Application of parabacteroides dielsii in preparation of medicine for treating sepsis

Abstract

The invention provides application of parabacteroides dieldrii in preparation of a medicine for treating sepsis, and belongs to the technical field of microorganisms. According to the application disclosed by the invention, the sepsis is treated by utilizing the parabiosis dielsii ATCC: 8503, the treatment effect of the parabiosis dielsii ATCC: 8503 is detected, and a result proves that after the parabiosis dielsii ATCC: 8503 is given, the survival rate of mice with sepsis can be obviously increased, the contents of ALT, AST, BUN and Creatinine in serum can be reduced, meanwhile, the wet / dry weight ratio of lung is reduced, and the liver, lung and kidney injury caused by sepsis is effectively relieved.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, and in particular to an application of Parabacteroides distichous in preparing a medicine for treating sepsis. Background Art

[0002] Sepsis is a life-threatening organ dysfunction or failure caused by the host's dysregulated immune response to pathogen infection. It can lead to multiple organ damages. For example, (1) sepsis can damage the patient's alveoli and bronchi, which in turn affects respiratory function, resulting in pulmonary symptoms such as dyspnea, chest tightness, and cough. In severe cases, it may develop into acute respiratory distress syndrome. (2) The inflammatory response caused by sepsis can damage liver cells, leading to weakened liver function, which may induce symptoms such as loss of appetite, nausea, retching, aversion to oil, and fatigue. Liver damage is an independent risk factor for multiple organ dysfunction and high mortality in patients with sepsis. Liver dysfunction can occur in the early stages of sepsis. Studies have shown that the incidence of sepsis-related liver dysfunction and liver failure is 34% to 46% and 1.3% to 22%. If sepsis patients are combined with liver dysfunction or liver failure, their mortality rate is as high as 54% to 68%. (3) Sepsis may cause glomerulonephritis, decreased blood perfusion, etc., which in turn damage kidney function and may increase the risk of renal failure. The proportion of sepsis-induced acute kidney injury (SAKI) is as high as 26% to 50%, and it has a unique pathophysiological mechanism. Every year, 48.9 million people suffer from sepsis worldwide, and the related death toll exceeds 11 million, accounting for 19.7% of the global death toll. It has become one of the major diseases that seriously threaten human health.

[0003] Parabacteroides distasonis (P. distasonis) is a Gram-negative, non-spore-forming, anaerobic, rod-shaped bacterium belonging to the genus Parabacteroides and one of the core flora of the human body. Correlation analysis showed that its content was significantly negatively correlated with disease states such as obesity, non-alcoholic fatty liver disease, and diabetes, suggesting that it may play a positive regulatory role in glucose and lipid metabolism. However, the role and mechanism of P. distasonis in sepsis have not been reported. Summary of the invention

[0004] The object of the present invention is to provide an application of Parabacteroides distichous in the preparation of a medicament for treating sepsis, wherein Parabacteroides distichous can alleviate liver, lung and kidney damage caused by sepsis.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The present invention provides an application of Parabacteroides distasonis in preparing a medicine for treating sepsis, characterized in that the standard strain number of the Parabacteroides distasonis is P. distasonis ATCC: 8503.

[0007] Furthermore, the Parabacteroides distichum can alleviate liver, lung and kidney damage caused by sepsis.

[0008] The present invention also provides a drug for treating sepsis, wherein the drug comprises Parabacteroides distasonis and / or its metabolites, and the standard strain number of the Parabacteroides distasonis is P. distasonis ATCC: 8503.

[0009] Furthermore, the drug also includes pharmaceutically acceptable excipients.

[0010] Furthermore, the dosage form of the pharmaceutical preparation is any one of a liquid preparation, a solid preparation, and a semisolid preparation.

[0011] The beneficial effects of the present invention compared with the prior art are:

[0012] The present invention utilizes Parabacteroides distasonis P.distasonis ATCC: 8503 to treat sepsis, and detects the therapeutic effect of Parabacteroides distasonis P.distasonis ATCC: 8503. The results show that administration of Parabacteroides distasonis ATCC: 8503 can significantly improve the survival rate of septic mice, reduce serum ALT, AST, BUN and Creatinine contents, and reduce the lung wet / dry weight ratio, thereby effectively alleviating sepsis-induced liver, lung and kidney damage. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0014] Figure 1 This is the result of Parabacteroides distichous improving the survival rate of septic mice. The data were analyzed by Log-rank (Mantel-Cox) test. * indicates that the difference compared with the LPS group is statistically significant p < 0.05;

[0015] Figure 2This is a graph showing the results of Example 2 of the present invention in which Parabacteroides distichous alleviates LPS-induced sepsis liver, lung, and kidney damage in mice, wherein A and B are graphs showing the results of ALT and AST levels in serum of each group; C and D are graphs showing the results of BUN and Creatinine levels in serum of each group; and E is a graph showing the results of lung wet / dry weight ratios of each group; * indicates that the difference compared with the LPS group is statistically significant, p<0.05. DETAILED DESCRIPTION

[0016] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0017] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0018] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0019] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to the skilled artisan. The present invention description and examples are exemplary only.

[0020] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0021] Example 1

[0022] Example 1 of the present invention detected the effect of Parabacteroides distichous on the survival rate of mice with sepsis induced by lipopolysaccharide (LPS), and the specific steps were as follows:

[0023] (1) Experimental Materials

[0024] Experimental animals

[0025] Twenty-four male C57BL / 6J mice aged 6-8 weeks and weighing 18-22 g were selected for the experiment. They were purchased from the Animal Center of Nanfang Hospital of Southern Medical University and raised in the SPF-level Animal Experiment Department of Nanfang Hospital of Southern Medical University. All operations involved in the animal breeding process were approved by the Ethics Committee and met the requirements of animal ethics.

[0026] Reagents and instruments

[0027] Parabacteroides distasonis (ATCC: 8503, Beina Cell Microbial Collection Management Center); CLARIOstar all-function multifunctional microplate reader (BMG, Germany); blood agar plate (Guangdong Huankai Microbiological Technology Co., Ltd.); round-bottom vertical anaerobic culture bag; anaerobic gas production bag (Guangdong Huankai Microbiological Technology Co., Ltd.); isoflurane (Reward Life Science Technology Co., Ltd.); physiological saline (Shijiazhuang Siyao Co., Ltd.); phosphate buffered saline (PBS) pH 7.4 buffer (Gibco).

[0028] (2) Experimental methods and results

[0029] 1) Animal experiments:

[0030] Sepsis model induced by intraperitoneal injection of LPS: 6-8 week old C57BL / 6J mice were randomly divided into Sham group, LPS group, and LPS group pretreated with Parabacteroides distasonis by intraperitoneal gavage (LPS+P.distasonis).

[0031] Sham group: mice were intraperitoneally injected with PBS solution;

[0032] LPS group: mice were intraperitoneally injected with 15 mg / kg LPS solution;

[0033] LPS+P.distasonis group: mice were treated with 200μL P.distasonis by oral gavage for 7 consecutive days, and then injected with 15mg / kg LPS solution intraperitoneally;

[0034] 2) Survival rate statistics

[0035] The death time points of the mice in the above groups were observed and recorded, and the survival time of the mice in each group was counted respectively.

[0036] Results Figure 1 , Figure 1 This is a graph showing the results of P. distasonis bacteria improving the survival rate of septic mice; Figure 1The symbols in the middle mean: data were analyzed using Log-rank (Mantel-Cox) test, * indicates that the difference compared with the I / R group is statistically significant p<0.05, Figure 1 The results showed that treatment with P. distasonis bacteria could significantly increase the survival time of septic mice.

[0037] Example 2

[0038] Example 2 of the present invention detected the therapeutic effect of Parabacteroides dissimilar on LPS-induced extraintestinal organ liver, lung, and kidney damage in mice with sepsis. The specific steps are as follows:

[0039] (1) Experimental Materials

[0040] Experimental animals

[0041] Twenty-four male C57BL / 6J mice aged 6-8 weeks and weighing 18-22 g were selected for the experiment. They were purchased from the Animal Center of Nanfang Hospital of Southern Medical University and raised in the SPF-level Animal Experiment Department of Nanfang Hospital of Southern Medical University. All operations involved in the animal breeding process were approved by the Ethics Committee and met the requirements of animal ethics.

[0042] Reagents and instruments

[0043] Parabacteroides distasonis (ATCC: 8503, Beijing Cell Microbial Collection Management Center); CLARIOstar all-function multifunctional microplate reader (BMG, Germany); blood plate (Guangdong Huankai Microbiological Technology Co., Ltd.); round-bottom vertical anaerobic culture bag; anaerobic gas production bag (Guangdong Huankai Microbiological Technology Co., Ltd.); isoflurane (Reward Life Science Technology Co., Ltd.); physiological saline (Shijiazhuang Siyao Co., Ltd.); phosphate buffered saline (phosphate buffersaline, PBS) pH 7.4 buffer (Gibco); Alanine aminotransferase (ALT) detection kit (Nanjing Jiancheng Bioengineering Institute); Aspartate aminotransferase (AST) detection kit (Nanjing Jiancheng Bioengineering Institute); Blood urea nitrogen BUN detection kit (Nanjing Jiancheng Bioengineering Institute); Blood creatinine Creatinine detection kit (Nanjing Jiancheng Bioengineering Institute).

[0044] (2) Experimental methods and results

[0045] 1) Animal experiments:

[0046] Sepsis model induced by intraperitoneal injection of LPS: 6-8 week old C57BL / 6J mice were randomly divided into Sham group, LPS group, and LPS group pretreated with Parabacteroides distasonis by intraperitoneal gavage (LPS+P.distasonis).

[0047] Sham group: mice were intraperitoneally injected with PBS solution, and samples were collected 12 hours later;

[0048] LPS group: mice were intraperitoneally injected with 15 mg / kg LPS solution, and samples were collected 12 h later;

[0049] LPS+P.distasonis group: mice were treated with 200μL P.distasonis by intragastric administration for 7 consecutive days, and then injected with 15mg / kg LPS solution intraperitoneally, and samples were collected 12h later;

[0050] 2) Detection of ALT and AST levels in serum

[0051] The serum was obtained after centrifugation of blood collected from the dissected mouse heart, and the procedure was performed according to the instructions of the mouse serum ALT and AST detection kit.

[0052] 3) Detection of BUN and Creatinine levels in serum

[0053] The serum was obtained after centrifugation of blood collected from the dissected mouse heart, and the procedure was performed according to the instructions of the mouse serum BUN and Creatinine detection kit.

[0054] 4) Lung wet / dry weight ratio detection method

[0055] The whole lung of the mouse was taken out and the wet weight was measured. The lung was then placed in a 65°C incubator to dry. After 24 hours, the lung was taken out and the dry weight was measured to calculate the lung wet / dry weight ratio.

[0056] Figure 2 AD results showed that compared with the Sham group, the ALT, AST, BUN and Creatinine levels in the serum of the LPS group mice were significantly increased; compared with the LPS group, the ALT, AST, BUN and Creatinine levels in the serum were significantly reduced after P. distasonis pretreatment. Figure 2 E The results showed that compared with the Sham group, the lung wet / dry weight ratio of mice in the LPS group increased significantly; compared with the LPS group, the lung wet / dry weight ratio was significantly reduced after pretreatment with P. distasonis bacteria. The above experimental results indicate that P. distasonis bacteria can alleviate the liver, kidney and lung damage induced by LPS sepsis in mice.

[0057] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A use of Parabacteroides distichous in the preparation of a drug for treating sepsis, characterized in that: The standard strain number of the Parabacteroides distichous is P. distasonis ATCC:8503.

2. The use of Parabacteroides distichoui in the preparation of a drug for treating sepsis according to claim 1, characterized in that: The Parabacteroides distichum can alleviate the liver, lung and kidney damage caused by sepsis.

3. A drug for treating sepsis, characterized in that: The drug includes Parabacteroides distichous and / or its metabolites, and the standard strain number of the Parabacteroides distichous is P. distasonis ATCC:8503.

4. The drug for treating sepsis according to claim 3, characterized in that: The drug also includes pharmaceutically acceptable excipients.

5. The drug for treating sepsis according to claim 4, characterized in that: The dosage form of the pharmaceutical preparation is any one of a liquid preparation, a solid preparation, and a semisolid preparation.

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