CN114292787A

CN114292787A - Lactobacillus plantarum P101 for preventing liver and kidney injury and application thereof

Info

Publication number: CN114292787A
Application number: CN202111657079.1A
Authority: CN
Inventors: 许恒毅; 王梦琦; 胥晓薇; 刘善级; 徐媛媛; 张锦峰; 袁宏斌; 聂鹏辉
Original assignee: Nanchang University
Current assignee: Nanchang University
Priority date: 2021-12-31
Filing date: 2021-12-31
Publication date: 2022-04-08
Anticipated expiration: 2041-12-31
Also published as: CN114292787B

Abstract

The invention discloses lactobacillus plantarum P101 for preventing liver and kidney injury and application thereof, and belongs to the technical field of microorganisms. The lactobacillus plantarum P101 is separated from pickled Chinese cabbage self-made by farmers in Jian city in Jiangxi province, the preservation number is CCTCC M2021108, and the preservation place is the China center for type culture preservation in Wuhan city. The lactobacillus plantarum P101 is gram-positive bacteria, has good acid-resistant and cholate-resistant characteristics and higher antioxidant activity, and is applied to carbon tetrachloride (CCl)₄) LureIn liver and kidney injury, pathological injury can be relieved, the activity of antioxidant enzyme can be improved, and CCl can be prevented to a certain extent₄The liver and kidney are damaged. The lactobacillus plantarum P101 strain has high safety and good probiotic performance, and can prevent CCl₄The application of the medicine for treating the liver and kidney injuries plays a beneficial role.

Description

Lactobacillus plantarum P101 for preventing liver and kidney injury and application thereof

Technical Field

The invention belongs to the technical field of microorganisms, and particularly relates to lactobacillus plantarum P101 for preventing liver and kidney injuries and application thereof.

Background

Acute or chronic damage to the liver and kidneys caused by drug or toxin exposure is a common health problem. Because the blood flow of liver and kidney is rich, oxygen consumption is large, tissue metabolic rate is high, and the action of various enzymes is active, some medicines or metabolites thereof are easy to damage liver and kidney. CCl₄Is a potential toxic substance, can be converted into trichloromethyl free radical in liver and kidney cell microsomes, thereby causing cell membrane peroxidation and liver and kidney injury.

Probiotics are closely related to human life, widely exist in nature, and are applied to various fermented foods, such as vegetables, meat, dairy products and the like. The probiotics exist in the gastrointestinal tract of a human body, are responsible for regulating the balance of intestinal microorganisms, controlling the endotoxin level, enhancing the immunity of the organism and removing harmful substances, and have great significance to the health of the organism. At present, probiotics are widely applied to the industries of food, industry, medical care and the like. Research shows that the probiotics also play an active role in protecting remote organs, such as preventing liver and kidney injuries and reducing the occurrence of cardiovascular and cerebrovascular diseases, so that the probiotics are adopted to prevent CCl₄The induced liver and kidney injury becomes possible. The probiotics adopted by the invention is lactobacillus plantarum which is widely existed in fermented food, is one of accepted probiotics strains, belongs to gram-positive bacteria, is mostly facultative anaerobic, has the optimal growth temperature of 30-37 ℃ and the optimal growth pH of about 6.5.

Through searching, the following patent publications which use active substances to relieve liver injury are found and related to the patent application of the invention:

use of enzymatic extract of Eupolyphaga Seu Steleophaga in preparation of medicine for preventing and/or treating CCl₄The invention relates to an application of a ground beetle enzymolysis extract in preventing CCl (hepatitis C virus), belonging to the technical field of medicines (CN106309502B)₄Application in treating liver injury. Mainly relates to a method for extracting ground beetle powder, and the ground beetle enzymolysis extract can obviously reduce CCl₄Can be used for treating mouse liver cell injury, and enhancing oxidation resistance of mice with acute liver injury. For CCl₄Active substances are mostly adopted to intervene in the caused liver injury, and the lactobacillus plantarum used in the invention has stronger innovation in relieving acute liver and kidney injuries.

The prior patents mostly focus on the therapeutic effect of probiotics on established liver damage. The lactobacillus plantarum P101 adopts a prevention mode to improve the capability of an organism to cope with acute injury, comprehensively evaluates two main metabolic organs, namely liver and kidney of the organism, and is not limited to the effect on a single organ. In summary, the present patent application is substantially different from other patent publications.

Disclosure of Invention

The invention aims to provide Lactobacillus plantarum P101(Lactobacillus plantarum P101) for preventing liver and kidney injury and application thereof; the lactobacillus plantarum P101 disclosed by the invention has good probiotic performance, good acid-resistant and cholate-resistant characteristics and high antioxidant activity.

The invention relates to a lactobacillus plantarum P101 for preventing liver and kidney injury, wherein the lactobacillus plantarum P101 is preserved in China center for type culture Collection (university of Wuhan) in Wuhan City at 1 month and 19 days 2021, and the preservation numbers are as follows: CCTCC M2021108.

Further, the lactobacillus plantarum P101 is isolated from a sauerkraut made by farmer.

The lactobacillus plantarum P101 can grow on MRS agar plate culture medium, is gram-positive bacillus, is facultative anaerobic and does not produce spores. In order to maintain the excellent performance of the lactobacillus plantarum, the lactobacillus plantarum is preserved in MRS mixed preservation solution containing 25% of glycerin and is frozen at the temperature of minus 80 ℃.

The invention also aims to provide application of the lactobacillus plantarum P101 in preparing medicines, health products, fermentation products or probiotic preparations for preventing or relieving liver and kidney injuries.

Further, the liver and kidney injury is caused by CCl₄Induced liver and kidney damage.

Furthermore, the number of viable bacteria used by the lactobacillus plantarum P101 in the application is 10⁹CFU/mL。

The invention is characterized in that lactobacillus plantarum P101 is intragastrically injected to Kunming mice for 21 days after 2 hours of last intragastrically-infusing₄And (3) inducing a liver and kidney injury model. Research results show that the lactobacillus plantarum P101 can remarkably reduce the liver and kidney injuries of mice.

Compared with the prior art, the invention has the beneficial effects that:

the lactobacillus plantarum P101 is a probiotic strain with excellent performance and higher safety, and can effectively prevent or relieve CCl₄Induced liver and kidney injury, and has high research value and application.

Drawings

FIG. 1 shows the organ coefficients of liver and kidney in mice (different letters indicate significant difference, p < 0.05).

FIG. 2 is a graph of the ratio of the activity level of alkaline phosphatase (ALP) to the activity level of aspartate aminotransferase/glutamate pyruvate transaminase (AST/ALT) in the serum of mice tested in vivo (significant differences are indicated between different letters, p < 0.05).

Figure 3 is the Creatinine (CRE) and Uric Acid (UA) levels in the serum of the mice in vivo experiments (significant differences are indicated between different letters, p < 0.05).

FIG. 4 is a graph of the level of Catalase (CAT) activity in experimental liver homogenates in mice (significant difference indicated by different letters, p < 0.05).

FIG. 5 is an experimental liver slice observation in mice.

FIG. 6 is an experimental in vivo kidney section view of mice.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be clearly and completely described below with reference to the embodiments. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by manufacturers, and are all conventional products sold in the market.

Example (b): prevention of CCl₄The resulting damage to the liver and kidney

After 21 Kunming mice with the age of 6 weeks are adaptively bred for 1 week, the Kunming mice are randomly divided into three groups: control group, model group and prevention group, wherein the control group and the model group are intragastrically filled with 100 μ L phosphate buffer solution every day, and the prevention group is intragastrically filled with 100 μ L (10 μ L) phosphate buffer solution every day⁹CFU/mL) suspension for 21 days. After 2h of last gastric lavage, 3mL/kg BW CCl is injected into abdominal cavity of model group and prevention group₄Olive oil (20% v/v), control group was injected intraperitoneally with the same amount of olive oil. Sacrificed after 16h fasting and blood, liver and kidney were collected for subsequent physicochemical analysis.

The liver and kidney weights were recorded, and the organ coefficient was calculated as organ weight (mg)/body weight (g). The liver and kidney were fixed using 4% paraformaldehyde solution and entrusted to the company for paraffin embedding and hematoxylin & eosin (H & E) staining. Collecting blood of mouse, standing for more than 6h, centrifuging for 10min at 5,000r/min, and repeating once to obtain mouse serum. The level of indices of liver and kidney function in serum were determined using the kit. Mixing liver tissue with normal saline at a ratio of 1:9(w/v), grinding, preparing into 10% tissue homogenate, centrifuging at 5,000r/min for 5min, collecting supernatant, and determining CAT activity of liver tissue with kit.

The experimental results are as follows: as shown in FIG. 1, the liver and kidney organ coefficients of the model group were significantly increased as compared with those of the control group, and the results showed that CCl was significantly increased₄Possibly resulting in tissue swelling. Thereby preventing mice in the groupThe organ coefficient tended to the control group, indicating that the intervention of lactobacillus plantarum P101 can alleviate tissue swelling.

From the results in FIG. 2, CCl is shown₄The ALP activity in the serum is obviously improved, and the AST/ALT ratio in the serum of the model group is obviously reduced, which indicates that the liver function is damaged. And the serum ALP activity and AST/ALT activity ratio of the mice prevented by the lactobacillus plantarum P101 are reduced back to the level equivalent to that of the control group.

From the results of fig. 3, it can be seen that CRE and UA in the control group excrete normally and are lower in serum, while CRE and UA in the serum of the model group mice are significantly increased, indicating renal dysfunction and metabolic imbalance, thereby causing CRE and UA in serum to accumulate. And the prevention of the decrease of CRE and UA contents in the serum of mice shows that the lactobacillus plantarum P101 has the function of preventing renal function injury and metabolic disturbance.

FIG. 4 shows that CAT activity in liver tissue of the model group is significantly decreased, CCl₄Resulting in a decrease in the antioxidant capacity of the liver. Lactobacillus plantarum P101 prevented significant return of CAT activity in liver tissue of the group to the level of normal control group.

As shown in FIG. 5, the control group of hepatocytes had normal structure and close arrangement, while the model group of hepatocytes had blurred edges, infiltrated inflammatory cells, and exhibited massive vacuolization of cytoplasm. After the lactobacillus plantarum P101 is prevented, the structure of liver cells is complete, the vacuolation phenomenon is relieved, and the pathological symptoms of the lactobacillus plantarum P101 are obviously improved compared with those of a model group.

As shown in fig. 6, the model group showed decrease of glomeruli, structural destruction, degeneration, vacuolization, swelling, necrosis and shedding of tubular cells, increase of interstitial space, interstitial bleeding and inflammatory cell infiltration. After the lactobacillus plantarum P101 is prevented, inflammatory cell infiltration and interstitial bleeding are still accompanied, but the injury is slight, the structures of glomeruli and renal tubules are complete, the cells are arranged orderly, and gaps are reduced.

The results show that the lactobacillus plantarum P101 can prevent CCl to a certain extent₄Induced liver and kidney damage of mice.

Specifically, the following are mentioned: the probiotics disclosed in the prior patents are mainly used for treating nonalcoholic fatty liver, hepatitis B cirrhosis, uremia or kidney stonesThe disease damage model has beneficial effects, only aims at a single organ or a specific disease model, and has certain limitation. Otherwise, the patent is based on CCl₄The damage model, Lactobacillus plantarum P101, can alleviate liver and kidney damage. The model injury can widely represent the injury of an organism caused by an exogenous poison, the liver and the kidney are the primary metabolic organs of the organism for dealing with the exogenous substances, and the model injury has important significance for evaluating the injury of the two organs. Therefore, the lactobacillus plantarum P101 has the practical significance of most possibly preventing and/or reducing the damage of environmental toxicants to the organism.

Finally, it should be noted that: the above examples are only intended to illustrate the technical solutions of the present application and not to limit them; although the present application has been described in detail with reference to preferred embodiments, those of ordinary skill in the art will understand that: modifications to the embodiments of the present application or equivalent replacements of some technical features may still be made, which should all be covered by the scope of the technical solution claimed in the present application.

Claims

1. The lactobacillus plantarum P101 for preventing liver and kidney injury is characterized in that the lactobacillus plantarum P101 is preserved with the preservation number of CCTCC M2021108, and the preservation place is the China center for type culture Collection in Wuhan City.

2. Use of lactobacillus plantarum P101 according to claim 1 for the preparation of a medicament for preventing or alleviating liver and kidney damage.

3. Use of lactobacillus plantarum P101 according to claim 1 for the preparation of a health product for preventing or alleviating liver and kidney damage.

4. Use of lactobacillus plantarum P101 according to claim 1 for the preparation of a fermented product or probiotic preparation for preventing or alleviating liver, kidney damage.

5. A Lactobacillus plantarum P101 for use according to any one of claims 2-4, wherein the liver-kidney injury is carbon tetrachloride-induced.

6. Use of Lactobacillus plantarum P101 according to any one of claims 2-4, wherein Lactobacillus plantarum P101 used in the use has a viable count of 10⁹CFU/mL。