5 Myalgia Relief Testing: At the end of the experiment, mice were fasted for 12 hours, blood was collected from the orbits and quickly centrifuged, and serum was collected. The muscle damage marker creatine kinase (CK) and muscle metabolic disorder marker lactate dehydrogenase (LDH) were detected according to the kit instructions. The results are shown in Table 3.

[0036] Add enzyme reagent to start the reaction, immediately monitor the absorbance change at 340nm wavelength using a spectrophotometer, calculate the absorbance decrease rate (ΔA / min), and substitute it into the calibration curve provided by the kit. Add NAD⁺ to start the reaction, immediately monitor the absorbance change at 340nm wavelength, and calculate the absorbance rising rate (ΔA / min). ε is the molar absorption coefficient of NADH, which is 6.22×10 3 L·mol -1 ・cm -1 .

[0037] Table 3 Hyperlipidemia caused by obesity can cause muscle soreness through lipotoxicity, oxidative stress and inflammation. As shown in Table 2, the levels of creatine kinase CK and lactate dehydrogenase LDH in the obesity model group increased significantly, indicating that the integrity of the muscle cell membrane was damaged and energy metabolism was disordered. The CK and LDH levels of the lemon fruit pulp alcohol water extract in each dose group (10-20 mg / kg) were significantly reduced in mice fed a high-fat diet, showing a dose-dependent effect, indicating that the muscle soreness caused by hyperlipidemia may be alleviated. It is worth noting that the CK and LDH levels of the positive drug 10 mg / kg were the highest. This is because atorvastatin blocks cholesterol synthesis by inhibiting HMG-CoA reductase, but its accompanying side effect of muscle cell necrosis leads to abnormally elevated CK and LDH levels. The CK and LDH levels of the lemon fruit pulp alcohol water extract in each dose group (10-20 mg / kg) Unlike atorvastatin, it does not interfere with the cholesterol metabolism pathway. The natural antioxidant ingredients it contains can directly protect muscle cells, so there is no myalgia side effect. This is probably because the alcohol-water extract of lemon pulp may activate the Nrf2 pathway, enhance superoxide dismutase (SOD) activity, scavenge ROS (reduce oxidative damage), and thus stabilize the muscle cell membrane structure and antagonize lipotoxicity and inflammatory damage. Compared with statins, the extract provides a safer alternative, especially for patients with obesity and muscle pain.

[0038] The number of devices and processing scales described herein are intended to simplify the description of the present invention. Applications, modifications, and variations of the present invention will be readily apparent to those skilled in the art.

[0039] Although the embodiments of the present invention have been disclosed above, they are not limited to the applications listed in the description and implementation methods. They can be fully applied to various fields suitable for the present invention. For those familiar with the art, additional modifications can be easily implemented. Therefore, without departing from the general concept defined by the claims and the scope of equivalents, the present invention is not limited to the specific details and examples shown and described herein.

Claims

1. Application of lemon fruit pulp alcohol water extract in the preparation of drugs for improving dyslipidemia.

2. The use according to claim 1, characterized in that Dyslipidemia is manifested as abnormal plasma lipid parameters caused by obesity.

3. The use according to claim 1, characterized in that The drug downregulates the levels of inflammatory factors in subjects with dyslipidemia caused by obesity.

4. The use according to claim 2, characterized in that The drug downregulates the levels of biomarkers of myocyte damage and metabolic abnormalities in subjects with dyslipidemia caused by obesity.

5. The use according to claim 1, wherein The kaffir lime dregs alcohol-water extract is prepared by the following steps: adding the dregs after squeezing the juice from fresh kaffir lime fruits to an ethanol aqueous solution with a volume fraction of 95% at a mass volume ratio of 1 kg / 4-5 L, heating to boiling, reflux extraction for 3-4 hours, filtering to remove the pomace, and volatilizing the extract until there is no alcohol taste to obtain an extract concentrate, which is the kaffir lime dregs alcohol-water extract.

6. The use according to claim 1, wherein The medicine is a pharmaceutical composition, which comprises the kaffir lime fruit meal alcohol water extract and a pharmaceutically acceptable carrier, and the kaffir lime fruit meal alcohol water extract serves as the only active ingredient of the pharmaceutical composition.

7. The use according to claim 6, characterized in that The pharmaceutically acceptable carrier includes one or more of a filler, a binder, a disintegrant, a lubricant and a flavoring agent.

8. The use according to claim 1, wherein The drug is prepared into a pharmaceutically acceptable dosage form.

9. The use according to claim 8, characterized in that The dosage forms include pills, tablets, powders, capsules, granules, powders, pellets, drops, sprays, injections, suspensions, ointments, gels, and suppositories.

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