CN117224558B - Application of RNA editing enzyme inhibitor in preparation of medicines for preventing and/or treating hepatic fibrosis - Google Patents
Application of RNA editing enzyme inhibitor in preparation of medicines for preventing and/or treating hepatic fibrosis Download PDFInfo
- Publication number
- CN117224558B CN117224558B CN202311052459.1A CN202311052459A CN117224558B CN 117224558 B CN117224558 B CN 117224558B CN 202311052459 A CN202311052459 A CN 202311052459A CN 117224558 B CN117224558 B CN 117224558B
- Authority
- CN
- China
- Prior art keywords
- liver
- fibrosis
- preventing
- aza
- liver fibrosis
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 206010019668 Hepatic fibrosis Diseases 0.000 title claims abstract description 19
- 239000003814 drug Substances 0.000 title claims abstract description 19
- 238000010357 RNA editing Methods 0.000 title abstract description 25
- 230000026279 RNA modification Effects 0.000 title abstract description 25
- 229940125532 enzyme inhibitor Drugs 0.000 title abstract description 21
- 239000002532 enzyme inhibitor Substances 0.000 title abstract description 21
- 229940079593 drug Drugs 0.000 title abstract description 6
- 238000002360 preparation method Methods 0.000 title description 9
- 208000019425 cirrhosis of liver Diseases 0.000 claims abstract description 57
- 210000005228 liver tissue Anatomy 0.000 claims abstract description 27
- 230000014509 gene expression Effects 0.000 claims abstract description 21
- 210000004024 hepatic stellate cell Anatomy 0.000 claims abstract description 21
- 239000003550 marker Substances 0.000 claims abstract description 16
- 102000008186 Collagen Human genes 0.000 claims abstract description 11
- 108010035532 Collagen Proteins 0.000 claims abstract description 11
- 229920001436 collagen Polymers 0.000 claims abstract description 11
- 230000008021 deposition Effects 0.000 claims abstract description 10
- 230000001575 pathological effect Effects 0.000 claims abstract description 7
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 6
- 239000000243 solution Substances 0.000 claims description 12
- OAUKGFJQZRGECT-UUOKFMHZSA-N 8-Azaadenosine Chemical compound N1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OAUKGFJQZRGECT-UUOKFMHZSA-N 0.000 claims description 7
- 210000000651 myofibroblast Anatomy 0.000 claims description 6
- 239000003795 chemical substances by application Substances 0.000 claims description 5
- 208000019423 liver disease Diseases 0.000 claims description 5
- 102100033601 Collagen alpha-1(I) chain Human genes 0.000 claims description 4
- 108010029483 alpha 1 Chain Collagen Type I Proteins 0.000 claims description 4
- 230000020411 cell activation Effects 0.000 claims description 3
- 238000006243 chemical reaction Methods 0.000 claims description 3
- 239000007928 intraperitoneal injection Substances 0.000 claims description 3
- 239000000443 aerosol Substances 0.000 claims description 2
- 230000037396 body weight Effects 0.000 claims description 2
- 239000002775 capsule Substances 0.000 claims description 2
- 239000006071 cream Substances 0.000 claims description 2
- 230000005764 inhibitory process Effects 0.000 claims description 2
- 239000007924 injection Substances 0.000 claims description 2
- 238000002347 injection Methods 0.000 claims description 2
- 238000010255 intramuscular injection Methods 0.000 claims description 2
- 239000007927 intramuscular injection Substances 0.000 claims description 2
- 238000010253 intravenous injection Methods 0.000 claims description 2
- 230000005976 liver dysfunction Effects 0.000 claims description 2
- 239000006210 lotion Substances 0.000 claims description 2
- 239000003094 microcapsule Substances 0.000 claims description 2
- 239000002674 ointment Substances 0.000 claims description 2
- 239000000843 powder Substances 0.000 claims description 2
- 239000007921 spray Substances 0.000 claims description 2
- 238000010254 subcutaneous injection Methods 0.000 claims description 2
- 239000007929 subcutaneous injection Substances 0.000 claims description 2
- 239000000829 suppository Substances 0.000 claims description 2
- 229940098465 tincture Drugs 0.000 claims description 2
- 241000699670 Mus sp. Species 0.000 abstract description 27
- 108090000623 proteins and genes Proteins 0.000 abstract description 16
- 102000004169 proteins and genes Human genes 0.000 abstract description 13
- 230000000694 effects Effects 0.000 abstract description 12
- 230000003908 liver function Effects 0.000 abstract description 10
- 230000004913 activation Effects 0.000 abstract description 5
- 241001465754 Metazoa Species 0.000 abstract description 4
- 230000001413 cellular effect Effects 0.000 abstract description 4
- 230000008859 change Effects 0.000 abstract description 4
- 238000013518 transcription Methods 0.000 abstract description 4
- 230000035897 transcription Effects 0.000 abstract description 4
- 230000002159 abnormal effect Effects 0.000 abstract description 3
- 101000865408 Homo sapiens Double-stranded RNA-specific adenosine deaminase Proteins 0.000 description 15
- 102100029791 Double-stranded RNA-specific adenosine deaminase Human genes 0.000 description 13
- 210000004185 liver Anatomy 0.000 description 12
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 9
- 239000004006 olive oil Substances 0.000 description 9
- 235000008390 olive oil Nutrition 0.000 description 9
- 108020004999 messenger RNA Proteins 0.000 description 8
- 102000003855 L-lactate dehydrogenase Human genes 0.000 description 7
- 108700023483 L-lactate dehydrogenases Proteins 0.000 description 7
- 206010016654 Fibrosis Diseases 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 5
- 210000002966 serum Anatomy 0.000 description 5
- 238000010186 staining Methods 0.000 description 5
- 230000004761 fibrosis Effects 0.000 description 4
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 4
- 230000002829 reductive effect Effects 0.000 description 4
- 238000013424 sirius red staining Methods 0.000 description 4
- 101150020966 Acta2 gene Proteins 0.000 description 3
- 238000011740 C57BL/6 mouse Methods 0.000 description 3
- 206010061218 Inflammation Diseases 0.000 description 3
- 206010067125 Liver injury Diseases 0.000 description 3
- 241000699666 Mus <mouse, genus> Species 0.000 description 3
- 101150093908 PDGFRB gene Proteins 0.000 description 3
- 108010051742 Platelet-Derived Growth Factor beta Receptor Proteins 0.000 description 3
- 102000018967 Platelet-Derived Growth Factor beta Receptor Human genes 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 230000004054 inflammatory process Effects 0.000 description 3
- 238000010172 mouse model Methods 0.000 description 3
- 230000035790 physiological processes and functions Effects 0.000 description 3
- 238000002560 therapeutic procedure Methods 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 102000007469 Actins Human genes 0.000 description 2
- 108010085238 Actins Proteins 0.000 description 2
- 102100036475 Alanine aminotransferase 1 Human genes 0.000 description 2
- 108010082126 Alanine transaminase Proteins 0.000 description 2
- 108010003415 Aspartate Aminotransferases Proteins 0.000 description 2
- 102000004625 Aspartate Aminotransferases Human genes 0.000 description 2
- 108010022452 Collagen Type I Proteins 0.000 description 2
- 102000012422 Collagen Type I Human genes 0.000 description 2
- 101710164680 Platelet-derived growth factor receptor beta Proteins 0.000 description 2
- 102100026547 Platelet-derived growth factor receptor beta Human genes 0.000 description 2
- 230000005856 abnormality Effects 0.000 description 2
- 125000000477 aza group Chemical group 0.000 description 2
- 231100000012 chronic liver injury Toxicity 0.000 description 2
- 230000034994 death Effects 0.000 description 2
- 231100000517 death Toxicity 0.000 description 2
- 239000000835 fiber Substances 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 230000017074 necrotic cell death Effects 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 210000002460 smooth muscle Anatomy 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- 238000001356 surgical procedure Methods 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 206010003827 Autoimmune hepatitis Diseases 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 206010057573 Chronic hepatic failure Diseases 0.000 description 1
- 241001550206 Colla Species 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 208000010334 End Stage Liver Disease Diseases 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 206010019280 Heart failures Diseases 0.000 description 1
- 206010019663 Hepatic failure Diseases 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 206010064390 Tumour invasion Diseases 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 230000002300 anti-fibrosis Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 230000030741 antigen processing and presentation Effects 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 230000009400 cancer invasion Effects 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 208000011444 chronic liver failure Diseases 0.000 description 1
- 230000007882 cirrhosis Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 230000013020 embryo development Effects 0.000 description 1
- 231100000584 environmental toxicity Toxicity 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 230000005182 global health Effects 0.000 description 1
- 231100000753 hepatic injury Toxicity 0.000 description 1
- 210000003494 hepatocyte Anatomy 0.000 description 1
- 230000005746 immune checkpoint blockade Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000007903 liver failure Diseases 0.000 description 1
- 231100000835 liver failure Toxicity 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 230000002107 myocardial effect Effects 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 235000019198 oils Nutrition 0.000 description 1
- 238000012261 overproduction Methods 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 210000003240 portal vein Anatomy 0.000 description 1
- 230000001124 posttranscriptional effect Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000008844 regulatory mechanism Effects 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 231100000241 scar Toxicity 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 208000037816 tissue injury Diseases 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
- 239000012224 working solution Substances 0.000 description 1
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to an application of an RNA editing enzyme inhibitor in preparing a medicament for preventing and/or treating hepatic fibrosis, belonging to the technical field of biological medicines. The invention proves that the RNA editing enzyme inhibitor 8-Aza can inhibit the expression of hepatic fibrosis marker molecule protein at the cellular level and the animal level for the first time, thereby inhibiting the activation of hepatic stellate cells. The RNA editing enzyme inhibitor 8-Aza can inhibit the liver fibrosis level of mice induced by CCl 4, including improving abnormal liver function, inhibiting transcription and expression of liver fibrosis related genes of liver tissues, obviously reducing collagen deposition of liver tissues and change of pathological structures of the liver tissues, and has obvious liver fibrosis resisting effect.
Description
Technical Field
The invention relates to the technical field of biological medicines, in particular to application of an RNA editing enzyme inhibitor in preparation of a medicine for preventing and/or treating liver fibrosis.
Background
Liver fibrosis is an abnormal repair response of chronic liver injury caused by a variety of causes, such as environmental toxicity, infection, metabolic causes, or other genetic factors including autoimmune hepatitis, can lead to chronic liver injury inflammation and fibrosis. The disease is mainly characterized by diffuse overproduction and deposition of extracellular matrix in the liver. Whereas activation or transformation of hepatic stellate cells into myofibroblasts is the primary driver of the disease. As the normal structure and physiological function of liver tissue is gradually destroyed, scar tissue gradually replaces liver parenchyma and further progresses to cirrhosis, liver failure or cancer, ultimately leading to death of the patient.
At present, various chronic liver diseases become global health burden; about 200 tens of thousands of deaths are caused annually, of which about 100 tens of thousands die from liver cirrhosis-related complications. At present, surgery and liver transplantation are the only opportunities for long-term survival in patients with severe liver disease. However, the availability of a suitable source of liver and the overall cost of surgery limit the clinical utility of this protocol. In addition, liver fibrosis often exhibits active progression once initiated, other therapies, such as long-term antiviral therapy and anti-liver fibrosis therapy directed to fibrous tissue proliferation and degradation, may have an ameliorating effect early in the disease but fail to prevent fibrosis progression in the late stages. Thus, the search for therapeutic alternatives and the search for new preventive strategies is particularly urgent.
A-to-I RNA editing is one of the most common types of RNA editing in mammals, and is widely involved in a variety of gene regulatory mechanisms at the transcriptional and posttranscriptional levels. RNA editing enzyme ADAR1 is one of the family members of proteins that catalyze the occurrence of such RNA editing patterns. Studies in ADAR1 mutant mice indicate that ADAR1 is critical for a variety of physiological functions, including embryonic development, immune response, and B and T lymphocyte development. Under physiological conditions, ADAR1 can prevent not only liver injury and liver fibrosis caused by inflammation, but also heart failure caused by myocardial cell inflammation. Under pathological conditions, the existing research proves that the up-regulation of ADAR1 expression in tumor cells can promote the formation and self-renewal of tumor stem cells. The deletion of tumor cell ADAR1 overcomes resistance to PD-1 checkpoint blockade caused by inactivation of cellular antigen presentation. The use of the inhibitor 8-azaadenosine (8-Azaadenosine, 8-Aza) can reduce tumor invasion, but no report has been found on the treatment of liver fibrosis with the RNA editing enzyme inhibitor 8-Aza.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide the application of the RNA editing enzyme inhibitor in preparing medicaments for preventing and/or treating liver fibrosis.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
in a first aspect, the present invention provides the use of an RNA editing enzyme inhibitor for the preparation of a medicament for the prevention and/or treatment of liver fibrosis.
In a second aspect, the invention provides the use of an RNA editing enzyme inhibitor in the manufacture of a medicament for reversing liver fibrosis.
The invention discovers that the RNA editing enzyme inhibitor can effectively treat hepatic fibrosis at the cellular level and the animal level for the first time, and discovers that the hepatic fibrosis marker molecule protein expression level of hepatic stellate cells is inhibited by adopting the RNA editing enzyme inhibitor 8-Aza to treat human hepatic stellate cells; the mouse model with liver fibrosis treated by RNA editing enzyme inhibitor 8-Aza has the effects of reversing liver fibrosis, and the mouse liver function index is found to be normal, liver tissue collagen deposition and liver tissue pathological structure reduction are achieved.
As a preferred embodiment of the use according to the invention, the RNA editing enzyme inhibitor is an A-to-I RNA editing enzyme inhibitor.
As a preferred embodiment of the use according to the invention, the RNA editing enzyme inhibitor is 8-azaadenosine.
As a preferred embodiment of the use of the present invention, the agent for preventing and/or treating hepatic fibrosis can inhibit activation of hepatic stellate cells and/or transformation to myofibroblasts; the inhibition of hepatic stellate cell activation and/or conversion to myofibroblasts is to inhibit expression of the marker molecules COL1A1, pdgfrβ and α -SMA.
As a preferred embodiment of the use of the present invention, the agent for preventing and/or treating liver fibrosis can improve liver dysfunction, inhibit collagen deposition in liver tissue and change in pathological structure of liver tissue.
As a preferred embodiment of the use according to the present invention, the drug for preventing and/or treating liver fibrosis is in the form of at least one of a capsule, a tablet, an oral preparation, a microcapsule preparation, an injection, a suppository, a spray, an ointment, a gel, a solution, a powder, a lotion, a tincture, an oil, a cream and an aerosol.
As a preferred embodiment of the use of the present invention, the agent for preventing and/or treating liver fibrosis is used for at least one of intravenous injection, intramuscular injection, subcutaneous injection and intraperitoneal injection.
As a preferred embodiment of the use according to the present invention, the agent for preventing and/or treating liver fibrosis is administered intraperitoneally at a dose of 2mg per 1kg body weight of the organism. In the experimental process, the RNA editing enzyme inhibitor with the concentration of 2mg/kg can obviously reduce liver function indexes of mice with liver fibrosis, reduce liver tissue collagen deposition and change of pathological structures of liver tissues, and effectively relieve liver fibrosis of the mice.
In a third aspect, the invention provides a medicament for treating liver fibrosis, the medicament comprising an RNA editing enzyme inhibitor and a pharmaceutically acceptable carrier.
As a preferred embodiment of the use according to the invention, the RNA editing enzyme inhibitor is 8-azaadenosine.
Compared with the prior art, the invention has the beneficial effects that:
The invention proves that the RNA editing enzyme inhibitor 8-Aza can inhibit the expression of hepatic fibrosis marker molecule protein at the cellular level and the animal level for the first time, thereby inhibiting the activation of hepatic stellate cells. The 8-Aza can inhibit the liver fibrosis level of mice induced by CCl 4, including improving abnormal liver function, inhibiting transcription and expression of liver fibrosis related genes of liver tissues, obviously reducing collagen deposition of the liver tissues and change of pathological structures of the liver tissues, and has obvious anti-liver fibrosis effect.
Drawings
FIG. 1 is a graph showing the analysis results of ADAR1 mRNA expression levels in the normal healthy group and the liver cirrhosis group in experimental example 1;
FIG. 2 shows protein expression levels of COL1A1, PDGFR beta and alpha-SMA, which are markers of hepatic fibrosis of hepatic stellate cells treated with 8-Aza in Experimental example 2;
FIG. 3 is a schematic diagram of a liver fibrosis mouse model and a dosing regimen in Experimental example 3;
FIG. 4 shows the results of liver function index detection of mice in each experimental group in experimental example 3, wherein the liver function index comprises glutamic pyruvic transaminase (ALT), glutamic oxaloacetic transaminase (AST) and Lactate Dehydrogenase (LDH) in serum, wherein A is AST, B is ALT, and C is LDH;
FIG. 5 shows the results of HE staining, masson staining and sirius red staining of the liver of mice of each experimental group in experimental example 3;
FIG. 6 shows mRNA transcription levels of hepatic fibrosis marker molecules of mice in each experimental group in experimental example 3, wherein A is type I collagen (Colla 1), B is platelet-derived growth factor receptor beta (Pdgfr beta), and C is alpha smooth muscle actin (Acta 2);
FIG. 7 shows the expression level of a hepatic fibrosis marker a-SMA (Acta 2 gene-encoded protein) protein in mice of each experimental group in experimental example 3;
In the above graph, "+" "indicates that there is a statistical difference between the two groups," + "" is p < 0.05, "p < 0.01," p < 0.001, "p < 0.0001.
Detailed Description
For a better description of the objects, technical solutions and advantages of the present invention, the present invention will be further described with reference to the following specific examples.
The reagents and consumables used in the following examples were all commercially available without any particular explanation.
In the following experimental examples, 8-azaadenosine (hereinafter, referred to as 8-Aza) was used as an RNA editing enzyme inhibitor.
8-Aza was supplied by Tocres under the trade designation 6868/10.
Hepatic stellate cells are human hepatic stellate cell line LX2, purchased from cell banks of institute of biochemistry and cell biology, academy of sciences of china (Shanghai, china).
C57BL/6 male mice were supplied by Guangdong Kangdong Biotechnology Co.
Experimental example 1 analysis of liver tissue ADAR1 Gene expression in patients with liver cirrhosis
After liver fibrosis occurs in patients, liver cirrhosis is liable to develop as the normal structure and physiological functions of liver tissue are destroyed. To investigate the relationship between ADAR1 and liver cirrhosis, in the data of ADAR1 mRNA expression levels of liver tissues of liver cirrhosis patients (origin GEO Dataset public sequencing platform (GEO 14323)), sequencing results of ADAR1 genes in normal healthy control group and liver tissues of liver cirrhosis patients were selected, respectively, and statistical analysis was performed, and the results are shown in table 1 and fig. 1.
TABLE 1 analysis of the expression level of ADAR1 in the normal healthy group and the liver cirrhosis group
As shown in table 1 and fig. 1, the mRNA level of ADAR1 was higher in the liver cirrhosis group than in the normal healthy group, indicating that ADAR1 was associated with liver cirrhosis, and since liver fibrosis is a necessary link for various chronic liver diseases to progress to end-stage liver diseases such as liver cirrhosis, blocking or reversing the progress of liver fibrosis is advantageous for delaying or even reversing the progress of liver cirrhosis, and thus the following experiments consider whether or not the use of ADAR1 inhibitors can be used for treating liver fibrosis.
Experimental example 28-Effect of Aza on hepatic stellate cells
Activation or differentiation of hepatic stellate cells into myofibroblasts is a core pathogenesis of hepatic fibrosis, so this experiment treats hepatic stellate cells by 8-Aza, and observes changes in hepatic stellate cells to evaluate the effect of 8-Aza on hepatic stellate cells.
(1) Preparation of the reagent:
8-Aza was dissolved in DMSO to prepare a 10mM stock solution.
(2) The experimental contents are as follows:
Human hepatic stellate cell line LX2 was inoculated with DMEM high sugar medium containing 10% fetal bovine serum and cultured overnight at 37 ℃ under 5% co 2.
The total protein was extracted and the expression levels of hepatic fibrosis marker type I collagen (COL 1A 1), platelet-derived growth factor receptor beta (PDGFR beta) and alpha smooth muscle actin (alpha-SMA) in the cells were detected by Western Blot (WB) by adding 8-Aza to a cell culture medium to a final concentration of 8-Aza of 10. Mu.M, culturing for 24 hours and 48 hours, and collecting the cells, and the results are shown in FIG. 2.
As shown in FIG. 2, the expression of hepatic fibrosis marker molecules COL1A1, PDGFR beta and alpha-SMA was inhibited in the 8-Aza-treated hepatic stellate cells compared to the hepatic stellate cells without the 8-Aza treatment, indicating that 8-Aza has an effect of inhibiting the expression of hepatic fibrosis marker molecules in the hepatic stellate cells.
Experimental example 38-Effect of Aza on liver fibrosis mice
To investigate the effect of 8-Aza on liver fibrosis mice, the effect of 8-Aza on liver fibrosis treatment was assessed by animal experiments.
(1) Preparation of reagents
At the time of use, CCl 4 was formulated as a 20% (v/v) solution with olive oil.
8-Aza was dissolved in DMSO and prepared as a 25mg/ml stock solution. At the time of use, the solution was diluted 100 times with PBS and prepared as 0.25mg/ml working solution.
(2) Preparation of liver fibrosis mouse model
The C57BL/6 male mice of 6-8 weeks of age were selected and given 20% CCl 4 solution, 20% CCl 4 solution was given to the mice by intraperitoneal injection at a dose of 0.5ml/kg, twice weekly for 6 weeks.
(3) Drug treatment
C57BL/6 male mice were randomly divided into 4 groups, olive oil group (5), CCl 4 group (6), PBS group (6) and 8-Aza group (6). Except for the olive oil group, the rest 3 groups were given 20% ccl 4 solution. After 4 weeks of CCl 4 solution administration, olive oil and CCl 4 groups were not subjected to any treatment while modeling, PBS group was given by weight (containing 1% DMSO, v/v), 8-Aza group was given by weight 8-Aza to a final concentration of 2mg/kg, 3 times per week for 2 weeks, and the dosing regimen is shown in FIG. 3.
(4) Sampling and detection
After 24h of the last administration, mice were anesthetized, blood was collected from the fundus venous plexus, placed in a refrigerator at 4℃overnight, blood samples were centrifuged at 2500rpm at 4℃for 15min, the supernatant serum was aspirated, and after 10-fold dilution with PBS, the liver function indexes glutamic pyruvic transaminase (ALT), glutamic oxaloacetic transaminase (AST) and Lactate Dehydrogenase (LDH) were detected, the results are shown in FIG. 4 and Table 2.
The abdominal cavity of the mice after blood withdrawal was opened and pre-chilled PBS was slowly perfused through the portal vein in an amount of 10 ml/mouse. Then, the left lobe of the liver is immersed in tissue fixing liquid to prepare paraffin sections, and HE staining, masson staining and sirius red staining are carried out. And extracting RNA from part of livers to detect mRNA levels of marker molecules Col1al, pdgfrβ and Acta2 of the liver tissue fibrosis of the mice, and extracting protein from part of livers to detect the expression level of alpha-SMA (Acta 2 encoding protein) of the liver tissue fibrosis marker molecules of the mice. HE staining, masson staining and sirius red staining results are shown in FIG. 5, hepatic fibrosis marker mRNA transcription level results are shown in FIG. 6, and hepatic fibrosis marker protein expression level results are shown in FIG. 7.
TABLE 2 results of liver function index detection in serum of each experimental group
As shown in table 2 and fig. 4, the ALT/AST/LDH activity was significantly increased in the serum of the cci 4 group mice compared to the olive oil group; compared with PBS group, serum ALT/AST/LDH activity of 8-Aza mice is obviously reduced, which indicates that 8-Aza can improve liver function abnormality caused by CCl 4, namely 8-Aza can improve liver function abnormality caused by liver fibrosis.
As shown in fig. 5, the results of HE staining of liver tissue of mice show that liver hepatocytes of olive oil group are normal in structure and liver lobule is normal in structure, and no damage is seen; the liver lobules of CCl 4 group and PBS group are seriously damaged, the structure is disordered, obvious swelling and deformation are caused, and sheet necrosis exists; compared with PBS group, the 8-Aza group has obviously reduced sheet necrosis area and obvious edema improvement. The results of Masson staining and sirius red staining of mouse liver tissue showed that olive oil group liver tissue had no collagen deposition, and that there was a large amount of collagen deposition in CCl 4 group and PBS group. The 8-Aza group showed significantly less collagen fibers than the PBS group, but some of them were still seen to have significantly improved collagen fibers. The above staining results show that 8-Aza can significantly improve the liver fibrosis degree of mice induced by CCl 4, namely 8-Aza has the effect of improving the liver fibrosis degree.
As shown in fig. 6, mRNA levels of the Pdgfr β, colla1 and Acta2 genes were significantly up-regulated in the liver tissue of the cci 4 and PBS groups of mice compared to the olive oil group; compared with the PBS group, the mRNA levels of Pdgfr β, collal and Acta2 genes of the liver tissue of the 8-Aza group mice are obviously reduced, and the difference is statistically significant. As shown in fig. 7, compared with olive oil group, the liver fibrosis marker molecules alpha-SMA protein expression levels of CCl 4 group and PBS group are obviously increased; compared with the PBS group, the expression quantity of the hepatic fibrosis marker molecule protein alpha-SMA of the 8-Aza group is obviously reduced.
The results show that the 8-Aza can obviously relieve liver tissue injury of mice, reduce liver collagen deposition, reverse liver fibrosis symptoms of the mice and has obvious anti-fibrosis effect.
Finally, it should be noted that the above embodiments are only for illustrating the technical solution of the present invention and not for limiting the scope of the present invention, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solution of the present invention may be modified or substituted equally without departing from the spirit and scope of the technical solution of the present invention.
Claims (7)
- Use of 8-aza-adenosine in preparing medicament for preventing and/or treating liver fibrosis.
- Application of 8-aza-adenosine in preparing medicine for reversing hepatic fibrosis.
- 3. The use according to claim 1, wherein the agent for preventing and/or treating liver fibrosis inhibits hepatic stellate cell activation and/or conversion to myofibroblasts; the inhibition of hepatic stellate cell activation and/or conversion to myofibroblasts is to inhibit expression of the marker molecules COL1A1, pdgfrβ and α -SMA.
- 4. The use according to claim 1, wherein the medicament for preventing and/or treating liver fibrosis is capable of improving liver dysfunction, inhibiting collagen deposition in liver tissue and altering pathological structure of liver tissue.
- 5. The use according to claim 1, wherein the medicament for preventing and/or treating liver fibrosis is in the form of at least one of a capsule, a tablet, a microcapsule, an injection, a suppository, a spray, an ointment, a gel, a solution, a powder, a lotion, a tincture, an oil, a cream and an aerosol.
- 6. The use according to claim 1, wherein the medicament for preventing and/or treating liver fibrosis is for at least one of intravenous injection, intramuscular injection, subcutaneous injection and intraperitoneal injection.
- 7. The use according to claim 6, wherein the medicament for preventing and/or treating liver fibrosis is administered intraperitoneally at a dose of 2mg per 1kg body weight of the organism.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311052459.1A CN117224558B (en) | 2023-08-21 | 2023-08-21 | Application of RNA editing enzyme inhibitor in preparation of medicines for preventing and/or treating hepatic fibrosis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311052459.1A CN117224558B (en) | 2023-08-21 | 2023-08-21 | Application of RNA editing enzyme inhibitor in preparation of medicines for preventing and/or treating hepatic fibrosis |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN117224558A CN117224558A (en) | 2023-12-15 |
| CN117224558B true CN117224558B (en) | 2024-05-17 |
Family
ID=89097556
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311052459.1A Active CN117224558B (en) | 2023-08-21 | 2023-08-21 | Application of RNA editing enzyme inhibitor in preparation of medicines for preventing and/or treating hepatic fibrosis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117224558B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2006203699A1 (en) * | 2000-02-10 | 2006-09-21 | New York University | Adenosine A2A receptor antagonists for treating and preventing hepatic fibrosis, cirrhosis and fatty liver |
| CN107875164A (en) * | 2017-11-16 | 2018-04-06 | 中国药科大学 | A kind of medicine for treating liver fibrosis |
-
2023
- 2023-08-21 CN CN202311052459.1A patent/CN117224558B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2006203699A1 (en) * | 2000-02-10 | 2006-09-21 | New York University | Adenosine A2A receptor antagonists for treating and preventing hepatic fibrosis, cirrhosis and fatty liver |
| CN107875164A (en) * | 2017-11-16 | 2018-04-06 | 中国药科大学 | A kind of medicine for treating liver fibrosis |
Non-Patent Citations (2)
| Title |
|---|
| ADAR1-mediated RNA editing is a novel oncogenic process in thyroid cancer and regulates miR-200 activity.;Julia, Ramírez-Moya;Allison R, Baker;Frank J, Slack;Pilar, Santisteban;Oncogene;20201231;第39卷(第18期);3738-3753 * |
| Hepatitis B virus evades immune recognition via RNA adenosine deaminase ADAR1-mediated viral RNA editing in hepatocytes;Liyuan Wang;Cellular & Molecular Immunology;20210712;第18卷;1871-1882 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN117224558A (en) | 2023-12-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Li et al. | Dexmedetomidine attenuates myocardial ischemia‐reperfusion injury in diabetes mellitus by inhibiting endoplasmic reticulum stress | |
| CN113307845B (en) | Polypeptide for promoting hepatocyte proliferation and/or inhibiting hepatocyte apoptosis and application thereof | |
| CN114306296A (en) | Application of disulfiram in preparation of medicine for treating membranous nephropathy | |
| CN112972447B (en) | Application of CaMK II inhibitor in preparation of medicine for preventing and/or treating acute pancreatitis | |
| Zhang et al. | Update on the sources, pharmacokinetics, pharmacological action, and clinical application of anisodamine | |
| Fan et al. | Celastrol relieves myocardial infarction-induced cardiac fibrosis by inhibiting NLRP3 inflammasomes in rats | |
| CN117224558B (en) | Application of RNA editing enzyme inhibitor in preparation of medicines for preventing and/or treating hepatic fibrosis | |
| CN108434139A (en) | Hypoxia inducible factor prolyl hydroxylase activity inhibitor is preparing the application in preventing acute kidney injury drug | |
| CN109288827B (en) | Application of ceramidase inhibitor D-e-MAPP in preparation of medicine for improving acute pancreatitis | |
| KR102206017B1 (en) | Pharmaceutical composition for treating skin fibrosis comprising PARP1 inhibitor | |
| Weng et al. | PER2 regulates reactive oxygen species production in the circadian susceptibility to ischemia/reperfusion injury in the heart | |
| CN108836965A (en) | Application of the niacinamide composition in preparation treatment Sorafenib hand-foot skin reaction drug | |
| CN113388615A (en) | MiRNA for preventing and/or treating acute pancreatitis and pharmaceutical application thereof | |
| Vaccaro et al. | Is spontaneous bacterial peritonitis an inducer of vasopressin analogue side-effects? A case report | |
| CN108635571B (en) | Application of irisin in preparation of medicine for preventing and treating severe acute pancreatitis | |
| CN112168966A (en) | Application of interfering NLRP3 inflammasome in preparing medicine for treating novel coronavirus pneumonia | |
| CN113350362A (en) | Application of vitamin D receptor agonist in preparation of medicine for preventing and/or treating acute pancreatitis | |
| CN109276567B (en) | Application of 7-hydroxycoumarin in preparation of medicine for treating acute kidney injury | |
| TWI607751B (en) | Statin compounds for the treatment of gastric cancer | |
| CN110917351A (en) | Use of MBD2 inhibitors for the prevention and treatment of fibrotic diseases | |
| CN112294961B (en) | ACP5 inhibitors and their use in the prevention and treatment of fibrotic diseases | |
| CN114907287B (en) | Compound for treating kidney injury | |
| CN108815512B (en) | Anticancer medicine and its application | |
| CN108685896B (en) | Application of oroxylin A in preparation of medicine for treating and/or preventing chronic peripheral vascular occlusive diseases | |
| CN109700807B (en) | Application of compound RH-1402 in preparation of acute kidney injury resistant medicine |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |