Composition for improving type 2 diabetes mellitus complicated with depression as well as preparation method and application of composition

Abstract

The invention provides a composition for improving type 2 diabetes mellitus complicated with depression as well as a preparation method and application of the composition, and relates to the technical field of medicines. The composition comprises the following components: a coptis chinensis extract, a hawthorn leaf extract, L-carnitine, acetyl L-carnitine and inulin. The coptis chinensis is subjected to first-time enzymolysis by using a compound enzyme 1, and then is subjected to second-time enzymolysis by using a compound enzyme 2; and after enzymolysis, performing ultrasonic-assisted extraction by using an ethanol solution to obtain the coptis chinensis extract. Carrying out first enzymolysis on the hawthorn by using a compound enzyme 3, and then carrying out second enzymolysis by using a compound enzyme 4; and after enzymolysis, carrying out microwave-assisted extraction by using an ethanol solution to obtain the hawthorn leaf extract. The composition has a synergistic effect of all the components in the aspects of improving the glucose metabolism function and depression behavioristics, can effectively reduce fasting blood glucose, insulin resistance indexes and glycosylated hemoglobin, increase the sweet water preference rate and shorten the immobility time of forced swimming and tail suspension experiments, and has a better effect of treating diabetes mellitus complicated with depression.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a composition for improving type 2 diabetes combined with depression, and a preparation method and application thereof. Background Art

[0002] Type 2 diabetes combined with depression is a comorbid syndrome characterized by abnormal glucose metabolism and persistent affective disorders. Clinically, it is mainly accompanied by the characteristics of mental disorders such as depression, and its pathological mechanism involves insulin resistance, etc. A large amount of epidemiological data confirms that there is a close and complex bidirectional relationship between the two diseases. On the one hand, the risk of depression in diabetic patients is significantly higher than that in the general population, with a prevalence of about twice that of the latter. On the other hand, depression itself is also an independent risk factor for type 2 diabetes, and the depressive state may promote the occurrence and development of type 2 diabetes through various pathways. Because the course of the disease is long and prone to recurrence, it seriously affects the quality of life and compliance of patients. Therefore, there is an urgent clinical need to prevent and treat type 2 diabetes combined with depression.

[0003] The pathogenesis of type 2 diabetes mellitus with depression remains unclear. Current clinical treatment is primarily based on multidimensional symptomatic intervention, but a unified approach targeting the core pathological mechanisms is still lacking. For depressive symptoms, serotonin reuptake inhibitors (SRIs) and serotonin and norepinephrine reuptake inhibitors (SNRIs) are often used as first-line medications for patients with diabetes mellitus and depression and anxiety. However, not all treatments sufficient to alleviate depressive symptoms can control blood glucose levels. For example, sertraline has been shown to alleviate depression without affecting blood glucose. Serotonin reuptake inhibitors, such as fluoxetine, can improve depressive-like behaviors in diabetic patients without affecting glucose homeostasis, but they are risk factors for certain diabetic complications. Therefore, the development of novel anti-depressant drugs for type 2 diabetes mellitus with depression is of great significance.

[0004] Acetyl-L-carnitine (ALC) exhibits rapid, robust, and long-lasting antidepressant effects in spontaneously depressed Flinders sensitive line (FSL) rats, genetic models of depression, and mice exposed to chronic, unpredictable stress. FSL rats exhibit sleep, immune, and neurochemical changes, as well as behavioral changes similar to those seen in patients with depression. Prior art only discloses the effects of ALC alone, without considering whether ALC can interact with other components to achieve synergistic effects.

[0005] Chinese invention patent CN101378734A discloses a micro-tablet-based pharmaceutical preparation that includes hundreds of ingredients, including berberine, hawthorn, L-carnitine, and acetyl-L-carnitine. The prepared pharmaceutical preparation has a certain effect on treating diabetes symptoms. However, the invention contains many ingredients, which are simply the accumulation of the effects of multiple ingredients, without considering the application of the overall composition in treating type 2 diabetes combined with depression. Chinese invention patent CN120022335A discloses an oral liquid for treating metabolic syndrome and its preparation method, comprising the following raw materials by weight: 12-18 parts of selenium-enriched yeast-modified astragalus, 10-14 parts of ultrafinely ground fermented coptis root, 14-18 parts of gynostemma pentaphyllum, 12-16 parts of hawthorn, 8-12 parts of oriental rhizome, 10-13 parts of turmeric, 6-8 parts of cinnamon extract, 3-6 parts of coenzyme Q10 microspheres, 5-10 parts of eucommia leaf extract, and 3-7 parts of nuciferine extract. The oral solution of this invention can effectively improve insulin resistance and reduce oxidative damage, playing a key role in controlling blood sugar and blood lipids and preventing complications in patients with metabolic syndrome. However, this technology does not disclose the effect of the components synergistically improving type 2 diabetes combined with depression.

[0006] In view of this, in order to address the deficiencies of the existing technology, further synergistic efficacy research is needed on the ingredients, in order to obtain a pharmaceutical composition for improving type 2 diabetes combined with depression in which the components cooperate with each other and enhance synergy. Summary of the Invention

[0007] One of the purposes of the present invention is to provide a pharmaceutical composition for improving type 2 diabetes combined with depression, comprising components of coptis chinensis extract, hawthorn leaf extract, L-carnitine, acetyl-L-carnitine and inulin; the composition has a synergistic effect of the components in improving glucose metabolism function and depressive behavior, can effectively reduce fasting blood sugar, insulin resistance index and glycated hemoglobin, increase sugar water preference rate, reduce immobility time in forced swimming and tail suspension tests, and has a better therapeutic effect.

[0008] A second object of the present invention is to provide a method for preparing the above-mentioned composition. In the above-mentioned composition, the Rhizoma Coptidis extract and the Crataegus Leaf extract are obtained by specific enzymatic hydrolysis and auxiliary extraction, which significantly improves the effect of the composition on improving type 2 diabetes combined with depression.

[0009] The third purpose of the present invention is to use the composition in the preparation of a drug for improving type 2 diabetes combined with depression.

[0010] In order to achieve the above object, the present invention adopts the following technical solutions: In a first aspect, the present invention provides a composition for improving type 2 diabetes combined with depression, comprising components: coptis root extract, hawthorn leaf extract, L-carnitine, acetyl-L-carnitine and inulin.

[0011] Preferably, the composition comprises the following components in parts by weight: 2-6 parts of coptis root extract, 1-5 parts of hawthorn leaf extract, 3-8 parts of L-carnitine, 2-10 parts of acetyl-L-carnitine and 10-20 parts of inulin.

[0012] Further preferably, the composition comprises the following components, in parts by weight: 3 parts of coptis root extract, 2 parts of hawthorn leaf extract, 5 parts of L-carnitine, 5 parts of acetyl-L-carnitine and 15 parts of inulin.

[0013] Preferably, the preparation method of the Coptidis Rhizome extract comprises the steps of: performing a first enzymatic hydrolysis of the Coptidis Rhizome using compound enzyme 1, and then performing a second enzymatic hydrolysis using compound enzyme 2; performing ultrasonic-assisted extraction with an ethanol solution after enzymatic hydrolysis, heating and inactivating the enzyme; and ultrafiltration and resin purification and separation to obtain the Coptidis Rhizome extract.

[0014] Further preferably, before enzymatic hydrolysis, the coptis root is crushed to a particle size of 20-60 mesh; more preferably 40 mesh.

[0015] Further preferably, before enzymatic hydrolysis, the coptis root is mixed with water at a solid-liquid ratio of 1:10-20 g / mL.

[0016] More preferably, the first enzymatic hydrolysis is carried out under the following conditions: adjusting the pH to 5-6, and performing enzymatic hydrolysis at 48-52° C. and 100-300 rpm with stirring for 2.5-5 hours.

[0017] More preferably, the first enzymatic hydrolysis is carried out under the following conditions: adjusting the pH to 5.5, and performing enzymatic hydrolysis at 50° C. and 200 rpm with stirring for 3.5 hours.

[0018] More preferably, the second enzymatic hydrolysis is carried out under the following conditions: adjusting the pH to 4.2-5, and performing enzymatic hydrolysis at 56-63° C. and 100-300 rpm for 1-2.5 hours.

[0019] More preferably, the second enzymatic hydrolysis is carried out under the following conditions: adjusting the pH to 4.5, and performing enzymatic hydrolysis at 60° C. and 200 rpm with stirring for 1.5 hours.

[0020] Further preferably, in the ultrasonic-assisted extraction, the volume concentration of ethanol is 70-80%, and the ultrasonic-assisted extraction is performed at 25-35 Hz and 300-500 W for 20-40 min.

[0021] Further preferably, in the ultrasonic-assisted extraction, the volume concentration of ethanol is 75%, and the ultrasonic-assisted extraction is performed at 30 Hz and 400 W for 30 min.

[0022] Further preferably, the molecular cut-off of the ultrafiltration is 8-12 kDa; further preferably, it is 10 kDa.

[0023] Further preferably, in the resin purification and separation, AB-8 macroporous adsorption resin is used for separation, and water and 75% ethanol are used for elution in sequence, with an elution flow rate of 1-3 BV / h, and the 75% ethanol eluate is collected.

[0024] More preferably, in the resin purification and separation, AB-8 macroporous adsorption resin is used for separation, and water and 75% ethanol are used for elution in sequence, with an elution flow rate of 2BV / h, and the 75% ethanol eluate is collected.

[0025] Most preferably, the preparation method of the Coptidis rhizome extract comprises the steps of: drying and crushing the Coptidis rhizome, passing through a 40-mesh sieve, and fully mixing the powder with water at a material-liquid ratio of 1:10 g / mL; adding complex enzyme 1 to the mixture, adjusting the pH to 5.5, and stirring at 50°C and 200 rpm for 3.5 hours; adjusting the pH to 4.5, adding complex enzyme 2, and stirring at 60°C and 200 rpm for 1.5 hours; after the enzymatic hydrolysis is completed, adding 95% (v / v) ethanol to make the final ethanol concentration of the feed liquid reach 75%, and performing ultrasonic-assisted extraction for 30 minutes at 30 kHz and 400 W (the temperature of the ice water bath is controlled at ≤50 ℃); then the temperature was raised to 95℃ and maintained for 15 minutes to completely inactivate all enzymes and terminate the reaction; after cooling to room temperature, the mixture was centrifuged at 5000 rpm / min for 15 minutes, the supernatant was collected, and ultrafiltration was performed through a 10 kDa molecular cut-off membrane; the filtrate was collected and concentrated under reduced pressure at 60℃ to a thick paste-like extract, the extract was mixed with water, the pH was adjusted to 5.5, and separated using AB-8 macroporous adsorption resin, and eluted with 5 times the column volume of deionized water and 75% ethanol at a flow rate of 2 BV / h, the 75% ethanol eluate was collected, and the filtrate was concentrated under reduced pressure at 60℃ to a thick paste-like extract, and freeze-dried to obtain the Coptidis rhizome extract.

[0026] Further preferably, the complex enzyme 1 comprises: cellulase, pectinase, tannase and papain.

[0027] More preferably, the complex enzyme 1 comprises, by weight, 0.5-1.5 parts of cellulase, 0.4-1 parts of pectinase, 0.2-0.8 parts of tannase and 0.2-0.4 parts of papain.

[0028] More preferably, the complex enzyme 1 comprises, by weight, 0.5 parts of cellulase, 0.4 parts of pectinase, 0.2 parts of tannase and 0.4 parts of papain; or the complex enzyme 1 comprises, by weight, 1.5 parts of cellulase, 1 part of pectinase, 0.8 parts of tannase and 0.4 parts of papain.

[0029] Further preferably, the added amount of the complex enzyme 1 is 3-5% of the mass of Coptis chinensis.

[0030] More preferably, the added amount of the complex enzyme 1 is 3.7% of the mass of Coptis chinensis.

[0031] Further preferably, the complex enzyme 2 comprises: acid protease and β-glucosidase.

[0032] More preferably, the complex enzyme 2 comprises, by weight, 0.2-0.4 parts of acid protease and 0.3-0.8 parts of β-glucosidase.

[0033] More preferably, the complex enzyme 2 comprises, by weight, 0.2 parts of acid protease and 0.8 parts of β-glucosidase; or the complex enzyme 2 comprises, by weight, 0.2 parts of acid protease and 0.3 parts of β-glucosidase.

[0034] Further preferably, the added amount of the complex enzyme 2 is 0.8-1.5% of the mass of Coptis chinensis.

[0035] More preferably, the added amount of the complex enzyme 2 is 1.1% of the mass of Coptis chinensis.

[0036] Preferably, the preparation method of the hawthorn leaf extract comprises the steps of: performing a first enzymatic hydrolysis on the hawthorn using compound enzyme 3, and then performing a second enzymatic hydrolysis using compound enzyme 4; after the enzymatic hydrolysis, performing microwave-assisted extraction with an ethanol solution and heating to inactivate the enzyme; and ultrafiltration and resin purification and separation to obtain the hawthorn leaf extract.

[0037] Further preferably, the hawthorn leaves are crushed before enzymatic hydrolysis, and the crushed particle size is 20-60 mesh; more preferably 50 mesh.

[0038] Further preferably, before enzymatic hydrolysis, the hawthorn leaves and water are mixed at a solid-liquid ratio of 1:10-20 g / mL; even more preferably, before enzymatic hydrolysis, the hawthorn leaves and water are mixed at a solid-liquid ratio of 1:12 g / mL; More preferably, the first enzymatic hydrolysis is carried out under the following conditions: adjusting the pH to 5-6, and performing enzymatic hydrolysis at 52-58° C. and 100-300 rpm with stirring for 2.5-5 hours.

[0039] More preferably, the first enzymatic hydrolysis is carried out under the following conditions: adjusting the pH to 5.5, and performing enzymatic hydrolysis at 55° C. and 200 rpm with stirring for 3.5 hours.

[0040] More preferably, the second enzymatic hydrolysis is carried out under the following conditions: adjusting the pH to 4.2-5, and performing enzymatic hydrolysis at 56-63° C. and 100-300 rpm for 1-2.5 hours.

[0041] More preferably, the second enzymatic hydrolysis is carried out under the following conditions: adjusting the pH to 4.5, and performing enzymatic hydrolysis at 60° C. and 200 rpm with stirring for 1.5 hours.

[0042] Further preferably, in the microwave-assisted extraction, the volume concentration of ethanol is 60-70%, and the microwave-assisted extraction is performed at 400-600W and 60-70°C for 8-20min.

[0043] Further preferably, in the microwave-assisted extraction, the volume concentration of ethanol is 65%, and the microwave-assisted extraction is performed at 500W and 65°C for 10 minutes.

[0044] Further preferably, the molecular cut-off of the ultrafiltration is 8kDa-12kDa molecular weight; further preferably, it is 10kDa.

[0045] Further preferably, in the resin purification and separation, AB-8 type macroporous adsorption resin is used for separation, and water, 30% ethanol, and 70% ethanol are used for elution in sequence, with an elution flow rate of 1-3 BV / h, and 75% ethanol eluate is collected.

[0046] More preferably, in the resin purification and separation, AB-8 macroporous adsorption resin is used for separation, and water, 30% ethanol, and 70% ethanol are used for elution in sequence, with an elution flow rate of 2BV / h, and the 70% ethanol eluate is collected.

[0047] Most preferably, the preparation method of the hawthorn leaf extract comprises the steps of: drying the hawthorn leaves and crushing them, passing them through a 50-mesh sieve, and thoroughly mixing the powder with deionized water at a material-liquid ratio of 1:12 g / mL; adding complex enzyme 3 to the mixture, adjusting the pH to 5.5, and stirring at 55°C and 200 rpm for enzymolysis for 3.5 hours; adjusting the pH to 4.5, adding complex enzyme 4, and stirring at 60°C and 200 rpm for enzymolysis for 1.5 hours; after the enzymolysis is completed, adding 95% (v / v) ethanol to make the final ethanol concentration of the feed liquid reach 65%, performing microwave-assisted extraction at a microwave power of 500 W and a temperature of 65°C for 10 minutes, then heating to 95°C and maintaining for 15 minutes to completely inactivate all enzymes and terminate the reaction; after cooling to room temperature, stirring at 5000 rpm / min, centrifuge for 15 min, take the supernatant, and ultrafilter through a 10 kDa molecular cutoff membrane, collect the filtrate, and concentrate under reduced pressure at 60 ° C to a thick paste-like extract. Add an appropriate amount of deionized water to the extract, separate it with AB-8 type macroporous adsorption resin, and elute with 5 times the column volume of deionized water, 30% ethanol, and 70% ethanol at a flow rate of 2 BV / h. Collect the 70% ethanol eluate, concentrate under reduced pressure at 60 ° C to a thick paste-like extract, and freeze-dry to obtain the hawthorn leaf extract.

[0048] Further preferably, the complex enzyme 3 comprises: cellulase, pectinase, tannase and papain.

[0049] More preferably, the complex enzyme 3 comprises, by weight, 0.5-1.5 parts of cellulase, 0.4-1 parts of pectinase, 0.2-0.8 parts of tannase and 0.2-0.4 parts of papain.

[0050] More preferably, the complex enzyme 3 comprises, by weight, 0.5 parts of cellulase, 0.4 parts of pectinase, 0.2 parts of tannase and 0.4 parts of papain; or the complex enzyme 3 comprises, by weight, 1.5 parts of cellulase, 1 part of pectinase, 0.8 parts of tannase and 0.4 parts of papain.

[0051] Further preferably, the added amount of the complex enzyme 3 is 1-3% of the mass of the hawthorn leaves.

[0052] More preferably, the added amount of the complex enzyme 3 is 1.5% of the mass of the hawthorn leaves.

[0053] Further preferably, the complex enzyme 4 comprises: acid protease and β-glucosidase.

[0054] More preferably, the complex enzyme 4 comprises, by weight, 0.2-0.4 parts of acid protease and 0.3-0.8 parts of β-glucosidase.

[0055] More preferably, the composite enzyme 4 comprises, by weight, 0.2 parts of acid protease and 0.8 parts of β-glucosidase; or the composite enzyme 4 comprises, by weight, 0.2 parts of acid protease and 0.3 parts of β-glucosidase.

[0056] Further preferably, the added amount of the complex enzyme 4 is 0.3-0.8% of the mass of the hawthorn leaves.

[0057] More preferably, the added amount of the complex enzyme 4 is 0.5% of the mass of the hawthorn leaves.

[0058] In a second aspect, the present invention provides a method for preparing the above-mentioned composition, comprising the steps of: mixing a coptis root extract, a hawthorn leaf extract, L-carnitine, acetyl-L-carnitine and inulin to obtain the composition.

[0059] Preferably, the preparation method of the Coptidis Rhizome extract comprises the steps of: performing a first enzymatic hydrolysis of the Coptidis Rhizome using compound enzyme 1, and then performing a second enzymatic hydrolysis using compound enzyme 2; performing ultrasonic-assisted extraction with an ethanol solution after enzymatic hydrolysis, heating and inactivating the enzyme; and ultrafiltration and resin purification and separation to obtain the Coptidis Rhizome extract.

[0060] Further preferably, before enzymatic hydrolysis, the coptis root is crushed to a particle size of 20-60 mesh; more preferably 40 mesh.

[0061] Further preferably, before enzymatic hydrolysis, the coptis root is mixed with water at a solid-liquid ratio of 1:10-20 g / mL.

[0062] More preferably, the first enzymatic hydrolysis is carried out under the following conditions: adjusting the pH to 5-6, and performing enzymatic hydrolysis at 48-52° C. and 100-300 rpm with stirring for 2.5-5 hours.

[0063] More preferably, the first enzymatic hydrolysis is carried out under the following conditions: adjusting the pH to 5.5, and performing enzymatic hydrolysis at 50° C. and 200 rpm with stirring for 3.5 hours.

[0064] More preferably, the second enzymatic hydrolysis is carried out under the following conditions: adjusting the pH to 4.2-5, and performing enzymatic hydrolysis at 56-63° C. and 100-300 rpm for 1-2.5 hours.

[0065] More preferably, the second enzymatic hydrolysis is carried out under the following conditions: adjusting the pH to 4.5, and performing enzymatic hydrolysis at 60° C. and 200 rpm with stirring for 1.5 hours.

[0066] Further preferably, in the ultrasonic-assisted extraction, the volume concentration of ethanol is 70-80%, and the ultrasonic-assisted extraction is performed at 25-35 Hz and 300-500 W for 20-40 min.

[0067] Further preferably, in the ultrasonic-assisted extraction, the volume concentration of ethanol is 75%, and the ultrasonic-assisted extraction is performed at 30 Hz and 400 W for 30 min.

[0068] Further preferably, the molecular cut-off of the ultrafiltration is 8kDa-12kDa molecular weight; further preferably, it is 10kDa.

[0069] Further preferably, in the resin purification and separation, AB-8 macroporous adsorption resin is used for separation, and water and 75% ethanol are used for elution in sequence, with an elution flow rate of 1-3 BV / h, and the 75% ethanol eluate is collected.

[0070] More preferably, in the resin purification and separation, AB-8 macroporous adsorption resin is used for separation, and water and 75% ethanol are used for elution in sequence, with an elution flow rate of 2BV / h, and the 75% ethanol eluate is collected.

[0071] Most preferably, the preparation method of the Coptidis rhizome extract comprises the steps of: drying and crushing the Coptidis rhizome, passing through a 40-mesh sieve, and fully mixing the powder with water at a material-liquid ratio of 1:10 g / mL; adding complex enzyme 1 to the mixture, adjusting the pH to 5.5, and stirring at 50°C and 200 rpm for 3.5 hours; adjusting the pH to 4.5, adding complex enzyme 2, and stirring at 60°C and 200 rpm for 1.5 hours; after the enzymatic hydrolysis is completed, adding 95% (v / v) ethanol to make the final ethanol concentration of the feed liquid reach 75%, and performing ultrasonic-assisted extraction for 30 minutes at 30 kHz and 400 W (the temperature of the ice water bath is controlled at ≤50 ℃); then the temperature was raised to 95℃ and maintained for 15 minutes to completely inactivate all enzymes and terminate the reaction; after cooling to room temperature, the mixture was centrifuged at 5000 rpm / min for 15 minutes, the supernatant was collected, and ultrafiltration was performed through a 10 kDa molecular cut-off membrane; the filtrate was collected and concentrated under reduced pressure at 60℃ to a thick paste-like extract, the extract was mixed with water, the pH was adjusted to 5.5, and separated using AB-8 macroporous adsorption resin, and eluted with 5 times the column volume of deionized water and 75% ethanol at a flow rate of 2 BV / h, the 75% ethanol eluate was collected, and the filtrate was concentrated under reduced pressure at 60℃ to a thick paste-like extract, and freeze-dried to obtain the Coptidis rhizome extract.

[0072] Further preferably, the complex enzyme 1 comprises: cellulase, pectinase, tannase and papain.

[0073] More preferably, the complex enzyme 1 comprises, by weight, 0.5-1.5 parts of cellulase, 0.4-1 parts of pectinase, 0.2-0.8 parts of tannase and 0.2-0.4 parts of papain.

[0074] More preferably, the complex enzyme 1 comprises, by weight, 0.5 parts of cellulase, 0.4 parts of pectinase, 0.2 parts of tannase and 0.4 parts of papain; or the complex enzyme 1 comprises, by weight, 1.5 parts of cellulase, 1 part of pectinase, 0.8 parts of tannase and 0.4 parts of papain.

[0075] Further preferably, the added amount of the complex enzyme 1 is 3-5% of the mass of Coptis chinensis.

[0076] More preferably, the added amount of the complex enzyme 1 is 3.7% of the mass of Coptis chinensis.

[0077] Further preferably, the complex enzyme 2 comprises: acid protease and β-glucosidase.

[0078] More preferably, the complex enzyme 2 comprises, by weight, 0.2-0.4 parts of acid protease and 0.3-0.8 parts of β-glucosidase.

[0079] More preferably, the complex enzyme 2 comprises, by weight, 0.2 parts of acid protease and 0.8 parts of β-glucosidase; or the complex enzyme 2 comprises, by weight, 0.2 parts of acid protease and 0.3 parts of β-glucosidase.

[0080] Further preferably, the added amount of the complex enzyme 2 is 0.8-1.5% of the mass of Coptis chinensis.

[0081] More preferably, the added amount of the complex enzyme 2 is 1.1% of the mass of Coptis chinensis.

[0082] Preferably, the preparation method of the hawthorn leaf extract comprises the steps of: performing a first enzymatic hydrolysis on the hawthorn using compound enzyme 3, and then performing a second enzymatic hydrolysis using compound enzyme 4; after the enzymatic hydrolysis, performing microwave-assisted extraction with an ethanol solution and heating to inactivate the enzyme; and ultrafiltration and resin purification and separation to obtain the hawthorn leaf extract.

[0083] Further preferably, the hawthorn leaves are crushed before enzymatic hydrolysis, and the crushed particle size is 20-60 mesh; more preferably 50 mesh.

[0084] Further preferably, before enzymatic hydrolysis, the hawthorn leaves and water are mixed at a solid-liquid ratio of 1:10-20 g / mL; even more preferably, before enzymatic hydrolysis, the hawthorn leaves and water are mixed at a solid-liquid ratio of 1:12 g / mL; More preferably, the first enzymatic hydrolysis is carried out under the following conditions: adjusting the pH to 5-6, and performing enzymatic hydrolysis at 52-58° C. and 100-300 rpm with stirring for 2.5-5 hours.

[0085] More preferably, the first enzymatic hydrolysis is carried out under the following conditions: adjusting the pH to 5.5, and performing enzymatic hydrolysis at 55° C. and 200 rpm with stirring for 3.5 hours.

[0086] More preferably, the second enzymatic hydrolysis is carried out under the following conditions: adjusting the pH to 4.2-5, and performing enzymatic hydrolysis at 56-63° C. and 100-300 rpm for 1-2.5 hours.

[0087] More preferably, the second enzymatic hydrolysis is carried out under the following conditions: adjusting the pH to 4.5, and performing enzymatic hydrolysis at 60° C. and 200 rpm with stirring for 1.5 hours.

[0088] Further preferably, in the microwave-assisted extraction, the volume concentration of ethanol is 60-70%, and the microwave-assisted extraction is performed at 400-600W and 60-70°C for 8-20min.

[0089] Further preferably, in the microwave-assisted extraction, the volume concentration of ethanol is 65%, and the microwave-assisted extraction is performed at 500W and 65°C for 10 minutes.

[0090] Further preferably, the molecular cut-off of the ultrafiltration is 8-12 kDa; further preferably, it is 10 kDa.

[0091] Further preferably, in the resin purification and separation, AB-8 type macroporous adsorption resin is used for separation, and water, 30% ethanol, and 70% ethanol are used for elution in sequence, with an elution flow rate of 1-3 BV / h, and 75% ethanol eluate is collected.

[0092] More preferably, in the resin purification and separation, AB-8 macroporous adsorption resin is used for separation, and water, 30% ethanol, and 70% ethanol are used for elution in sequence, with an elution flow rate of 2BV / h, and the 70% ethanol eluate is collected.

[0093] Most preferably, the preparation method of the hawthorn leaf extract comprises the steps of: drying the hawthorn leaves and crushing them, passing them through a 50-mesh sieve, and thoroughly mixing the powder with deionized water at a material-liquid ratio of 1:12 g / mL; adding complex enzyme 3 to the mixture, adjusting the pH to 5.5, and stirring at 55°C and 200 rpm for enzymolysis for 3.5 hours; adjusting the pH to 4.5, adding complex enzyme 4, and stirring at 60°C and 200 rpm for enzymolysis for 1.5 hours; after the enzymolysis is completed, adding 95% (v / v) ethanol to make the final ethanol concentration of the feed liquid reach 65%, performing microwave-assisted extraction at a microwave power of 500 W and a temperature of 65°C for 10 minutes, then heating to 95°C and maintaining for 15 minutes to completely inactivate all enzymes and terminate the reaction; after cooling to room temperature, stirring at 5000 rpm / min, centrifuge for 15 min, take the supernatant, and ultrafilter through a 10 kDa molecular cutoff membrane, collect the filtrate, and concentrate under reduced pressure at 60 ° C to a thick paste-like extract. Add an appropriate amount of deionized water to the extract, separate it with AB-8 type macroporous adsorption resin, and elute with 5 times the column volume of deionized water, 30% ethanol, and 70% ethanol at a flow rate of 2 BV / h. Collect the 70% ethanol eluate, concentrate under reduced pressure at 60 ° C to a thick paste-like extract, and freeze-dry to obtain the hawthorn leaf extract.

[0094] Further preferably, the complex enzyme 3 comprises: cellulase, pectinase, tannase and papain.

[0095] More preferably, the complex enzyme 3 comprises, by weight, 0.5-1.5 parts of cellulase, 0.4-1 parts of pectinase, 0.2-0.8 parts of tannase and 0.2-0.4 parts of papain.

[0096] More preferably, the complex enzyme 3 comprises, by weight, 0.5 parts of cellulase, 0.4 parts of pectinase, 0.2 parts of tannase and 0.4 parts of papain; or the complex enzyme 3 comprises, by weight, 1.5 parts of cellulase, 1 part of pectinase, 0.8 parts of tannase and 0.4 parts of papain.

[0097] Further preferably, the added amount of the complex enzyme 3 is 1-3% of the mass of the hawthorn leaves.

[0098] More preferably, the added amount of the complex enzyme 3 is 1.5% of the mass of the hawthorn leaves.

[0099] Further preferably, the complex enzyme 4 comprises: acid protease and β-glucosidase.

[0100] More preferably, the complex enzyme 4 comprises, by weight, 0.2-0.4 parts of acid protease and 0.3-0.8 parts of β-glucosidase.

[0101] More preferably, the composite enzyme 4 comprises, by weight, 0.2 parts of acid protease and 0.8 parts of β-glucosidase; or the composite enzyme 4 comprises, by weight, 0.2 parts of acid protease and 0.3 parts of β-glucosidase.

[0102] Further preferably, the added amount of the complex enzyme 4 is 0.3-0.8% of the mass of the hawthorn leaves.

[0103] More preferably, the added amount of the complex enzyme 4 is 0.5% of the mass of the hawthorn leaves.

[0104] In the final aspect, the invention provides the use of the above composition in the preparation of a drug for improving type 2 diabetes combined with depression.

[0105] In the present invention, the active ingredients in the Coptis chinensis extract are alkaloid components such as berberine, coptisine, and jatrorrhizine, which have the effects of lowering blood sugar and improving depression. However, single use is prone to gastrointestinal irritation, and long-term use can lead to side effects such as constipation.

[0106] The active ingredients in hawthorn leaf extract are flavonoids such as vitexin and isovitexin, which have the effects of lowering blood sugar and improving insulin resistance.

[0107] L-carnitine is an amino acid-like substance with multiple functions in the body, playing a crucial role in regulating lipid metabolism. Modern pharmacology suggests that L-carnitine supplementation can improve glucose metabolism in patients with type 2 diabetes and prevent complications such as mental disorders.

[0108] Acetyl-L-carnitine is an acetylated derivative of L-carnitine. It can easily penetrate the blood-brain barrier after oral administration and has a neuroprotective effect.

[0109] Inulin, a natural soluble dietary fiber, has the potential to regulate intestinal flora and improve metabolism. Studies have shown that inulin can promote gastrointestinal motility and bowel movements, and is used as a prebiotic to promote the proliferation of probiotics such as Bifidobacterium.

[0110] In the composition of the present invention, the above ingredients work synergistically to bring about significant effects of improving sugar metabolism function and depressive behavior.

[0111] The beneficial effects of the present invention are: 1. The present invention utilizes the combination of coptis root extract, hawthorn leaf extract, L-carnitine, acetyl-L-carnitine and inulin to play a synergistic role in improving type 2 diabetes combined with depression, can effectively improve the glucose metabolism function and the behavioral symptoms of depressed mice, and has a better therapeutic effect.

[0112] 2. The coptis root extract and hawthorn leaf extract of the present invention are obtained by specific enzymatic hydrolysis and auxiliary extraction. The above extracts interact with L-carnitine, acetyl-L-carnitine and inulin, which significantly improves the effect of improving type 2 diabetes combined with depression. DETAILED DESCRIPTION

[0113] The following non-limiting examples are provided to enable those skilled in the art to more fully understand the present invention, but are not intended to limit the present invention in any way. The following is merely an illustrative description of the scope of the present invention, and those skilled in the art may make various changes and modifications to the present invention based on the disclosed content, which should also fall within the scope of the present invention.

[0114] The present invention will be further described below by way of specific examples. The various chemical reagents used in the examples of the present invention were obtained through conventional commercial channels unless otherwise specified. Unless otherwise specified, the contents described below are all by weight. Unless otherwise specified, it is understood that the experiments were conducted at room temperature.

[0115] Cellulase (50 U / mg, Catalog No. A62048), pectinase (30 U / mg, Catalog No. A91700), and papain (800 U / mg, Catalog No. B99890) were purchased from Beijing Inokai Technology Co., Ltd. Tannase (100 U / mg, Catalog No. 113164A) was purchased from Adamas Life, and acid protease (700 U / mg, Catalog No. P832411) and β-glucosidase (10-30 U / mg, Catalog No. G824159) were purchased from Macklin. Products from different manufacturers did not significantly affect the efficacy.

[0116] In an embodiment of the present invention, the preparation method of the Coptidis Rhizoma extract is: Coptis chinensis was dried and ground, passed through a 40-mesh sieve, and thoroughly mixed with deionized water at a material-to-liquid ratio of 1:10 (g / mL). Subsequently, complex enzyme 1 was added to the mixture, the pH was adjusted to 5.5, and enzymatic hydrolysis was carried out at 50°C and 200 rpm for 3.5 hours. After completion, the pH was adjusted to 4.5, and complex enzyme 2 was added. Enzymatic hydrolysis was carried out at 60°C and 200 rpm for 1.5 hours. After the enzymatic hydrolysis was completed, 95% (v / v) ethanol was added to a final ethanol concentration of 75%. Ultrasonic extraction was performed at 30 kHz and 400 W for 30 minutes (with an ice-water bath controlled at ≤50°C). The temperature was then raised to 95°C and maintained for 15 minutes to completely inactivate all enzymes, terminating the reaction. After cooling to room temperature, the mixture was centrifuged at 5000 rpm for 15 minutes. The supernatant was collected and ultrafiltered through a 10 kDa molecular weight cutoff membrane. The filtrate was collected and concentrated under reduced pressure at 60° C. to a thick paste-like extract. An appropriate amount of deionized water was added to the extract to dissolve it or form a uniform suspension. The pH was adjusted to 5.5, and the extract was separated using an AB-8 macroporous adsorption resin. The extract was eluted with 5 times the column volume of deionized water and 75% ethanol at a flow rate of 2 BV / h, and the 75% ethanol eluate was collected and concentrated under reduced pressure at 60° C. to a thick paste-like extract. The extract was freeze-dried into powder to obtain the Coptis chinensis extract.

[0117] The compound enzyme 1 is composed of 1.5% cellulase, 1% pectinase, 0.8% tannase and 0.4% papain based on the weight of the coptis chinensis powder. The compound enzyme 2 is composed of 0.2% acid protease and 0.8% β-glucosidase based on the weight of the coptis chinensis powder.

[0118] In an embodiment of the present invention, the preparation method of hawthorn leaf extract is: Hawthorn leaves were dried and crushed, passed through a 50-mesh sieve, and thoroughly mixed with deionized water at a material-to-liquid ratio of 1:12 (g / mL). Subsequently, compound enzyme 3 was added to the mixture, the pH was adjusted to 5.5, and enzymatic hydrolysis was carried out at 55°C and 200 rpm for 3.5 hours. After completion, the pH was adjusted to 4.5, and compound enzyme 4 was added. Enzymatic hydrolysis was carried out at 60°C and 200 rpm for 1.5 hours. After the enzymatic hydrolysis was completed, 95% (v / v) ethanol was added to bring the final ethanol concentration of the feed solution to 65%. Microwave-assisted extraction was performed at a microwave power of 500 W and a temperature of 65°C for 10 minutes. The temperature was then raised to 95°C and maintained for 15 minutes to completely inactivate all enzymes and terminate the reaction. After cooling to room temperature, centrifuge at 5000 rpm / min for 15 minutes, take the supernatant, and ultrafilter through a 10 kDa molecular cutoff membrane, collect the filtrate, and concentrate under reduced pressure at 60°C to a thick paste-like extract, add an appropriate amount of deionized water to the extract to dissolve it or form a uniform suspension, separate it with AB-8 type macroporous adsorption resin, and elute with 5 times the column volume of deionized water, 30% ethanol, and 70% ethanol at a flow rate of 2 BV / h, collect the 70% ethanol eluate, concentrate under reduced pressure at 60°C to a thick paste-like extract, and freeze-dry it into powder to obtain the hawthorn leaf extract.

[0119] The compound enzyme 3 is composed of 0.5% cellulase, 0.4% pectinase, 0.2% tannase and 0.4% papain by weight of the hawthorn leaf powder. The compound enzyme 4 is composed of 0.2% acid protease and 0.3% β-glucosidase by weight of the hawthorn leaf powder.

[0120] Example 1 A composition for improving type 2 diabetes combined with depression comprises the following components in parts by weight: 3 parts of coptis root extract, 2 parts of hawthorn leaf extract, 5 parts of L-carnitine, 5 parts of acetyl-L-carnitine, and 15 parts of inulin. The components are mixed to obtain a composition.

[0121] Example 2 A composition for improving type 2 diabetes combined with depression, comprising the following components by weight: 2 parts of coptis root extract, 1 part of hawthorn leaf extract, 3 parts of L-carnitine, 2 parts of acetyl-L-carnitine, and 10 parts of inulin. The components are mixed to obtain a composition.

[0122] Example 3 A composition for improving type 2 diabetes combined with depression comprises the following components in parts by weight: 6 parts of coptis root extract, 5 parts of hawthorn leaf extract, 8 parts of L-carnitine, 10 parts of acetyl-L-carnitine, and 20 parts of inulin. The components are mixed to obtain a composition.

[0123] Example 4 A composition for improving type 2 diabetes combined with depression, comprising the following components by weight: 2 parts of coptis root extract, 1 part of hawthorn leaf extract, 8 parts of L-carnitine, 10 parts of acetyl-L-carnitine, and 15 parts of inulin. The components are mixed to obtain a composition.

[0124] Example 5 A composition for improving type 2 diabetes combined with depression, comprising the following components by weight: 6 parts of coptis root extract, 5 parts of hawthorn leaf extract, 3 parts of L-carnitine, 2 parts of acetyl-L-carnitine, and 20 parts of inulin. The components are mixed to obtain a composition.

[0125] Comparative Example 1 The difference from Example 1 is that the composition has different components. The composition comprises, by weight, 5 parts of L-carnitine, 5 parts of acetyl-L-carnitine, and 15 parts of inulin. The rest is the same as in Example 1.

[0126] Comparative Example 2 The difference from Example 1 is that the composition has different components. The composition comprises 3 parts of Coptis chinensis extract and 2 parts of Crataegus pinnatifida leaf extract by weight. The rest is the same as Example 1.

[0127] Comparative Example 3 The difference from Example 1 is that the components of the composition are different. The components of the composition, by weight, are: 3 parts of Coptis chinensis extract, 2 parts of Crataegus pinnatifida leaf extract, 5 parts of L-carnitine, 5 parts of acetyl-L-carnitine, 15 parts of inulin, 1.5 parts of Hypericum perforatum extract, 3.5 parts of Cornus officinalis extract, and 3 parts of ginseng extract.

[0128] The preparation methods of Hypericum perforatum extract, Cornus officinalis extract and Panax ginseng extract were prepared by referring to the contents of Example 1 of Chinese invention patent CN109481611B.

[0129] Comparative Example 4 The difference from Example 1 is that the components of the composition are different, and the Coptidis rhizome extract is replaced by Curcuma longa extract.

[0130] The composition comprises the following components in parts by weight: 3 parts of turmeric extract, 2 parts of hawthorn leaf extract, 5 parts of L-carnitine, 5 parts of acetyl-L-carnitine and 15 parts of inulin.

[0131] The preparation method of turmeric extract refers to the preparation method of coptis chinensis extract, except that coptis chinensis is replaced by turmeric.

[0132] Comparative Example 5 The difference from Example 1 is that the weight ratios of the components in the composition are different.

[0133] The composition comprises the following components in parts by weight: 8 parts of coptis root extract, 6 parts of hawthorn leaf extract, 10 parts of L-carnitine, 12 parts of acetyl-L-carnitine and 5 parts of inulin.

[0134] Comparative Example 6 The difference from Example 1 is that the preparation method of the Coptidis Rhizoma extract is different.

[0135] In the preparation method of the Coptis chinensis extract of this comparative example, compound enzyme 1 is not used for enzymatic hydrolysis, and other parameters are the same as those in Example 1.

[0136] The preparation method of Coptis chinensis extract is as follows: Coptis chinensis was dried and ground, passed through a 40-mesh sieve, and thoroughly mixed with deionized water at a solid-liquid ratio of 1:10 (g / mL). The mixture was then adjusted to pH 4.5, and complex enzyme 2 was added. Enzymatic hydrolysis was carried out at 60°C and 200 rpm for 1.5 hours. After enzymatic hydrolysis, 95% (v / v) ethanol was added to a final ethanol concentration of 75%. Ultrasonic extraction was then performed at 30 kHz and 400 W for 30 minutes (with an ice-water bath controlled at ≤50°C). The temperature was then raised to 95°C and maintained for 15 minutes to completely inactivate all enzymes and terminate the reaction. After cooling to room temperature, the mixture was centrifuged at 5000 rpm for 15 minutes. The supernatant was collected and ultrafiltered through a 10 kDa molecular weight cutoff membrane. The filtrate was collected and concentrated under reduced pressure at 60° C. to a thick paste-like extract. An appropriate amount of deionized water was added to the extract to dissolve it or form a uniform suspension. The pH was adjusted to 5.5, and the extract was separated using an AB-8 macroporous adsorption resin. The extract was eluted with 5 times the column volume of deionized water and 75% ethanol at a flow rate of 2 BV / h, and the 75% ethanol eluate was collected and concentrated under reduced pressure at 60° C. to a thick paste-like extract. The extract was freeze-dried into powder to obtain the Coptis chinensis extract.

[0137] The complex enzyme 2 is composed of 0.2% of acid protease and 0.8% of beta-glucosidase based on the weight of the coptis chinensis powder.

[0138] Comparative Example 7 The difference from Example 1 is that the preparation method of the Coptidis Rhizoma extract is different.

[0139] In the preparation method of the Coptis chinensis extract of this comparative example, compound enzyme 2 is not used for enzymatic hydrolysis, and other parameters are the same as those in Example 1.

[0140] The preparation method of Coptis chinensis extract is as follows: Coptis chinensis was dried and ground, passed through a 40-mesh sieve, and thoroughly mixed with deionized water at a material-to-liquid ratio of 1:10 (g / mL). Complex enzyme 1 was then added to the mixture, and the pH was adjusted to 5.5. Enzymatic hydrolysis was carried out at 50°C and 200 rpm for 3.5 hours. After the enzymatic hydrolysis, 95% (v / v) ethanol was added to a final ethanol concentration of 75%. Ultrasonic extraction was then performed at 30 kHz and 400 W for 30 minutes (with an ice-water bath controlled at ≤50°C). The temperature was then raised to 95°C and maintained for 15 minutes to completely inactivate all enzymes and terminate the reaction. After cooling to room temperature, the mixture was centrifuged at 5000 rpm for 15 minutes. The supernatant was collected and ultrafiltered through a 10 kDa molecular weight cutoff membrane. The filtrate was collected and concentrated under reduced pressure at 60° C. to a thick paste-like extract. An appropriate amount of deionized water was added to the extract to dissolve it or form a uniform suspension. The pH was adjusted to 5.5, and the extract was separated using an AB-8 macroporous adsorption resin. The extract was eluted with 5 times the column volume of deionized water and 75% ethanol at a flow rate of 2 BV / h, and the 75% ethanol eluate was collected and concentrated under reduced pressure at 60° C. to a thick paste-like extract. The extract was freeze-dried into powder to obtain the Coptis chinensis extract.

[0141] The complex enzyme 1 is composed of 1.5% of cellulase, 1% of pectinase, 0.8% of tannase and 0.4% of papain based on the weight of coptis root powder.

[0142] 1. Synergistic efficacy detection of the combination The compositions prepared in the above examples and comparative examples were tested for their efficacy.

[0143] The dosage is 2 g / kg, administered once daily, and the solvent is a deionized water solution containing 0.5% sodium carboxymethyl cellulose.

[0144] (1) Establishment and drug administration of a mouse model of type 2 diabetes combined with depression A total of 140 male, SPF-qualified C57BL / 6J mice, 6-7 weeks old, weighing 20-25g, were selected. After 3 days of acclimation, the model mice were fed a 60 kcal% high-fat diet. After 5 weeks, they were intraperitoneally injected with streptozotocin (50 mg / kg) prepared in 0.1 mol / L sterile citrate buffer daily for 4 consecutive days. A normal control group was fed a standard diet and, after 5 weeks, intraperitoneally injected with sterile citrate buffer (pH 4.4). Body weights of the model mice were measured at 0, 1, 2, and 3 weeks after streptozotocin injection. Fasting blood glucose was measured by tail tip blood sampling after a 6-hour fast. Mice with fasting blood glucose levels ≥11.1 mmol / L for 3 consecutive weeks were selected and further screened for depression using a sugar preference test.

[0145] The CUMS modeling method is as follows: model mice are randomly given two of the following stimuli every day, including: stimulation 1. plantar electrical stimulation (0.8 mA, stimulation 10 seconds, rest 20 seconds, a total of 5 minutes); stimulation 2. cage tilt (12 hours); stimulation 3. wetting the bedding (24 hours); stimulation 4. cold stimulation (4°C, 20 minutes); stimulation 5. environmental day and night reversal (24 hours); stimulation 6. cage vibration (200 rpm, 10 minutes); stimulation 7. behavioral restraint (4 hours); stimulation 8. social isolation, solitary housing (24 hours); stimulation 9. noise exposure (80 decibels, 2 hours).

[0146] The normal control group consisted of 10 rats per group. After the model was established, the rats were divided into the model control group, Example 1-5 groups, and Comparative Example 1-7 groups, with 10 rats per group. All treatment groups were gavage-treated, while the normal control group and the model control group were gavage-treated with 0.5% sodium carboxymethylcellulose in normal saline once daily for 4 consecutive weeks. After 6 hours of fasting, blood was collected from the tip of the tail to measure fasting blood glucose and fasting insulin, followed by behavioral testing experiments.

[0147] (2) Behavioral testing Behavioral testing experiments were carried out in sequence, including sugar water preference test, tail suspension test and forced swim test, and the data were analyzed using SMARTV3.0 software.

[0148] The sugar preference test was performed as follows: 72 hours before the experiment, a bottle of water containing 1% sucrose and a bottle of pure water (both with the same apparent weight) were placed on either side of the mouse cage. 24 hours later, the bottles were inverted to accommodate the trained mice. After the 48 hours of acclimation training described above, all mice were deprived of water for 24 hours. The bottles of water containing 1% sucrose and pure water were reconfigured and randomly placed on either side of the mouse cage. After the mice had been drinking water for 24 hours, the remaining drinking water was weighed, and sucrose consumption was calculated using the following formula: Sugar water preference rate (%) = sucrose water consumption (g) / (sucrose water consumption (g) + pure water consumption (g)) × 100%.

[0149] The tail suspension test was performed as follows: The mouse was fixed head-down on a suspension device using medical tape, 1 cm from the tip of the tail. The immobility time (s) during the last 4 minutes of the 6-minute experimental period was recorded for each mouse.

[0150] The forced swim test was performed as follows: One day before the experiment, each mouse was acclimated to swimming in a swimming apparatus (10 cm diameter, 25 cm height, water temperature 25 ± 1°C) for 15 minutes. Twenty-four hours after the experiment, the immobility time (s) during the last 3 minutes of the 5-minute experimental period was recorded for each mouse.

[0151] (3) Fasting blood glucose, insulin resistance index and glycated hemoglobin testing After blood was collected from the tip of the tail, fasting blood glucose and fasting insulin were measured using a blood glucose meter after fasting for 6 hours, and the insulin resistance index (IRI) was calculated using the following formula: Insulin resistance index = [fasting blood glucose (mmol / L) × fasting insulin (μU / mL)] / 22.5.

[0152] Blood was collected from the tip of the tail and capillary electrophoresis was used to quantitatively analyze glycated hemoglobin after 6 h of fasting.

[0153] (4) Test results Table 1 Behavioral test results

[0154] Note: ### represents the comparison between the model control group and the normal control group. ### P <0.001; *** Comparison between Example 1 to Example 5 groups and the model control group is shown. *** P <0.001; Comparative Examples 1-7 and Example 1, △ P <0.05, △△ P <0.01, △△△ P <0.001.

[0155] Table 2 Fasting blood glucose and insulin resistance index test results

[0156] Note: ### represents the comparison between the model control group and the normal control group. ### P <0.001; *** Comparison between Example 1 to Example 5 groups and the model control group is shown. *** P <0.001; Comparative Examples 1-7 and Example 1, △ P <0.05, △△ P <0.01, △△△ P <0.001.

[0157] The results of each group after administration are shown in Table 1-Table 2. The sugar preference rate of the model control group was significantly lower than that of the normal control group ( P<0.001), fasting blood glucose, insulin resistance index, glycosylated hemoglobin and other glucose metabolism indicators, as well as the immobility time in the tail suspension test and forced swimming test were significantly higher than those in the normal control group ( P <0.001), indicating that the type 2 diabetes mellitus combined with depression mouse model was successfully established, and depressive-like behaviors were obvious.

[0158] From the results of Example 1 to Example 5, it can be seen that compared with the model control group, the glucose metabolism indicators of Example 1 to Example 5 groups were significantly reduced, the sugar preference rate was significantly increased, and the immobility time was significantly decreased, indicating that the combined use of each component in the composition of the present application can improve the glucose metabolism function and depressive symptoms of the model; the reduction in glucose metabolism indicators and immobility time of Example 1 to Example 5 groups exceeded the sum of the reductions in Comparative Example 1 and Comparative Example 2, and the increase in the sugar preference rate exceeded the sum of the increase in Comparative Example 1 and Comparative Example 2. Therefore, the technical effect after the overall use of the composition is superior to the sum of the technical effects of improving glucose metabolism after partial use of the composition.

[0159] From the results of Comparative Example 1-Comparative Example 2, it can be seen that the deletion of the extracts of Coptis chinensis and Hawthorn leaf lacks direct intervention and neuroprotection on the insulin signaling pathway, glucose uptake and utilization, resulting in loss of effect; the deletion of L-carnitine, acetyl-L-carnitine and inulin cannot improve mitochondrial function and intestinal flora, thereby indirectly reducing insulin sensitivity, inhibiting peripheral metabolic improvement and brain energy supply, and the extracts of Coptis chinensis and Hawthorn leaf alone are not enough to completely improve the various indicators of the model. Therefore, the various components of the composition are indispensable, and changing the components of the composition will significantly reduce the hypoglycemic and depression treatment effects of mice with type 2 diabetes and depression.

[0160] The results of Comparative Example 3 indicate that, referring to Chinese invention patent CN109481611B, the addition of extracts from Hypericum perforatum, Cornus officinalis, and Panax ginseng may have antagonized the relevant pathways of action of the extracts from Coptis chinensis and Crataegus pinnatifida, causing interference with non-target pathways and offsetting the therapeutic effect. Therefore, only the composition containing the specific five ingredients of the present invention, which works in concert with each other, exhibits the significant improvement effect of the present invention in mice with diabetes and depression. Adding other components significantly reduces the hypoglycemic therapeutic effect in mice with type 2 diabetes and depression.

[0161] The results of Comparative Example 4 show that, referring to Chinese invention patent CN109481611B, when turmeric was substituted for coptis chinensis, while similar to coptis chinensis in terms of anti-inflammatory and hypoglycemic activity, turmeric was weaker than coptis chinensis, resulting in insufficient hypoglycemic and antidepressant effects. Therefore, replacing the components of the composition significantly reduced the hypoglycemic therapeutic effect in mice with type 2 diabetes and depression.

[0162] The results of Comparative Example 5 indicate that excessive amounts of coptis root and hawthorn leaves can produce certain side effects. Excess carnitine may be converted to trimethylamine-N-oxide (TMAO) by intestinal bacteria, while insufficient inulin prevents effective regulation of intestinal flora, further leading to a loss of efficacy. Therefore, using the composition outside the ratios of Examples 1-5 significantly reduced the hypoglycemic effect in mice with type 2 diabetes and depression.

[0163] From the results of Comparative Examples 6 and 7, it can be seen that the preparation method of the extract does not use complex enzyme 1 or 2 for enzymatic hydrolysis, the composition of the product obtained by extraction changes, and the composition will significantly reduce the hypoglycemic and depression treatment effects of mice with type 2 diabetes and depression.

[0164] 2. Safety Testing of Composition Acute toxicity test: Adult C57BL / 6J mice, SPF grade, weighing 20-25 g, were selected and randomly divided into 6 groups (10 mice in each group, half male and half female). One group served as a blank control group and was gavaged with a deionized water solution containing 0.5% sodium carboxymethyl cellulose; the remaining groups were gavaged with the drug composition (Example 1-5 groups, dosage: 5 g / kg).

[0165] The mice were observed for 14 consecutive days, including their spirit, appetite, drinking water and activities, and the poisoning status, death time and number of deaths were recorded.

[0166] The results showed that all groups of mice had no abnormalities after administration and no acute poisoning symptoms, indicating that the components of the compositions prepared in Examples 1 to 5 were safe and had no toxic reactions.

[0167] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A composition for improving type 2 diabetes combined with depression, characterized in that: The invention comprises the following ingredients: coptis root extract, hawthorn leaf extract, L-carnitine, acetyl-L-carnitine and inulin.

2. The composition according to claim 1, characterized in that The invention comprises the following components in parts by weight: 2-6 parts of coptis root extract, 1-5 parts of hawthorn leaf extract, 3-8 parts of L-carnitine, 2-10 parts of acetyl-L-carnitine and 10-20 parts of inulin.

3. The composition according to claim 2, characterized in that The composition comprises the following components in parts by weight: 3 parts of coptis root extract, 2 parts of hawthorn leaf extract, 5 parts of L-carnitine, 5 parts of acetyl-L-carnitine and 15 parts of inulin.

4. The composition according to any one of claims 1 to 3, characterized in that The preparation method of the coptis chinensis extract comprises the following steps: performing a first enzymatic hydrolysis on the coptis chinensis using a composite enzyme 1, and then performing a second enzymatic hydrolysis using a composite enzyme 2; performing ultrasonic-assisted extraction with an ethanol solution after the enzymatic hydrolysis, heating and inactivating the enzyme; and ultrafiltration and resin purification and separation to obtain the coptis chinensis extract.

5. The composition according to claim 4, characterized in that The first enzymatic hydrolysis is carried out under the following conditions: adjusting the pH to 5-6, stirring at 48-52°C and 100-300 rpm for 2.5-5 hours; The specific conditions of the second enzymatic hydrolysis are as follows: adjusting the pH to 4.2-5, stirring and enzymatic hydrolysis at 56-63° C. and 100-300 rpm for 1-2.5 hours.

6. The composition according to claim 4, characterized in that In the ultrasonic-assisted extraction, the volume concentration of ethanol is 70-80%, and the ultrasonic-assisted extraction is performed at 25-35 Hz and 300-500 W for 20-40 minutes.

7. The composition according to claim 4, characterized in that The compound enzyme 1 comprises cellulase, pectinase, tannase and papain, and the addition amount of the compound enzyme 1 is 3-5% of the mass of the coptis chinensis; The compound enzyme 2 comprises acidic protease and beta-glucosidase, and the added amount of the compound enzyme 2 is 0.8-1.5% of the mass of the coptis chinensis.

8. The composition according to any one of claims 1 to 3, characterized in that The preparation method of the hawthorn leaf extract comprises the following steps: performing a first enzymatic hydrolysis on hawthorn using a composite enzyme 3, and then performing a second enzymatic hydrolysis using a composite enzyme 4; performing microwave-assisted extraction with an ethanol solution after the enzymatic hydrolysis, heating and inactivating the enzyme; and ultrafiltration and resin purification and separation to obtain the hawthorn leaf extract.

9. The method for preparing the composition according to any one of claims 1 to 8, characterized in that: The method comprises the steps of: mixing coptis root extract, hawthorn leaf extract, L-carnitine, acetyl-L-carnitine and inulin to obtain a composition.

10. Use of the composition according to any one of claims 1 to 8 or the composition prepared by the preparation method according to claim 9 in preparing a drug for improving type 2 diabetes combined with depression.

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