Lactobacillus paracasei fermented turmeric for mediating metabolic diseases

Abstract

The invention provides a Lactobacillus paracasei fermented turmeric for regulating metabolic diseases. Specifically, the Lactobacillus paracasei fermented turmeric obtained by the method disclosed by the invention contains components such as water-soluble curcuminoids and can effectively inhibit fat accumulation and fat generation in an individual. Therefore, by administering an effective amount of the Lactobacillus paracasei fermented turmeric disclosed by the invention to an individual, the effect of preventing or treating metabolic diseases induced by a high-fat diet can be effectively achieved.

Description

Technical Field

[0001] The invention relates to the use of fermentation products, in particular to Lactobacillus paracasei fermented turmeric for regulating metabolic diseases. Background Art

[0002] The number of obese people is increasing year by year, mainly due to poor eating and exercise habits. In addition to causing metabolic abnormalities in the body, obesity can also lead to other metabolic diseases such as diabetes and non-alcoholic fatty liver disease. Currently, the World Health Organization, the United States, Canada, etc. have declared obesity a chronic disease.

[0003] The main active ingredients in the rhizome of turmeric (Curcuma longa) are curcuminoids (also known as curcumin, turmeric extract), which belong to the class of natural polyphenols. Curcuminoids include curcumin, demethoxycurcumin and bis-demethoxycurcumin. Turmeric has been used as a traditional herbal medicine in many Asian countries, and studies have found that it has antioxidant, anti-inflammatory and anti-cancer properties. Although many studies have pointed out that curcumin has strong biological activity, the characteristics of the structure of curcumin lead to problems such as poor water solubility, poor absorption, rapid metabolism, and low bioavailability. Although coating technology or nanotechnology is currently used to overcome the problems of poor absorption and low water solubility of curcumin to improve the lack of utilization in the body, the above technology still enables the processed curcumin to truly exert its efficacy in the body. Summary of the invention

[0004] The main purpose of the present invention is to provide a Lactobacillus paracasei fermented turmeric for regulating metabolic diseases. The turmeric is obtained by the special fermentation method disclosed by the present invention, has active ingredients such as water-soluble curcuminoids, and can effectively improve the bioavailability and absorption rate of curcumin, and further can achieve the effect of preventing or improving metabolic diseases by regulating fat metabolism and blood sugar metabolism in an organism.

[0005] Therefore, in order to achieve the above-mentioned purpose, the present invention provides a use of fermented turmeric for regulating metabolic diseases, wherein the fermented turmeric is prepared by fermenting turmeric with Lactobacillus paracasei, that is, by administering an effective amount of the fermented turmeric disclosed by the present invention or a composition containing the effective amount of fermented turmeric to an individual, metabolic diseases or symptoms caused by a high-fat diet can be effectively improved or prevented.

[0006] Among them, the Lactobacillus paracasei is classified and named Lacticaseibacillus paracasei MC1-40; the preservation date is April 28, 2022; the preservation unit is the German Microbial Culture Collection Center; address: Germany; the preservation number is: DSM 34254.

[0007] In an embodiment of the present invention, the fermented turmeric contains water-soluble curcuminoids.

[0008] Specifically, one embodiment of the present invention provides a use of the fermented turmeric in preparing a weight loss composition.

[0009] Another embodiment of the present invention provides a use of fermented turmeric in preparing a composition for treating and / or preventing diseases associated with metabolic imbalance.

[0010] Among them, the metabolic imbalance disease is caused by a high-fat diet, such as non-alcoholic fatty liver disease, hyperglycemia, etc.

[0011] The present invention also provides a fermented turmeric composition, comprising an effective amount of fermented turmeric, wherein the fermented turmeric is prepared by fermenting turmeric with Lactobacillus paracasei, and the Lactobacillus paracasei is classified and named Lacticaseibacillus paracasei MC1-40; the preservation date is April 28, 2022; the preservation unit is the German Microbiological Culture Collection Center; the address is: Germany; the preservation number is: DSM 34254.

[0012] The beneficial effects of the present invention are:

[0013] The invention provides a Lactobacillus paracasei fermented turmeric for regulating metabolic diseases and uses thereof. The turmeric is obtained by a special fermentation method disclosed by the invention and has active ingredients such as water-soluble curcuminoids, and can effectively improve the bioavailability and absorption rate of curcumin, and further can achieve the effect of preventing or improving metabolic diseases by regulating fat metabolism and blood sugar metabolism in an organism. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 The results are of analyzing the intracellular oil droplet content after HepG2 cells were cultured under different conditions.

[0015] Figure 2 The figure shows the weight changes of mice in each group during the experimental period in Example 2.

[0016] Figure 3A These are the results of H&E staining of liver tissues of mice in each group in Example 2.

[0017] Figure 3BThis is the result of analyzing the triglyceride content in the liver tissue of each group of mice in Example 2.

[0018] Figure 4 These are the results of H&E staining of gonadal adipose tissue of each group of mice in Example 2.

[0019] Figure 5A This is the result of analyzing the expression level of SirT1 in the liver tissue of each group of mice in Example 2.

[0020] Figure 5B This is the result of analyzing the expression of PGC-1α in the liver tissue of each group of mice in Example 2.

[0021] Fig. 6A To analyze the blood glucose results of each group of mice in Example 2 subjected to an oral glucose tolerance test.

[0022] Figure 6B For analysis Fig. 6A The results of measuring the blood glucose area under the curve of each group of mice.

[0023] Fig. 7A This is the result of analyzing the expression of GluT2 in the liver tissue of each group of mice in Example 2.

[0024] Figure 7B This is the result of analyzing the expression of pAkt / Akt in the liver tissue of each group of mice in Example 2. DETAILED DESCRIPTION

[0025] The invention provides a Lactobacillus paracasei fermented turmeric for regulating metabolic diseases, wherein the Lactobacillus paracasei fermented turmeric disclosed in the invention contains water-soluble curcuminoids and has the ability to inhibit fat accumulation and fat generation. Therefore, by administering an effective amount of the Lactobacillus paracasei fermented turmeric disclosed in the invention or a composition containing the Lactobacillus paracasei fermented turmeric disclosed in the invention to an individual, metabolic diseases induced by a high-fat diet, such as non-alcoholic fatty liver disease, obesity, hyperglycemia, diabetes, etc., can be effectively prevented or treated.

[0026] Furthermore, the Lactobacillus paracasei fermented turmeric disclosed in the present invention is obtained by fermenting turmeric with Lactobacillus paracasei, wherein the so-called Lactobacillus paracasei refers to a strain belonging to the species Lactobacillus paracasei or classified as the species Lactobacillus paracasei, Lacticaseibacillus paracasei MC1-40 (deposited in the German Collection of Microorganisms on April 28, 2022, with the deposit number: DSM34254); generally speaking, the fermentation reaction conditions include a pH value of 4.0-6.0, a fermentation temperature of 35-40°C, and a fermentation time of 21-36 hours, and the culture medium used is a culture medium that is well known to technicians in the technical field to which the present invention belongs and can be used to cultivate lactic acid bacteria or the growth of Lactobacillus paracasei.

[0027] The so-called "composition" of the present invention refers to an article containing 0.1% to 100% of the Lactobacillus paracasei fermented turmeric disclosed in the present invention, which can be a food, a medicine, a nutritional supplement, and can be prepared into different forms according to needs, including but not limited to powder, liquid, tablet, pill and capsule.

[0028] In order to illustrate the technical features and effects of the present invention, several embodiments will be described below with reference to the accompanying drawings.

[0029] Lacticaseibacillus paracasei MC1-40 used in the following examples were deposited in the German Collection of Microorganisms (Deutsche.20Sammlung von Mikroorganismen und Zellkulturen GmbH·DSMZ) on April 28, 2022, with the deposit number: DSM 34254.

[0030] The cells used in the following examples, such as HepG2, are biological materials that can be easily obtained by those skilled in the art, and therefore do not need to be preserved.

[0031] Example 1: Preparation of fermented turmeric

[0032] The method comprises the following steps: using a culture medium containing 1-2% glucose, 1-2% peptone, 0.01-0.08% magnesium sulfate and the like to culture a seed bacteria of Lacticaseibacillus paracasei MC1-40; when the lactic acid bacteria in the culture medium reach a logarithmic growth phase, inoculating the lactic acid bacteria into a main fermentation culture medium for main fermentation; the main fermentation culture medium contains 2-4% glucose, 2-6% peptone, 0.01-0.08% magnesium sulfate and 3% turmeric powder; the fermentation pH is controlled at 4.0-6.0, the fermentation temperature is 35-40° C. and the fermentation time is 21-36 hours; and after the fermentation is completed, drying is performed to obtain the fermented turmeric powder.

[0033] The unfermented turmeric powder and fermented turmeric liquid were extracted with water and methanol as extraction solvents, respectively, to obtain turmeric methanol extract, turmeric water extract, fermented turmeric methanol extract and fermented turmeric water extract. The methanol extract and water extract were analyzed by HPLC, wherein the HPLC analysis conditions were as follows: the mobile phase was 2% acetic acid solution and acetonitrile (6:4 (v / v)); the analytical column was C18; the detection wavelength was 420nm; the column temperature was 35℃; the flow rate was 2mL / min; and the sample injection volume was 10μL.

[0034] The results of HPLC analysis show that the total curcuminoid content in the turmeric methanol extract is 0.51%, of which BDMC (bisdemthoxycurcumin) is 0.13%, DMC (demethoxycurcumin) is 0.18%, and curcumin is 0.20%; the curcuminoid content in the fermented turmeric methanol extract is 0.39%, of which BDMC is 0.11%, DMC is 0.16%, and curcumin is 0.12%; while curcuminoids were not detected in the turmeric water extract, but the curcuminoid content in the fermented turmeric water extract was 0.12%.

[0035] The above results show that curcumin in the unfermented turmeric powder is insoluble in water due to its hydrophobic properties, and its content is almost undetectable in the aqueous solution; while the fermented turmeric obtained by fermenting turmeric with lactic acid bacteria such as L. paracasei contains 0.12% curcuminoids in water after analysis. In other words, the fermented turmeric and its fermented composition disclosed in the present invention can effectively improve the lack of low water solubility.

[0036] Example 2: Cell test

[0037] HepG2 cells (6×10 4 Cells / well) were cultured in a 96-well plate for 24 hours, wherein the culture medium was DMEM culture medium containing 10% fetal bovine serum; the culture conditions were 37°C and 5% carbon dioxide; when the cells were cultured to about 70% to 80% confluent, blank culture medium, DMSO, 16mM FFA (free fatty acids, hereinafter referred to as oleic acid), 16mM FFA and curcumin standard (25μM, dissolved in DMSO), 16mM FFA and unfermented turmeric aqueous solution (25μM), 16mM FFA and fermented turmeric aqueous solution (25μM) were added respectively, and cultured for another 24 hours, wherein the fermented turmeric aqueous solution was prepared by fermented turmeric prepared by the method disclosed in Example 1 and water. After the culture was completed, the cells in each well were subjected to Oil Red O staining to analyze the intracellular oil droplet staining, and the results were as follows: Figure 1shown.

[0038] Depend on Figure 1 It can be seen from the results that HepG2 cells will produce obvious oil droplets after being treated with oleic acid. Although the amount of fat accumulation in liver cells can be reduced after treatment with curcumin standard and unfermented turmeric aqueous solution, the effect is not as good as the effect of the fermented turmeric aqueous solution disclosed in the present invention on inhibiting the formation of oil droplets in liver cells. The fermented turmeric aqueous solution is significantly better than the unfermented turmeric aqueous solution or the turmeric standard. The above results show that the fermented turmeric produced by the fermentation disclosed in the present invention can indeed effectively enhance the effect of turmeric and its active ingredients in inhibiting oil production, which means that the fermented turmeric disclosed in the present invention has the ability to regulate fat production in liver cells, thereby achieving the effect of inhibiting or slowing down the occurrence of non-alcoholic fatty liver or fat metabolism-related diseases.

[0039] Example 3: Animal Experiment

[0040] Six-week-old C57BL / 6 male black mice were divided into four groups and fed for 17 weeks under the following conditions:

[0041] Group 1: normal diet (5001M feed) and normal drinking water.

[0042] Group 2: 50% high-fat feed and normal drinking water.

[0043] Group 3: 50% high-fat feed, normal drinking water, 5% of feed weight of unfermented turmeric powder and inactivated bacteria (Lacticaseibacillus paracasei MC1-40).

[0044] Group 4: 50% high-fat feed, normal drinking water, 5% of the feed weight of fermented turmeric powder and inactivated bacteria (Lacticaseibacillus paracasei MC1-40), wherein the fermented turmeric powder was prepared by referring to the method disclosed in Example 1.

[0045] During the experiment, the weight changes of mice in each group were recorded. Figure 2 shown.

[0046] Depend on Figure 2 The results showed that the weight of mice in group 2 was significantly higher than that of mice in group 1, indicating that administration of high-fat diet would indeed increase the weight of mice; the weight of mice in group 3 and group 4 decreased compared with that of mice in group 2, among which, the weight gain of mice in group 4 was significantly slower than that of mice in group 3, and the weight of mice in group 4 was almost the same as that of mice in group 1.

[0047] Depend on Figure 2The results show that the fermented turmeric disclosed in the present invention can effectively alleviate obesity caused by a high-fat diet, and its effect of inhibiting weight gain is significantly better than that of unfermented turmeric; in other words, the fermented turmeric disclosed in the present invention can indeed inhibit or slow down the weight gain induced by a high-fat diet in the body, so as to effectively achieve the effect of weight loss and prevention of metabolic-related diseases.

[0048] Example 4: Tissue Section

[0049] After the mice in each group of Example 3 were sacrificed, their liver tissues were taken, paraffin-embedded and sectioned, and then H&E stained. The triglyceride content in the liver of each group was analyzed. The results are as follows: Figure 3A and Figure 3B As shown; the gonadal fat tissue of each group of mice was taken for tissue sectioning and H&E staining. The results are shown in Figure 4 shown.

[0050] Depend on Figure 3A The results show that the cells in the liver tissue of the first group of mice are neatly arranged and there is no pathological phenomenon; the liver tissue of the second group of mice has obvious white fat vacuoles (arrows in the figure), which shows lipid accumulation; the white fat vacuoles in the liver tissue of the third and fourth groups of mice are significantly lower than that of the second group of mice. Figure 3B The results showed that compared with the first group of mice, the triglyceride content in the liver of the second group of mice was significantly increased; while the triglyceride content in the liver of the fourth group of mice was significantly lower than that of the second group of mice. Figure 4 The results showed that the size of gonadal adipocytes in group 2 mice was significantly larger than that in group 1 mice, while the size of gonadal adipocytes in group 4 mice was significantly smaller than those in groups 2 and 3 mice.

[0051] From Figure 3 and Figure 4 The results show that a high-fat diet does cause fat accumulation in the liver and other tissues, leading to individual obesity and the occurrence of lipid metabolism-related diseases such as non-alcoholic fatty liver disease. By administering the fermented turmeric disclosed in the present invention to an individual, the fat accumulation and triglyceride content in the tissues induced by a high-fat diet can be effectively improved, which means that the fermented turmeric disclosed in the present invention can prevent or treat lipid metabolism-related diseases caused by a high-fat diet, such as non-alcoholic fatty liver disease, obesity, etc.

[0052] Example 5: Protein Expression

[0053] After the mice in each group in Example 3 were sacrificed, their liver tissues were taken out, and the expressions of SirT1 and PGC-1α in the liver tissues of the mice in each group were analyzed by Western blot and commercially available ELISA kit. The results are as follows: Figure 5A and Figure 5B shown.

[0054] Depend on Figure 5A and Figure 5B The results showed that the expression of Sirt1 and PGC-1α in the liver tissue of group 4 mice was higher than that of group 2 and group 3 mice. Since Sirt1 is a major regulator of mitochondrial biogenesis, it will promote the expression of PGC-1α involved in energy metabolism. Figure 5A and Figure 5B The results show that the fermented turmeric disclosed in the present invention can promote fatty acid oxidation by enhancing the expression of Sirt1 and PGC-1α, which means that the fermented turmeric disclosed in the present invention can effectively reduce the probability of obesity or related metabolic diseases induced by a high-fat diet.

[0055] Example 6: Analysis of blood sugar regulation ability

[0056] In Example 3, each group of mice was subjected to an oral glucose tolerance test (OGTT) at the 16th week of the experiment. The fasting blood glucose levels of each group of mice were first measured, and then each group of mice was given a glucose solution of 2 g / kg body weight by tube feeding. The blood glucose levels were measured at 30, 60, 90, 120 and 180 minutes after feeding. The results are shown in Table 2. Fig. 6A As shown, the area under the curve of OGTT was analyzed, and the results were as follows Figure 6B shown.

[0057] After the experiment in Example 2 was completed, liver tissues of mice in each group were taken and analyzed by Western blot and commercially available ELISA kit for the expression of proteins such as GluT2 and pAkt / Akt in the liver tissues of mice in each group. The results are as follows: Fig. 7A and Figure 7B shown.

[0058] Depend on Fig. 6A The results showed that at the 0th minute, the fasting blood glucose values ​​of mice in group 2 were higher than those of mice in group 1 and reached statistical significance. At 30 minutes after tube feeding with glucose solution, the blood glucose concentration of mice in group 2 began to rise. At 60, 90 and 120 minutes, the blood glucose concentration of mice in group 2 gradually decreased but was still higher than that of mice in group 1 and reached statistical significance at 90 minutes. The blood glucose concentration of mice in group 4 was lower than that of mice in group 2 at 30 minutes after tube feeding with glucose solution. At 60, 90 and 120 minutes after tube feeding with glucose solution, the blood glucose concentrations of mice in groups 3 and 4 were not significantly different from those of mice in group 2. Figure 6B The results showed that the mice in group 2 had a higher AUC, while the AUC of mice in group 4 was significantly lower than that of mice in group 2.

[0059] Furthermore, by Fig. 7A and Figure 7BThe results show that the fermented turmeric disclosed in the present invention can increase the value of pAkt / Akt and significantly increase the expression of glucose transporter Glut2.

[0060] The results of FIG. 6 and FIG. 7 show that a high-fat diet slows down the rate of blood sugar decrease in an oral glucose tolerance test, but administration of the fermented turmeric disclosed in the present invention can effectively improve the rapid increase in blood sugar within 30 minutes of the diet. In other words, the fermented turmeric disclosed in the present invention has a better ability to metabolize glucose and regulate blood sugar, so as to stabilize the blood sugar induced by a high-fat diet, thereby avoiding the occurrence of diseases related to poor blood sugar metabolism.

Claims

1. A use of fermented turmeric in preparing a weight loss composition, characterized in that: The fermented turmeric is prepared by fermenting turmeric with Lactobacillus casei; Among them, the fermented turmeric contains water-soluble curcuminoids; The Lactobacillus paracasei is classified and named Lacticaseibacillus paracasei; the preservation date is April 28, 2022; the preservation unit is the German Microbiological Culture Collection Center; the address is Germany; the preservation number is: DSM 34254.

2. A use of fermented turmeric in preparing a composition for treating and / or preventing diseases related to metabolic imbalance caused by a high-fat diet, characterized in that: The fermented turmeric is prepared by fermenting turmeric with Lactobacillus casei; Among them, the Lactobacillus paracasei is classified and named Lacticaseibacillus paracasei; the preservation date is April 28, 2022; the preservation unit is the German Microbiological Culture Collection Center; the address is Germany; the preservation number is: DSM 34254; The fermented turmeric contains water-soluble curcuminoids.

3. The use according to claim 2, characterized in that: The disease associated with metabolic imbalance is obesity.

4. The use according to claim 2, characterized in that: The disease associated with metabolic imbalance is non-alcoholic fatty liver disease.

5. The use according to claim 2, characterized in that: The disease associated with metabolic imbalance is hyperglycemia.

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