JPH04208226A - Lipid metabolic improver - Google Patents

Abstract

PURPOSE:To obtain a safe lipid metabolic improver, containing a hot water extraction residue of lactic acid bacilli as an active ingredient, having inhibitory action on blood serum lipid rise, hepatic lipid rise, etc., and useful for preventing arteriosclerosis and other diseases caused by accumulation of cholesterol. CONSTITUTION:A lipid metabolic improver is obtained by culturing lactic acid bacilli [e.g. preferably Lactobacillus.casei YIT9018 strain (FERM P-665)] in a corn steep liquor culture medium composed of 5% glucose and 14% corn steep liquor, providing microbial cells, then suspending the microbial cells in water at 20-80g/l concentration, allowing the suspension to stand at 50-70 deg.C for 1-5hr, keeping the suspension at 100-121 deg.C for 10-30min and using the resultant extraction residue as an active ingredient.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to a lipid metabolism improving agent having an effect of suppressing serum lipid increase, a hepatic lipid increase suppressing effect, a sterol excretion promoting effect, and the like.

[Conventional technology]

In recent years, the mortality rate due to circulatory diseases has been increasing year by year, and when combined with diseases caused by arteriosclerosis such as myocardial infarction and cerebral infarction, they are the leading cause of death among adults.

Although there are various causes of arteriosclerosis, an increase in serum lipids, especially serum cholesterol levels, is considered to be one of the most important risk factors.

Genetic diseases are the primary cause of elevated serum cholesterol levels. In this case, for seriously ill patients, drugs such as cholesterol synthesis inhibitors, nicomol, clofibrate, ion exchange resins, and anabolic steroids are used at the same time as dietary therapy; sex,
It has side effects such as gastrointestinal disorders and carcinogenicity.

Another major cause of elevated serum cholesterol levels is excessive fat intake due to recent changes in dietary habits, and this is becoming more noticeable among young adults. Dietary hypercholesterolemia does not usually lead to severe hypercholesterolemia, but the problem is that cholesterol gradually accumulates in blood vessels from a young age, leading to arteriosclerosis in adulthood. When combined with hypertriglyceridemia, there is a risk of myocardial infarction or cerebral infarction. For this type of hyperlipidemia, dietary therapy that limits fat intake to an appropriate range is more important than drug therapy, which has many problems such as side effects. However, dietary restrictions are difficult to implement strictly because they cause mental pain and take away the enjoyment of eating, and their effectiveness is often limited.

Excessive cholesterol in serum accumulates on the walls of blood vessels, leading to arteriosclerosis, but once cholesterol is taken into the liver, it is converted to bile acids without being accumulated there, or released into the intestinal tract as is. If the blood and feces are excreted from the body, the pool of cholesterol in the body will also be reduced, and arteriosclerosis caused by the above mechanism will be prevented. Therefore,
Although it is expected to have not only the effect of lowering serum cholesterol levels, but also the effect of lowering liver cholesterol levels and the effect of promoting bile acid excretion, to date, there has been no known substance that exhibits these three effects alone.

JP-A-59-80610 discloses that a substance obtained by removing components with a molecular weight of about 3,500 or less from a proteolytic enzyme-treated product of a microorganism of the genus Streptococcus has cholesterol-lowering activity, and JP-A-61-80610 discloses that a substance obtained by removing components with a molecular weight of about 3,500 or less has cholesterol-lowering activity. No. 109729 discloses that microorganisms belonging to the genus Bifidobacterium or Lactobacillus have a cholesterol-lowering effect, and JP-A-62-258323 discloses that microorganisms belonging to the genus Bifidobacterium or Bifidobacterium have a cholesterol-lowering effect. Although it has been described that the microorganisms to which they belong have an effect of suppressing the increase in serum cholesterol, there is no description that they have any more effective effect on improving lipid metabolism.

[Problem to be solved by the invention]

In view of the above-mentioned problems, the purpose of the present invention is to comprehensively treat dietary hypercholesterolemia by suppressing the increase in serum lipids and liver lipids associated with high-fat food intake, and promoting the excretion of sterols into feces. The object of the present invention is to provide a lipid metabolism improving agent that is effective in preventing the effects of cancer and is safe even when ingested for a long period of time.

[Means to solve the problem]

The present invention, which has succeeded in achieving the above objects, uses as an active ingredient a hot water extraction residue of Lactobacillus or a product obtained by further treating the hot water extraction residue with a proteolytic enzyme. The present invention provides a lipid metabolism improving agent that also has effects such as promoting excretion.

I will first explain the method for producing the lipid metabolism improving agent of the present invention;
As the lactic acid bacterium, those belonging to the genus Lactobacillus are suitable, and among them, Lactobacillus casei is preferred. A preferred specific example is Lactobacillus casei strain YI79018 (Feikoken Bacterium No. 665).

The Lactobacillus used in the lipid metabolism improving agent of the present invention may be cultured in any medium by a conventional method, but considering the use, 5% glucose and 14% corn syrup
It is preferable to use a cornstap liquor culture medium consisting of:

Bacterial cells are collected from the culture medium using a conventional method and washed. continue,
The bacterial cells were suspended in water at a concentration of 20 to 80 g/l, and
After being allowed to stand at 70°O for 1-5 hours, it may be held at 100-121°C for 10-30 minutes, but the step of leaving it at 50-70'O can be omitted.

After the treatment, the extraction residue is collected by a conventional method for bacterial cell separation such as centrifugation and freeze-dried. The dried product can be used as it is as the first lipid metabolism improving agent of the present invention.

If this is further treated with proteolytic enzymes,
The cells are treated with a proteolytic enzyme capable of decomposing cell proteins, such as actinase or trypsin, for an appropriate period of time until most of the cell proteins are hydrolyzed, and then fractionated based on molecular weight to remove protein hydrolysates. Fractionation is carried out by ultrafiltration, dialysis, which can remove fractions containing protein decomposition products with a molecular weight of 50,000 or less.
This is carried out using means such as molecular sieves. The high molecular weight fraction containing the active ingredient is concentrated under reduced pressure and then lyophilized for use in the second lipid metabolism improving agent of the present invention.

The lipid metabolism improving agent according to the present invention not only suppresses the rise in serum cholesterol levels but also suppresses the rise in liver lipids,
In addition, since it promotes the excretion of sterol into feces, it is extremely effective in preventing diseases caused by cholesterol accumulation, such as arteriosclerosis. Proteolytic enzyme-treated products are particularly excellent in these effects.

The lipid metabolism improving agent of the present invention can be orally administered as it is or mixed with excipients and other auxiliary agents commonly used in pharmaceutical manufacturing. Since it is mostly tasteless and odorless, it can be added to any food or drink and administered on a daily basis.

〔Example〕

Production Example 1 Lactobacillus casei YIT9018 was inoculated into corn staple liquor medium 400a and cultured at 35°C for 2 hours while controlling the medium pH to 6.0. after that,
The bacterial cells were collected by centrifugation, washed, and dried as 2
．． 4 kg of bacterial cells were obtained.

The bacterial cells were suspended in 481i water and left at 55°C for 2 hours while controlling the pH to 7. Then 100°C
The mixture was heated for 10 minutes to elute soluble components. The soluble fraction was removed by centrifugation, and the bacterial cell hot water extraction residue (hereinafter referred to as
1.5 kg (dry weight) of formulation A) was obtained.

Production Example 2 300 g of the bacterial cell hot water extraction residue obtained in Production Example 1 was
/15M phosphate buffer (pH 11,6) 3Q and homogenized using a high-pressure homogenizer, then 12g of protease actinase (Kaken Pharmaceutical Co., Ltd.) was added and the mixture was heated at 50°C for 16 hours. Made it react. Note that toluene was added as a preservative to prevent spoilage during the reaction.

After the reaction is completed, fractions with a molecular weight of 50,000 or less are removed by ultrafiltration, and then freeze-dried to obtain a dried product (hereinafter referred to as Formulation B) 1.
00g was obtained.

Test Example 1 Four-week-old male SD rats were divided into three groups of six animals per group, and each group was allowed to freely consume the feed shown in Table 1, and kept for two weeks. Note that all rats were previously fed with a standard diet for a control group for one week.

Table 1 Control group ■ Control group ■ Test group feed ingredients (standard diet) (high-fat diet) (formulation A additive diet)
Casein 20 20 20 Lard 99 Soybean oil 1011 Mineral mixture 4 4 4 Vitamin mixture 1 1 1 Choline chloride
0.15 0.15 0.15 Cholesterol -0,50,5 Na Cholate O,1250,12
5 Filter paper powder 5 5 1 Formulation A4 Sucrose 59.85 59.225 5
9.225 total 100 100
1oO (Note) Mineral mixture, vitamin mixture: For the Herb mixture group, the feed intake for the next two weeks was 235 ± 1 for the control group.
8g1 control group■ 233±22g1 test group 247±2
5g, and there was no difference between the groups. Thereafter, after fasting for 18 hours, blood was collected from the abdominal aorta under anesthesia. After blood collection, the liver was removed while being perfused with physiological saline, immediately cooled on ice, weighed, and stored frozen.

In addition, feces were collected for 2 days before fasting.

The collected blood was centrifuged to collect serum, and serum lipids were analyzed. A dried liver sample was extracted with a chloroform-methanol mixture (2:1), and liver lipids were analyzed. After drying, the feces were extracted with hot ethanol and the extract was analyzed.

The analysis results are as shown in Tables 2 to 4, and in the control group ■, total cholesterol levels and arteriosclerotic index increased by feeding a high-fat diet supplemented with cholesterol; Even on a high-fat diet, in the test group to which Formulation A was added, increases in numerical values in all of the above analytical items were significantly suppressed. Furthermore, serum triglyceride values also tended to decrease in the test group.

Table 2 Serum lipids Note: Values are standard deviation above the mean. There is a significant difference between questions a and b and c and d at a risk rate of 5% (the same applies to each table below).

Table 3 Liver Lipid Table 4 Fecal Lipid Test Example 2 A test similar to Test Example 1 was conducted using 1 group of 8 4-week-old male SD rats. However, Formulation B according to Production Example 2 was used instead of Formulation A as the lipid metabolism improving agent added to the feed of the test group.

The feed intake for the next two weeks was 232 ± 24 g for the standard diet control group (■) and 229 ± 22 g for the high-fat diet control group (■).
It was 239±16 g in the g1 test group, and there was no difference between each group.

Analysis results of serum lipids, liver lipids, and fecal lipids are shown in Table 5.
As shown in 7, the total cholesterol level, VLDL + LDL cholesterol level, and arteriosclerotic index increased in the control group ■ by feeding a high-fat diet with added cholesterol, but even with the same high-fat diet, formulation B increased. In the test group, increases in all of the above analysis items were significantly suppressed. Furthermore, serum triglyceride values also tended to slightly decrease in the test group.

Table 5 Serum Lipids Table 6 Liver Lipids Table 7 Fecal Lipids [Effects of the Invention] As mentioned above, the lipid metabolism improving agent of the present invention, which is composed of a hot water extract of Lactobacillus or a protease-treated product thereof, significantly improves serum lipids. It exhibits an effect of suppressing the rise in cholesterol levels, an effect of suppressing the rise of liver lipids, and an effect of promoting the excretion of cholesterol and bile acids into the feces, and is extremely effective in preventing arteriosclerosis and other diseases caused by cholesterol accumulation.

Moreover, Lactobacillus is a non-pathogenic and safe bacterium, as is clear from the fact that it has been used for a long time in the production of lactic acid fermented foods. No deaths were observed even at a dose of 8 g/kg, and there is no safety problem even when administered for a long period of time.

Therefore, the lipid metabolism improving agent of the present invention is extremely advantageous, as it is suitable for use as an orally administered medicine, and also for daily intake by mixing with food to help prevent arteriosclerosis and promote health.

Claims (3)

[Claims] (1) A lipid metabolism improving agent containing a hot water extraction residue of Lactobacillus as an active ingredient. (2) A lipid metabolism improving agent containing a lactic acid bacterium hot water extraction residue treated with a proteolytic enzyme as an active ingredient. (3) The lipid metabolism improving agent according to claim 1 or 2, wherein the lactic acid bacterium is Lactobacillus casei.