WO2019074213A1 - Lactobacillus paracasei lm1010 strain and composition for prevention or treatment of obesity or diabetes, comprising same - Google Patents

Abstract

The present invention relates to Lactobacillus paracasei LM1010 (accession number KCCM12309P) and a composition for prevention and treatment of obesity or diabetes, comprising the same. More specifically, the Lactobacillus paracasei LM1010 strain of the present invention reduces not only adipocytes, triglycerides, subcutaneous fat, and cholesterol, but also regulates the gene expression of an obesity-related biomarker, thus exhibiting the effect of preventing and treating obesity. Concurrently, the strain has the effect of preventing and treating diabetes by reducing blood glucose levels and insulin levels to restrain insulin resistance.

Description

Lactobacillus paracasei LM1010 strain and a composition for preventing and treating obesity or diabetes comprising the same

The present invention relates to a Lactobacillus casei para LM1010 (Lactobacillus paracasei LM1010) strain and the prevention and treatment of obesity or diabetes in the composition comprising the same. Specifically, the Lactobacillus paracase LM1010 strain of the present invention reduces fat cells, triglycerides, subcutaneous fat and cholesterol, thereby preventing and treating obesity, and at the same time decreasing blood glucose concentration and insulin concentration It has the effect of preventing and treating diabetes by inhibiting insulin resistance. In addition, Lactobacillus para casei LM1010 (Lactobacillus paracasei LM1010) is PPARγ, C / EBPα, FAS, etc. inhibit the expression of a biomarker associated with local differentiation and appetite regulating hormone leptin, obesity-related gene SCD1, FAS, expression of SREBP1 .

Recently, as the lifestyle such as diet and eating habits are rapidly westernized due to the improvement of income level and industrial development, chronic diseases and adult diseases are rapidly increasing. One of the causes is known as obesity.

Obesity is an energy metabolic disorder caused by an imbalance between energy intake and energy consumption, and is consequently defined as an accumulation of excess fat in fat cells.

Obesity is a chronic disease that is a problem all over the world. Obesity, unlike other diseases, is characterized not only by external problems but also by related diseases such as metabolic diseases, hypertension, diabetes, hyperlipidemia, arteriosclerosis, ischemic heart disease, fatty liver, gallstone disease and the like.

Although obesity is a serious disease in itself, it also causes cosmetic problems, so many obesity remedies have been developed in various countries around the world in an effort to develop various obesity remedies. So far, obesity remedies have been divided into obesity treatment of obesity-depressant type and obesity treatment of lipolysis inhibitor-type.

Although the obesity treatment agents of the appetite-decreasing agent family have an advantage of being excellent in the weight-reducing effect, the obesity treatment agents of the appetite-decreasing agent series have an action mechanism to reduce the food intake by acting on the central nervous system, There is a crucial problem that the effect does not persist or causes serious side effects in long-term use.

Fenfluramine, sibutramine and rimonabant have been commercialized as an early appetite suppressant, but they have been discontinued due to serious mental illness problems such as heart disease, hypertension, increased heart attack, depression and suicide.

Orlistat (brand name: Xenical), a lipolytic inhibitor, is inactivated by irreversible binding to pancreatic lipase, a lipolytic enzyme of the pancreas that breaks down triglycerides, thereby reducing the absorption of triglycerides and cholesterol, It acts as an anti-obesity mechanism.

Therefore, taking orlistat reduces fat breakdown, so the fat consumed as a food is not absorbed into the body and is discharged, reducing the amount of fat absorbed into the body and reducing weight. However, in spite of this efficacy, orlistat has side effects such as abdominal pain, diarrhea, inhibition of absorption of fat soluble vitamin, and it is reported that severe liver damage occurs when taking organs for a long time, and discussions on the safety of orlistat are underway.

Since most of the currently marketed drugs for treatment of obesity have serious side effects, the social demand for the treatment of obesity that can effectively treat obesity without side effects is very high. Therefore, various studies are being carried out in order to develop a therapeutic agent for obesity which can treat obesity without side effects in various countries around the world. However, such a therapeutic agent for obesity has not been developed and marketed to date.

On the other hand, there is a possibility of using the lactic acid bacteria having excellent safety as a therapeutic agent because they are not absorbed into the body and exist in symbiosis with the human body while staying in the intestinal space.

Lactobacillus acts on the digestive system of the human body to decompose fibrous and complex proteins into important nutrients. As such, probiotics are collectively referred to as living microorganisms that improve the intestinal microbiological environment of a host in the gastrointestinal tract of an animal, including humans, and have a beneficial effect on the health of the host.

Probiotics are recognized as having the ability to regulate metabolism and immunological function. Representative probiotics include Lactobacillus sp. And Lactococcus sp. Strains. Among them, Lactobacillus sp. Strains have been known to play an important role in maintaining a healthy digestive organs and vaginal environment as a major member of normal microbial communities in the intestines of the human body.

In this regard, Korean Patent Registration No. 1679045 (published on November 23, 2016) discloses LMT1-30 strains of Lactobacillus paracasei strains effective for the treatment of obesity and Korean Patent Publication No. 2015-0118084 Discloses a composition for improving obesity comprising Lactobacillus paracasei, and Korean Patent Laid-Open Publication No. 2016-0098955 (published on Aug. 18, 2016) discloses a composition for improving obesity comprising lactobacillus paracase Although the JBD305 strain has been disclosed, no strain of Lactobacillus paracase has been disclosed which is effective in inhibiting adipocyte differentiation, inhibiting appetite, and improving insulin resistance.

Accordingly, the present inventors have found that Lactobacillus paracasei LM1010 inhibits mRNA and protein expression of genes involved in the induction of adipocyte differentiation, thereby ultimately inhibiting adipocyte differentiation and fat accumulation, as well as inhibiting appetite And insulin resistance at the same time, thereby completing the present invention.

The inventors of the present invention have made extensive studies to discover probiotics having an excellent anti-obesity effect, and as a result, confirmed that the Lactobacillus paracasei LM1010 strain has an excellent anti-obesity effect.

Surprisingly, the strain LM1010 not only attains inhibition of adipocyte differentiation, inhibits fat accumulation and regulates appetite, but also has a remarkable effect of reducing insulin resistance.

The inventors of the present invention found that Lactobacillus paracase LM1010 inhibits adipocyte differentiation and inhibits fat accumulation as well as reduces appetite and reduces the efficiency of diets, Prevention and treatment.

In addition, the present inventors have confirmed that the Lactobacillus paracase LM1010 strain decreases blood glucose concentration and insulin concentration, thereby reducing insulin resistance that can occur with obesity.

In the present invention, the Lactobacillus paracase LM1010 strain inhibits differentiation into adipocytes and fatty accumulation in the lipid precursor cells, and reduces various kinds of action mechanisms, enzymes and hormones such as body weight, abdominal fat and blood cholesterol And gene expression not only in obesity but also in the treatment and prevention of hyperlipidemia, metabolic diseases and diabetes.

The present invention also provides a composition for preventing and treating Lactobacillus paracase LM1010 and obesity, overweight, hyperlipidemia, metabolic diseases and diabetes comprising the same.

The present invention relates to a Lactobacillus paracasei LM1010 deposited with the Korean Culture Center of Microorganisms (KCCM) under accession number KCCM 12309P.

The Lactobacillus paracase LM1010 of the present invention was deposited in Korea on September 13, 2017 as KFCC11726P at the Korean Microorganism Conservation Center and was deposited on KCCM 12309P at the Korean Microorganism Conservation Center on September 05, 2018 It was an international deposit.

The present invention relates to a Lactobacillus paracasei LM1010 strain (KCCM12309P) having an efficacy for preventing or treating obesity and diabetes at the same time.

The present invention relates to a pharmaceutical composition for preventing or treating obesity or diabetes, which comprises Lactobacillus paracase LM1010 strain (KCCM12309P), wherein said Lactobacillus paracase LM1010 strain simultaneously exhibits lipolysis inhibition and an appetite regulating effect in a small intestine cell or digestive tract Or pharmaceutical compositions for therapeutic use.

At this time, the lipolysis inhibition can be achieved by inhibiting the gene expression of the obesity-related biomarker, and the appetite regulation can be achieved by decreasing the secretion of leptin hormone.

In addition, the pharmaceutical composition can prevent or treat diabetes due to obesity or obesity by reducing the insulin resistance of the Lactobacillus paracase strain LM1010.

The present invention is directed to a method of treating obesity or diabetes comprising administering to a subject in need of treatment of obesity or diabetes an effective amount of a Lactobacillus paracase strain LM1010.

At this time, the Lactobacillus paracase LM1010 strain can simultaneously exhibit the lipolysis inhibitory effect and the appetite regulating effect in the small intestine cell or digestive tract to be administered.

The lipolysis inhibition can also be achieved by inhibiting the gene expression of an obesity-related biomarker, and the appetite regulation can be achieved by decreasing the secretion of leptin hormone.

In addition, the Lactobacillus paracase strain LM1010 can reduce insulin resistance in the subject to be administered.

The present invention relates to a pharmaceutical composition for preventing or treating obesity or diabetes, which comprises Lactobacillus paracase LM1010 strain (KCCM12309P), wherein said Lactobacillus paracase LM1010 strain simultaneously exhibits a lipolysis inhibitory effect and an appetite control effect in a small intestine cell or digestive tract Prevention or amelioration, wherein the food composition may be a health functional food, a dairy product, a fermented product or a food additive.

The present invention relates to a pharmaceutical composition for preventing or treating obesity or diabetes, which comprises Lactobacillus paracase LM1010 strain (KCCM12309P), wherein said Lactobacillus paracase LM1010 strain simultaneously exhibits a lipolysis inhibitory effect and an appetite control effect in a small intestine cell or digestive tract Prevention or amelioration of an animal feed composition.

The Lactobacillus paracase LM1010 strain of the present invention inhibits adipocyte differentiation and intracellular fat accumulation, and has an excellent anti-obesity effect.

Further, the Lactobacillus paracase LM1010 strain of the present invention is effective for preventing or treating metabolic diseases caused by obesity or obesity and hyperlipemia due to reduction of body fat amount, decrease of total cholesterol, decrease of plasma triglyceride and liver triglyceride .

In addition, the Lactobacillus paracase LM1010 strain of the present invention improves insulin resistance through reduction of glucose and insulin concentrations in blood, thereby exhibiting the preventive or therapeutic effect of diabetes caused by obesity and obesity.

In addition, the Lactobacillus paracase LM1010 strain of the present invention suppresses the appetite by reducing the synthesis of leptin, which is an appetite regulating hormone, and inhibits the synthesis of Stearoyl-CoA desaturase-1 (SCD1) It inhibits gene expression of the enzyme Fatty-acid Synthase (FAS) and Sterol regulatory element-binding transcription factor 1 (SREBP1), a binding protein enzyme that regulates lipid synthesis, resulting in regulation of various enzymes and hormones Thereby preventing or treating the disease.

FIG. 1 shows the results of staining of differentiated adipocytes with Oil-red-O by treating LM1010 culture extract 1%, 10% treated group and control PBS with 3T3-L1 lipid precursor cells, Represents the number of spheres (arrows).

Fig. 2 shows the triglyceride in 3T3-L1 adipocytes of day 9 differentiation treated with LM1010 cultured extract 1% treated group, 10% treated group and control PBS, respectively.

FIG. 3 is a real-time PCT result of the cDNA obtained using the cDNA Synthesis Kit (TAKARA, 6110A) on the RNA extracted from the lipid precursor cells in which the differentiation was completed. As a control group, PBS treated group and Lactobacillus paracase LM1010 strain And the relative mRNA expression of the obesity-related biomarker in the 10% treated group.

FIG. 4 shows a comparison of weight gain rates of the first to third groups.

FIG. 5 shows the number of adipose tissues and adipocyte size stained with H & E stain in each group in epididymal fat.

Figure 6 shows the weights of liver, spleen, and kidneys in each group.

FIG. 7 shows the results of observing the presence or absence of macro fat vacuole in 20% (A, B, C) and 40 magnifications (D, E, F) by staining liver with H & F stain.

FIG. 8 shows the results of measurement of expression of SCD1, FAS, and SREBP1 genes in liver of each group.

Obesity is a metabolic disorder caused by an imbalance between the consumption of food and energy consumption, which means that the body is over-accumulated. In addition, obesity is closely related to insulin resistance, glucose tolerance, hyperlipidemia, and metabolic diseases. Therefore, due to obesity, hyperlipemia, metabolic diseases and diabetes can occur.

Hyperlipidemia is a state in which many lipid substances are present in the blood, accumulating in the blood vessels and causing inflammation, resulting in cardiovascular diseases. Hyperlipidemia is often caused by increased lipid levels in the blood due to genetic factors. ≪ / RTI > and other causes such as diabetes and the like.

Metabolic diseases consist of many metabolic diseases, many of which promote the onset of atherosclerosis and increase the risk of developing cardiovascular disease. Although the mechanism of action of metabolic diseases is not fully understood, most patients with metabolic diseases have increased insulin levels and have insulin resistance.

Diabetes is a type of metabolic disease that occurs due to abnormality in glucose metabolism. It is characterized by hyperglycemia with elevated blood glucose concentration, which causes various symptoms and signs and excretes glucose from the urine. Although it is known that 30 ~ 70% of diabetic outcomes are determined by genetic influences, the rapid increase in the number of type 2 diabetes recently in Korea is closely related to the westernization of lifestyle and the increase of obese patients .

The present invention relates to a composition for preventing or treating obesity or obesity-induced diseases, characterized in that the LM1010 strain of Lactobacillus paracase simultaneously inhibits the differentiation into adipocytes and the effect of controlling appetite.

The 'inhibition of differentiation into adipocytes' can be achieved by inhibiting the expression of genes such as PPARγ, C / EBPα, and FAS that are involved in the differentiation process into adipocytes.

PPARγ and C / EBPα induce lipid differentiation as a transcription factor associated with energy metabolism, and FAS plays an important role in the synthesis of unsaturated fatty acids and the uptake of fatty acids.

The 'appetite control' can be achieved by decreasing the secretion of leptin hormone.

The present invention also relates to a composition for preventing or treating diabetes caused by obesity or obesity, wherein the LM1010 strain of Lactobacillus paracase is reduced in insulin resistance.

Insulin resistance refers to the inability of the cells to burn glucose effectively because of insulin's ability to lower blood sugar, and obesity directly affects insulin resistance. As insulin resistance increases, glucose production in the liver is not regulated, glucose is not used in the muscle, and blood sugar is converted to fat in the body, which prevents accumulation. If the insulin resistance is high, the human body produces a large amount of insulin, which can lead to hypertension, hyperlipidemia, diabetes and the like.

Insulin resistance is produced by genetic reasons and various environmental factors, and environmental factors include lack of exercise, obesity and excessive calorie intake.

Insulin is responsible for glucose tolerance, and if insulin does not function, glucose tolerance occurs and impaired glucose tolerance occurs. This is considered to be the entire stage of diabetes, and a decrease in tolerance means a disorder of insulin secretion or function or an excess of antagonistic hormone.

The composition of the present invention can be used in medicines, health functional foods, dairy products, fermented products, food additives, animal feeds and the like.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are for further illustrating the present invention, and the scope of the present invention is not limited by these examples.

[Example 1]

Inhibition of adipocyte differentiation and fat accumulation inhibition in 3T3-L1 adipocytes

In order to confirm that the Lactobacillus paracase LM1010 strain deposited with the Korean Culture Center of Microorganisms (KCCM) under the accession number KCCM12309P inhibits adipocyte differentiation and inhibits intracellular fat accumulation in lipid precursor cells, The following experiment was conducted.

First, 3T3-L1 adipose precursor cells were treated with LM1010 culture extracts 1% and 10%, and PBS as a control, respectively. The differentiated adipose cells were stained with oil-red-O, and after 1 hour, they were stained with a microscope The cells were observed.

As shown in FIG. 1, it was confirmed that the number of lipids (arrows) in the adipocytes significantly decreased in the treatment group treated with the LM1010 culture extract, as compared with the control group supplemented with PBS instead of the LM1010 culture extract. The same experiment was repeated three times.

In particular, in the 10% LM1010 treated group, the number of adipocytes and lipids was significantly reduced compared with the 1% LM1010 treated group, and the adipocyte differentiation was apparently varied depending on the concentration.

Therefore, the LM1010 culture extract has an effect of inhibiting the differentiation of adipocytes and the accumulation of intracellular fat, and the higher the content of the extract, the more excellent the effect is.

[Example 2]

3T3-L1 triglyceride degradation promotion and accumulation inhibitory action in adipocytes

To measure the amount of intracellular triglycerides (TG, triglyceride) accumulated in the differentiation process of adipocytes, 1% and 10% of the extracts of LM1010 strain were added and cultured, and PBS was used instead of the strain extract as a control And cultured. Adipose cells on day 9 of differentiation were harvested and used.

The harvested cells were washed three times with PBS, disrupted, centrifuged at 15,000 rpm at 4 ° C, and the supernatant was used as a sample. 10 μL of the sample and 150 μL of the enzyme reagent were added, and the reaction was allowed to proceed at room temperature for 15 minutes, and the absorbance was measured at 530 nm.

The results of measurement of triglyceride in adipocytes are shown in Table 1 and FIG.

	Control group		LM1010 1% treated group		LM1010 10% treated group
Triglyceride (mg / dL)	42.68	38.45	5.75
Decrease relative to control (%)	0	14.1	86.61

As shown in Table 1, in the 1% LM1010 treated group and the 10% treated group, the neutral fat reduction was 14.1% and 86.61%, respectively, as compared with the control group.

Therefore, it can be seen that the LM1010 culture extract promotes the decomposition of triglyceride in adipocytes and inhibits the accumulation.

[Example 3]

Inhibitory effect of 3T3-L1 adipocyte-associated biomarkers on gene expression

Real-time PCR was performed to determine whether Lactobacillus paracase inhibits the expression of representative biomarkers involved in the differentiation of adipocytes into adipocytes.

The lipid precursor cells that had been differentiated were harvested with trizol, extracted with RNA, and cDNA complementary to RNA was obtained using cDNA Synthesis Kit (TAKARA, 6110A). Real-time PCR was performed using the synthesized cDNA. The results are shown in FIG.

FIG. 3 compares the relative mRNA expression of obesity-related biomarkers in a group treated with PBS as a control group and a group treated with 10% Lactobacillus paracase LM1010 strain.

In Table 2, PPARγ, C / EBPα, and FAS genes involved in the differentiation of 3T3-L1 into adipocytes were used as biomarkers and GAPDH as an internal control.

PPARγ and C / EBPα induce lipid differentiation as a transcription factor associated with energy metabolism, and FAS plays a role in increasing unsaturated fatty acid synthesis and fatty acid influx.

role	gene	Accession No.
	Glyceraldehyde 3-phosphate dehydrogenase (GAPDH)	BC083080
Fat synthesis	Fatty acid synthase (FAS)	NM_007988
Energy metabolism regulation	Peroxisome proliferator activated receptor gamma 2 (PPARgamma)	NM_011146
	CCAAT / enhancer binding protein alpha (C / EBPa)	BC058161

As can be seen in FIG. 3, when Lactobacillus paracase LM1010 was treated with 3T3-L1 adipocyte differentiation, PPARγ, C / EBPα, and FAS expression levels were 66.6%, 81.75% 72.89%. ≪ / RTI > As a result, it was confirmed that the LM1010 strain of Lactobacillus paracase inhibited the expression of biomarkers related to lipid differentiation, and it was confirmed that the lipid differentiation was inhibited by the lipid.

[Example 4]

Experiment of anti-obesity effect in experimental animals

In order to confirm the anti-obesity effect in mice, 5-week-old C57BL6 / J mice were weighed for 1 week and divided into 3 groups.

(HFD, 5.2 kcal / g) in group 2, high fat diet (HFD) and high fat diet (HFD) in group 1, LM1010 (10 ⁹ CFU / mouse) was administered orally five times a week. Weights were measured weekly and the amount of diets fed was analyzed. At week 9, the major organs were extracted and weighed.

Decrease in weight gain

0 Weight of the body weight was measured at 100% with respect to time, and is shown in FIG. As shown in FIG. 4, in the third group (HFD and LM1010 administration group), there was a difference in the weight gain rate from the first group to the second group (HFD).

At week 7, the weight gain of the third group was 151.06%, the weight gain of the second group was 159.44%, and the weight gain of the third group was 7% lower than that of the second group. In the 8th week, the weight gain rate of the third group was 153.82%, compared with 7th week, the weight increase rate of the second group was 171.46% . Particularly, at week 8, weight gain in group 3 decreased by 17% more than group 2.

As shown in Table 3, the weight gain of the third group was 11.42 and the weight gain of the second group was 14.84, which was about 20% or more.

As a result, compared with group 2, which received high-fat diet, and group 3, which received LM1010 concurrently with high fat diet, the difference in body weight gain started to change from 1 week later, .

	Group 1	Group 2	Group 3
7 Weight gain (%)	124.82	159.44	151.06
8 Weight gain (%)	123.92	171.46	153.82
Weight gain (g) for 8 weeks	4.86	14.84	11.42

Reduction of food efficiency

Weight gain, weekly dietary intake, calorie intake, and diet efficiency for 8 weeks are shown in Table 4 below.

	Group 1	Group 2	Group 3
Weight gain (g)	4.86	14.84	11.42
Dietary intake (g / mouse per week)	18.38	16.61	14.13
Calorie intake (kcal / mouse per week)	56.99	87.05	74.03
Eating efficiency (weight gain / dietary intake)	0.03	0.11	0.10

As shown in Table 4, when the dietary intake per calorie and the calorie intake were compared, it was confirmed that the third group was decreased as compared with the second group. As a result of calculating the dietary efficiency by weight gain and dietary intake, it was 0.11 in the second group, but 0.10 in the third group, indicating that the weight gain was significantly lower than the dietary intake.

Reduction of subcutaneous, epididymal and brown adipose tissue

The amount of subcutaneous fat, epididymal fat and brown adipose tissue for 8 weeks was measured and shown in Table 5 below.

	Group 1	Group 2	Group 3
Subcutaneous fat (g)	0.35	1.296	0.828
Epididymis fat (g)	0.55	2.28	1.577
Brown adipose tissue (g)	0.062	0.144	0.082

As shown in Table 5 and FIG. 5, it was confirmed that the body fat of the third group was significantly reduced compared with the body fat of the second group.

In addition, epididymal fat was stained with H & E stain, and the results are shown in FIG. As shown in FIG. 5, the size of adipocytes was the largest in the second group, and the size of the adipocytes in the third group was reduced to less than half of that of the second group.

Weight loss of soy sauce

The weights of liver, spleen and kidney for 8 weeks were measured and are shown in Table 6 below.

	Group 1	Group 2	Group 3
Weight of soy sauce (g)	1.12	1.508	1.078
Weight of spleen (g)	0.072	0.076	0.070
Height of the kidney (g)	0.3	0.34	0.330

As shown in Table 6, the weight of the liver of the third group was significantly decreased as compared with that of the second group. The spleen and kidney weights did not differ significantly in each group.

In the case of obesity, it is known that fat metabolism in the body occurs abnormally and lipid component accumulates in the liver, thus increasing liver weight. As shown in Table 6 and FIG. 6, in the group administered with LM1010 strain (Group 3), liver weight was reduced to a normal level, and LM1010 strain was involved in lipid metabolism in the liver and inhibited fatty accumulation of hepatocytes It can be seen that the effect is.

Fatty liver reduction

The liver was stained with H & E stain and the presence or absence of macro fat vacuole (X20 magnification and X40 magnification) was observed and is shown in FIG.

As shown in FIG. 7, macro fat vacuole was observed in the liver tissues of the second group (arrow), and macro fat vacuole was not observed in the liver tissues of the first and third groups.

As a result, it can be seen that the LM1010 strain has a fatty liver inhibiting effect.

Reduce blood glucose, triglycerides and cholesterol levels

8 weeks of blood glucose, triglyceride and cholesterol levels were measured and are shown in Table 7 below.

	Group 1	Group 2	Group 3
Glucose (mg / dL)	228.7	320.1	243.8
Triglyceride (mg / dL)	147	165	83
Cholesterol (mg / dL)	136	218	176

In the case of obesity, insulin resistance occurs, resulting in elevated blood insulin levels, resulting in a temporary increase in glucose levels due to metabolism. As shown in Table 7, the blood glucose level of the third group was lower than that of the second group, indicating that the LM1010 strain also had a positive effect on insulin resistance.

In addition, blood triglyceride and cholesterol levels were found to be lower in the third group than in the second group, and the triglyceride levels were lower than those in the first group without the high fat diet.

Reduced levels of insulin and leptin

The levels of insulin and leptin for 8 weeks were measured and are shown in Table 8 below.

	Group 1	Group 2	Group 3
Insulin (ng / ml)	≪ 0.1	1.00	0.63
Leptin (ng / ml)	0.43	1.74	1.18

As mentioned above, in the case of obesity, the insulin resistance is increased and the insulin secretion is increased, and the insulin concentration in the third group is remarkably decreased compared with the insulin concentration in the second group.

Thus, it can be seen that the LM1010 strain has a remarkable effect on the improvement of insulin resistance through reduction of blood glucose concentration and insulin concentration.

In addition, leptin is an appetite-regulating hormone secreted from adipocytes. When fat cells accumulate in adipose tissue and the size of adipocytes increases, the synthesis of leptin is promoted and the concentration of leptin increases.

In the experiment on the concentration of the blood triglyceride, the concentration of triglyceride in the third group was significantly lower than that of the second group. In other words, it was found that the concentration of leptin in the third group was significantly lower than that of the second group because of the decrease in triglyceride and fat cells in the second group.

Obesity related gene expression changes

Liver fat was extracted from mice fed with LM1010 strain for 8 weeks and RNA was extracted using Trizol (Thermo scientific, USA). Subsequently, DNA complementary to RNA was obtained using the PrimeScript ^™ 1 ^st strand cDNA synthesis kit (TAKARA, Japan), and the expression of SCD1, FAS, and SREBP1 genes related to lipid oxidation using SYBR green (TAKARA, Japan) Were confirmed by real-time PCR.

Stearoyl-CoA desaturase-1 (SCD1) is an enzyme that mediates the synthesis of monounsaturated fatty acids. It is highly expressed in obese individuals and is closely related to dysfunction of fat metabolism in genetically obese or type 2 diabetic patients .

Fatty-acid Synthase (FAS) is a fatty acid synthase that is essential for the production of intracellular fat. The increase in the expression of this gene can be interpreted as an increase in intracellular lipid synthesis.

Sterol regulatory element-binding transcription factor 1 (SREBP1) is a binding protein enzyme that regulates lipid synthesis. It is a gene that affects lipid synthesis, along with the two genes listed above. .

Table 9 below shows the primers used in the experiment, and the mRNA expression of the 36B4 gene is an internal control.

number	gene	primer	sequence
One	36B4	Forward	CGT CCT CGT TGG AGT GACE
		Reverse	CGG TGC GTC AGG GAT TG
2	SCD1	Forward	TGG GTT GGC TGC TTG TG
		Reverse	GCG TGG GCA GGA TGA AG
3	FAS	Forward	GCC CAG CTT CCA TCT TTA CT
		Reverse	CAG GAT GTT GTG GTT TAT CCG C
4	SREBP1C	Forward	TAG TCC GAA GCC GGG TGG GCG CCG GCG CCA T
		Reverse	GAT GTC GTT CAA AAC CGC TGT GTG TCC AGT TC

As shown in FIG. 8, expression of SCD1, FAS, and SREBP1 genes in liver was examined. As a result, expression level was decreased in the third group as compared with the second group. The average value of 2 ^ -ddCt is shown in Table 10 using 36B4.

	Group 1	Group 2	Group 3
SCD1	One	2.238	1.134
FAS	One	1.779	1.084
SREBP1	One	2.052	1.056

Thus, it can be seen that the strain LM1010 has an effect of inhibiting the expression of an obesity-inducing gene.

[Amended by Rule 26, 27.09.2018]

Claims (9)

1. Lactobacillus paracasei LM1010 strain (KCCM12309P).
2. Lactobacillus paracase LM1010 strain (KCCM12309P) which has the efficacy to prevent or treat obesity and diabetes at the same time.
3. A prophylactic or therapeutic agent for obesity or diabetes, comprising Lactobacillus paracasei LM1010 strain (KCCM12309P), wherein said Lactobacillus paracase LM1010 strain simultaneously exhibits lipolysis inhibition and an appetite regulating effect in a small intestine cell or digestive tract A pharmaceutical composition.
4. The pharmaceutical composition for preventing or treating obesity or diabetes according to claim 3, wherein the lipolysis inhibition is achieved by inhibiting gene expression of an obesity-related biomarker.
5. The pharmaceutical composition for preventing or treating obesity or diabetes according to claim 3, wherein the appetite control is achieved by reducing the secretion of leptin hormone.
6. The pharmaceutical composition according to claim 3, wherein the Lactobacillus paracase LM1010 strain reduces insulin resistance. The pharmaceutical composition for preventing or treating diabetes caused by obesity or obesity.
7. A method for preventing or improving obesity or diabetes, which comprises Lactobacillus paracasei LM1010 strain (KCCM12309P), wherein said Lactobacillus paracase LM1010 strain simultaneously exhibits lipolysis inhibition and an appetite control effect in a small intestine cell or digestive tract Food composition.
8. The food composition according to claim 7, wherein the food composition is a health functional food, a dairy product, a fermented product, or a food additive.
9. (KCCM12309P), wherein said Lactobacillus paracase LM1010 strain simultaneously exhibits lipolysis inhibition and an appetite regulating effect in a small intestine cell or digestive tract, wherein said Lactobacillus paracase LM1010 strain ≪ / RTI >

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