CN114292787B - Lactobacillus plantarum P101 for preventing liver and kidney injury and application thereof - Google Patents
Lactobacillus plantarum P101 for preventing liver and kidney injury and application thereof Download PDFInfo
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- CN114292787B CN114292787B CN202111657079.1A CN202111657079A CN114292787B CN 114292787 B CN114292787 B CN 114292787B CN 202111657079 A CN202111657079 A CN 202111657079A CN 114292787 B CN114292787 B CN 114292787B
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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 farmhouse-made pickled Chinese cabbage in Jian City of Jiangxi province, the preservation number is CCTCC M2021108, and the preservation place is China center for type culture Collection of Wuhan City. The lactobacillus plantarum P101 is gram positive bacteria, has good acid resistance and bile salt resistance and higher antioxidant activity, and can be used in carbon tetrachloride (CCl) 4 ) In induced liver and kidney injury, the preparation can relieve pathological injury, promote antioxidant enzyme activity, and prevent CCl to a certain extent 4 The liver and kidney are damaged. The lactobacillus plantarum P101 strain has higher safety and good probiotics performance, and the strain is used for preventing CCl 4 The application of the resulting liver and kidney injury plays a beneficial role.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to lactobacillus plantarum P101 for preventing liver and kidney injury 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 of the abundant blood flow of liver and kidney, high oxygen consumption, high tissue metabolism rate, active action of various enzymes and extremely easy damage of liver and kidney of some medicines or metabolites thereof. CCl (CCl) 4 Is a potential toxic substance which can be converted into trichloromethyl radical in liver and kidney cell microsomes, thereby leading to peroxidation of cell membranes and damage of the liver and the kidney.
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. Probiotics are also present in human gastrointestinal tract, are responsible for regulating intestinal microbial balance, controlling endotoxin level, enhancing organism immunity, removing harmful substances, and have great significance on organism health. At present, probiotics have been widely used in the industries of food, industry, medical care, and the like. Research shows that probiotics also play a positive role in protecting remote organs, such as preventing liver and kidney injury and reducing cardiovascular and cerebrovascular diseases, so that the probiotics are adopted to prevent CCl 4 Induced liver and kidney damage is possible. The probiotics adopted by the invention are lactobacillus plantarum which are widely existing in fermented foods, are one of accepted probiotic bacteria, belong to gram-positive bacteria, are mostly facultative anaerobic, have an optimal growth temperature of 30-37 ℃ and an optimal growth pH of about 6.5.
By searching, the following patent publications are found, which are relevant to the present patent application, for alleviating liver damage by using active substances:
preparation of ground beetle enzymolysis extract for preventing and/or treating CCl 4 The invention belongs to the technical field of medicines, and relates to an application of an enzymatic extract of ground beetle in preventing CCl (common cold herb) in medicines for treating liver injury (CN 106309502B) 4 Application in liver injury. Mainly relates to a method for extracting ground beetle powder, and the ground beetle enzymolysis extract can obviously reduce CCl 4 Injury to mouse liver cells enhances the antioxidant capacity of mice with acute liver injury. For CCl 4 The liver injury caused by the method is mostly interfered by active substances, and the patent uses lactobacillus plantarum to relieve acute liver and kidneyThe damage has stronger innovation.
The prior art has focused on the therapeutic effects of probiotics on established liver lesions. The lactobacillus plantarum P101 adopts a prevention mode to improve the capability of an organism to cope with acute injury, comprehensively evaluates liver and kidney of two main metabolic organs of the organism, and is not limited to the effect on a single organ. In summary, the present patent application has substantial differences from other patent publications.
Disclosure of Invention
The invention aims to provide a lactobacillus plantarum P101 for preventing liver and kidney injuryLactobacillus plantarumP101) and its use; the lactobacillus plantarum P101 has good probiotics, good acid resistance and bile salt resistance and high antioxidant activity.
The invention relates to a lactobacillus plantarum P101 for preventing liver and kidney injury, wherein the lactobacillus plantarum P101 is stored in the China center for type culture Collection (university of Wuhan) of Wuhan in 2021, and the storage number is: cctccc M2021108.
Further, the lactobacillus plantarum P101 was isolated from farmhouse-made pickled Chinese cabbage.
The lactobacillus plantarum P101 can grow on an MRS agar plate culture medium, is gram-positive bacillus, is facultative anaerobic and does not produce spores. To maintain the excellent performance of the lactobacillus plantarum of the present invention, it was stored in an MRS mixed preservation solution containing 25% glycerol and frozen at-80 ℃.
Another object of the invention is to provide the use of Lactobacillus plantarum P101 for the manufacture of a medicament for preventing or alleviating carbon tetrachloride-induced liver and kidney injury.
Another object of the invention is to provide the use of lactobacillus plantarum P101 for the preparation of a health care product for the assisted protection of carbon tetrachloride-induced liver injury.
Further, the viable count of Lactobacillus plantarum P101 used in the application is 10 9 CFU/mL。
The invention infuses the lactobacillus plantarum P101 into the Kunming mice for 21 days, and then the CCl is injected into the abdominal cavity after the last stomach infusion of 2 h 4 And (5) inducing a liver and kidney injury model. The research result shows that the lactobacillus plantarum P101 can obviously reduce liver and kidney injury of mice.
Compared with the prior art, the invention has the beneficial effects that:
the lactobacillus plantarum P101 is a probiotic with excellent performance and higher safety, and can effectively prevent or reduce CCl 4 The induced liver and kidney injury has higher research value and application.
Drawings
Figure 1 is a graph of liver and kidney organ coefficients of in vivo experiments in mice (significant differences are shown between different letters,p<0.05）。
FIG. 2 is a graph showing the ratio of alkaline phosphatase (ALP) activity level to glutamic oxaloacetic transaminase/glutamic pyruvic transaminase (AST/ALT) activity level in the serum of the in vivo experiment of mice (the difference between the different letters indicates that there is a significant difference,p<0.05）。
figure 3 is a graph showing the presence of significant differences between Creatinine (CRE) and Uric Acid (UA) levels in serum from experiments in mice,p<0.05）。
figure 4 is a graph of Catalase (CAT) activity levels in liver homogenates from in vivo experiments in mice (significant differences are shown between the different letters,p<0.05）。
fig. 5 is an in vivo experimental liver slice observation of mice.
Fig. 6 is an in vivo experimental kidney section observation 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 in the following examples. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are commercially available and conventional products, and manufacturers are not identified.
Examples: prevention of CCl 4 Resulting liver and kidney injury
After 1 week of adaptive feeding of 21 6 week old Kunming mice, three groups were randomized: control, model and prophylaxis groups, control and modelGroups were filled with 100. Mu.L of phosphate buffer daily, and groups were prevented from being filled with 100. Mu.L (10) 9 CFU/mL) for 21 days. After the last gastric lavage 2 h, 3 mL/kg BW CCl was injected intraperitoneally into the model and prophylactic groups 4 Olive oil (20% v/v), the control group was intraperitoneally injected with an equal amount of olive oil. After fasting 16 h, the blood, liver and kidneys were collected for subsequent physicochemical analysis.
Liver and kidney weights were recorded, and organ coefficient=organ weight (mg)/body weight (g) was calculated. Livers and kidneys were fixed using 4% paraformaldehyde solution and the company was entrusted with paraffin embedding and hematoxylin & eosin (H & E) staining. The blood of the mice is collected and kept still for more than 6 h, and the blood is centrifuged for 10 min under the condition of 5,000 r/min, and the blood is repeated once to obtain the serum of the mice. The level of liver function and kidney function indicators in serum was determined using the kit. Liver tissue is evenly mixed with physiological saline solution 1:9 (w/v) and then ground to prepare 10% tissue homogenate, the tissue homogenate is centrifuged for 5 min under the condition of 5,000 r/min, the supernatant is collected, and the CAT activity of the liver tissue is measured by using the kit.
Experimental results: as shown in FIG. 1, the model group showed a significant increase in liver and kidney organ coefficients compared to the control group, and this result indicated that CCl 4 Possibly resulting in tissue swelling. Whereas the organ coefficients of the mice in the prophylaxis group tended to the control group, indicating that the intervention of lactobacillus plantarum P101 was able to alleviate tissue swelling.
As can be seen from the results of FIG. 2, CCl 4 Resulting in a significant increase in ALP activity in serum and a significant decrease in AST/ALT ratio in model group serum, indicating impaired liver function. The ALP activity and AST/ALT activity ratio of the mice serum prevented by the lactobacillus plantarum P101 are reduced to be equivalent to those of the control group.
As can be seen from the results of fig. 3, the control group had normal CRE and UA excretion, the serum content was low, and the model group mice had significantly increased serum CRE and UA content, indicating renal dysfunction, metabolic disorder, and thus accumulation of CRE and UA in serum. The decrease of CRE and UA content in serum of mice in the prevention group shows that the lactobacillus plantarum P101 has the function of preventing kidney function injury and metabolic disorder.
The results in fig. 4 show that CAT viability was significantly reduced in liver tissue of the model group,CCl 4 resulting in a decrease in the antioxidant capacity of the liver. Lactobacillus plantarum P101 the CAT activity in liver tissue of the preventive group was significantly increased to the level of the normal control group.
As shown in fig. 5, the control group hepatocytes were normally structured and closely arranged, while the model group hepatocytes were blurred in edge, and inflammatory cells were infiltrated, and a large amount of cytoplasmic vacuolation occurred. After the lactobacillus plantarum P101 is prevented, the structure of liver cells is complete, cavitation phenomenon is reduced, and pathological symptoms of the lactobacillus plantarum P101 are obviously improved compared with a model group.
As shown in fig. 6, the model group showed reduced glomeruli, structural destruction, occurrence of tubular cell degeneration, vacuolation, swelling, necrosis and shedding, increased gap, interstitial hemorrhage and inflammatory cell infiltration. After the prevention of lactobacillus plantarum P101, although it is still accompanied with inflammatory cell infiltration and interstitial bleeding, the injury is slight, and the glomerulus and tubule structure are complete, the cells are orderly arranged, and the gap is reduced.
The results show that the lactobacillus plantarum P101 can prevent CCl to a certain extent 4 Induced liver and kidney injury in mice.
Specifically described are: the probiotics disclosed in the prior patent publication mainly play a beneficial role in specific disease damage models such as non-alcoholic fatty liver, hepatitis B liver cirrhosis, uremia or kidney stones, and have certain limitation only aiming at a single organ or specific disease model. Differently, this patent is based on CCl 4 The injury model, lactobacillus plantarum P101 can relieve liver and kidney injury. The model damage can widely represent damage to organisms caused by exogenous poison, and the liver and the kidney are primary metabolic organs of the organisms for coping with exogenous substances, and have important significance in evaluating the damage of the two organs. Therefore, the lactobacillus plantarum P101 is very likely to prevent and/or reduce the damage of environmental poison to the organism, and the method has practical significance.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present application and not for limiting the same; although the present application has been described in detail with reference to preferred embodiments, those of ordinary skill in the art will appreciate that: modifications may be made to the specific embodiments of the present application or equivalents may be substituted for part of the technical features, which are all included in the scope of the technical solutions claimed herein.
Claims (4)
1. Lactobacillus plantarum strain for preventing liver and kidney injuryLactobacillus plantarum) The lactobacillus plantarum P101 is characterized in that the preservation number of the lactobacillus plantarum P101 is CCTCC M2021108, and the preservation place is China center for type culture collection of Wuhan City.
2. Use of lactobacillus plantarum P101 as claimed in claim 1 in the manufacture of a medicament for preventing or alleviating carbon tetrachloride-induced liver and kidney damage.
3. Use of lactobacillus plantarum P101 as claimed in claim 1 for the manufacture of a health care product for the assisted protection of carbon tetrachloride-induced liver injury.
4. Use of lactobacillus plantarum P101 according to claim 2 or 3, characterized in that the number of viable bacteria used in lactobacillus plantarum P101 in said use is 10 9 CFU/mL。
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| CN112553115B (en) * | 2020-12-23 | 2022-09-16 | 南昌大学 | Application of lactobacillus plantarum ZDY2013 in preparation of products for relieving kidney injury |
| CN114164149B (en) * | 2021-11-29 | 2023-10-03 | 南昌大学 | Lactobacillus plantarum P101 for relieving obesity and lead toxicity and application thereof |
| CN115418332A (en) * | 2022-09-02 | 2022-12-02 | 重庆第二师范学院 | Lactobacillus plantarum capable of preventing and improving chemical liver injury |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102994422A (en) * | 2012-11-12 | 2013-03-27 | 北京和美科健生物技术有限责任公司 | Application of lactobacillus plantarum P-8 in improvement of alcoholic liver injury |
| CN112553115A (en) * | 2020-12-23 | 2021-03-26 | 南昌大学 | Application of lactobacillus plantarum ZDY2013 in preparation of products for relieving kidney injury |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102994422A (en) * | 2012-11-12 | 2013-03-27 | 北京和美科健生物技术有限责任公司 | Application of lactobacillus plantarum P-8 in improvement of alcoholic liver injury |
| CN112553115A (en) * | 2020-12-23 | 2021-03-26 | 南昌大学 | Application of lactobacillus plantarum ZDY2013 in preparation of products for relieving kidney injury |
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