WO2022007408A1 - 肌苷在制备抗感染药物中的应用 - Google Patents

肌苷在制备抗感染药物中的应用 Download PDF

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WO2022007408A1
WO2022007408A1 PCT/CN2021/077156 CN2021077156W WO2022007408A1 WO 2022007408 A1 WO2022007408 A1 WO 2022007408A1 CN 2021077156 W CN2021077156 W CN 2021077156W WO 2022007408 A1 WO2022007408 A1 WO 2022007408A1
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inosine
complement
mice
content
expression
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李惠
彭博
彭宣宪
蒋明
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Sun Yat Sen University
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
    • A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
    • A61K31/7064—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines
    • A61K31/7076—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines containing purines, e.g. adenosine, adenylic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents

Definitions

  • the invention belongs to the technical field of biomedicine. More specifically, it relates to the application of inosine in the preparation of anti-infective drugs.
  • Inosine also known as inosine, inosine, etc.
  • Inosine is a nucleoside compound composed of inosine and ribose.
  • Inosine is a normal component of the human body and is the precursor of adenine, which can directly enter the body cells through the cell membrane and participate in normal metabolism in the body, such as protein synthesis, nucleic acid metabolism, energy metabolism, etc.
  • inosine is often used as a coenzyme drug, mainly for the adjuvant treatment of acute and chronic hepatitis, and also for the treatment of leukopenia, thrombocytopenia, heart failure, central retinitis, and optic atrophy.
  • Bacterial infection is a disease caused by pathogens or opportunistic pathogens invading the host to grow and multiply, producing toxins and other metabolites.
  • the main clinical drugs for the treatment of host infection are antibiotics, which can act on specific links of the bacterial DNA, RNA and protein synthesis system, interfere with the metabolic function of the bacteria, thereby hindering the life activities of the bacteria or inhibiting the growth of bacteria, such as ⁇ -lactam antibiotics (penicillins, cephalosporins, etc.), fluoroquinolone antibiotics (ciprofloxacin, ofloxacin, etc.), macrolide antibiotics (roxithromycin, clarithromycin), aminoglycosides Antibiotics (Amikacin, etc.), etc.
  • ⁇ -lactam antibiotics penicillins, cephalosporins, etc.
  • fluoroquinolone antibiotics ciprofloxacin, ofloxacin, etc.
  • macrolide antibiotics roxithromycin, clar
  • the commonly used antibiotics in the prior art mainly inhibit the growth and reproduction of bacteria by interfering with the metabolism of bacteria.
  • bacteria will gradually develop drug resistance, which makes the treatment of bacterial infections more difficult. . Therefore, it is more important to provide a drug that can improve the host's resistance to bacterial infection.
  • the technical problem to be solved by the present invention is to overcome the deficiencies and deficiencies of the prior art lacking drugs for improving the host's resistance to bacterial infection, and to provide a brand-new application of inosine in the preparation of a drug for improving the host's resistance to bacterial infection.
  • the purpose of the present invention is to provide the application of inosine in the preparation of anti-infective drugs.
  • Another object of the present invention is to provide an anti-infective drug containing an effective amount of inosine.
  • inosine can combine with proline hydroxylase to regulate cytokines in the body, such as IL-1 ⁇ factor and complement 3C.
  • cytokines in the body such as IL-1 ⁇ factor and complement 3C.
  • proline hydroxylase to regulate cytokines in the body, such as IL-1 ⁇ factor and complement 3C.
  • HIF-1 ⁇ hypoxia-inducible factor
  • the inosine includes inosine, pharmaceutically acceptable salts and esters of inosine, selectively substituted analogs, inosine stereoisomers, geometric isomers, hydrates, solvates or pharmaceutically acceptable inosines. Accepted salt or prodrug.
  • the present invention also provides an anti-infective drug containing an effective amount of inosine.
  • the medicine is an oral dosage form or an injection dosage form.
  • the medicament can also be used in veterinary treatment of pets, introduced breed animals and farm animals, including mammals, rodents, and the like. Examples of other animals include horses, dogs, cats, and the like.
  • the research results of the present invention show that inosine can be combined with proline hydroxylase to degrade hypoxia-inducible factor (HIF-1 ⁇ ) protein, regulate IL-1 ⁇ factor and complement 3C in the body, and make the effects of LPS stimulation or bacterial infection.
  • the content of IL-1 ⁇ factor in the organisms under the condition is significantly reduced, and the content of complement 3C is significantly increased, which improves the body's own resistance, thereby improving the survival rate of infected organisms, and has broad application prospects in the treatment of anti-infection.
  • Fig. 1 is a statistical graph of the survival rate data of inosine-improved bacteria-infected mice in Example 1 of the present invention, wherein A-LPS stimulation, B-Escherichia coli Y17 infection, C-Edwardia lentus EIB202 infection, D-algal lysis Vibrio VA.
  • Fig. 2 is a statistical graph of the effect of inosine on the content of IL-1 ⁇ factor and complement C3 in the blood of mice in Example 1 of the present invention.
  • Example 3 is a statistical diagram of the content of IL-1 ⁇ factor and complement C3 in the blood of dead and surviving mice in Example 1 of the present invention.
  • Figure 4 is a statistical graph of the effect of exogenous inosine on the expression of il1b and complement c3 genes in mouse macrophage RAW264.7 in Example 2 of the present invention.
  • Figure 5 is a statistical graph of the effect of different concentrations of exogenous inosine on the expression of il1b and complement c3 genes in mouse macrophage RAW264.7 in Example 2 of the present invention.
  • Figure 6 is a statistical graph of the effect of exogenous inosine on the expression of il1b and complement c3 genes in mouse macrophage Raw264.7-ASC in Example 2 of the present invention.
  • Example 7 is a statistical diagram of the binding situation of inosine and proline hydroxylase in Example 3 of the present invention.
  • Example 8 is a statistical graph of the effect of inosine on the expression of hypoxia-inducible factor HIF-1 ⁇ protein in Example 3 of the present invention.
  • Figure 9 is a statistical graph of the effects of exogenous inosine and proline hydroxylase inhibitors on the expression of i11b and c3 genes in Example 3 of the present invention.
  • Figure 10 is a statistical graph of the effects of exogenous inosine and PHD gene-deficient cells on the expression of i11b and c3 genes in Example 3 of the present invention.
  • Fig. 11 is a statistical graph showing the effect of exogenous addition of inosine on the survival rate of fish infected by Edwardsiella lentus in Example 4 of the present invention.
  • Figure 12 is a statistical graph of the effect of exogenous inosine on the expression of il1b and complement c3 genes infected by Edwardsiella lentus in Example 4 of the present invention.
  • the present invention is further described below with reference to the accompanying drawings and specific embodiments, but the embodiments do not limit the present invention in any form.
  • the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the technical field.
  • IL-1 ⁇ factor and complement C3 are proteins
  • il1b and complement c3 in italics are genes.
  • Example 1 Inosine improves the resistance of mice to bacterial infection
  • mice (5-8 weeks old, body weight 21 ⁇ 3g) were fed for a week to adapt to the experimental environment and then randomly divided into 4 groups (1 group for LPS and 3 kinds of bacteria), with 40 mice in each group.
  • 20 mice were injected intramuscularly with inosine at a dose of 300 mg/kg, and the other 20 mice were intramuscularly injected with the same volume of normal saline.
  • four groups of mice were stimulated with LPS or infected with three kinds of bacteria by intraperitoneal injection.
  • the doses were: LPS 75mg/kg, Edwardella lentus EIB202 was 5 ⁇ 10 7 CFU/piece, Vibrio alginolyticus VA was 3 ⁇ 10 8 CFU/piece, and human E.
  • coli Y17 was 2 ⁇ 10 6 CFU/piece . Observed for 7 days, counted the death of mice, and calculated the survival rate of mice. The calculation formula is: number of mice surviving/total number of mice ⁇ 100%, and the results are shown in Figure 1.
  • the survival rate of the mice injected with inosine was significantly higher than that of the control group mice without inosine injection, indicating that inosine can improve the survival rate of bacteria-infected mice.
  • the specific situation is as follows: the survival rate of mice in the LPS group after inosine injection was 80%, while the survival rate of mice in the LPS group without inosine injection was only 25%, and the survival rate increased by 55%, which is a significant difference. (Fig. 1A); the survival rate of mice in the group infected with E. coli Y17 after injection of inosine was 80%, while the survival rate of mice in the group infected with E.
  • coli Y17 without injection of inosine was only 40%, and the survival rate improved 40%, a significant difference (Fig. 1B); the survival rate of mice infected with E. lentus EIB202 after inosine injection was 70%, while the survival rate of mice in the E. lentus infection group without inosine injection was only 30%, the survival rate increased by 40%, with a clear difference ( Figure 1C); the survival rate of mice in the group infected with V. alginolyticus after injection of inosine was 80%, while the survival rate of V. alginolyticus without injection of inosine was 80%. The survival rate of mice in the group was only 50%, and the survival rate increased by 30%, with a clear difference (Fig. 1D).
  • mice were randomly divided into 4 groups (1 each for LPS and 3 bacteria) with 20 mice in each group.
  • 10 mice were injected intramuscularly with inosine at a dose of 300 mg/kg, and the other 10 mice were injected with the same volume of normal saline.
  • 4 groups of mice were stimulated with LPS or infected with 3 kinds of bacteria by intraperitoneal injection, respectively.
  • the doses were as follows: LPS was 5 mg/kg, and the infection doses of the three kinds of bacteria were 1 ⁇ 10 4 CFU/only. 2 hours after LPS stimulation, 12 hours after V. alginolyticus and Y17 infection, and 16 hours after EIB202 infection, the blood of each mouse was collected by orbital blood sampling, and the contents of IL-1 ⁇ factor and complement C3 were determined by ELISA kit. See Figure 2 for the results.
  • the specific conditions are as follows: the blood IL-1 ⁇ factor content of mice in the LPS group after inosine injection was 222.43 pg/mL, and the complement C3 content was 81.6 ⁇ g/mL, while the blood IL-1 ⁇ factor content of mice in the non-inosine injection group was 716.53 pg/mL, the content of complement C3 was 32.43 ⁇ g/mL, the content of IL-1 ⁇ factor decreased by 3.22 times, while the content of complement C3 increased by 2.52 times, there was a significant difference between the two groups; The blood IL-1 ⁇ factor content of the mice in the Y17 infection group was 180.54 pg/mL, and the complement C3 content was 96.39 ⁇ g/mL, while the blood IL-1 ⁇ factor content of the mice in the non-inosine injection group was 511.08 pg/mL, and the complement C3 content was 511.08 pg/mL.
  • the content of IL-1 ⁇ factor decreased by 2.83 times, while the content of complement C3 increased by 3.16 times, and there was a significant difference between the two groups; the blood IL-1 ⁇ of the mice in the group infected with Vibrio alginolyticus after injection of inosine
  • the factor content was 48.68 pg/mL
  • the complement C3 content was 68.57 ⁇ g/mL
  • the blood IL-1 ⁇ factor content of the mice without inosine injection was 322.15 pg/mL
  • the complement C3 content was 26.29 ⁇ g/mL
  • the IL-1 ⁇ content was 26.29 ⁇ g/mL.
  • the content of factor decreased by 6.62 times, while the content of complement C3 increased by 2.61 times, and there was a significant difference between the two groups; the blood IL-1 ⁇ factor content of mice in the group infected with E.
  • the C3 content was 65.65 ⁇ g/mL, while the blood IL-1 ⁇ factor content of the mice without inosine injection was 424.23 pg/mL, the complement C3 content was 42.02 ⁇ g/mL, the IL-1 ⁇ factor content decreased by 4.72 times, and the complement C3
  • the content increased by 1.56 times, and there was a significant difference between the two groups.
  • mice were randomly divided into 2 groups: 20 mice in the Vibrio alginolyticus challenge group and 10 mice in the saline control group.
  • the intraperitoneal challenge dose of Vibrio alginolyticus was 3 ⁇ 10 8 CFU/vibration, and the control group was intraperitoneally injected with the same dose of normal saline.
  • the orbital blood was collected, and each mouse and the corresponding blood samples were marked, and the samples were stored at -80°C for later IL-1 ⁇ and C3 content determination.
  • the mice were then observed for 7 days, and the survival of each mouse was recorded. It was found that 12 of the 20 challenged mice survived and 8 died.
  • the IL-1 ⁇ factor and complement C3 contents of each mouse were measured by ELISA, and the IL-1 ⁇ factor and complement C3 contents of the dead and surviving mice were divided into one group respectively, and the results were statistically obtained in Figure 3.
  • the average content of IL-1 ⁇ factor in the blood of the dead mice was 360.11pg/mL, while the average content of IL-1 ⁇ factor in the blood of the survival group mice was 290.06pg/mL, and the content of IL-1 ⁇ factor in the blood of the dead mice was 290.06pg/mL. significantly higher than surviving mice.
  • the mean content of complement C3 in the blood of dead mice was 9.29 ⁇ g/mL, while the mean content of complement C3 in the blood of mice in the survival group was 15.36 ⁇ g/mL.
  • the content of complement C3 in the blood of dead mice was significantly lower than that of surviving mice.
  • Example 2 The effect of exogenous inosine on the expression of il1b and complement c3 genes in mouse macrophages
  • RAW264.7 cells were removed from the liquid nitrogen, rapidly dissolved into 37 °C water bath pot and cultured in a CO 2 incubator with low glucose DMEM (containing 10% FBS). Change the fluid after two days. When the cell growth density reached about 80-90%, the cells were digested with 0.0025% EDTA, and then the cells were added to a 6-well plate at an amount of 1 ⁇ 10 6 /well for plating.
  • the plated cells were placed after a CO 2 incubator for 12 hours, the volume of the cell culture medium at a concentration of inosine, 5mM added control group and the like. After further culturing for 12 hours, stimulated with 100ng/ml LPS for 2, 8 and 24 hours, the cells were collected and total RNA was extracted. 4 replicates per condition. Fluorescence quantitative PCR was used to detect the expression of il1b and complement c3 genes.
  • the primers for the two genes are: il1b (upstream: tcaggcaggcagtatcactc, downstream: gaggatgggctcttcttcaa), complement c3 (upstream: catcctgcactcaggtagtg, downstream: gcattagatccctgagtgac). The results are shown in Figure 4.
  • mouse macrophages show the same response to LPS stimulation as mice after being exposed to exogenous inosine, that is, inosine can inhibit the expression of il1b in mouse macrophages, while promoting mouse macrophages. expression of c3.
  • inosine was added at concentrations of 0.63mM, 1.25mM, 2.5mM, 5mM and 10mM, respectively.
  • the control group was only added with cell culture medium, with 4 replicates in each group. .
  • Incubation was continued for an additional 12 hours, followed by stimulation with 100 ng/ml LPS for 8 hours. Cells were collected and RNA was extracted for each sample. Fluorescence quantitative detection of the expression of il1b and complement c3 after LPS stimulation under the condition of exogenous addition of different concentrations of inosine, see Figure 5 for the statistics of the results.
  • the specific situation is: compared with the group without inosine, il1b decreased by 2.988 times after adding 0.625mM inosine, and complement c3 increased by 1.076 times; after adding 1.25mM inosine, il1b decreased by 4.506 times, and increased by 1.385 times after c3; After adding 2.5mM inosine, il1b decreased by 4.875 times, and c3 increased by 1.441 times; after adding 5mM inosine, il1b decreased by 7.672 times, and c3 increased by 2.060 times; after adding 10mM inosine, il1b decreased by 13.803 times, and c3 increased by 2.099 times, there is a significant difference between them.
  • the Raw264.7-ASC cells were cultured with reference to the procedure of 2.1, and the cells were plated in a 6-well plate at a number of 1 ⁇ 10 6 per well.
  • the plated cells were placed in an incubator for 12 hours, and inosine at a concentration of 5 mM was added, and the control group was added with cell culture medium, with 4 replicates in each group.
  • the cells were stimulated with 100 ng/ml LPS for 8 hours, and 10 ⁇ M nigericin was added for 2 hours, and the supernatant culture medium was collected.
  • the contents of IL-1 ⁇ factor and complement C3 in the supernatant were determined by ELISA, and the results are shown in FIG. 6 .
  • Example 3 Exogenous inosine regulates the expression of il1b and c3 by binding to proline hydroxylase and affecting the stability of hypoxia-inducible factor
  • LPS-induced elevation of il1b is related to hypoxia-inducible factor (HIF-1 ⁇ ) (Palsson-Mcdermott et al. Cell metabolism. 2015, 21(1): 65-80; Sumbayev VV. FEBS letters .2008,582(2):319-326); proline hydroxylase (PHD) can degrade HIF-1 ⁇ protein and destroy its stability (Couvelar et al.Clinical Cancer Research.2008,14(20):6634 -6639).
  • HIF-1 ⁇ hypoxia-inducible factor
  • PTD proline hydroxylase
  • the regulation of the expression of il1b and c3 by inosine may be related to proline hydroxylase, therefore, the present invention detected the relationship between inosine and proline hydroxylase, as well as hif-1 ⁇ , i11b and c3 in cells after LPS stimulation expression.
  • DMSO concentration of 430 ⁇ M
  • NT dye concentration of 430 ⁇ M
  • purified proline hydroxylase PHD protein concentration 10 ⁇ M
  • tagged protein His tag negative control
  • inosine was added at concentrations of 0.6 mM, 2.5 mM and 10 mM, and the control group was added with cell culture medium. After culturing for another 12 hours, the cells were stimulated with 100ng/ml LPS for 8 hours, and the cells were collected respectively, and their total proteins were extracted and subjected to SDS-PAGE electrophoresis. After the total protein was electroporated onto PVDF membrane, it was incubated with rabbit anti-hypoxia-inducible factor HIF-1 ⁇ antibody (Abcam Biological Company, USA) and mouse anti-rabbit secondary antibody (Xiamen Bosheng Biological Co., Ltd.) to detect hypoxia-inducible factor. The expression of protein HIF-1 ⁇ , the results are shown in Figure 8.
  • hypoxia-inducible factor HIF-1 ⁇ protein gradually decreased, and even could not be detected.
  • the details are as follows: After RAW264.7 cells were stimulated by LPS, the expression of hypoxia-inducible factor protein was the highest, while the expression of hypoxia-inducible factor protein decreased after adding 0.6 and 2.5 mM inosine. Factor proteins are not even expressed.
  • the expression of il1b in Raw264.7 cells was increased after LPS stimulation, while inosine inhibited the expression of il1b by a factor of 12.
  • adding the proline hydroxylase inhibitor DMOG promoted the expression of il1b up to 3.3 times; while adding the inhibitor and adding inosine, the expression of il1b decreased.
  • the amino acid hydroxylase inhibitor DMOG was 0.5 mM, the addition of inosine was 89% of that without addition of inosine.
  • the expression of c3 in Raw264.7 cells also increased after LPS stimulation, and the addition of inosine further increased the expression of il1b by 1.66 times.
  • the expression of c3 was inhibited; while adding the inhibitor and inosine together, the expression of c3 was up-regulated.
  • the dose of DMOG was 0.5 mM
  • the addition of inosine was 1.9 times that of the non-added inosine.
  • Example 4 Inosine improves fish resistance to Edwardsiella lentus infection
  • the experimental fish (crucian carp, grass carp and tilapia) were fed in the laboratory for a week to adapt to the experimental environment, and then randomly divided into 4 groups.
  • intramuscular injection was used to inject 10 ⁇ L of inosine per fish ((25 ⁇ g, 50 ⁇ g and 100 ⁇ g for crucian carp and grass carp, 10 ⁇ g, 20 ⁇ g and 40 ⁇ g for tilapia), twice a day. The same volume was injected in the control group.
  • intramuscular injection of Edwardsiella lentus (1 ⁇ 10 5 CFU/crucian carp, 1 ⁇ 10 6 CFU/grass carp, 1 ⁇ 10 5 CFU/tilapia) was used. Observed continuously for 15 days, and counted dead fish , and calculate the survival rate, see Figure 11 for the statistics of the results.
  • the survival rate of crucian carp, grass carp and tilapia infected with Edwardsiella lentus gradually increased.
  • the survival rate of crucian carp was 40% without injection of inosine, 57% after injection of 25 ⁇ g inosine, 73% after injection of 50 ⁇ g inosine, and 77% after injection of 100 ⁇ g inosine %, the survival rate increased by 17-37%.
  • grass carp was 40% without injection of inosine, 43% after injection of 25 ⁇ g inosine, 53% after injection of 50 ⁇ g inosine, 80% after injection of 100 ⁇ g inosine, surviving rate increased by 3-40%.
  • Tilapia survived 60% without inosine injection, 63% with 10 ⁇ g inosine, 70% with 20 ⁇ g inosine, and 77% with 40 ⁇ g inosine , survival increased by 3-17%.
  • Each fish was divided into 2 groups, the E. lentus infection group and the inosine+Ed. lentus infection group.
  • intramuscular injection was used to inject 10 ⁇ L of inosine into each fish (the content was 50 ⁇ g respectively), and the control group was injected with the same volume of normal saline. 2 times a day. The control group was injected with the same volume of normal saline.
  • Edwardella lentus was injected intramuscularly (1 ⁇ 10 4 CFU/crucian carp, 1 ⁇ 10 5 CFU/grass carp, 1 ⁇ 10 4 CFU/tilapia).
  • il1b1 upstream atgcgctgctcaacttcat, downstream ctggcccttattttgttgag
  • il1b2 upstream caaagcgatcctcttcattt, downstream attcgggtcatcagttttaa
  • C3 upstream tggggatggatctgaaca, downstream tgcccatgatgaggtacga).
  • Grass carp il1b (upstream gccatttccaagagtaatct, downstream aacggattcaaagtgttatta), C3 (upstream atacatcggcatcactgaac, downstream cccaaacggatattatgaag).
  • Tilapia il1b (upstream aaggcacaaacctctatctg, downstream tgtcgcgtttgtagaagaga), c3 (upstream atgttaccttaacacccaag, downstream gctctccatacaggtacctg), see Figure 12 for the statistics of the results.
  • inosine promotes the expression of c3 and inhibits the expression of il1b.
  • grass carp il1b decreased by 2 times after inosine injection, and complement c3 increased by 3 times.
  • il1b1 decreased by 3.2 times
  • il1b2 decreased by 5 times
  • complement c3 increased by 2.5 times after inosine injection.
  • il1b decreased 4-fold and complement c3 increased 3.9-fold after inosine injection.

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Abstract

肌苷在制备抗感染的药物中的应用。肌苷可以与脯氨酸羟化酶结合，降解缺氧诱导因子蛋白(HIF-1α)蛋白，调节机体中的IL-1β因子、补体3C，使在LPS刺激或细菌感染影响下的生物体的IL-1β因子含量显著降低，补体3C含量显著增加，提高机体自身的抵抗力，从而提高被感染生物的存活率。

Description

肌苷在制备抗感染药物中的应用 技术领域
本发明属于生物医药技术领域。更具体地，涉及肌苷在制备抗感染药物中的应用。
背景技术
肌苷(Inosine)，也称为次黄苷、次黄嘌呤核苷等，是由次黄嘌呤与核糖结合而成的核苷类化合物。肌苷是人体的正常成分，为腺嘌呤的前体，能直接透过细胞膜进入体细胞，参与体内的正常代谢，如蛋白质合成、核酸代谢、能量代谢等。临床上肌苷常用作辅酶类药物，主要用于急、慢性肝炎的辅助治疗，同时也可以用于治疗白细胞减少、血小板减少、心力衰竭、中心视网膜炎、视神经萎缩等。
细菌感染是致病菌或条件致病菌侵入宿主中生长繁殖，产生毒素和其他代谢产物所引起的疾病。目前临床上治疗宿主感染的药物主要为抗生素，可作用于细菌的DNA、RNA和蛋白质合成系统的特定环节，干扰细菌的代谢作用，从而妨碍细菌的生命活动或抑制细菌生长，如β-内酰胺类抗生素(青霉素类、头孢菌素类等)、氟喹诺酮类抗生素(环丙沙星、氧氟沙星等)、大环内酯类抗生素(罗红霉素、克拉霉素)、氨基糖苷类抗生素(丁氨卡那霉素等)等。但是现有技术常用抗生素主要是通过干扰细菌的代谢来抑制细菌的生长繁殖，而细菌在生长繁殖的过程中在抗生素的选择压力下，会逐渐产生耐药性，使得针对细菌感染的治疗更加困难。因此，提供一种提高宿主抵抗细菌感染的药物，具有更加重要的意义。
发明内容
本发明要解决的技术问题是克服现有技术缺少提高宿主抵抗细菌感染药物的缺陷和不足，提供一种全新的肌苷在制备提高宿主抵抗细菌感染药物中的应用。
本发明的目的是提供肌苷在制备抗感染药物中的应用。
本发明另一目的是提供一种含有有效量肌苷的抗感染药物。
本发明上述目的通过以下技术方案实现：
本发明研究发现，肌苷可以与脯氨酸羟化酶结合，调节机体中的细胞因子，如IL-1β因子、补体3C。实验证明，生物体在LPS刺激或细菌感染影响下，细胞因子IL-1β因子含量会显著增加，补体3C含量则显著降低；摄入肌苷后，肌苷可以与脯氨酸羟化酶结合，可以降解缺氧诱导因子(HIF-1α)蛋白，显著抑制IL-1β因子含量增加，同时促进补体3C含量增加，提高机体自身的抵抗力，从而提高被感染生物的存活率。
进一步地，所述肌苷包括肌苷，肌苷的可药用的盐和酯，选择性取代的类似物，肌苷立体异构体、几何异构体、水合物、溶剂化物或药学上可接受的盐或前药。
另外的，本发明还提供了一种含有有效量肌苷的抗感染药物。
进一步地，所述药物为口服剂型或注射剂型。
更进一步地，所述药物除了对人类治疗有益以外，还可应用于兽医治疗宠物、引进品种的动物和农场的动物，包括哺乳动物，啮齿类动物等等。另外一些动物的实例包括马、狗和猫等。
进一步地，实验证明所述药物也可以应用于鱼类感染的治疗，显著提高感染鱼类的存活率。
本发明具有以下有益效果：
本发明研究结果显示，肌苷可以与脯氨酸羟化酶结合，降解缺氧诱导因子(HIF-1α)蛋白，调节机体中的IL-1β因子、补体3C，使在LPS刺激或细菌感染影响下的生物体的IL-1β因子含量显著降低，补体3C含量显著增加，提高机体自身的抵抗力，从而提高被感染生物的存活率，在抗感染的治疗中具有广阔的应用前景。
附图说明
图1为本发明实施例1中肌苷提高细菌感染小鼠的存活率数据统计图，其中，A-LPS刺激，B-大肠杆菌Y17感染，C-迟缓爱德华氏菌EIB202感染，D-溶藻弧菌VA。
图2为本发明实施例1中肌苷对小鼠血液中IL-1β因子、补体C3含量影响的数据统计图。
图3为本发明实施例1中死亡和存活小鼠血液中IL-1β因子、补体C3含量统计图。
图4为本发明实施例2中外源肌苷对小鼠巨噬细胞RAW264.7的il1b和补体c3基因表达影响的数据统计图。
图5为本发明实施例2中不同浓度外源肌苷对小鼠巨噬细胞RAW264.7的il1b和补体c3基因表达影响的数据统计图。
图6本发明实施例2中外源肌苷对小鼠巨噬细胞Raw264.7-ASC的il1b和补体c3基因表达影响的数据统计图。
图7为本发明实施例3中肌苷与脯氨酸羟化酶的结合情况统计图。
图8为本发明实施例3中肌苷对缺氧诱导因子HIF-1α蛋白表达影响的数据统计图。
图9为本发明实施例3中外源肌苷与脯氨酸羟化酶抑制剂对i11b和c3基因表达影响的数据统计图。
图10为本发明实施例3中外源肌苷与PHD基因缺失细胞对i11b和c3基因表达影响的数据统计图。
图11为本发明实施例4中外源添加肌苷对鱼类迟缓爱德华氏菌感染的存活率影响的数据统计图。
图12为本发明实施例4中外源肌苷对鱼类迟缓爱德华氏菌感染的il1b和补体c3基因表达影响的数据统计图。
具体实施方式
以下结合说明书附图和具体实施例来进一步说明本发明，但实施例并不对本发明做任何形式的限定。除非特别说明，本发明采用的试剂、方法和设备为本技术领域常规试剂、方法和设备。
其中，IL-1β因子、补体C3为蛋白质，斜体的il1b和补体c3为基因。
除非特别说明，以下实施例所用试剂和材料均为市购。
实施例1 肌苷提高小鼠对细菌感染的抵抗能力
1.1肌苷提高细菌感染小鼠的存活率
从-80℃冰箱取出保存的细菌甘油菌株，全部加入到100mL的LB培养基中(250mL培养瓶)，恒温(溶藻弧菌和迟缓爱德华氏菌为30℃，大肠杆菌Y17 为37℃)200rpm培养。待细菌生长至OD600约1.0时，离心收集细菌，弃上清。用生理盐水洗涤3次备用。
Balb/c小鼠(5-8周龄，体重21±3g)先喂养一周使其适应试验环境后，随机分成4组(LPS和3种细菌各为1组)，每组40只小鼠。每组中20只小鼠按照300mg/kg剂量采用肌肉注射方式注射肌苷，另外20只小鼠肌肉注射相同体积的生理盐水。3小时后，4组小鼠分别以腹腔注射方式进行LPS刺激或3种细菌感染。剂量分别为：LPS 75mg/kg，迟缓爱德华氏菌EIB202为5×10 7CFU/只、溶藻弧菌VA为3×10 8CFU/只、人源大肠杆菌Y17为2×10 6CFU/只。观察7天，统计小鼠的死亡情况，并计算小鼠存活率。计算公式为：小鼠存活数量/小鼠总数×100％，结果参见图1。
由图可见，注射肌苷小鼠组，其存活率明显高于未注射肌苷的对照组小鼠，说明肌苷可以提高细菌感染小鼠的存活率。具体情况如下：注射肌苷后再注射LPS组的小鼠存活率为80％，而未注射肌苷仅注射LPS组的小鼠存活率仅为25％，存活率提高了55％，具有明显差异(图1A)；注射肌苷后再用大肠杆菌Y17感染组小鼠的存活率为80％，而未注射肌苷仅大肠杆菌Y17感染组小鼠的存活率仅为40％，存活率提高了40％，具有明显差异(图1B)；注射肌苷后再用迟缓爱德华菌EIB202感染组的小鼠存活率为70％，而未注射肌苷仅迟缓爱德华菌感染组的小鼠存活率仅为30％，存活率提高了40％，具有明显差异(图1C)；注射肌苷后再用溶藻弧菌感染组的小鼠存活率为80％，而未注射肌苷仅溶藻弧菌感染组的小鼠存活率仅为50％，存活率提高了30％，具有明显差异(图1D)。
1.2肌苷对小鼠血液中IL-1β因子、补体C3含量的影响
现有技术研究发现，炎症因子紊乱会引起自身免疫性疾病导致死亡(Shaw et al.Trends in Molecular Medicine,2011,17(2):57-64；Mao et al.Frontiers in Immunology,2018.9:2566；Gabay et al.Nature Reviews Rheumatology,2010,6(4):232-241)，血液中的IL-1β因子和补体C3含量都与机体免疫功能、细菌感染后机体的存活率密切相关。
将80只小鼠随机分成4组(LPS和3种细菌各1组)，每组20只。每组中10只小鼠按照300mg/kg剂量采用肌肉注射方式注射肌苷，另外10只小鼠注射相同体积的生理盐水。3小时后，4组小鼠分别以腹腔注射进行LPS刺激或3种细菌感染。剂量分别为：LPS是5mg/kg，3种细菌感染剂量都为1×10 4CFU/只。 LPS刺激后2小时，溶藻弧菌和Y17感染后12小时，EIB202感染后16小时，采用眼眶取血方式采集每只小鼠血液，用ELISA试剂盒测定IL-1β因子和补体C3的含量，结果参见图2。
从图中可以看出，小鼠注射肌苷组，不管是LPS刺激还是大肠杆菌Y17、溶藻弧菌VA或迟缓爱德华菌EIB202感染，其血液中IL-1β因子含量都显著低于未注射肌苷组；而补体C3含量却显著高于未注射肌苷组。具体情况如下：注射肌苷后再注射LPS组小鼠血液IL-1β因子含量为222.43pg/mL，补体C3含量为81.6μg/mL，而未注射肌苷组小鼠血液IL-1β因子含量为716.53pg/mL，补体C3的含量为32.43μg/mL，IL-1β因子含量降低了3.22倍，而补体C3含量增加了2.52倍，两组间均具有明显差异；注射肌苷后再用大肠杆菌Y17感染组小鼠血液IL-1β因子含量为180.54pg/mL，补体C3含量为96.39μg/mL，而未注射肌苷组小鼠血液IL-1β因子含量为511.08pg/mL，补体C3含量为30.54μg/mL，IL-1β因子含量降低了2.83倍，而补体C3含量增加了3.16倍，两组间均具有明显差异；注射肌苷后再用溶藻弧菌感染组小鼠血液IL-1β因子含量为48.68pg/mL，补体C3含量为68.57μg/mL，而未注射肌苷组小鼠血液IL-1β因子含量为322.15pg/mL，补体C3的含量为26.29μg/mL，IL-1β因子含量降低了6.62倍，而补体C3含量增加了2.61倍，两组间均具有明显差异；注射肌苷后再用迟缓爱德华菌感染组小鼠血液IL-1β因子含量为89.97pg/mL，补体C3含量为65.65μg/mL，而未注射肌苷组小鼠血液IL-1β因子含量为424.23pg/mL，补体C3含量为42.02μg/mL，IL-1β因子含量降低了4.72倍，而补体C3含量增加了1.56倍，两组间均具有明显差异。
这些结果说明，肌苷通过降低宿主血液IL-1β因子的含量，升高补体C3的含量，从而提高了细菌感染后小鼠的存活率。
1.3死亡和存活小鼠血液中IL-1β因子、补体C3含量的检测
将30只昆明小鼠随机分成2组：溶藻弧菌攻毒组20只，生理盐水对照组10只。溶藻弧菌腹腔攻毒剂量为3×10 8CFU/只，对照组腹腔注射同等剂量生理盐水。6小时后采用眼眶取血，并将每只小鼠及相应的血液样本做好标记，样品-80℃保存用于后期IL-1β和C3含量测定。然后继续观察小鼠7天，记录每只小鼠的存活情况。结果发现，20只攻毒小鼠中有12只存活，8只死亡。采用ELISA方法测定每只小鼠IL-1β因子和补体C3含量，并将死亡和存活小鼠的IL-1β因 子和补体C3含量各自归为1组，将结果统计得图3。
由图可见，死亡小鼠血液中IL-1β因子含量均值为360.11pg/mL，而存活组小鼠血液中IL-1β因子含量均值为290.06pg/mL，死亡小鼠血液中IL-1β因子含量显著高于存活小鼠。死亡小鼠血液中补体C3含量均值为9.29μg/mL，而存活组小鼠血液中补体C3含量均值为15.36μg/mL，死亡小鼠血液中补体C3含量显著低于存活小鼠。
这些结果说明，IL-1β因子和补体C3的含量高低与细菌感染后小鼠的存活和死亡有关，IL-1β因子含量过高或补体C3含量过低的小鼠易死亡。
实施例2 外源肌苷对小鼠巨噬细胞il1b和补体c3基因表达的影响
2.1外源肌苷对小鼠巨噬细胞RAW264.7的il1b和补体c3基因表达的影响
将RAW264.7细胞从液氮中取出，迅速放入37℃水浴锅中溶解，用低糖DMEM(含10％FBS)在CO 2培养箱中培养。两天后换液。当细胞生长密度达到80～90％左右时，用0.0025％EDTA消化细胞，再以1×10 6/每孔的数量将细胞加入6孔板中进行铺板。
将铺好的细胞放入CO 2培养箱中培养12小时后，加入浓度为5mM的肌苷，对照组加入等体积的细胞培养基。再继续培养12小时后，用100ng/ml LPS刺激2、8和24小时，分别收集细胞，提取总RNA。每个条件4个重复。采用荧光定量PCR检测il1b和补体c3基因的表达情况。这2个基因引物分别为：il1b(上游：tcaggcaggcagtatcactc，下游：gaggatgggctcttcttcaa)，补体c3(上游：catcctgcactcaggtagtg，下游：gcattagatccctgagtgac)，结果参见图4。
由图可见，小鼠巨噬细胞受到LPS刺激后，外源添加肌苷与不添加肌苷比较，il1b表达下降，而c3表达增加。具体情况如下：LPS刺激后2小时，添加肌苷的细胞il1b表达下降3.05倍，而c3表达基本不变；LPS刺激后8小时，添加肌苷的细胞il1b表达下降5.15倍，而c3表达增加1.81倍；LPS刺激后24小时，添加肌苷的细胞il1b表达下降1.98倍，而c3表达增加1.81倍。
这些结果说明小鼠巨噬细胞在受到外源肌苷作用后，对LPS刺激表现出和小鼠同样的反应，即肌苷可以抑制小鼠巨噬细胞il1b的表达，而促进小鼠巨噬细胞c3的表达。
2.2不同浓度外源肌苷对小鼠巨噬细胞RAW264.7的il1b和补体c3基因表达的影响
将铺好的RAW264.7细胞放入培养箱培养12小时后，分别加入浓度为0.63mM、1.25mM、2.5mM、5mM和10mM的肌苷，对照组仅加细胞培养基，每组4个重复。再继续培养12小时，然后用100ng/ml的LPS刺激8小时。收集细胞，提取每个样品的RNA。荧光定量检测外源添加不同浓度肌苷情况下，LPS刺激后il1b和补体c3的表达情况，结果统计参见图5。
由图可见，小鼠巨噬细胞受到LPS刺激后，外源添加肌苷与不添加肌苷比较，il1b表达降低，且随着肌苷浓度的增加而逐渐下降；补体c3表达上升，且随着添加肌苷浓度的增加而逐渐上升。具体情况为：相对于不添加肌苷组，加入0.625mM肌苷后il1b下降了2.988倍，补体c3上升了1.076倍；加入1.25mM肌苷后il1b下降了4.506倍，c3后上升1.385了倍；加入2.5mM肌苷后il1b下降了4.875倍，c3上升了1.441倍；加入5mM肌苷后il1b下降了7.672倍，c3上升了2.060倍；加入10mM肌苷后il1b下降了13.803倍，c3上升了2.099倍，相互间具有显著差异。
2.3外源肌苷对小鼠巨噬细胞Raw264.7-ASC的IL-1β因子和补体C3的影响
参考2.1的过程培养Raw264.7-ASC细胞，以1×10 6/每孔的数量将细胞加入6孔板中进行铺板。将铺好的细胞放入培养箱培养12小时，加入浓度为5mM的肌苷，对照组加入细胞培养基，每组4个重复。再继续培养12小时后，用100ng/ml的LPS刺激8小时，加入10μM的尼日利亚菌素培养2小时，收集上清培养液。ELISA测定上清中的IL-1β因子和补体C3的含量，结果参见图6。
由图可见，仅LPS刺激细胞，IL-1β蛋白的分泌量2919pg/mL，而添加肌苷再受到LPS刺激的细胞，IL-1β蛋白的分泌量降低到1007pg/mL，下降了2.9倍。而补体C3蛋白的分泌情况正相反。仅LPS刺激细胞，补体C3分泌量为8.32μg/mL，而添加肌苷再受到LPS刺激的细胞，补体C3分泌量为19.9μg/mL，上升了2.4倍。相互间具有显著差异。
这些结果说明，外源肌苷可使受到LPS刺激的小鼠巨噬细胞il1b基因表达下降，IL-1β蛋白分泌减少，而补体c3基因表达上升，补体C3蛋白分泌增加。
实施例3 外源肌苷通过与脯氨酸羟化酶结合影响缺氧诱导因子的稳定性来调节il1b和c3的表达
现有技术研究发现，LPS引起的il1b的升高与缺氧诱导因子(HIF-1α)有关(Palsson-Mcdermott et al.Cell metabolism.2015,21(1):65-80；Sumbayev VV. FEBS letters.2008,582(2):319-326)；脯氨酸羟化酶(PHD)可以降解HIF-1α蛋白，破坏其稳定性(Couvelard et al.Clinical Cancer Research.2008,14(20):6634-6639)。肌苷调节il1b和c3的表达可能与脯氨酸羟化酶有关，因此，本发明检测了肌苷和脯氨酸羟化酶间的关系，以及LPS刺激后细胞中hif-1α，i11b和c3的表达情况。
3.1肌苷与脯氨酸羟化酶的相互作用
将30μLDMSO加入NT染料(浓度为430μM)染料中，完全溶解后，用DMSO稀释至30μM后，将纯化的脯氨酸羟化酶PHD蛋白(浓度10μM)或者标签蛋白His tag(阴性对照)与NT染料等体积混匀(总体积200μL)，室温孵育30min。然后加入分离柱中，当最后一滴液体从分离柱中流出后，加入PBS磷酸盐缓冲液，在柱子底部收集蛋白样品，至600μL左右即停止收集，然后用NT.LabelFree微量热泳动(MST)分子间相互作用分析仪(德国Nano Temper公司)检测染料和蛋白结合的丰度来判断蛋白是否结合至染料上。将结合了染料的PHD蛋白加到200μL离心管中，进行梯度稀释。在每个离心管中加入500uM肌苷，室温孵育30min，Nano Temper上机检测，结果参见图7。
由图可见，阴性对照标签蛋白His tag与肌苷不结合，而脯氨酸羟化酶可以与肌苷结合，其结合常数Kd值为15.5mM。
3.2肌苷对缺氧诱导因子HIF-1α蛋白表达的影响
将铺好的RAW264.7细胞放入培养箱培养12小时后，加入浓度为0.6mM、2.5mM和10mM的肌苷，对照组加入细胞培养基。再继续培养12小时后，用100ng/ml LPS刺激8小时，分别收集细胞，提取其总蛋白质，进行SDS-PAGE电泳。将总蛋白质电转至PVDF膜上后，用兔抗缺氧诱导因子HIF-1α抗体(美国Abcam生物公司)和鼠抗兔二抗抗体(厦门博生生物有限公司)进行孵育，检测缺氧诱导因子蛋白HIF-1α表达情况，结果参见图8。
由图可见，随着添加肌苷浓度的增加，缺氧诱导因子HIF-1α蛋白的表达逐渐下降，甚至无法检测到。具体情况如下：RAW264.7细胞受到LPS刺激后，缺氧诱导因子蛋白的表达最高，而添加0.6和2.5mM肌苷后，缺氧诱导因子蛋白的表达下降，添加10mM肌苷后，缺氧诱导因子蛋白甚至不表达。
3.3外源肌苷与脯氨酸羟化酶结合对i11b和c3基因表达的影响
(1)将Raw264.7细胞，以1×10 6/每孔数量铺板到6孔板中，放入细胞培养 箱培养12小时。加入0.005mM、0.05mM和0.5mM的脯氨酸羟化酶抑制剂DMOG处理2小时后，再加入5mM的肌苷，对照组加入细胞培养基。再继续培养12小时后，用100ng/ml的LPS刺激8小时。每组4个重复。分别收集细胞，提取总RNA。荧光定量PCR检测il1b和c3基因的表达情况，结果见图9。
由图可见，Raw264.7细胞受到LPS刺激后il1b表达升高，而肌苷抑制了il1b表达，抑制倍数为12倍。Raw264.7细胞受到LPS刺激后，加入脯氨酸羟化酶抑制剂DMOG，则促进il1b的表达，最高可达3.3倍；而加入抑制剂同时再加入肌苷，则il1b的表达降低，当脯氨酸羟化酶抑制剂DMOG为0.5mM时，加入肌苷是未加肌苷的89％。
同时，Raw264.7细胞受到LPS刺激后c3表达也升高，再加入肌苷使il1b表达继续增加1.66倍。Raw264.7细胞受到LPS刺激后，如加入脯氨酸羟化酶抑制剂DMOG，则抑制c3表达；而加入抑制剂同时再加入肌苷，则c3的表达上调，当脯氨酸羟化酶抑制剂DMOG为0.5mM时，加入肌苷是未加肌苷的1.9倍。
(2)将Raw264.7-PHD -/-细胞(PHD基因缺失细胞)以1×10 6/每孔数量铺板到6孔板中，放入细胞培养箱培养12小时。加入5mM的肌苷，对照组加入细胞培养基，每组4个重复。继续培养12小时后，用100ng/ml的LPS刺激8小时，收集细胞，提取每个样品的总RNA。荧光定量PCR检测il1b和c3的表达情况，结果参见图10。
由图可见，脯氨酸羟化酶PHD缺失后，il1b表达增加，而补体c3表达下降，而脯氨酸羟化酶PHD缺失株添加肌苷后，肌苷对il1b表达的抑制效果和对补体c3表达的促进效果均消失。
上述结果说明，如果脯氨酸羟化酶PHD受到了抑制或者细胞缺少这个酶，那么细胞受到LPS刺激后，即使再加入肌苷，由于肌苷与脯氨酸羟化酶PHD结合减少，或者无法与脯氨酸羟化酶PHD结合，从而不能影响缺氧诱导因子的稳定性，这样就无法抑制il1b表达和促进c3表达。也就是说明肌苷首先必须与脯氨酸羟化酶PHD结合，才能影响缺氧诱导因子的稳定性，从而实现对il1b和c3表达的调节。
实施例4 肌苷提高鱼类对迟缓爱德华菌感染的抵抗能力
4.1外源添加肌苷提高鱼类对迟缓爱德华氏菌感染的存活率影响
将试验用鱼(鲫鱼，草鱼和罗非鱼)在实验室喂养一周，使其适应试验环境 后，各随机分成4组。先采用肌肉注射方式每尾鱼注射体积10μL的肌苷((鲫鱼和草鱼剂量分别为25μg、50μg和100μg，罗非鱼剂量分别为10μg、20μg和40μg)，每天2次。对照组注射相同体积的生理盐水。3天后，采用肌肉注射迟缓爱德华氏菌(1×10 5CFU/鲫鱼，1×10 6CFU/草鱼，1×10 5CFU/罗非鱼)。连续观察15天，统计死亡鱼的数量，并计算存活率，结果统计参见图11。
由图可知，随着肌苷注射量的增加，鲫鱼、草鱼和罗非鱼感染迟缓爱德华氏菌后存活率逐渐增加。具体为：鲫鱼在未注射肌苷的情况下，存活率为40％，注射25μg肌苷后存活率为57％，注射50μg肌苷后存活率为73％，注射100μg肌苷后存活率为77％，存活率提高了17-37％。草鱼在未注射肌苷的情况下，存活率为40％，注射25μg肌苷后存活率为43％，注射50μg肌苷后存活率为53％，注射100μg肌苷后存活率为80％，存活率提高了3-40％。罗非鱼在未注射肌苷的情况下，存活率为60％，注射10μg肌苷后存活率为63％，注射20μg肌苷后存活率为70％，注射40μg肌苷后存活率为77％，存活率提高了3-17％。
4.2外源肌苷对鱼类感染迟缓爱德华氏菌后的免疫因子影响
将每种鱼各分组2组，迟缓爱德华氏菌感染组，肌苷+迟缓爱德华氏菌感染组。先采用肌肉注射方式每尾鱼注射10μL的肌苷(含量分别为50μg)，对照组注射相同体积的生理盐水。每天2次。对照组注射相同体积的生理盐水。三天后，采用肌肉注射迟缓爱德华氏菌(1×10 4CFU/鲫鱼，1×10 5CFU/草鱼，1×10 4CFU/罗非鱼)。6小时后，收集鱼的脾脏样品，每组4个生物学重复，荧光定量PCR检测免疫基因il1b和c3的表达情况。引物如下：鲫鱼：il1b1(上游atgcgctgctcaacttcat，下游ctggcccttattttgttgag)，il1b2(上游caaagcgatcctcttcattt，下游attcgggtcatcagttttaa)，C3(上游tggggatggatctgaaaca，下游tgcccatgatgaggtacga)。草鱼：il1b(上游gccatttccaagagtaatct，下游aacggattcaaaagtgttatta)，C3(上游atacatcggcatcactgaac，下游cccaaacggatattatgaag)。罗非鱼：il1b(上游aaggcacaaacctctatctg，下游tgtcgcgtttgtagaagaga)，c3(上游atgttaccttaacacccaag，下游gctctccatacaggtacctg)，结果数据统计参见图12。
由图可见，肌苷促进c3表达，而抑制il1b表达。具体情况为：相对于单独注射细菌组，草鱼在注射肌苷后il1b下降了2倍，补体c3上升了3倍。鲫鱼在注射肌苷后il1b1下降了3.2倍，il1b2下降了5倍，补体c3上升了2.5倍。罗非鱼在注射肌苷后il1b下降了4倍，补体c3上升了3.9倍。
这些结果说明，肌苷可提高宿主抗细菌感染在鱼类模型上也是适用的。
上述实施例为本发明较佳的实施方式，但本发明的实施方式并不受上述实施例的限制，其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化，均应为等效的置换方式，都包含在本发明的保护范围之内。

Claims (10)

  1. 肌苷在制备抗感染药物中的应用。
  2. 根据权利要求1所述应用，其特征在于，所述肌苷可以与脯氨酸羟化酶结合调节细胞因子。
  3. 根据权利要求2所述应用，其特征在于，所述细胞因子为IL-1β因子。
  4. 根据权利要求3所述应用，其特征在于，所述肌苷可以抑制所述IL-1β因子含量增加。
  5. 根据权利要求2所述应用，其特征在于，所述细胞因子为补体3C。
  6. 根据权利要求5所述应用，其特征在于，所述肌苷可以促进所述补体3C含量增加。
  7. 根据权利要求1～6任一所述应用，其特征在于，所述抗感染药物可以提高被感染生物的存活率。
  8. 根据权利要求7所述应用，其特征在于，所述肌苷包括肌苷，肌苷的可药用的盐和酯，选择性取代的类似物，肌苷立体异构体、几何异构体、水合物、溶剂化物或药学上可接受的盐或前药。
  9. 一种抗感染药物，其特征在于，含有有效量的肌苷。
  10. 根据权利要求9所述药物，其特征在于，所述药物为口服剂型或注射剂型。
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