阅读说明：本技术 一种干预慢性肾炎及尿毒症的长双歧杆菌膳食组合及应用 (Bifidobacterium longum dietary composition for intervening chronic nephritis and uremia and application thereof ) 是由 宋锦安 于 2021-01-18 设计创作，主要内容包括：本发明涉及保健食品领域,具体的讲是一种干预慢性肾炎及尿毒症的长双歧杆菌膳食组合及应用,包括100mL长双歧杆菌BL-G301益生菌牛乳和40g谷物粉压片,制备方法为将-80℃冷冻保藏的益生菌菌粉,取出,温度至室温后,将益生菌菌粉加至巴氏灭菌牛乳中；将长双歧杆菌BL-G301益生菌牛乳的制备方法制备获得物和谷物粉压片一同作为膳食搭配,改善人体肠道内的氧化应激,降低人体内炎症因子和尿毒素水平。本发明与现有技术相比,采用益生菌干预慢性肾炎,可以降低慢性肾炎患者氧化应激水平；改善慢性肾炎患者炎症因子水平和尿毒素水平；改善粪便pH值,提升粪便有机酸含量,肠道菌群更加健康,有益菌增多,有害菌减少,菌群结构更利于肠道健康。(The invention relates to the field of health food, in particular to a bifidobacterium longum dietary composition for intervening chronic nephritis and uremia and application thereof, wherein the bifidobacterium longum dietary composition comprises 100mL of bifidobacterium longum BL-G301 probiotic cow milk and 40G of cereal powder tablet, and the preparation method comprises the steps of taking out the probiotic powder which is frozen and preserved at the temperature of-80 ℃, and adding the probiotic powder into pasteurized cow milk after the temperature is up to the room temperature; the product obtained by the preparation method of the bifidobacterium longum BL-G301 probiotic cow milk and the grain powder tablet are used together as a diet, so that the oxidative stress in the intestinal tract of a human body is improved, and the levels of inflammatory factors and uremia in the human body are reduced. Compared with the prior art, the invention adopts probiotics to intervene in chronic nephritis, and can reduce the oxidative stress level of chronic nephritis patients; improving the level of inflammatory factors and the level of uremic toxins in patients with chronic nephritis; the pH value of the excrement is improved, the organic acid content of the excrement is increased, intestinal flora is healthier, beneficial bacteria are increased, harmful bacteria are reduced, and the flora structure is more beneficial to intestinal health.)

1. A bifidobacterium longum dietary composition for intervening chronic nephritis and uremia is characterized in that: comprises 100mL of bifidobacterium longum BL-G301 probiotic cow milk and 40G of cereal powder tablets.

2. The dietary composition of Bifidobacterium longum for the intervention of chronic nephritis and uremia according to claim 1, wherein: the preparation method of the bifidobacterium longum BL-G301 probiotic cow milk comprises the following steps:

step 1: taking out the Bifidobacterium longum BL-G301 probiotic powder which is frozen and preserved at the temperature of minus 80 ℃ for standby, wherein the viable count in the Bifidobacterium longum BL-G301 probiotic powder is more than or equal to 1 multiplied by 10¹⁰ CFU/g；

Step 2: after the temperature is reduced to room temperature, the probiotic bacteria powder is added into pasteurized milk until the viable count is 7.4 multiplied by 10⁸± 5.4 × 10⁸ CFU/100 mL。

3. The dietary composition of Bifidobacterium longum for the intervention of chronic nephritis and uremia according to claim 1, wherein: the cereal flour flakes comprise 10% white sugar and 0.5% sodium chloride.

4. The dietary composition of Bifidobacterium longum for the intervention of chronic nephritis and uremia according to claim 2, wherein: the preparation method of the bifidobacterium longum BL-G301 probiotic powder comprises the following steps:

step 1: inoculating a Bifidobacterium longum strain subjected to freezing preservation at-80 ℃ in a basal culture medium, and culturing at 37 ℃ for 12h to obtain an activated seed culture medium;

step 2: respectively inoculating the activated seed culture medium in a fermentation culture medium in an inoculation amount of 2-5%, and culturing at 37 ℃ for 8-10 h to obtain fermented bacterial liquid;

and step 3: collecting bacterial liquid, and centrifuging by adopting a refrigerated centrifuge, wherein the centrifugation conditions are as follows: the temperature is 4 ℃, the rotating speed is 6000r/min, the time is 10min, and after centrifugation, the bacterial sludge and the protective agent are mixed according to the proportion of 1: 2, emulsifying, and carrying out vacuum freeze drying for 12 hours at the cold trap temperature of a freeze dryer of minus 80 ℃ to obtain bacterial powder;

and 4, step 4: heating the solid culture medium to completely melt, cooling to about 50 ℃, taking 0.5 mL of diluent with proper dilution gradient in a sterilized culture dish, pouring BMD culture medium, mixing uniformly, taking 3 parallel samples in each gradient, carrying out anaerobic culture at 37 ℃ for 48 hours, and counting the total number of colonies.

5. The dietary composition of Bifidobacterium longum for the intervention of chronic nephritis and uremia according to claim 4, wherein: the basic culture medium in the step 1 comprises the following components: 20 g/L of lactose, 10g/L of peptone, 10g/L of beef extract powder, 5 g/L, L g/L of yeast extract-1 g/L of cysteine, 2 g/L of dipotassium phosphate, 2 g/L of diamine hydrogen citrate, 5 g/L of sodium acetate, 0.25 g/L of magnesium sulfate heptahydrate, 0.05 g/L of manganese sulfate monohydrate and 801 g/L of tween-801.

6. The dietary composition of Bifidobacterium longum for the intervention of chronic nephritis and uremia according to claim 4, wherein: the fermentation medium in the step 2 comprises the following components: 20 g/L of glucose, 10g/L of peptone, 10g/L of beef extract powder, 5 g/L of yeast extract, 2 g/L of dipotassium phosphate, 2 g/L of dihydrodiamine citrate, 5 g/L of sodium acetate, 4.92 g/L, L g/L of magnesium sulfate heptahydrate-1 g/L of cysteine, 0.4 g/L of tryptophan, 0.4 g/L of aspartic acid, 0.05 g/L of manganese sulfate monohydrate and 801 g/L of tween.

7. The dietary composition of Bifidobacterium longum for the intervention of chronic nephritis and uremia according to claim 4, wherein: the protective agent in the step 3 comprises the following components: 110g/L of skimmed milk powder, 40 g/L of sucrose and 20 g/L of monosodium glutamate.

8. The dietary composition of Bifidobacterium longum for the intervention of chronic nephritis and uremia according to claim 4, wherein: the BMD culture medium in the step 4 comprises the following components: 5 g/L of lactose, 10g/L of tryptone, 4 g/L of beef extract powder, 3 g/L of yeast extract, 4 g/L of sodium chloride, 1 g/L of cysteine hydrochloride, 25 mg/L of mupirocin lithium salt and 801 g/L of tween-801; 0.8 g/L agar powder.

9. The dietary composition of Bifidobacterium longum for the intervention of chronic nephritis and uremia according to claim 1, wherein: the Bifidobacterium longum BL-G301 strain has the preservation places as follows: china Center for Type Culture Collection (CCTCC), the preservation number of the strain is CCTCC NO: m2013689, the preservation time is as follows: 12 and 23 in 2013.

10. The application of bifidobacterium longum dietary composition for intervening chronic nephritis and uremia is characterized in that: the product obtained by the preparation method of the bifidobacterium longum BL-G301 probiotic cow milk according to claims 1-8 and cereal powder tablets are matched as a meal together, so that the oxidative stress in the intestinal tract of a human body is improved, and the levels of inflammatory factors and uremic toxins in the human body are reduced.

Technical Field

The invention relates to the field of health-care food, in particular to a bifidobacterium longum dietary composition for intervening chronic nephritis and uremia and application thereof.

Background

Chronic nephritis has become a major public health problem worldwide based on morbidity and prevalence, which has increased by about 5% over the last 4 years, with an overall average prevalence of 13.4%.

Chronic nephritis promotes the accumulation of organic residues in the body which can lead to a range of negative body effects including exacerbating the inflammatory state, oxidative stress, cardiovascular dysfunction and risk of death. There is currently no good drug control for chronic nephritis, and in order to control this accumulation, it is common to intervene by restricting the intake of certain food groups, such as fruits, vegetables and cereals, to individuals suffering from chronic nephritis. Intake limitations on these foods often affect the supplementation of micronutrients, dietary fibres and phytochemicals with antioxidant and anti-inflammatory activity. Research shows that the supplement of the probiotic preparation can improve the antioxidant and anti-inflammatory activity of a host and is beneficial to the absorption of the nutrition of the host, but the technology and the application of the probiotic-related preparation for intervening chronic nephritis and uremia are lacked at present.

Therefore, it is necessary to design a bifidobacterium longum dietary composition for intervening chronic nephritis and uremia and an application thereof.

Disclosure of Invention

The invention breaks through the difficult problems in the prior art and designs the bifidobacterium longum dietary composition for intervening chronic nephritis and uremia and the application thereof.

In order to achieve the purpose, the invention designs a bifidobacterium longum dietary composition for intervening chronic nephritis and uremia and application thereof, wherein the bifidobacterium longum dietary composition comprises 100mL of bifidobacterium longum BL-G301 probiotic cow milk and 40G of cereal powder tablets.

The preparation method of the bifidobacterium longum BL-G301 probiotic cow milk comprises the following steps:

step 1: taking out the Bifidobacterium longum BL-G301 probiotic powder which is frozen and preserved at the temperature of minus 80 ℃ for standby, wherein the viable count in the Bifidobacterium longum BL-G301 probiotic powder is more than or equal to 1 multiplied by 10¹⁰ CFU/g；

Step 2: after the temperature is reduced to room temperature, the probiotic bacteria powder is added into pasteurized milk until the viable count is 7.4 multiplied by 10⁸± 5.4 × 10⁸ CFU/100 mL。

The cereal flour flakes comprise 10% white sugar and 0.5% sodium chloride.

The preparation method of the bifidobacterium longum BL-G301 probiotic powder comprises the following steps:

step 1: inoculating a Bifidobacterium longum strain subjected to freezing preservation at-80 ℃ in a basal culture medium, and culturing at 37 ℃ for 12h to obtain an activated seed culture medium;

step 2: respectively inoculating the activated seed culture medium in a fermentation culture medium in an inoculation amount of 2-5%, and culturing at 37 ℃ for 8-10 h to obtain fermented bacterial liquid;

and step 3: collecting bacterial liquid, and centrifuging by adopting a refrigerated centrifuge, wherein the centrifugation conditions are as follows: the temperature is 4 ℃, the rotating speed is 6000r/min, the time is 10min, and after centrifugation, the bacterial sludge and the protective agent are mixed according to the proportion of 1: 2, emulsifying, and carrying out vacuum freeze drying for 12 hours at the cold trap temperature of a freeze dryer of minus 80 ℃ to obtain bacterial powder;

and 4, step 4: heating the solid culture medium to completely melt, cooling to about 50 ℃, taking 0.5 mL of diluent with proper dilution gradient in a sterilized culture dish, pouring BMD culture medium, mixing uniformly, taking 3 parallel samples in each gradient, carrying out anaerobic culture at 37 ℃ for 48 hours, and counting the total number of colonies.

The basic culture medium in the step 1 comprises the following components: 20 g/L of lactose, 10g/L of peptone, 10g/L of beef extract powder, 5 g/L, L g/L of yeast extract-1 g/L of cysteine, 2 g/L of dipotassium phosphate, 2 g/L of diamine hydrogen citrate, 5 g/L of sodium acetate, 0.25 g/L of magnesium sulfate heptahydrate, 0.05 g/L of manganese sulfate monohydrate and 801 g/L of tween-801.

The fermentation medium in step 2 comprises the following components: 20 g/L of glucose, 10g/L of peptone, 10g/L of beef extract powder, 5 g/L of yeast extract, 2 g/L of dipotassium phosphate, 2 g/L of dihydrodiamine citrate, 5 g/L of sodium acetate, 4.92 g/L, L g/L of magnesium sulfate heptahydrate-1 g/L of cysteine, 0.4 g/L of tryptophan, 0.4 g/L of aspartic acid, 0.05 g/L of manganese sulfate monohydrate and 801 g/L of tween.

The protective agent in the step 3 comprises the following components: 110g/L of skimmed milk powder, 40 g/L of sucrose and 20 g/L of monosodium glutamate.

BMD medium composition in step 4: 5 g/L of lactose, 10g/L of tryptone, 4 g/L of beef extract powder, 3 g/L of yeast extract, 4 g/L of sodium chloride, 1 g/L of cysteine hydrochloride, 25 mg/L of mupirocin lithium salt and 801 g/L of tween-801; 0.8 g/L agar powder.

Bifidobacterium longum BL-G301 strain is classified as Bifidobacterium longum BL-G301

(Bifidobacterium longum BL-G301) with the preservation unit: china Center for Type Culture Collection (CCTCC) with the preservation unit address as follows: the preservation number of the Wuhan university in Wuhan, China is CCTCC NO: m2013689, the preservation time is as follows: 12 and 23 in 2013.

The product obtained by the preparation method of the bifidobacterium longum BL-G301 probiotic cow milk and the grain powder tablet are used together as a diet, so that the oxidative stress in the intestinal tract of a human body is improved, and the levels of inflammatory factors and uremia in the human body are reduced.

Compared with the prior art, the invention adopts probiotics to intervene in chronic nephritis, and can reduce the oxidative stress level of chronic nephritis patients; improving the level of inflammatory factors and the level of uremic toxins in patients with chronic nephritis; the pH value of the excrement is improved, the organic acid content of the excrement is increased, intestinal flora is healthier, beneficial bacteria are increased, harmful bacteria are reduced, and the flora structure is more beneficial to intestinal health.

Drawings

FIG. 1 is a bar graph of the concentration of markers of oxidative stress in the blood of a dry prognosis patient of the invention.

FIG. 2 is a bar graph of the concentration of inflammatory markers in the blood of a patient following an intervention of the present invention.

FIG. 3 is a bar graph of the concentration of uremic toxins in the blood of patients following an intervention of the present invention.

FIG. 4 is a bar graph of pH and concentration of organic acids in stool samples from patients following intervention in the present invention.

Detailed Description

The invention will now be further described.

The Bifidobacterium longum strain BL-G301 used in the present invention is classified as Bifidobacterium longum BL-G301

(Bifidobacterium longum BL-G301) with the preservation unit: china Center for Type Culture Collection (CCTCC) with the preservation unit address as follows: the preservation number of the Wuhan university in Wuhan, China is CCTCC NO: m2013689, the preservation time is as follows: 12 and 23 in 2013.

The invention designs a bifidobacterium longum dietary composition for intervening chronic nephritis and uremia, which comprises 100mL of bifidobacterium longum BL-G301 probiotic milk and 40G of cereal powder tablets.

The preparation method of the bifidobacterium longum BL-G301 probiotic cow milk comprises the following steps:

step 1: taking out the Bifidobacterium longum BL-G301 probiotic powder which is frozen and preserved at the temperature of minus 80 ℃ for standby, wherein the viable count in the Bifidobacterium longum BL-G301 probiotic powder is more than or equal to 1 multiplied by 10¹⁰ CFU/g；

Step 2: after the temperature is reduced to room temperature, the probiotic bacteria powder is added into pasteurized milk until the viable count is 7.4 multiplied by 10⁸± 5.4 × 10⁸ CFU/100 mL。

The cereal flour flakes comprise 10% white sugar and 0.5% sodium chloride.

The preparation method of the bifidobacterium longum BL-G301 probiotic powder comprises the following steps:

step 1: inoculating a Bifidobacterium longum strain subjected to freezing preservation at-80 ℃ in a basal culture medium, and culturing at 37 ℃ for 12h to obtain an activated seed culture medium;

step 2: respectively inoculating the activated seed culture medium in a fermentation culture medium in an inoculation amount of 2-5%, and culturing at 37 ℃ for 8-10 h to obtain fermented bacterial liquid;

and step 3: collecting bacterial liquid, and centrifuging by adopting a refrigerated centrifuge, wherein the centrifugation conditions are as follows: the temperature is 4 ℃, the rotating speed is 6000r/min, the time is 10min, and after centrifugation, the bacterial sludge and the protective agent are mixed according to the proportion of 1: 2, emulsifying, and carrying out vacuum freeze drying for 12 hours at the cold trap temperature of a freeze dryer of minus 80 ℃ to obtain bacterial powder;

and 4, step 4: heating the solid culture medium to completely melt, cooling to about 50 ℃, taking 0.5 mL of diluent with proper dilution gradient in a sterilized culture dish, pouring BMD culture medium, mixing uniformly, taking 3 parallel samples in each gradient, carrying out anaerobic culture at 37 ℃ for 48 hours, and counting the total number of colonies.

The basic culture medium in the step 1 comprises the following components: 20 g/L of lactose, 10g/L of peptone, 10g/L of beef extract powder, 5 g/L, L g/L of yeast extract-1 g/L of cysteine, 2 g/L of dipotassium phosphate, 2 g/L of diamine hydrogen citrate, 5 g/L of sodium acetate, 0.25 g/L of magnesium sulfate heptahydrate, 0.05 g/L of manganese sulfate monohydrate and 801 g/L of tween-801.

The fermentation medium in step 2 comprises the following components: 20 g/L of glucose, 10g/L of peptone, 10g/L of beef extract powder, 5 g/L of yeast extract, 2 g/L of dipotassium phosphate, 2 g/L of dihydrodiamine citrate, 5 g/L of sodium acetate, 4.92 g/L, L g/L of magnesium sulfate heptahydrate-1 g/L of cysteine, 0.4 g/L of tryptophan, 0.4 g/L of aspartic acid, 0.05 g/L of manganese sulfate monohydrate and 801 g/L of tween.

The protective agent in the step 3 comprises the following components: 110g/L of skimmed milk powder, 40 g/L of sucrose and 20 g/L of monosodium glutamate.

BMD medium composition in step 4: 5 g/L of lactose, 10g/L of tryptone, 4 g/L of beef extract powder, 3 g/L of yeast extract, 4 g/L of sodium chloride, 1 g/L of cysteine hydrochloride, 25 mg/L of mupirocin lithium salt and 801 g/L of tween-801; 0.8 g/L agar powder.

The bifidobacterium longum BL-G301 probiotic cow milk and the grain powder tablet are used together as a diet, so that the oxidative stress in the intestinal tract of a human body is improved, and the levels of inflammatory factors and uremia in the human body are reduced.

The embodiments of the present invention will be further described with reference to examples.

Example 1:

taking the prepared bifidobacterium longum BL-G301 probiotic cow milk and sorghum powder extruded sheets as experimental groups, and matching 100mL of probiotic cow milk with 40G of sorghum powder extruded sheets every day; the control group consisted of pasteurized milk and extruded corn flour chips without probiotics, with 100mL of milk per day along with 40g of extruded corn flour chips.

58 volunteers with chronic nephritis were selected, participants aged over 18 years and subjected to three to four hemodialysis sessions per week for at least three months. The individuals were randomly divided into two groups, 29 individuals each for the experimental and control groups.

Blood samples were collected by qualified professionals prior to hemodialysis and immediately stored at-80 ℃.

Detecting the enzymatic activity (SOD), Malondialdehyde (MDA) and Total Antioxidant Capacity (TAC) of superoxide dismutase in the sample.

In connection with fig. 1, in terms of oxidative stress, the test group (SG) from which probiotic BL-G301 was obtained was intervened with a decrease in serum MDA (p < 0.05) and an increase in SOD and TAC (p < 0.05) compared to the Control Group (CG).

Example 2:

the prepared bifidobacterium longum BL-G301 probiotic cow milk and sorghum powder extruded sheet are matched as the diet of an experimental group, and cow milk with 100mL of probiotics is matched with 40G of sorghum powder extruded sheet every day; the control group consisted of pasteurized milk and extruded corn flour chips without probiotics, with 100mL of milk per day along with 40g of extruded corn flour chips.

58 volunteers with chronic nephritis were selected, participants aged over 18 years and subjected to three to four hemodialysis sessions per week for at least three months. The individuals were randomly divided into two groups, 29 individuals each for the experimental and control groups.

Blood samples were collected by qualified professionals prior to hemodialysis and immediately stored at-80 ℃.

The concentrations of IL-6, IL-10 and TNF- α and the concentration of C-reactive protein (CRP) in the sample are measured in response to the concentration of inflammatory factors.

In conjunction with FIG. 2, the concentration of CRP was reduced (p < 0.05) in the experimental group (SG) compared to the Control Group (CG); no obvious change among IL-6, IL-10 and TNF-alpha cytokines (p >)

0.05)。

Example 3:

the prepared bifidobacterium longum BL-G301 probiotic cow milk and sorghum powder extruded sheet are matched as the diet of an experimental group, and cow milk with 100mL of probiotics is matched with 40G of sorghum powder extruded sheet every day; the control group consisted of pasteurized milk and extruded corn flour chips without probiotics, with 100mL of milk per day along with 40g of extruded corn flour chips.

58 volunteers with chronic nephritis were selected, participants aged over 18 years and subjected to three to four hemodialysis sessions per week for at least three months. The individuals were randomly divided into two groups, 29 individuals each for the experimental and control groups.

Blood samples were collected by qualified professionals prior to hemodialysis and immediately stored at-80 ℃.

Three indicators representing the degree of uremia in the test sample were measured: serum indole sulfate, p-cresol sulfate and indole 3-acetic acid.

Referring to FIG. 3, compared to the Control Group (CG), the concentration of indole sulfate and p-cresol sulfate in serum was lower in the experimental group (SG) (p < 0.05); while the concentration of indole 3-acetic acid did not change significantly (p > 0.05).

Example 4

The prepared bifidobacterium longum BL-G301 probiotic cow milk and sorghum powder extruded sheet are matched as the diet of an experimental group, and cow milk with 100mL of probiotics is matched with 40G of sorghum powder extruded sheet every day; the control group consisted of pasteurized milk and extruded corn flour chips without probiotics, with 100mL of milk per day along with 40g of extruded corn flour chips.

58 volunteers with chronic nephritis were selected, participants aged over 18 years and subjected to three to four hemodialysis sessions per week for at least three months. The individuals were randomly divided into two groups, 29 individuals each for the experimental and control groups.

Stool samples were collected by volunteers using sterile bottles and the collected stool samples were stored by quick freezing at-18 °.

And (3) detecting the pH value of the fecal sample and the concentrations of organic acids such as acetic acid, propionic acid, butyric acid and the like.

Referring to fig. 4, the end point stool pH was lower (p < 0.05) for the experimental group (SG) and higher organic acid concentrations of acetic, propionic and butyric acids for the experimental group (SG) compared to the Control Group (CG).

The combination of the prepared product of the method for preparing the bifidobacterium longum BL-G301 probiotic cow milk and the cereal powder tablet are used together as a diet, so that the level of the toxin concentration of the uremia in blood of the chronic nephritis patients can be reduced, and the pH value of the feces of the chronic nephritis patients can be reduced and the organic acid concentration of the feces can be increased.