Application of lactobacillus plantarum GUANKE in prevention and treatment of renal injury

Abstract

The invention relates to the technical field of medicines, and discloses application of lactobacillus plantarum GUANKE in prevention and treatment of renal injury. A kidney injury mouse model is established through a ZEN infected mouse, and lactobacillus plantarum GUANKE powder is fed into the stomach of the kidney injury mouse, so that compared with a model group, the serum, kidney function, antioxidant enzyme activity and kidney related inflammation indexes of the mouse fed with the lactobacillus plantarum GUANKE are correspondingly improved; and the effect of high-dose treatment of the lactobacillus plantarum GUANKE is more remarkable than that of low-dose treatment of the lactobacillus plantarum GUANKE. In addition, the tissue pathological section of the kidney shows that compared with the inflammation, necrosis and lipid degeneration of the mouse tissue of the model group, the inflammation, necrosis and lipid degeneration of the mouse tissue are relieved after the treatment of the lactobacillus plantarum GUANKE. In conclusion, the lactobacillus plantarum GUANKE has the beneficial effects in treating the renal injury.

Description

Technical Field

[0001] The invention relates to the technical field of medicine, and in particular to application of Lactobacillus plantarum GUANKE in preventing and treating kidney damage. Background Art

[0002] Zearalenone (ZEN), also known as F-2 toxin, is a mycotoxin widely found in grains. It can cause oxidative damage to the kidneys and induce inflammation, posing a potential threat to animal and human health. Acute poisoning by zearalenone can cause muscle tremors and severe kidney damage in animals, and they may also suddenly fall to the ground and die; chronic poisoning by zearalenone can cause loss of appetite, severe fat loss, and poor growth. Therefore, in people's daily lives, it is necessary to prevent damage to the body caused by zearalenone. In addition, kidney damage can also occur during the treatment of anti-tumor drugs such as cisplatin. Kidney damage and other situations occur from time to time, and finding effective drugs to treat kidney damage has become an urgent problem to be solved.

[0003] There have been many studies in the prior art on the treatment of renal injury. For example, the patent with application publication number CN117503836A discloses the use of Quzhike and Quzhike extracts in the prevention and treatment of renal injury. The above patent extracts active drugs from Quzhike to treat the mouse model of acute renal injury caused by cisplatin, realizing the new use of Quzhike and extracts in the treatment of renal injury. However, the active drugs for the treatment of renal injury provided by the patent need to be extracted from Quzhike, the process is relatively complicated, and the efficacy of the extract will be limited by the extraction rate of the active drugs.

[0004] In order to provide more practical solutions for the treatment and relief of renal injury, more effective therapeutic drugs are still needed. Summary of the invention

[0005] In order to solve the above technical problems, the present invention provides the use of Lactobacillus plantarum GUANKE in preventing and treating kidney damage.

[0006] The specific technical scheme of the present invention is: In one aspect, the present invention provides use of Lactobacillus plantarum GUANKE in preparing a medicament for preventing and / or treating kidney injury.

[0007] Lactobacillus plantarum GUANKE, the strain preservation number is CGMCC NO.21720, the preservation date is January 22, 2021, the preservation classification name is Lactobacillus Plantarum GUANKE, and the preservation unit is the General Microbiology Center of China Microbiological Culture Collection Administration.

[0008] The present invention researchers establish renal injury mouse model by ZEN infection mice, and gavage plant lactobacillus GUANKE powder to renal injury mice, find that compared with the model group, the serum, renal function, antioxidant enzyme activity and kidney-related inflammation index of mice gavaged with plant lactobacillus GUANKE have been improved accordingly, and the effect of plant lactobacillus GUANKE high-dose treatment is more significant than the effect of plant lactobacillus GUANKE low-dose treatment. In addition, the histopathological sections of the kidney show that, compared to the inflammation, necrosis and lipid degeneration of the mouse tissue of the model group, plant lactobacillus GUANKE reduces inflammation, necrosis and lipid degeneration of the mouse tissue after treatment. In summary, show that plant lactobacillus GUANKE has a beneficial effect in treating renal injury.

[0009] As a preferred embodiment of the above application, the dosage form of the drug includes granules, ordinary tablets, dispersible tablets, effervescent tablets, orally disintegrating tablets, lozenges, chewable tablets, capsules, soft capsules, microcapsules, pills, powders, pellets, sustained-release preparations, controlled-release preparations, oral liquid preparations and injections.

[0010] Optionally, the renal injury is acute renal injury.

[0011] Optionally, the kidney damage is caused by zearalenone.

[0012] In another aspect, the present invention provides a composition for treating renal injury, wherein the active ingredient of the composition is Lactobacillus plantarum GUANKE.

[0013] As a preferred embodiment of the above composition, the renal injury is acute renal injury.

[0014] As a preferred embodiment of the above composition, the renal damage is caused by zearalenone.

[0015] Preferably, the above composition comprises one or more pharmaceutically acceptable excipients.

[0016] As a preferred embodiment of the above composition, the composition is prepared into granules, ordinary tablets, dispersible tablets, effervescent tablets, orally disintegrating tablets, lozenges, chewable tablets, capsules, soft capsules, microcapsules, pills, powders, pellets, sustained-release preparations, controlled-release preparations, oral liquid preparations or injections.

[0017] Compared with the prior art, the present invention has the following technical effects: 1. The present invention establishes a mouse model of renal injury by infecting mice with ZEN, and gavages Lactobacillus plantarum GUANKE powder to mice with renal injury. It is found that compared with the model group, the serum, renal function, antioxidant enzyme activity and kidney-related inflammation index of mice gavaged with Lactobacillus plantarum GUANKE have been improved accordingly, and the effect of high-dose treatment of Lactobacillus plantarum GUANKE is more significant than the effect of low-dose treatment of Lactobacillus plantarum GUANKE. In addition, the histopathological sections of the kidney show that inflammation, necrosis and lipid degeneration of mouse tissues are reduced after treatment of Lactobacillus plantarum GUANKE compared to inflammation, necrosis and lipid degeneration of mouse tissues in the model group. In summary, the beneficial effect of Lactobacillus plantarum GUANKE in treating renal injury.

[0018] 2. At the same time, as a probiotic, Lactobacillus plantarum GUANKE has less side effects on the body. Experiments have shown that compared with the blank control group, the body weight and kidney weight of mice gavaged with GUANKE did not change, which shows that GUANKE bacteria is harmless to mice. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 The statistical results of body weight measurement of mice in 5 experimental groups; Figure 2 The statistical results of kidney weights of mice in the five experimental groups are shown; Figure 3 The results of the detection of BUN concentration in the serum of mice in the five experimental groups; Figure 4 The results of the detection of UA concentration in the serum of mice in the five experimental groups; Figure 5 The results of the detection of CRE concentration in the serum of mice in the five experimental groups; Figure 6 The results of the detection of GSH activity in the kidneys of mice in the five experimental groups; Figure 7 The results of the detection of CAT activity in the kidneys of mice in the five experimental groups; Figure 8 The results of the detection of SOD activity in the kidneys of mice in the five experimental groups; Fig. 9 The results of the detection of MDA content in the kidneys of mice in the five experimental groups; Fig.10 The results of the detection of TNF-α content in the kidneys of mice in the five experimental groups; Fig.11 The results of the detection of IL-1β content in the kidneys of mice in the five experimental groups; Fig.12 The results of the detection of IL-6 content in the kidneys of mice in the five experimental groups; Fig.13 HE staining images of kidneys of mice in the five experimental groups. DETAILED DESCRIPTION

[0020] The present invention will be further described below in conjunction with embodiments. Those of ordinary skill in the art will be able to implement the present invention based on these descriptions. In addition, the embodiments of the present invention involved in the following description are generally only embodiments of a part of the present invention, rather than all embodiments. Therefore, based on the embodiments in the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative work should belong to the scope of protection of the present invention.

[0021] The plant lactobacillus GUANKE used in the embodiment of the present invention has a strain deposit number of CGMCC NO.21720, a deposit date of January 22, 2021, and a deposit classification named Lactobacillus Plantarum GUANKE. The deposit unit is the General Microbiological Center of the China Microbiological Culture Collection Administration Committee, and the address is the Institute of Microbiology, Chinese Academy of Sciences, No. 3, No. 1, Beichen West Road, Chaoyang District, Beijing, Postal Code: 100101. This strain is a public strain. The above-mentioned plant lactobacillus GUANKE is fermented and freeze-dried to obtain GUANKE powder.

[0022] Example 1 Establishment of mouse model and drug administration (1) Experimental preparation Before the experiment, 50 newly purchased 7-week-old BALB / C female mice were fed for 7 days of adaptation. Then, according to the random principle, these mice were randomly divided into 5 experimental groups, with 5 mice in each group. The experimental groups included Control group, GUANKE group, ZEN group, ZEN+LG group, and ZEN+HG group.

[0023] (2) Model establishment and drug administration The total drug administration of the 5 experimental groups is as follows: Control group: The animals were fed with the same amount of saline solution as the other groups every day during the experiment, for a total of 28 days; GUANKE group: GUANKE powder was administered intragastrically every day during the experiment, with a dosage of 1×10 10 CFU / mouse, total administration for 28 days; ZEN group: During the experiment, ZEN was administered intragastrically every day at a dose of 40 mg / kg. Four hours after oral administration of ZEN, PBS was administered intragastrically again at the same volume as that of the ZEN+LG group and ZEN+HG group. The administration lasted for 28 days in total. ZEN+LG group: ZEN was administered orally every day during the experiment at a dose of 40 mg / kg, and GUANKE (administered at a dose of 1×10 6 CFU / mouse), and the drug was administered for 28 days; ZEN+HG group: ZEN was administered orally every day during the experiment at a dose of 40 mg / kg, and GUANKE (administered at a dose of 1×10 10 CFU / mouse), and the drug was administered for 28 days in total.

[0024] The specific experimental process is: Day 1 to 27: During the 28-day test period, the animals were provided with adequate food and water and were free to eat. The mental state, behavior, diet, weight changes and deaths of mice in each group were observed and recorded every day. The mice in the five experimental groups were dosed daily according to the above-mentioned dosing conditions.

[0025] Day 28: After fasting for 24 hours, the mice in each group were weighed, the eyeballs were removed and blood was collected, and the serum was separated after standing at room temperature for 4 hours. The mice were killed by dislocation of the neck, the kidneys were removed and rinsed with PBS, wiped dry and weighed, and the kidney organ index was calculated, and the kidney surface was checked for organic lesions. The right kidney was taken for routine histopathological testing, and the left kidney was taken for molecular level related index testing.

[0026] Example 2 Histopathological study 1. Weight and organ indicators On the 28th day, the weight of mice in the five experimental groups was measured. Figure 1 , the statistical results of mouse kidney weight are shown in Figure 2 The body weight and kidney weight of mice can directly reflect the health status of mice.

[0027] The results of weighing the body weight and kidney weight of the mice showed that compared with the Control group, the body weight and kidney weight of the mice in the GUANKE group did not change; compared with the Control group, the body weight and kidney weight of the mice in the ZEN group decreased by 28% (P<0.01) and 33.5% (P<0.01), respectively; compared with the ZEN group, the body weight and kidney weight of the mice in the ZEN+LG group increased by 12.7% (P<0.05) and 32.2% (P<0.01), respectively; compared with the ZEN group, the body weight and kidney weight of the mice in the ZEN+HG group increased by 27% (P<0.01) and 38.2% (P<0.01), respectively; compared with the ZEN group, the body weight and kidney weight of the mice in the ZEN+HG group increased by 11.3% (P<0.01) and 8.8% (P<0.05), respectively. The above results indicate that GUANKE bacteria are harmless to mice, while ZEN is harmful to mice. ZEN reduces the body weight and kidney weight of mice, while GUANKE bacteria treatment reverses this damage, and within a certain range, the effect becomes better as the dose of GUANKE bacteria increases.

[0028] 2. Serological indicators On the 28th day, the concentrations of BUN, UA and CRE in the serum of mice were detected. The results of the BUN, UA and CRE concentrations of mice in the five experimental groups were shown in Table 1. Figure 3 , Figure 4 , Figure 5 . BUN, UA and CRE are substances metabolized in the kidneys and are important indicators for measuring the degree of kidney damage. In a healthy state, the concentrations of BUN, UA and CRE in the blood are relatively low; when the kidneys are damaged, the metabolism of BUN, UA and CRE in the blood will decrease, thereby increasing the levels of BUN, UA and CRE in the blood.

[0029] The results of BUN, UA and CRE concentrations in mice showed that compared with the Control group, the levels of BUN, UA and CRE in the serum of mice in the GUANKE group did not change; the levels of BUN, UA and CRE in the serum of mice in the ZEN group increased by 112% (P<0.01), 143% (P<0.01) and 71.2% (P<0.01), respectively. Compared with the ZEN group, the levels of BUN, UA and CRE in the serum of mice in the ZEN+LG group decreased by 8.2%, 14.8% and 0.4%, respectively; compared with the ZEN group, the levels of BUN, UA and CRE in the serum of mice in the ZEN+HG group decreased by 32.3% (P<0.01), 33.1% (P<0.01) and 33.2% (P<0.01), respectively. Compared with the ZEN+LG group, the levels of BUN, UA and CRE in the serum of mice in the ZEN+HG group were down-regulated by 22.3% (P<0.05), 15.9% (P<0.05) and 32.7% (P<0.05), respectively. The above results show that ZEN can cause kidney damage in mice, and GUANKE powder can treat kidney damage caused by ZEN, and the effect is better with the increase of dose within a certain range.

[0030] 3. Oxidative stress indicators On the 28th day, the activities of GSH, CAT, SOD and the content of MDA in the kidneys of mice were detected. The results of the activities of GSH, CAT, SOD and the content of MDA were shown in Table 1. Figure 6 , Figure 7 , Figure 8 , Fig. 9 .

[0031] The results of the test on the activities of GSH, CAT, SOD and the content of MDA in the kidneys of mice showed that compared with those of the Control group, there was no difference in the activities of GSH, CAT, SOD and the content of MDA in the kidneys of mice in the GUANKE group; compared with the Control group, the activities of GSH, CAT and SOD in the kidneys of mice in the ZEN group were downregulated by 62.2% (P<0.01), 55.1% (P<0.01) and 22.3% (P<0.01), and the content of MDA was upregulated by 123.4% (P<0.01). Compared with the ZEN group, the activities of GSH, CAT and SOD in the kidneys of mice in the ZEN+LG group were increased by 22.8%, 25.9% and 5.8%, and the content of MDA was decreased by 29.2% (P<0.05); compared with the ZEN group, the activities of GSH, CAT and SOD in the kidneys of mice in the ZEN+HG group were increased by 88.4% (P<0.01), 95% (P<0.01) and 17.1% (P<0.05), and the content of MDA was decreased by 124.8% (P<0.01). Compared with the ZEN+LG group, the activities of GSH, CAT and SOD in the kidneys of mice in the ZEN+HG group were increased by 21.2% (P<0.05), 24.3% (P<0.05) and 10.5%, and the content of MDA was decreased by 41.2% (P<0.05). The above results show that ZEN treatment increases free radicals in the kidneys of mice and generates lipid peroxidation products. GUANKE bacteria reduces the oxidative stress damage caused by ZEN to the kidneys, and the effect is better with increasing dose within a certain range.

[0032] 4. Inflammatory factor indicators On the 28th day, the levels of TNF-α, IL-1β and IL-6 in the kidneys of mice were detected. The results of TNF-α, IL-1β and IL-6 content detection were shown in Table 1. Fig.10 , Fig.11 , Fig.12 .

[0033] The results of TNF-α, IL-1β and IL-6 content detection in mouse kidneys showed that compared with the Control group, there was no difference in the levels of TNF-α, IL-1β and IL-6 in the kidneys of mice in the GUANKE group; compared with the Control group, the levels of TNF-α, IL-1β and IL-6 in the kidneys of mice in the ZEN group were increased by 99.5% (P<0.01), 150.8% (P<0.01) and 171.8% (P<0.01). Compared with the ZEN group, the levels of TNF-α, IL-1β and IL-6 in the kidneys of mice in the ZEN+LG group were downregulated by 22.4%, 39.8% (P<0.05), and 46.1% (P<0.05); compared with the ZEN group, the levels of TNF-α, IL-1β and IL-6 in the kidneys of mice in the ZEN+HG group were downregulated by 65.7% (P<0.01), 79.9% (P<0.05), and 144.7% (P<0.05). Compared with the ZEN+LG group, the levels of TNF-α, IL-1β and IL-6 in the kidneys of mice in the ZEN+HG group were downregulated by 34.6%, 28.6%, and 64.5%. The above results indicate that ZEN treatment increased the levels of pro-inflammatory factors and decreased the levels of anti-inflammatory factors in the kidneys of mice. GUANKE bacteria reduced the inflammatory damage to the kidneys caused by ZEN, and the effect became better with the increase of dose within a certain range.

[0034] 5. Pathological indicators On the 28th day, the right kidneys of mice in each experimental group were removed, fixed with 10% formalin, embedded, sliced, and stained with hematoxylin-eosin (HE). The damage of the kidneys was then observed under an electron microscope. The HE staining images of the right kidneys of mice in each group are shown in Fig.13 HE staining of sections can most intuitively reveal the morphology of tissue damage. By observing HE staining of kidney sections, the damage to the kidney can be clearly observed.

[0035] Depend on Fig.13It can be seen from the kidney sections of the Control group and the GUANKE group that the renal capsule has a regular shape (black arrow), and the gaps between the proximal tubules (red arrow) and the distal tubules (blue arrow) are obvious; while the renal capsule of the ZEN group has an irregular shape (black arrow), and the gaps between the proximal tubules (red arrow) and the distal tubules (blue arrow) are not obvious, and a large number of inflammatory cells can be seen infiltrating, and the kidney is seriously damaged; the renal capsule of the ZEN+LG group has a more regular shape (black arrow), and the gaps between the proximal tubules (red arrow) and the distal tubules (blue arrow) are more obvious, and a small number of inflammatory cells infiltrate; the renal capsule of the ZEN+HG group has a regular shape (black arrow), and the gaps between the proximal tubules (red arrow) and the distal tubules (blue arrow) are obvious. The above results show that ZEN can cause kidney damage in mice, and GUANKE bacteria can treat kidney damage caused by ZEN, and the effect is better with the increase of dose within a certain range.

[0036] The raw materials and equipment used in the present invention, unless otherwise specified, are all commonly used raw materials and equipment in the art; the methods used in the present invention, unless otherwise specified, are all conventional methods in the art.

[0037] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any way. Any simple modification, change and equivalent transformation made to the above embodiment based on the technical essence of the present invention still falls within the protection scope of the technical solution of the present invention.

Claims

1. Application of Lactobacillus plantarum GUANKE in the preparation of a medicine for preventing and / or treating kidney damage.

2. The use according to claim 1, characterized in that: The dosage forms of the drug include granules, ordinary tablets, dispersible tablets, effervescent tablets, orally disintegrating tablets, lozenges, chewable tablets, capsules, soft capsules, microcapsules, pills, powders, drop pills, sustained-release preparations, controlled-release preparations, oral liquid preparations and injections.

3. The use according to claim 1, characterized in that: The renal injury is acute renal injury.

4. The use according to claim 1, characterized in that: The kidney damage is caused by zearalenone.

5. A composition for treating renal injury, characterized in that: The active ingredient of the composition is Lactobacillus plantarum GUANKE.

6. The composition according to claim 5, characterized in that: The renal injury is acute renal injury.

7. The composition according to claim 5, characterized in that: The kidney damage is caused by zearalenone.

8. The composition according to claim 5, characterized in that: The composition includes one or more pharmaceutically acceptable excipients.

9. The composition according to claim 5 or 8, characterized in that: The composition is prepared into granules, ordinary tablets, dispersible tablets, effervescent tablets, orally disintegrating tablets, lozenges, chewable tablets, capsules, soft capsules, microcapsules, pills, powders, drop pills, sustained-release preparations, controlled-release preparations, oral liquid preparations or injections.

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