CN114774302A - Lactobacillus plantarum CQPC02, and separation method and application thereof - Google Patents
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- CN114774302A CN114774302A CN202210224353.4A CN202210224353A CN114774302A CN 114774302 A CN114774302 A CN 114774302A CN 202210224353 A CN202210224353 A CN 202210224353A CN 114774302 A CN114774302 A CN 114774302A
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- 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
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses lactobacillus plantarum which is lactobacillus plantarum CQPC02, is preserved in China general microbiological culture Collection center of the Committee for culture Collection of microorganisms, and has a preservation number of CGMCC No. 14491. Also discloses application of the lactobacillus plantarum CQPC02 in preparing a product for preventing and/or treating lupus nephritis. The application observes the intervention effect of the lactobacillus plantarum CQPC02 on lupus nephritis by establishing an animal model. The effect of the lactobacillus plantarum CQPC02 on lupus nephritis is really realized by detecting related indexes and inflammation related cytokine levels in mouse serum, and the action mechanism of the lactobacillus plantarum CQPC02 is further deeply clarified by pathological observation and expression detection of tissue mRNA and protein genes.
Description
Technical Field
The invention relates to the technical field of microorganisms and biology, in particular to lactobacillus plantarum CQPC02 and a separation method and application thereof.
Background
Sichuan pickles are traditional Chinese naturally fermented vegetables. Its production history is about two thousand years. The long history of production and use makes it a Chinese marking food. The area for eating the pickle frequently exceeds 2000 million hectares. The Sichuan pickle is rich in nutrition and beneficial substances such as vitamins and mineral elements. Besides being used as a side dish, it can also be used with other foods to make delicious dishes. Due to the geographical environment of the Sichuan basin and the unique living habits of mixed life of multiple nationalities, the Sichuan naturally fermented pickle has special quality, and a large amount of fermentation microorganisms are generated in the natural fermentation process. Studies have shown that microorganisms found in naturally fermented kimchi are useful in the food industry, and they also exhibit various biological activities, including intestinal protection, weight loss, anti-inflammatory, and anti-oxidative effects. The microorganisms in the natural fermented pickle in Sichuan China are various, have good development and utilization values, and are potential probiotic resource sources.
Lupus nephritis is systemic lupus erythematosus, a complex immune disorder nephritis, and lupus nephritis often appears after renal failure, and the symptoms of glomerulonephritis appear after the onset of the disease. Lupus nephritis causes diseases such as immunity reduction, lymph node hyperplasia and glomerulonephritis, renal tissue can generate glomerular sclerosis or diffuse hyperplasia, metabolism and toxin expelling functions of a patient are seriously damaged, chronic renal failure is finally caused, and life is threatened in severe cases. Most lupus nephritis shows excessive activation of B cells and T cells after onset, the T cells are differentiated into helper T cells, and the helper T cells only play a role in an intermediate process in immunity, but more T cells can be developed into regulatory T cells capable of dissolving inflammation in the presence of probiotics, so that nephritis is directly interfered. The self-structural components of the probiotics can also stimulate and activate the immune system directly in an antigen mode, and play a role in discharging toxic substances in vivo and relieving inflammation. The phytoalkane can successfully construct a lupus nephritis model in experimental research, is generally agreed in scientific research, and has been used for testing the effect of medicaments and health-care food on lupus nephritis, and the phytoalkane can cause inflammation and strengthen immune reaction, so that T cells are highly activated, and the reactivity of B cells is greatly increased, thereby promoting animals to generate various autoantibodies, and further generating the lupus nephritis.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: lactobacillus plantarum is provided.
In order to solve the technical problems, the technical scheme is as follows: the lactobacillus plantarum is lactobacillus plantarum CQPC02, the strain is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No. 14491.
The lactobacillus plantarum has the following preservation information:
classification and naming of lactobacillus plantarum;
latin article name: lactobacillus plantarum;
the unit for preserving the biological material sample is full name: china general microbiological culture Collection center;
address: xilu No.1 Hospital No. 3, Beijing, Chaoyang, Beicheng;
the preservation date is as follows: collected by the 04 th collection in 08 months in 2017 and registered in a book; the preservation center tests the survival of the plants in 2017 at 04 th month 08;
the preservation number is as follows: CGMCC No. 14491.
The application also protects a method for separating and purifying lactobacillus plantarum, which comprises the following steps: respectively taking 1mL of sauerkraut water sample, and performing 10-fold gradient dilution to 10 with sterile physiological saline water-6Then take 10 out-4、10-5、 10-6Coating a plate by using 100 mu L of 3 gradient bacterial liquids, culturing at 37 ℃ for 24-48h, and observing and recording colony morphology; and selecting colonies with different forms on the plate for streaking separation, culturing at 37 ℃ for 48h, then selecting single colonies with different forms on the plate again for streaking separation, and repeating the steps for 2 to 3 times until pure single colonies with consistent forms are obtained.
The application also protects the application of the lactobacillus plantarum CQPC02 in preparing products for preventing and/or treating lupus nephritis.
The product is food, health product or medicine.
The application also protects a product for preventing and/or treating lupus nephritis, the active ingredient of the product is lactobacillus plantarum, the lactobacillus plantarum is lactobacillus plantarum CQPC02, and the preservation number of the lactobacillus plantarum is CGMCC number 14491.
The application also protects a product, the active ingredient of which is lactobacillus plantarum, the lactobacillus plantarum is lactobacillus plantarum CQPC02, and the preservation number is CGMCC No. 14491.
The product is food, health product or medicine.
Has the advantages that: the lactobacillus plantarum CQPC02 is a lactobacillus strain which is separated and identified by the research team from natural fermented yoghurt prepared by Tibetan herdsman families collected from Sichuan Hongyuan, has better in-vitro resistance basically, the survival rate in artificial gastric juice with the pH value of 3.0 is over 90 percent, and the growth efficiency in bile salt with the pH value of 0.3 percent is close to 70 percent.
The application observes the intervention effect of the lactobacillus plantarum CQPC02 on lupus nephritis by establishing an animal model. The effect of the lactobacillus plantarum CQPC02 on lupus nephritis is really realized by detecting related indexes and inflammation related cytokine levels in mouse serum, and the action mechanism of the lactobacillus plantarum CQPC02 is further deeply clarified by pathological observation and expression detection of mRNA and protein genes of tissues.
Drawings
FIG. 1H & E stained sections of mouse kidney tissue;
FIG. 2 mRNA expression in mouse kidney tissue.
Detailed Description
The process of the present invention is further illustrated by the following examples, which are not intended to limit the invention thereto.
A method for separating and purifying lactobacillus plantarum comprises the following steps: respectively taking 1mL of sauerkraut water sample, and performing 10-fold gradient dilution to 10 with sterile physiological saline water-6Then take 10 out-4、10-5、10-6Plating 3 gradient bacteria solution 100 μ L, culturing at 37 deg.C for 24-48h,observing and recording the colony morphology; and selecting colonies with different forms on the plate for streaking separation, culturing at 37 ℃ for 48h, then selecting single colonies with different forms on the plate again for streaking separation, and repeating the steps for 2 to 3 times until pure single colonies with consistent forms are obtained.
1 materials and methods
1.1 materials and reagents
The lactobacillus plantarum CQPC02 is separated and identified in family natural fermentation pickle in Chongqing southern quay, the strain is preserved in China general microbiological culture Collection center of the Committee for culture Collection of microorganisms, and the patent preservation number is CGMCC No. 14491.
SPF-grade six-week-old female C57BL/J6 mice with the weight of 23 +/-2 g are purchased from Chongqing medical university experimental animal centers, and the production license number is SCXK (Yu) 2018-0003. The animal experiment in the research is approved by animal experiment ethics committee in functional food synergistic innovation of Chongqing, and the approval number is 2021050007B.
Shanghai Merline Biochemical technologies, Inc. for Norphytane; the Total Protein (TP), Serum Creatinine (SCR), urea nitrogen (BUN), Total Cholesterol (TC), Triglyceride (TG) and Albumin (ALB) determination kit Nanjing institute of bioengineering; horseradish peroxidase, Sigma usa; shanghai enzyme-linked Biotechnology, Inc. of IL-6, IL-12, TNF-alpha and IFN-gamma detection kits; TRIzol reagent Invitrogen, usa; SYBR Green PCR Master Mix, qPCR primers, SDS-PAGE precast gel, primary antibody, secondary antibody, Thermo Fisher Scientific, USA; protein concentration measurement kit Bio-Rad, USA; the other reagents are all domestic analytical purifiers.
1.2 instruments and devices
BX43 microscope, olympus, japan; varioskan LUX multifunctional microplate reader, StoponePlus quantitative PCR instrument, iBright imaging System, Thermo Fisher Scientific Inc. USA.
1.3 methods
1.3.1 animal experiments
C57BL/J6 mice are bred in the environment with the temperature of 20 +/-1 ℃ and the humidity of 30-40%, the experimental mice can freely eat and drink water, and the adaptive feeding of the mice is formally carried out after 7 daysAnd (4) performing an experiment. 50 mice were randomly divided into 5 groups of 10 mice each, normal, model, drug positive control, LP-CQPC02 low concentration treated (LP-CQPC02-L) and LP-CQPC02 high concentration treated (LP-CQPC02-H) groups, respectively. The normal mice were injected with normal saline solution once and the other mice were injected with 0.5mL norphytane once after the first day of the experiment. The drug positive control group mice are dosed with 10mg/kg peridoterin solution every day, and the LP-CQPC02-L group and LP-CQPC02-H group mice are dosed with 10 every day8CFU/kg and 109CFU/kg dose gavage LP-CQPC02, prednisone and LP-CQPC02 for gavage duration 12. After 12 weeks, the mice were sacrificed by cutting their necks, and heart blood and internal organs of the mice were dissected and collected for measurement.
1.3.2 urine protein assay
After the start of the experiment, the mice were kept in metabolic cages every two weeks, and the urination of the mice in 1 day was collected and the amount of protein in the urine of the mice was measured within 24 hours using a total protein kit (Coomassie Brilliant blue method).
1.3.3 serum and tissue inflammatory cytokine assays
The whole blood of the mouse is collected by taking the blood from the heart, and then the whole blood is centrifugally separated for 10min at 1500rpm and 4 ℃, and the upper serum is removed for the experiment. In addition, 0.1g of kidney tissue was weighed, 0.9mL of physiological saline was added to the kidney tissue, and then the kidney tissue was homogenized at 4 ℃ and centrifuged (4000rpm, 10min) to obtain the supernatant for the experiment. And finally, measuring the levels of IL-6, IL-12, TNF-alpha and IFN-gamma inflammatory cytokines in the serum and the tissues according to a detection kit method.
1.3.4 serum SCr, BUN, TC, TG and ALB level determination
Mouse serum was collected by 1.3.3 method and the levels of SCr, BUN, TC, TG and ALB in the mouse serum were determined by the test kit method.
1.3.5 anti-dsDNA antibody assay
Mice blood was collected using orbital bleeds two weeks after the start of the experiment and then assayed for anti-dsDNA antibodies using a microtiter plate and indirect immunofluorescence assay after mouse serum was prepared as per 1.3.3.
1.3.6 tissue section Observation
After the mice were dissected, kidney tissues of the mice were fixed with 10% formalin. After dehydrating for 48H, the tissue samples were embedded with paraffin and sectioned, and finally stained with hematoxylin-eosin (H & E) fuel and examined for histopathological changes using an optical microscope.
1.3.7 qPCR experiment
0.9mL of physiological saline was added to 0.1g of mouse kidney tissue, and the tissue mixture was homogenized. RNA was then extracted from mouse kidney tissue using RNAzol (1.0 mL). The absorbance values of the extracted RNAs were measured at 260nm and 280nm, the RNA purity and concentration were calculated, and the RNA concentration was adjusted to 1. mu.g/. mu.L. After reverse transcription to generate cDNA, 1. mu.L of cDNA, 10. mu.L of SYBR Green PCR Master Mix, 7. mu.L of sterile distilled water and 1. mu.L of upstream and downstream primer solutions were mixed to prepare a reaction system solution. Followed by 60s at 95 ℃; at 95 ℃ for 15s, 40 cycles; at 55 ℃ for 30 s; 72 ℃ for 35 s; at 95 ℃ for 30 s; the reaction is carried out at 55 ℃ for 35s and 2 is used-ΔΔCtThe method quantitatively analyzes related genes, and GAPDH is used as an internal reference in the experiment (Table 1).
TABLE 1 primer sequences used in this experiment
1.4 statistical analysis
After the animal experiments were completed, all the indexes were subjected to three replicates, and the results obtained by the measurements were expressed as mean values while standard deviations (mean ± standard deviation) were noted. And then whether each group of index values obtained by adopting one-factor variance analysis has a significant difference on P <0.05 or not.
2 results and analysis
2.1 amount of protein in urine of mouse
During the experiment, the amount of protein in urine of normal group mice did not change significantly, but urine protein of other group mice increased with time. After 2 weeks, 7, 5 and 4 mice developed proteinuria and all model mice developed proteinuria under the action of LP-CQPC02-L, LP-CQPC02-H and prednisone. After 12 weeks, the urine protein content of lupus nephritis mice (model group) was higher than that of LP-CQPC02 and prednisone-treated mice and normal group mice (table 2). The urine protein output of mice treated with high concentrations of LP-CQPC02 and prednisone was closest to that of normal mice, and the urine protein of mice in the group LP-CQPC02-H was only higher than that of mice in the prednisone group.
TABLE 2 urine protein content of each group of mice during the experiment
Note: different letters indicate significant differences between groups at a P <0.05 level, the same letters indicate no significant differences, the following.
2.2 mouse serum and Kidney tissue IL-6, IL-12, TNF-alpha and IFN-gamma cytokine levels
As shown in tables 3 and 4, the serum and kidney tissues of mice in the normal group exhibited significantly lower levels of IL-6, IL-12, TNF- α and IFN- γ cytokines than those in the other groups (P < 0.05). IL-6, IL-12, TNF-alpha and IFN-gamma cytokine levels were significantly downregulated (P <0.05) in lupus nephritis mice (model group) after the action of LP-CQPC02 and prednisone versus model group mice, and the ability of high concentrations of LP-CQPC02(LP-CQPC02-H) and prednisone to downregulate these cytokine levels was stronger.
TABLE 3 mouse serum IL-6, IL-12, TNF-alpha and IFN-gamma levels
TABLE 4 mouse Kidney tissue IL-6, IL-12, TNF- α and IFN- γ levels
2.3 mouse serum SCR, BUN, TC, TG, TP and ALB levels
Serum levels of SCr, BUN, TC and TG were higher in the model group mice than in the other groups, while serum levels of SCr, BUN, TC and TG were lowest in the normal mice (P <0.05, table 5). Compared with the model group mice, the levels of SCr, BUN, TC and TG of the mice treated by LP-CQPC02 and prednisone are also reduced, but are higher than those of normal mice. LP-CQPC02 can approximate the levels of SCr, BUN, TC and TG in nephritis mice to normal levels. In addition, TP and ALB serum levels are in opposite trends, and the levels of each group are a normal group, a prednisone group, a LP-CQPC02-H group, a LP-CQPC02-H group and a control group from high to low.
TABLE 5 mouse serum SCR, BUN, TC, TG, TP and ALB levels
2.4 dsDNA positivity Rate
Autoantibodies dsDNA were detected by indirect immunofluorescence at weeks 2, 4, 6, 7, 10 and 12 post-treatment. The results show that all mice in the model group are positive from the end of 6 weeks, and the induction of lupus nephritis is successful. The mice in the LP-CQPC02-L group were tested to be positive at the end of 8 weeks, and the mice in the LP-CQPC02-H group and the prednisone group were tested to be positive at the end of 10 weeks, which means that LP-CQPC02 and prednisone slow down the rate of the mice to suffer from lupus nephritis, and the effects of LP-CQPC02 and prednisone are close (Table 6).
TABLE 6 dsDNA positivity rates of various groups of lupus nephritis mice
2.5 Kidney histopathological Observation
As shown in FIG. 1, the kidney tissues of the mice in the model group have serious pathological changes, a large number of glomeruli have irregular real forms, part of the glomeruli have rupture phenomena, and serious inflammatory cell infiltration phenomena occur among the tissues. The glomeruli and the cell structure of the mice in the normal group are complete, and LP-CQPC02 and prednisone can relieve kidney tissue pathological changes caused by lupus nephritis and relieve kidney group damage. Meanwhile, the high-concentration LP-CQPC02(LP-CQPC02-H) and prednisone have good effects, and can promote the tissue morphology of kidney tissues to be close to that of a normal group.
2.6 mRNA expression in mouse Kidney tissue
As shown in FIG. 2, the normal group of mouse kidney tissues showed the least expression of mRNA for NF-. kappa. B, TGF-. beta.1, VEGF, ICAM-1 and VCAM-1, and the most expression of I.kappa.B-. alpha.was observed. The expression of IkB-alpha is the weakest in the model group, and the expression of NF-k B, TGF-beta 1, VEGF, ICAM-1 and VCAM-1 is the strongest. Compared with the model group, the LP-CQPC02 and prednisone can remarkably up-regulate the expression of I kappa B-alpha of kidney tissues of the model group, down-regulate the expression of NF-kappa B, TGF-beta 1, VEGF, ICAM-1 and VCAM-1 (P <0.05), and the effects of the high-concentration LP-CQPC02(LP-CQPC02-H) and prednisone are stronger than those of the low-concentration LP-CQPC02(LP-CQPC 02-L).
Urinary protein plays an important role in the occurrence and development of kidney diseases, a large amount of proteinuria is one of main symptoms in complex comprehensive kidney diseases, and the occurrence of proteinuria is the clinical main manifestation of lupus nephritis. The lupus nephritis mice with the modeling result in the research also show proteinuria, and LP-CQPC02 and prednisone can reduce the protein amount in the urine of the lupus nephritis mice to play a role in relieving lupus nephritis, and the effect of LP-CQPC02 is increased along with the increase of the concentration. Serum creatinine and urea nitrogen are nitrogen-containing organic compounds that are the end products of protein metabolism. When kidney function is normal, these small molecules are filtered from the glomerulus. When the kidney is damaged, the filtration capacity of the glomerulus is reduced, and the serum creatinine and urea nitrogen content is increased, so that the increase of the serum creatinine and urea nitrogen level can be used as an index for clinical diagnosis of kidney damage. Excessive cholesterol and triglycerides are the cause of hyperlipidemia, and when renal disease worsens to some extent, the characteristics of hyperkalemia coexist. Thus, cholesterol and triglycerides can also be considered as indicators of reduced renal function and renal impairment. Nephrotic syndrome patients experience a significant reduction in total protein in serum due to long-term proteinuria. Albumin is the most common protein in serum and is commonly used in the treatment of severe disease, including edema due to renal disease, reduction in total protein and albumin content in the case of renal dysfunction in the clinic. Therefore, the maintenance of the content of the total serum protein and albumin is an important way for maintaining the normal renal function. In the research, LP-CQPC02 and prednisone also show the capability of inhibiting the increase of serum creatinine, urea nitrogen, cholesterol and triglyceride and the reduction of total protein and albumin caused by lupus nephritis, and play a role in protecting the kidney by regulating the relevant indexes of the nephropathy.
IL-12 plays an important role in autoimmune response of lupus nephritis, and the level of IL-12 is increased in the onset period of lupus nephritis. One of the characteristics of lupus nephritis is the appearance of a large amount of autoantibodies, IL-12 can promote the cells to directly produce the autoantibodies, and the increase of the IL-12 level further causes the large amount of the autoantibodies to be produced, thereby aggravating the disease. IFN-gamma is used as an inflammation medium and participates in the whole immune inflammation process of nephritis, and the IFN-gamma level of a patient with glomerulonephritis is clinically shown to be remarkably increased. After nephritis occurs, inflammatory-related cytokines are changed, and the contents of inflammatory-related cytokines such as IL-6, IL-12, TNF-alpha and IFN-gamma in blood are also significantly increased. In the research, the levels of inflammatory cytokines IL-6, IL-12, TNF-alpha and IFN-gamma of a lupus nephritis mouse are greatly increased, and LP-CQPC02 and prednisone can obviously inhibit the changes.
The essential process for autoreactive antibodies in lupus nephritis is the conversion to IgG by mutation and class. The massive deposition of IgG anti-dsDNA antibodies and immune complexes in plasma in glomeruli leads to kidney damage, which in turn causes inflammation leading to infiltration of inflammatory cells. In addition, a high concentration of dsDNA antibodies was found to occur almost exclusively in lupus nephritis, and the dsDNA antibodies showed specificity for lupus nephritis, and thus could be used as an index for diagnosing systemic lupus nephritis. Clinically, the kidney tissues of patients with lupus nephritis mostly have the phenomena of glomerular cell proliferation change, glomerular neutrophil infiltration and the like. The experimental indices of this study also confirm these manifestations, LP-CQPC02 and prednisone both marked the ability to inhibit the appearance of dsDNA antibodies and protect against pathological changes in renal tissue.
The NF-kB signal transduction pathway is involved in various pathological processes, so the signal pathway of the NF-kB signal transduction pathway is a potential target for intervention of drugs or bioactive substances. NF-. kappa.B is an important transcription factor that, when activated, activates a variety of immune and inflammation-related genes, including TNF, IL-1, IL-2, IL-6, IL-8, IL-10, adhesion molecules (ICAM-1, VCAM-1). NF- κ B is involved in a variety of pathophysiological processes and in the pathogenesis of renal disease. In nephritis, the body's inflammatory response is too intense, resulting in damage to the kidney's own tissues. In therapy, inhibition of NF- κ B activity may reduce inflammatory injury. The NF-. kappa.B activity is closely related to I.kappa.B-. alpha.activity. When the expression of I kappa B-alpha is weakened, NF-kappa B is separated from I kappa B-alpha, and the effects of promoting inflammation and damaging cells and tissues are achieved; when the expression of I kappa B-alpha is increased, the dissociated NF-kappa B can be rapidly combined with newly synthesized I kappa B, thereby reducing the activity of NF-kappa B and protecting organisms. Activation of NF-. kappa.B may cause inflammatory growth factors including TGF-. beta.1, and TGF-. beta.1 may activate NF-. kappa.B as a regulatory product through a positive feedback pathway. TGF-. beta.1 is the most important factor for promoting fibrosis in the body and is involved in organ fibrosis including the kidney, and TGF-. beta.1 is often abnormally expressed in renal diseases. TGF-. beta.1 in the kidney is secreted by podocytes, which secrete TGF-. beta.1 through a pre-endothelial association, and by mesangial cells (GMCs), which secrete TGF-. beta.1 upon stimulation by immunoglobulin (IgA). Glomerular endothelial cells are capable of TGF- β 1 secretion by stimulation with VEGF following renal damage. Abnormal expression of TGF-beta 1 and VEGF is a typical expression after lupus nephritis is developed, so that the two expressions can be regulated to effectively control the lupus nephritis and play a role in inhibiting inflammatory cytokines such as IL-6, IL-12, TNF-alpha and IFN-gamma. NF-kB is activated to enter nucleus and combine with target sequence and regulate the transcription activity of relevant genes, such as ICAM-1, VCAM-1, TNF-alpha, IL-6, etc. The enhancement and initiation of these factor genes comprises the NF-. kappa.B binding site. ICAM-1 and VCAM-1 are two adhesion factors, both belonging to the immunoglobulin superfamily. ICAM-1 and VCAM-1 mediate leukocyte adhesion to endothelial cells by binding to receptors on the leukocyte surface and subsequent leukocyte transfer across endothelial cells. The accumulation of leukocytes can block capillaries, and activated leukocytes can release a large amount of toxic substances to damage neurons and glial cells, thereby aggravating tissue damage and nephritis. In addition, the leukocytes also release some inflammatory mediators and cytokines, exacerbating the inflammatory response, and attracting more leukocytes into the tissue to form a malignant cycle.
The research observes a lactobacillus plantarum LP-CQPC02 newly found from naturally fermented pickled vegetables, and verifies the intervention effect of LP-CQPC02 on lupus nephritis through an animal model. The experimental result shows that LP-CQPC02 can relieve mouse serum and tissue inflammatory lesions caused by lupus nephritis, and particularly LP-CQPC02 can regulate TGF-beta 1 which is a marker of lupus nephritis. The existing lupus nephritis treatment medicines used clinically generally have certain side effects, prednisone serving as a medicine with small side effect in the research is used as a medicine positive control, the effect of LP-CQPC02 is found to be close to that of prednisone through comparison, and the lupus nephritis treatment medicines can be healthily intervened. Therefore, LP-CQPC02 has the function of being used as probiotics to intervene lupus nephritis.
Claims (7)
1. The lactobacillus plantarum is lactobacillus plantarum CQPC02, the strain is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No. 14491.
2. The method for separating and purifying Lactobacillus plantarum mentioned in the above, comprising the steps of: respectively taking 1mL of sauerkraut water sample, and performing 10-fold gradient dilution to 10 with sterile physiological saline water-6Then take 10 out-4、10-5、10-6Plating 100 mu L of 3 gradient bacteria liquid, culturing at 37 ℃ for 24-48h, and observing and recording colony morphology; and selecting colonies with different forms on the plate for streaking separation, culturing at 37 ℃ for 48h, then selecting single colonies with different forms on the plate again for streaking separation, and repeating the steps for 2 to 3 times until pure single colonies with consistent forms are obtained.
3. Application of lactobacillus plantarum CQPC02 in preparation of products for preventing and/or treating lupus nephritis.
4. Use according to claim 3, characterized in that: the product is food, health product or medicine.
5. The active ingredient of the product for preventing and/or treating lupus nephritis is lactobacillus plantarum CQPC02 with the preservation number of CGMCC No. 14491.
6. A product contains Lactobacillus plantarum as active ingredient, wherein the Lactobacillus plantarum is Lactobacillus plantarum CQPC02 with the preservation number of CGMCC No. 14491.
7. A product according to claim 5 or 6, characterized in that: the product is food, health product or medicine.
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| CN114854619A (en) * | 2022-03-09 | 2022-08-05 | 重庆第二师范学院 | Lactobacillus plantarum HFY09 and separation method and application thereof |
| CN114854619B (en) * | 2022-03-09 | 2023-09-29 | 重庆第二师范学院 | Lactobacillus plantarum HFY09 and separation method and application thereof |
| CN114854621A (en) * | 2022-03-10 | 2022-08-05 | 重庆第二师范学院 | Lactobacillus plantarum HFY15 and separation method and application thereof |
| CN114854621B (en) * | 2022-03-10 | 2023-09-29 | 重庆第二师范学院 | Lactobacillus plantarum HFY15 and separation method and application thereof |
| CN115505551A (en) * | 2022-11-23 | 2022-12-23 | 善恩康生物科技（苏州）有限公司 | Lactobacillus helveticus and application thereof in preventing or treating nephritis |
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