CN109620848B - Application of lactococcus lactis - Google Patents
Application of lactococcus lactis Download PDFInfo
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- CN109620848B CN109620848B CN201811621336.4A CN201811621336A CN109620848B CN 109620848 B CN109620848 B CN 109620848B CN 201811621336 A CN201811621336 A CN 201811621336A CN 109620848 B CN109620848 B CN 109620848B
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- 230000009401 metastasis Effects 0.000 claims abstract description 7
- 239000002246 antineoplastic agent Substances 0.000 claims abstract description 5
- 230000001737 promoting effect Effects 0.000 claims abstract description 5
- 229940041181 antineoplastic drug Drugs 0.000 claims abstract description 4
- 206010054094 Tumour necrosis Diseases 0.000 claims abstract description 3
- 239000003814 drug Substances 0.000 claims description 15
- 201000001441 melanoma Diseases 0.000 claims description 13
- 230000000259 anti-tumor effect Effects 0.000 claims description 10
- 238000002360 preparation method Methods 0.000 claims description 9
- 239000007924 injection Substances 0.000 claims description 2
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- 229940124597 therapeutic agent Drugs 0.000 claims 1
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- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
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- 235000021472 generally recognized as safe Nutrition 0.000 description 2
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- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/04—Antineoplastic agents specific for metastasis
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/21—Streptococcus, lactococcus
- A23V2400/231—Lactis
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- General Health & Medical Sciences (AREA)
- Mycology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Molecular Biology (AREA)
- Epidemiology (AREA)
- Oncology (AREA)
- Nutrition Science (AREA)
- Engineering & Computer Science (AREA)
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- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention belongs to the technical field of biology, and particularly discloses application of lactococcus lactis. The invention applies lactococcus lactis in preparing antitumor drugs or foods, and comprises the following steps: inhibiting tumor growth, prolonging life of tumor-bearing model, promoting tumor necrosis, and resisting tumor metastasis.
Description
Technical Field
The invention relates to the technical field of biological medicines, and in particular relates to application of lactococcus lactis in preparation of antitumor medicines or foods.
Background
Lactococcus lactis is a kind of probiotics with a large content in intestinal tracts, mainly exists in dairy products and plant products, and the safety index of lactococcus lactis evaluated by Food and Drug Administration (FDA) is Generally Recognized As Safe (GRAS), so that lactococcus lactis is widely applied to the food industry. Since its discovery and initial use in dairy fermentation, lactococcus lactis has also recently developed rapidly in biotechnological production.
Cancer is a common frequently-occurring disease seriously harming human health, and overcoming the tumor problem is always a hot problem of domestic and foreign research. To date, anti-tumor methods include drug therapy, surgical resection, radiation therapy, chemotherapy, and the like. In the medical treatment, a plurality of antitumor drugs are limited in use due to large toxic and side effects.
Disclosure of Invention
The invention aims to provide a lactococcus lactis which is applied to preparing a medicament or food for resisting tumors, inhibiting tumor growth, prolonging the life cycle of a tumor-bearing model, promoting large-area tumor necrosis and resisting tumor metastasis. The invention not only discovers that the lactococcus lactis can inhibit the growth of tumors, but also discovers that the lactococcus lactis can prolong the life cycle of a tumor-bearing model, can promote large-area necrosis of the tumors and can inhibit tumor metastasis.
In addition, the application of lactococcus lactis in preparing antitumor drugs or foods is also studied.
Preferably, the tumor comprises: liver cancer cell, pancreatic cancer cell, lung cancer cell, colorectal cancer cell, breast cancer cell, cervical cancer cell, melanoma and the like.
The application of lactococcus lactis in preparing anti-melanoma drugs or foods is researched.
Preferably, the anti-melanoma tumor comprises inhibiting the growth of a melanoma tumor, prolonging the survival of a melanoma tumor model, promoting the extensive necrosis of a melanoma tumor, and inhibiting tumor metastasis.
Preferably, the application of lactococcus lactis is to prepare the preparation in the form of injection, vein, oral or spray by using the conventional preparation technology.
Preferably, the application of lactococcus lactis in preparing an anti-tumor medicament or food which is combined with a conventional anti-tumor treatment medicament or food.
Preferably, the application of the lactococcus lactis is the application of preparing an anti-melanoma medicine or food which is combined with a conventional anti-melanoma medicine or food.
The beneficial effects of the invention are as follows:
(1) The invention discovers that the lactococcus lactis has obvious anti-tumor effect, and the method comprises the following steps: obviously inhibit the growth of tumors and effectively prolong the life cycle of tumor-bearing models; the result of the hematoxylin-eosin (HE) staining on the tumor tissue shows that the lactococcus lactis can promote the large-area necrosis of tumor cells, thereby inhibiting the tumor growth and inhibiting the tumor metastasis.
(2) The lactococcus lactis disclosed in this patent can be administered orally.
Drawings
FIG. 1 is a graph showing the volume change of melanoma in mice of different administration groups;
FIG. 2 shows the survival rate of mice in different administration groups;
FIG. 3 is a graph showing the results of HE staining of tumor sections (A: control group; B: experimental group).
Detailed Description
The present invention will be described in further detail with reference to the following embodiments.
Example 1
(1) Grouping and administration method
The invention respectively tries tumor models inoculated by liver cancer cells, pancreatic cancer cells, lung cancer cells, colorectal cancer cells, breast cancer cells, cervical cancer cells and melanoma cell strains, all show similar good anti-tumor effects, and only the research results of representative tumors are listed here.
The tumor volume of the mice is about 100mm 3 Tumor-bearing mice were randomly divided into 2 groups, which were a control group and an experimental group, respectively. Transferring the newly activated lactococcus lactis NZ9000 to a GM17 liquid culture medium, and performing anaerobic static culture at 30 ℃ overnight; the culture is continued by the transfer according to the proportion of 1; centrifuging the bacterial liquid at 5000rpm for 5min, discarding supernatant, collecting bacteria, washing with sterile physiological saline containing 5% glucose twice, resuspending bacteria, and adjusting final concentration of bacteria to 2 × 10 according to OD value 9 CFU/mL is ready for use. Experimental group Each mouse was gavaged daily and 100. Mu.L of freshly prepared bacterial liquid was obtained, i.e. the gavaged bacterial load of each mouse was 2X 10 8 CFU; control groups Each mouse was gavaged daily with 100. Mu.L of sterile saline containing 5% glucose. All mice were gavaged continuously for 15 days, every 2 days according to the tumor volume formula V =0.52 × length × width 2 Mouse tumor size data and mouse survival were recorded.
Mice were inoculated with 2X 10 5 Tumor black spot growth was seen on day 6 after each B16F1 melanoma cell, with a tumor model generation rate of 100%, and tumor growth of mouse B16F1 melanoma was recorded within 15 days of continuous administration per group as shown in FIG. 1.
As can be seen from fig. 1, analysis of the mouse melanoma tumor growth curve comparing the differences between the two groups using t-test according to GraphPad Prism7 statistical software, it can be found that the experimental group orally fed lactococcus lactis exhibited inhibition of tumor growth (P < 0.0001) compared to the control group, from which it can be concluded: the oral lactococcus lactis has a therapeutic effect on mouse melanoma.
The survival conditions of the melanoma-bearing mice recorded within 15 days after each group is continuously administrated are shown in figure 2, and it can be seen that the survival conditions of the melanoma-bearing mice are analyzed by a survival analysis method (Kaplan-Meier method) according to GraphPad Prism7 statistical software, the survival rate of the tumor-bearing mice in a control group can be prolonged by oral feeding of lactococcus lactis, the survival rate of the tumor-bearing mice in the control group is 25% after the mice are subjected to gastric lavage for 15 days, the survival rate of the mice in the experimental group is still more than 40%, and the effect of remarkably prolonging the survival rate of the mice in the oral feeding of lactococcus lactis is proved compared with that in the control group.
(2) HE staining of tumor sections
Taking out the tumor of the mouse in a sterile environment to prepare a paraffin section; sequentially placing paraffin sections into xylene I20min, xylene II 20min, anhydrous ethanol I5min, anhydrous ethanol II 5min and 75% alcohol 5min, and then washing with tap water for dewaxing; staining the slices in hematoxylin staining solution for 3-5min, washing with tap water, differentiating the differentiation solution, washing with tap water, returning blue to the blue solution, and washing with running water; the slices are dehydrated for 5min respectively by 85 percent and 95 percent gradient alcohol in sequence and are dyed for 5min in eosin dye solution; placing the slices in anhydrous ethanol I for 5min, anhydrous ethanol II for 5min, anhydrous ethanol III for 5min, xylene I for 5min and xylene II for 5min, sealing with neutral gum; microscopic examination, image acquisition and analysis, and the result is shown in fig. 3.
As can be seen from FIG. 3, the cell distribution of the tumor sections of the control group is relatively complete, and the tumor cells of the experimental group have a small part of necrosis, and the result proves that the oral lactococcus lactis can cause the necrosis of a part of tumor cells.
The invention also tries to apply other conventional probiotic preparation methods to the lactococcus lactis preparation, and different preparation forms have the effects of better resisting tumors, inhibiting the growth of the tumors, prolonging the life cycle of tumor-bearing models, promoting large-area necrosis of the tumors and inhibiting tumor metastasis; the invention also tries to combine the lactococcus lactis with the conventional anti-tumor medicament or treatment method to further improve the tumor treatment effect like the conventional anti-tumor medicament or food; the invention also tries to combine the lactococcus lactis with the conventional anti-melanoma treatment medicine or food, so that the anti-melanoma effect can be further improved.
The above-mentioned embodiments are intended to illustrate the objects, technical solutions and advantages of the present invention in further detail, and it should be understood that the above-mentioned embodiments are only exemplary embodiments of the present invention, and are not intended to limit the present invention, and any modifications, equivalents, improvements and the like made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (4)
1. The application of lactococcus lactis is characterized by being applied to the preparation of antitumor drugs;
the anti-tumor is one or more of inhibiting tumor growth, prolonging life cycle of a tumor-bearing model, promoting large-area tumor necrosis and resisting tumor metastasis;
the tumor is a melanoma tumor;
the lactococcus lactis is L.lactis NZ9000.
2. The use of lactococcus lactis according to claim 1, for the preparation of an antitumor agent for use in combination with a conventional antitumor therapeutic agent.
3. The use of lactococcus lactis according to claim 1, wherein the medicament is in the form of an injection, an oral preparation or a spray.
4. Use of lactococcus lactis according to claim 3, wherein the medicament is in the form of an intravenous formulation.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811621336.4A CN109620848B (en) | 2018-12-28 | 2018-12-28 | Application of lactococcus lactis |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811621336.4A CN109620848B (en) | 2018-12-28 | 2018-12-28 | Application of lactococcus lactis |
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| CN109620848A CN109620848A (en) | 2019-04-16 |
| CN109620848B true CN109620848B (en) | 2022-10-14 |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20140020685A (en) * | 2012-08-10 | 2014-02-19 | 한국생명공학연구원 | A novel lactococcus sp. strain and use thereof |
| CN104382948A (en) * | 2014-11-21 | 2015-03-04 | 江南大学 | Application method of lactococcus lactis in treating intestinal diseases |
| CN107058366B (en) * | 2017-03-03 | 2020-03-24 | 内蒙古自治区农牧业科学院 | Lactococcus lactis food-grade double-screening marker secretion expression vector, engineering strain containing vector, construction method and application thereof |
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2018
- 2018-12-28 CN CN201811621336.4A patent/CN109620848B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20140020685A (en) * | 2012-08-10 | 2014-02-19 | 한국생명공학연구원 | A novel lactococcus sp. strain and use thereof |
| CN104382948A (en) * | 2014-11-21 | 2015-03-04 | 江南大学 | Application method of lactococcus lactis in treating intestinal diseases |
| CN107058366B (en) * | 2017-03-03 | 2020-03-24 | 内蒙古自治区农牧业科学院 | Lactococcus lactis food-grade double-screening marker secretion expression vector, engineering strain containing vector, construction method and application thereof |
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