CN111012765A - Application of pseudopterogorgia-hancei alcohol as active substance in preparing medicine for treating cerebral apoplexy - Google Patents
Application of pseudopterogorgia-hancei alcohol as active substance in preparing medicine for treating cerebral apoplexy Download PDFInfo
- Publication number
- CN111012765A CN111012765A CN201911203933.XA CN201911203933A CN111012765A CN 111012765 A CN111012765 A CN 111012765A CN 201911203933 A CN201911203933 A CN 201911203933A CN 111012765 A CN111012765 A CN 111012765A
- Authority
- CN
- China
- Prior art keywords
- lemnalol
- pseudopterogorgia
- alcohol
- application
- inflammatory
- 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.)
- Pending
Links
- 239000003814 drug Substances 0.000 title claims abstract description 25
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 title claims abstract description 12
- 208000006011 Stroke Diseases 0.000 title claims description 31
- 206010008190 Cerebrovascular accident Diseases 0.000 title claims description 16
- 239000013543 active substance Substances 0.000 title abstract description 5
- 230000002490 cerebral effect Effects 0.000 title description 18
- 241001184986 Pseudopterogorgia Species 0.000 claims abstract description 10
- 229940079593 drug Drugs 0.000 claims description 9
- 208000029028 brain injury Diseases 0.000 claims description 5
- 239000000126 substance Substances 0.000 claims description 2
- 230000003247 decreasing effect Effects 0.000 claims 1
- LPXOPRGPLUWGKB-UHFFFAOYSA-N Lemnalol Natural products C1C(O)C(=C)C2C3(C)CCC(C(C)C)C2C31 LPXOPRGPLUWGKB-UHFFFAOYSA-N 0.000 abstract description 39
- LPXOPRGPLUWGKB-VRPMWHRCSA-N lemnalol Chemical compound C1[C@@H](O)C(=C)[C@@H]2[C@]3(C)CC[C@@H](C(C)C)[C@H]2[C@H]31 LPXOPRGPLUWGKB-VRPMWHRCSA-N 0.000 abstract description 39
- 241000700159 Rattus Species 0.000 abstract description 14
- 230000000694 effects Effects 0.000 abstract description 12
- 206010061218 Inflammation Diseases 0.000 abstract description 11
- 230000000324 neuroprotective effect Effects 0.000 abstract description 11
- 230000004054 inflammatory process Effects 0.000 abstract description 10
- 230000007246 mechanism Effects 0.000 abstract description 8
- 210000005036 nerve Anatomy 0.000 abstract description 5
- 239000004090 neuroprotective agent Substances 0.000 abstract description 5
- 230000002401 inhibitory effect Effects 0.000 abstract description 4
- 239000002547 new drug Substances 0.000 abstract description 3
- 238000007877 drug screening Methods 0.000 abstract description 2
- 241001596954 Larimichthys Species 0.000 abstract 1
- 230000014509 gene expression Effects 0.000 description 22
- 230000002757 inflammatory effect Effects 0.000 description 11
- 230000006378 damage Effects 0.000 description 10
- 108090000193 Interleukin-1 beta Proteins 0.000 description 9
- 102100029438 Nitric oxide synthase, inducible Human genes 0.000 description 9
- 101710089543 Nitric oxide synthase, inducible Proteins 0.000 description 9
- 206010008118 cerebral infarction Diseases 0.000 description 9
- 230000028709 inflammatory response Effects 0.000 description 9
- 210000000274 microglia Anatomy 0.000 description 9
- 238000011160 research Methods 0.000 description 9
- 102000003777 Interleukin-1 beta Human genes 0.000 description 8
- 208000027418 Wounds and injury Diseases 0.000 description 8
- 239000012634 fragment Substances 0.000 description 8
- 208000014674 injury Diseases 0.000 description 8
- 108090000623 proteins and genes Proteins 0.000 description 8
- 238000011144 upstream manufacturing Methods 0.000 description 8
- 108090001005 Interleukin-6 Proteins 0.000 description 7
- 208000026106 cerebrovascular disease Diseases 0.000 description 7
- 210000002540 macrophage Anatomy 0.000 description 7
- 108020004999 messenger RNA Proteins 0.000 description 7
- 238000000034 method Methods 0.000 description 7
- 230000001575 pathological effect Effects 0.000 description 7
- 230000010410 reperfusion Effects 0.000 description 7
- 102000004889 Interleukin-6 Human genes 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 6
- 230000005764 inhibitory process Effects 0.000 description 6
- 239000003550 marker Substances 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- GOZMBJCYMQQACI-UHFFFAOYSA-N 6,7-dimethyl-3-[[methyl-[2-[methyl-[[1-[3-(trifluoromethyl)phenyl]indol-3-yl]methyl]amino]ethyl]amino]methyl]chromen-4-one;dihydrochloride Chemical compound Cl.Cl.C=1OC2=CC(C)=C(C)C=C2C(=O)C=1CN(C)CCN(C)CC(C1=CC=CC=C11)=CN1C1=CC=CC(C(F)(F)F)=C1 GOZMBJCYMQQACI-UHFFFAOYSA-N 0.000 description 4
- 101150093802 CXCL1 gene Proteins 0.000 description 4
- 101150031350 Cxcl2 gene Proteins 0.000 description 4
- 208000032382 Ischaemic stroke Diseases 0.000 description 4
- 102100038280 Prostaglandin G/H synthase 2 Human genes 0.000 description 4
- 238000010171 animal model Methods 0.000 description 4
- 230000003110 anti-inflammatory effect Effects 0.000 description 4
- 210000005013 brain tissue Anatomy 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 201000010099 disease Diseases 0.000 description 4
- 238000011156 evaluation Methods 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- 230000004112 neuroprotection Effects 0.000 description 4
- 230000003040 nociceptive effect Effects 0.000 description 4
- 102000004127 Cytokines Human genes 0.000 description 3
- 108090000695 Cytokines Proteins 0.000 description 3
- 206010061216 Infarction Diseases 0.000 description 3
- 241000006758 Lemnalia Species 0.000 description 3
- 108050003267 Prostaglandin G/H synthase 2 Proteins 0.000 description 3
- 238000011529 RT qPCR Methods 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 101150088826 arg1 gene Proteins 0.000 description 3
- 239000002299 complementary DNA Substances 0.000 description 3
- 230000007547 defect Effects 0.000 description 3
- 230000007574 infarction Effects 0.000 description 3
- 239000007928 intraperitoneal injection Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 210000004498 neuroglial cell Anatomy 0.000 description 3
- 238000012827 research and development Methods 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 238000010839 reverse transcription Methods 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- VLXDPFLIRFYIME-QRTUWBSPSA-N (1S,2R,6R,7R,8S)-1,3-dimethyl-8-propan-2-yltricyclo[4.4.0.02,7]dec-3-ene Chemical compound C1C=C(C)[C@@H]2[C@@]3(C)CC[C@@H](C(C)C)[C@@H]2[C@H]31 VLXDPFLIRFYIME-QRTUWBSPSA-N 0.000 description 2
- 102000019034 Chemokines Human genes 0.000 description 2
- 108010012236 Chemokines Proteins 0.000 description 2
- 208000005189 Embolism Diseases 0.000 description 2
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 2
- 238000012404 In vitro experiment Methods 0.000 description 2
- 229940123915 Ion channel antagonist Drugs 0.000 description 2
- 244000198896 Lagerstroemia speciosa Species 0.000 description 2
- 101100260702 Mus musculus Tinagl1 gene Proteins 0.000 description 2
- 238000011530 RNeasy Mini Kit Methods 0.000 description 2
- 210000000683 abdominal cavity Anatomy 0.000 description 2
- 230000017531 blood circulation Effects 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 210000003169 central nervous system Anatomy 0.000 description 2
- 230000006735 deficit Effects 0.000 description 2
- 239000003651 drinking water Substances 0.000 description 2
- 235000020188 drinking water Nutrition 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 210000003194 forelimb Anatomy 0.000 description 2
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 2
- 210000004969 inflammatory cell Anatomy 0.000 description 2
- 238000010253 intravenous injection Methods 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- 230000000302 ischemic effect Effects 0.000 description 2
- 210000003657 middle cerebral artery Anatomy 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 229930014626 natural product Natural products 0.000 description 2
- 231100000915 pathological change Toxicity 0.000 description 2
- 230000036285 pathological change Effects 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 230000003827 upregulation Effects 0.000 description 2
- 230000000007 visual effect Effects 0.000 description 2
- 230000006269 (delayed) early viral mRNA transcription Effects 0.000 description 1
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 102000007469 Actins Human genes 0.000 description 1
- 108010085238 Actins Proteins 0.000 description 1
- 101100055072 Agrocybe aegerita Agr1 gene Proteins 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 201000006474 Brain Ischemia Diseases 0.000 description 1
- 102100032912 CD44 antigen Human genes 0.000 description 1
- 206010008092 Cerebral artery thrombosis Diseases 0.000 description 1
- 206010008120 Cerebral ischaemia Diseases 0.000 description 1
- 206010008132 Cerebral thrombosis Diseases 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 101000868273 Homo sapiens CD44 antigen Proteins 0.000 description 1
- 101000725401 Homo sapiens Cytochrome c oxidase subunit 2 Proteins 0.000 description 1
- 101001046686 Homo sapiens Integrin alpha-M Proteins 0.000 description 1
- 101000605127 Homo sapiens Prostaglandin G/H synthase 2 Proteins 0.000 description 1
- 101150109636 Inos gene Proteins 0.000 description 1
- 102100022338 Integrin alpha-M Human genes 0.000 description 1
- 201000001429 Intracranial Thrombosis Diseases 0.000 description 1
- 102000002274 Matrix Metalloproteinases Human genes 0.000 description 1
- 108010000684 Matrix Metalloproteinases Proteins 0.000 description 1
- 208000036110 Neuroinflammatory disease Diseases 0.000 description 1
- 102000011779 Nitric Oxide Synthase Type II Human genes 0.000 description 1
- 108010076864 Nitric Oxide Synthase Type II Proteins 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- 238000010222 PCR analysis Methods 0.000 description 1
- 101150000187 PTGS2 gene Proteins 0.000 description 1
- 206010033799 Paralysis Diseases 0.000 description 1
- 208000037273 Pathologic Processes Diseases 0.000 description 1
- 206010057249 Phagocytosis Diseases 0.000 description 1
- 238000010802 RNA extraction kit Methods 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 206010063837 Reperfusion injury Diseases 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 101150087698 alpha gene Proteins 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 238000000540 analysis of variance Methods 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 239000005557 antagonist Substances 0.000 description 1
- 230000006909 anti-apoptosis Effects 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 230000030741 antigen processing and presentation Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000008499 blood brain barrier function Effects 0.000 description 1
- 210000001218 blood-brain barrier Anatomy 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 230000001964 calcium overload Effects 0.000 description 1
- 210000001168 carotid artery common Anatomy 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- RNFNDJAIBTYOQL-UHFFFAOYSA-N chloral hydrate Chemical compound OC(O)C(Cl)(Cl)Cl RNFNDJAIBTYOQL-UHFFFAOYSA-N 0.000 description 1
- 229960002327 chloral hydrate Drugs 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 238000001647 drug administration Methods 0.000 description 1
- 238000009509 drug development Methods 0.000 description 1
- 230000002964 excitative effect Effects 0.000 description 1
- 230000003492 excitotoxic effect Effects 0.000 description 1
- 231100000063 excitotoxicity Toxicity 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 239000003527 fibrinolytic agent Substances 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 238000010191 image analysis Methods 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 238000011502 immune monitoring Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 208000028867 ischemia Diseases 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000031990 negative regulation of inflammatory response Effects 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- 230000003959 neuroinflammation Effects 0.000 description 1
- 230000002314 neuroinflammatory effect Effects 0.000 description 1
- 230000007658 neurological function Effects 0.000 description 1
- 230000016273 neuron death Effects 0.000 description 1
- 231100000189 neurotoxic Toxicity 0.000 description 1
- 230000002887 neurotoxic effect Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 229920001778 nylon Polymers 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000009054 pathological process Effects 0.000 description 1
- 230000008782 phagocytosis Effects 0.000 description 1
- 230000003285 pharmacodynamic effect Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000000770 proinflammatory effect Effects 0.000 description 1
- 230000004224 protection Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 239000002516 radical scavenger Substances 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000020341 sensory perception of pain Effects 0.000 description 1
- 229930004725 sesquiterpene Natural products 0.000 description 1
- 150000004354 sesquiterpene derivatives Chemical class 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
Images
Classifications
-
- 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
- A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Urology & Nephrology (AREA)
- Cardiology (AREA)
- Vascular Medicine (AREA)
- Heart & Thoracic Surgery (AREA)
- Epidemiology (AREA)
- Biomedical Technology (AREA)
- Neurology (AREA)
- Neurosurgery (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention provides an application of pseudopterogorgia pseudosciaena active substance (Lemnalol) in preparing a medicine for treating stroke, belonging to the field of pharmacy. The pseudopterogorgia alcohol (Lemnalol) has the obvious effects of protecting nerves and inhibiting inflammatory reaction on tMCAO rats and the like, and accords with the new guidance of the new drug screening in the world at present. Therefore, the application provides the possibility of the pseudopterogorgia pseudosciaenae alcohol (Lemnalol) serving as a novel neuroprotective agent and a potential neuroprotective mechanism, and provides a new experimental basis for clinical treatment application.
Description
Technical Field
The invention belongs to the field of pharmacy, and particularly relates to application of pseudopterogorgia alcohol in preparation of a medicine for reducing brain injury in stroke.
Background
Stroke, commonly known as stroke, is a disorder of cerebral blood circulation which is a sudden onset of disease, and is also the most serious complication of cerebrovascular disease. According to the third national cause of death investigation (2012) in China, the stroke fatality rate is ranked first and is as high as 22.45%, the disability rate is as high as 75%, and serious burden is brought to families and society.
Over the last 30 years, there were nearly 200 stroke drugs entering clinical trials worldwide. However, to date, only tissue-type fibrinolytic agents (tPA) have acquired the us FDA's clinical application approval. However, because tPA can only be used for cerebral thrombosis patients and has the defect of narrow treatment time window (the treatment is effective within 3 hours of the onset of the disease), the medicine has the treatment effect on 3-5 percent of cerebral apoplexy patients. However, the current research on the cerebral apoplexy treatment drugs discovers that the ion channel antagonist, the oxygen radical scavenger, the excitatory transmitter antagonist and the like are effective in basic research, but are really used for clinical discovery and have poor curative effect. For the reasons of clinical trial failure of the cerebral apoplexy medicaments, international pharmacians conduct multi-directional research, summarize possible reasons causing clinical trial failure, and propose to search medicaments with multiple neuroprotective effects such as anti-apoptosis, anti-oxidation and anti-inflammation and the like as research and development objects in the process of developing the cerebral apoplexy medicaments so as to improve the chance of developing the cerebral apoplexy medicaments successfully. Therefore, the active search for new drugs with multiple neuroprotective effects is the focus of drug development for the treatment of ischemic stroke, but no neuroprotective agent with a clear therapeutic effect clinically proven so far has been available.
Ischemic stroke causes a series of early events (from minutes to hours) such as excitotoxicity, oxidative stress, calcium overload and the like due to insufficient blood flow in local areas, so that glial cells are activated to secrete inflammatory mediators such as cytokines, chemokines, matrix metalloproteinases and the like, inflammatory cells derived from blood penetrate into cerebral ischemic areas, infiltrated leukocytes release the cytokines and the chemokines, glial cells are further activated, inflammatory signal cascade amplification reaction is generated, various cytotoxic components are released, blood brain barriers are damaged, and late events (from hours to days) such as immune injury, neuronal death and the like are further aggravated. Since patients often go to hospital for medical treatment hours after the onset of disease, the main pathological feature at this stage is inflammatory injury. Based on the discovery of the pathological mechanism and the failure of research and development of the medicines such as the ion channel antagonist and the like, the inhibition of inflammatory reaction after ischemic stroke has a longer treatment time window, and is considered to be an important direction for research and development of the cerebral stroke medicines.
Microglia is a main innate immune cell of the central nervous system and also a macrophage which plays a role in the nervous system and is responsible for immune monitoring, antigen presentation, phagocytosis, secretion of regulatory immune molecules and the like, CD11b is a marker of microglia and macrophages, and after being activated in inflammatory response after cerebral stroke, microglia/macrophages express Cxcl1 and Cxcl2, IL-1 β, IL-6, TNF- α, iNOS and other marker genes, among which IL-1 β is an important cytokine and plays a key role in neuroinflammation, while under pathological conditions, inflammatory cells, macrophages and glial cells and the like express inducible nitric oxide synthase (indoviborans, iNOS) in large quantities, thereby inducing the production of excessive pro-inflammatory NOS, which causes neurotoxic response in the central nervous system, so that iNOS is considered as an important factor affecting "pathological effect of inflammatory response" after pathological injury, and thus a critical inflammatory response is considered to be an important marker for the inhibition of nociceptive response of nociceptive injury, such as a further research on pathological changes of nociceptive response of nociceptive factors, including Cxcl 44, CD 493, CD44, and a further research on pathological changes of inflammatory response of nociception.
The marine environment is special and complex, and the survival and metabolic mode of marine organisms is special, so that many marine natural products have specific structures and important pharmacological effects, such as antibacterial, antiviral, antitumor, anti-inflammatory and the like, and are important sources of innovative medicines. Among them, the active substance derived from coralloid is one of the hot spots in the international marine natural product research, and many compounds with novel structures and remarkable biological activity are found in such marine animals. The object of the present invention, Lemnalol (pseudopterogous alcohol), a kind of sesquiterpene (see FIG. 1) of the type of an ylangene, was first extracted from the plant Lagerstroemia speciosa (Lemnalia tenuis Verseveldt), and then also extracted from 2 species of Taiwan Lagerstroemia speciosa (Lemnalia cervicorni and Lemnalia flava) of the genus. The chemical structure of Lemanlol is as follows:
since the inflammatory response traverses the entire pathological process of ischemic brain injury, inhibition of post-stroke inflammatory response provides a longer therapeutic window for the treatment of ischemic stroke, and particularly in clinical practice, inhibition of post-stroke inflammatory response is considered as a potential promising therapeutic approach since the majority of stroke patients are admitted several hours after onset, at which time inflammatory response has become the main pathological mechanism. The study of the invention finds that Lemnalol can effectively reduce infarct volume and neurological function defect on a rat cerebral central artery ischemia (MCAO) reperfusion model, obviously inhibit inflammatory reaction, improve survival rate and have a wider treatment time window (the drug administration is still effective within 24 hours for the first time). Recently, it has been discovered that up-regulation of inflammatory factor gene is one of the characteristics of gene expression change of stroke patients, and effective inhibition of inflammatory reaction is one of the important ways for neuroprotection. Effectively inhibit neuroinflammatory reaction and have obvious protective effect in the treatment of cerebral apoplexy. In addition, in vitro experiments of the invention also show that Lemnalol can effectively inhibit the expression of M1 type marker genes such as BV-2 microglia iNOS and COX2 stimulated by LPS, and can promote the expression of M2 type marker genes such as IL-10, Arg1 and the like, and the conversion of the functional state (M1/M2 type) of microglia/macrophage is a key mechanism influencing the subsequent damage of cerebral apoplexy and is considered as an important way for treating cerebral apoplexy. The invention discovers that Lemnalol has obvious effects of protecting nerves and inhibiting inflammatory reaction on tMCAO rats, and the like, and accords with new guidance of new drug screening in the world at present. Therefore, the research aims to provide the possibility of Lemnalol as a novel neuroprotective agent and a potential neuroprotective mechanism, and provide a new experimental basis for clinical treatment application.
Disclosure of Invention
The invention aims to provide application of pseudopterogorgia pseudosciaenae alcohol in preparation of a medicine for reducing brain injury in stroke, aims to provide the possibility of pseudopterogorgia pseudosciaenae alcohol (Lemnalol) serving as a novel neuroprotective agent and a potential neuroprotective mechanism, and provides a new experimental basis for clinical treatment and application.
In order to achieve the purpose, the invention adopts the following technical scheme:
the application of the pseudopterogorgia alcohol (Lemnalol) in preparing the medicine for reducing the brain injury in the stroke is as follows:
the invention has the advantages that:
the invention realizes the improvement of the neuroprotective performance, the pseudopterogorgia pseudosciaenae alcohol (Lemnalol) has obvious neuroprotective effect on cerebral apoplexy, the cerebral infarction volume is up to 30 percent relative to a modeling group, and the administration group is reduced to about 10 percent. The pharmacodynamic characteristics of the traditional Chinese medicine are more in accordance with clinical requirements, and patients can go to hospitals after having a disease for hours, and the main pathological feature at this stage is inflammatory injury. The combined medicine can effectively inhibit inflammatory reaction after cerebral arterial thrombosis, thereby having longer treatment time window. Is expected to be used as a novel neuroprotective agent.
Drawings
FIG. 1 is a graph of the cerebral infarct volume and volumetric ratio statistics for Lemnalol (LN) 2 hours after MCAO reperfusion, after 6 days of continuous intravenous injection (15 mg/kg/day); wherein (A) is the cerebral infarction volume; (B) the volume ratio of cerebral infarction is counted.
FIG. 2 is an analysis of the expression level of inflammatory factors.
FIG. 3 shows the effect of Lemnalol (LN) on the mRNA and protein expression levels of IL-6 and IL-1 β, where a is the IL-6 mRNA expression level, b is the IL-1 β mRNA expression level, c is the IL-6 protein expression level, and d is the IL-1 β protein expression level.
FIG. 4 is a graph of the effect of Lemnalol (LN) on the expression levels of iNOS and COX-2 mRNA; wherein a is the iNOS mRNA expression level; b is COX-2 mRNA expression level.
FIG. 5 shows the effect of Lemnalol (LN) on the expression of M2-related genes, such as IL-10 and Arg 1.
Detailed Description
Example 1
1 Material
1.1 healthy adults of laboratory animalsSPF grade SD rats 60, body mass (280 ± 20) g, purchased from shanghai slyke laboratory animals ltd, certification No.: 2007000509960, license number: SCXK 2007 and 0005, which is bred in SPF animal laboratory of the university of Fujian Chinese medicine laboratory animal center. Keeping the temperature and humidity constant for 24h, ensuring sufficient food and drinking water, simulating natural day and night change, giving illumination at 6:00-18:00, keeping the day at 18: 00-next day and keeping the day at 6:00 in a dark state, and carrying out adaptive feeding for one week.
1.2 Total RNA extraction Kit RNeasy Mini Kit (QIAGEN, Cat. No.: 74106) of main reagent and tested drug, RT Kit (Fermentas, Cat. No.: K1622) of reverse transcription Kit, fluorescent quantitative PCR Master Mix (Applied Biosystems, Cat. No.: R11022), PCR primers were synthesized by Shanghai Jieli bioengineering, Inc.
TNF- α gene primers:
the upstream sequence 5 '-GCCACCACGCTCTTC-TGTC-3';
the downstream sequence 5'-GCTACGGGCTTGTCACTCG-3' is a sequence of sequences,
the length of the PCR amplified fragment was 149 bp.
IL-6 gene primers:
upstream sequence 5'-AGACTTCACAGAGGATACCACCCAC-3';
the downstream sequence 5'-CAATCAGAATTGCCATTGCACAA-3' is a sequence of sequences,
the length of the PCR amplified fragment is 129 bp.
IL-1 β gene primer:
an upstream sequence 5'-ACAAGGAGAGACAAGCAACGACAA-3';
the downstream sequence of the flow line 5'-TTTCCATCTTCTTCTTTGGGTATTG-3',
the length of the PCR amplified fragment was 149 bp.
iNOS gene primers:
an upstream sequence 5'-CAGATCCCGAAACGCTACACTT-3';
the downstream sequence 5'-TGCGGCTGGACTTCTCACTC-3' is a sequence of sequences,
the PCR amplified fragment was 175bp in length.
Cxcl1 gene primer:
an upstream sequence 5'-CCACTCACCTGCTGCTACTCATT-3';
the downstream sequence 5'-GTATGTCTGGACCCATTCCTTCTTG-3' is a sequence of sequences,
the length of the PCR amplified fragment is 113 bp.
Cxcl2 gene primer:
an upstream sequence 5'-GCTCCCAGCCAGGTGTCATTTT-3';
the downstream sequence 5'-AAGACTCTCAGGCATTCAGTTCCAG-3' is a sequence of sequences,
the length of the PCR amplified fragment was 105 bp.
Arg1 gene primer:
an upstream sequence 5'-CGTTGACCTTGTCTTGTTTTGG-3';
the downstream sequence 5'-CTGGTTCTGTTCGGTTTGCTG-3' is a sequence of sequences,
the PCR amplified fragment was 138bp in length.
GAPDH primer upstream sequence 5'-CAACGGGAAACCCATCACCA-3';
the downstream sequence 5 '-ACGCCAGTAGACTC-CACGACAT-3',
the length of the PCR amplified fragment is 96 bp.
iNOS, Cox2 and Agr1 antibody (Santa Cruz); β -actin antibody was purchased from Biyun Biotech research institute.
2 method of experiment
2.1 MCAO animal model rat preoperative fasting 12 h, free drinking water, intraperitoneal injection of 10% chloral hydrate (300 mg. kg-1) anesthesia, supine position fixation, nylon monofilament line plug from the left common carotid artery to the middle cerebral artery initiation end. 2 h after embolism, the wire plug is pulled out and then is perfused for 24 h. Sham operated animals did not embolize middle cerebral arteries. During the procedure, the temperature of the animals was continuously monitored and maintained at 37 ℃. And (4) evaluating the nerve function damage degree according to a Longa evaluation method, wherein 3-4 grades are successful model making and can be used for experiments.
2.2 Experimental groups and dosing methods the experimental rats were divided into sham (sham), Model Control (MCAO), and Lemnalol dosing (MCAO + LN) (15 mg/kg/d) drugs, 6 per group. The Lemnalol administration group is subjected to reperfusion after 2 h of embolism, immediately subjected to intraperitoneal injection after nerve function injury evaluation, and is subjected to intraperitoneal injection of Lemnalol (15 mg/kg/d) or normal saline for 24h, wherein 3 animals in each group are randomly selected, and brain tissues are taken for inflammation index analysis. In addition, the drug is given every 24h, and after 6d, the head of each group of experimental animals is cut off and the brain is taken for TTC analysis.
2.3 statistics of rat neural function impairment score and cerebral infarction volume determination
Evaluation of nerve function impairment: according to Longa evaluation method: grade 0, no defect; grade 1, inability to extend contralateral forelimb; grade 2, contralateral forelimb flexion; level 3, slightly turning to the opposite side; grade 4, severe revolutions; grade 5, contralateral paralysis.
After 6 days of administration, rat brain tissue was collected, and coronal sections were prepared at the visual cross and 2mm before and after the visual cross, and incubated in TTC for 30min at 37 ℃ in the dark. Normal tissue was stained rose-red, while infarcted tissue was white. After staining, the tissue was photographed and infarct volume was calculated using image analysis.
2.4 fluorescent real-time quantitative PCR (RT-qPCR) for detecting gene expression of Cxcl1, Cxcl2, TNF- α, IL-6, IL-1 β, iNOS, etc
Extracting total RNA according to a QIAGEN RNeasy Mini Kit, carrying out reverse transcription by using a RT reverse transcription Kit of Fermentas to obtain cDNA, and establishing a qPCR system by using the cDNA as a template, wherein the system comprises 10 mu l of SYBR Select Master mix, 2 mu MF-primer 2 mu l, 2 mu M R-primer 2 mu l, 2 mu l of cDNA and 20 mu l of ddH2O 4 mu l; the reaction conditions are 50 ℃ for 2min, 95 ℃ for 10 min, 95 ℃ for 15 s, annealing at 58 ℃ for 15 s and polymerization at 72 ℃ for 40 cycles. The expression level of mRNA among the groups was calculated by the 2-. DELTA.Ct method using GAPDH as an internal reference.
2.5 cell culture
Mouse microglia BV2 was cultured in RPMI1640 medium containing 10% final fetal bovine serum and 1% Penicillin-Streptomycin diabody, at 37 ℃ in an incubator with 5% CO 2. When the cells exceeded 80% confluence, cell subcultures were performed.
2.6 ELISA kits (R & D Systems)
After the 96-well plate is coated overnight, the target inflammatory factor is detected by an antibody sandwich method. And (3) measuring the absorbance (A value) at the wavelength of 450 nm by a chromogenic reaction and a microplate reader, calculating the concentration of the sample, and quantitatively detecting the inflammatory factor.
2.7 statistical analysis was performed using SPSS 18.0 statistical software, data are expressed in x. + -.s, and differences between groups of data were analyzed using ANOVA method. 3 results
3.1 after reperfusion of MCAO rat for 2 hours, Lemnalol (15 mg/kg/d) or normal saline is injected into abdominal cavity for 6 days, and then the material is obtained for analysis, so that Lemnalol has neuroprotective effect with wider treatment time window.
The detection of TTC and the like on the affected side brain tissue shows that Lemnalol reduces the infarct volume of MCAO rats and has a wider treatment time window: lemnalol (LN) 6 days of continuous intravenous injection (15 mg/kg/day) was effective in reducing cerebral infarct volume 2 hours after MCAO reperfusion (FIG. 1A). TTC staining showed that the cerebral infarction volume ratio statistics showed that the administration after 2 hours after reperfusion was up to 30% of cerebral infarction volume relative to the model group, and the administration group was also reduced to around 10% (fig. 1B).
3.2 Lemnalol reduces the expression of a series of inflammatory factors in the affected side of the semi-brain tissue of MCAO reperfusion model rats.
After MCAO rats are perfused for 2 hours again, Lemnalol (15 mg/kg/d) or normal saline is injected into the abdominal cavity for 24 hours, then the materials are taken out, brain tissues are taken for subsequent analysis, and the reanalalol is found to be capable of inhibiting the expression levels of a series of inflammatory factors such as Cxcl1, Cxcl2, TNF- α -6, IL-1 β and the like through reanaltime PCR analysis, and the results are shown in figure 2.
3.3 BV-2 microglia inflammation model (100 ng LPS stimulation), and analyzed 24h after administration, Lemnalol effectively inhibits M1 type-related marker gene expression and has dose effect.
(1) Lemnalol effectively inhibited IL-6 and IL-1 β expression (FIG. 3);
(2) lemnalol was effective in inhibiting iNOS and COX-2 expression (FIG. 4);
3.4 BV-2 microglia inflammation model (100 ng LPS stimulation), analyzed 24h after administration, Lemnalol promoted the expression of M2-related genes like IL-10 and Arg1 with dose effect (FIG. 5).
4 conclusion
In conclusion, the study of Lemnalol on the neuroprotective and anti-inflammatory effects of stroke is based on in vivo and in vitro experiments. The study is established that Lemnalol has a remarkable neuroprotective effect on MCAO rats, and when an administration group of the MCAO rats after 2 hours of reperfusion injury is analyzed, Lemnalol shows a remarkable inflammation inhibition effect in the protection effect on cerebral ischemia injury, and directly participates in the regulation and control of inflammatory response in the treatment of cerebral apoplexy, and clinical investigation and research find that the up-regulation of inflammatory factor genes is one of the characteristics of gene expression change of patients with cerebral apoplexy, and the effective inhibition of inflammatory response is one of important ways for neuroprotection. Therefore, the anti-inflammatory action of Lemnalol after stroke is one of the neuroprotective action mechanisms. Based on the fact that the functional states of microglia/macrophages (M1 and M2) directly affect secondary injury after cerebral apoplexy, and the functional states are reported to have an important role in regulating neuroprotection in the literature, the change of the activity states of the macrophages in M1 and M2 can be an important mechanism of neuroprotection and anti-inflammatory action of Lemnalol.
The above description is only a preferred embodiment of the present invention, and all equivalent changes and modifications made in accordance with the claims of the present invention should be covered by the present invention.
SEQUENCE LISTING
<110> Fujian university of traditional Chinese medicine
<120> application of pseudopterogorgia lancifolium active substance sciadol in preparing medicine for treating cerebral apoplexy
<130>16
<160>16
<170>PatentIn version 3.3
<210>1
<211>19
<212>DNA
<213> Artificial sequence
<400>1
gccaccacgc tcttctgtc 19
<210>2
<211>19
<212>DNA
<213> Artificial sequence
<400>2
gctacgggct tgtcactcg 19
<210>3
<211>25
<212>DNA
<213> Artificial sequence
<400>3
agacttcaca gaggatacca cccac 25
<210>4
<211>23
<212>DNA
<213> Artificial sequence
<400>4
caatcagaat tgccattgca caa 23
<210>5
<211>24
<212>DNA
<213> Artificial sequence
<400>5
acaaggagag acaagcaacg acaa 24
<210>6
<211>25
<212>DNA
<213> Artificial sequence
<400>6
tttccatctt cttctttggg tattg 25
<210>7
<211>22
<212>DNA
<213> Artificial sequence
<400>7
cagatcccga aacgctacac tt 22
<210>8
<211>20
<212>DNA
<213> Artificial sequence
<400>8
<210>9
<211>23
<212>DNA
<213> Artificial sequence
<400>9
ccactcacct gctgctactc att 23
<210>10
<211>25
<212>DNA
<213> Artificial sequence
<400>10
gtatgtctgg acccattcct tcttg 25
<210>11
<211>22
<212>DNA
<213> Artificial sequence
<400>11
gctcccagcc aggtgtcatt tt 22
<210>12
<211>25
<212>DNA
<213> Artificial sequence
<400>12
aagactctca ggcattcagt tccag 25
<210>13
<211>22
<212>DNA
<213> Artificial sequence
<400>13
cgttgacctt gtcttgtttt gg 22
<210>14
<211>21
<212>DNA
<213> Artificial sequence
<400>14
ctggttctgt tcggtttgct g 21
<210>15
<211>20
<212>DNA
<213> Artificial sequence
<400>15
<210>16
<211>22
<212>DNA
<213> Artificial sequence
<400>16
acgccagtag actccacgac at 22
Claims (2)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201911203933.XA CN111012765A (en) | 2019-11-29 | 2019-11-29 | Application of pseudopterogorgia-hancei alcohol as active substance in preparing medicine for treating cerebral apoplexy |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201911203933.XA CN111012765A (en) | 2019-11-29 | 2019-11-29 | Application of pseudopterogorgia-hancei alcohol as active substance in preparing medicine for treating cerebral apoplexy |
Publications (1)
Publication Number | Publication Date |
---|---|
CN111012765A true CN111012765A (en) | 2020-04-17 |
Family
ID=70203751
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201911203933.XA Pending CN111012765A (en) | 2019-11-29 | 2019-11-29 | Application of pseudopterogorgia-hancei alcohol as active substance in preparing medicine for treating cerebral apoplexy |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN111012765A (en) |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101164540A (en) * | 2006-08-03 | 2008-04-23 | 中山大学 | Application of marine steroids compound in preparing medicine for treating neuronal damage |
TW200841875A (en) * | 2007-04-24 | 2008-11-01 | Univ Nat Sun Yat Sen | Lemnalol applying for treating inflammation and pain |
-
2019
- 2019-11-29 CN CN201911203933.XA patent/CN111012765A/en active Pending
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101164540A (en) * | 2006-08-03 | 2008-04-23 | 中山大学 | Application of marine steroids compound in preparing medicine for treating neuronal damage |
TW200841875A (en) * | 2007-04-24 | 2008-11-01 | Univ Nat Sun Yat Sen | Lemnalol applying for treating inflammation and pain |
Non-Patent Citations (2)
Title |
---|
EUGENIA GENTILE等: "Marine pharmacology: therapeutic targeting of matrix metalloproteinases in neuroinflammation", 《DRUG DISCOVERY TODAY》 * |
丁伟等: "《血性脑血管并研究新进展》", 31 July 2009, 中国海洋大学出版社 * |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Chaparro-Huerta et al. | Proinflammatory cytokines and apoptosis following glutamate-induced excitotoxicity mediated by p38 MAPK in the hippocampus of neonatal rats | |
Roddy et al. | Stanniocalcin-1 rescued photoreceptor degeneration in two rat models of inherited retinal degeneration | |
Mladenović et al. | 6-Hydroxydopamine increases the level of TNFα and bax mRNA in the striatum and induces apoptosis of dopaminergic neurons in hemiparkinsonian rats | |
Wan et al. | Molecular mechanism underlying the ability of caffeic acid to decrease uric acid levels in hyperuricemia rats | |
CA2629977C (en) | Extract of dioscorea sp. and the medical uses thereof | |
KR20180042217A (en) | Treatment method | |
EP3581198B1 (en) | Manganese-type high-stability superoxide dismutase for use in treating fatty liver | |
Li et al. | Recovery of post-stroke cognitive and motor deficiencies by Shuxuening injection via regulating hippocampal BDNF-mediated Neurotrophin/Trk Signaling | |
Maña et al. | Deleterious role of IFNγ in a toxic model of central nervous system demyelination | |
Bethea et al. | Ovarian steroids regulate gene expression in the dorsal raphe of old female macaques | |
Wu et al. | Glycyrrhizic acid protects juvenile epileptic rats against hippocampal damage through activation of Sirtuin3 | |
Ren et al. | Ginsenoside Rd attenuates mouse experimental autoimmune neuritis by modulating monocyte subsets conversion | |
EP2788016A1 (en) | Compositions for preventing or treating adverse reactions of egfr inhibition | |
Liu et al. | Novel synergistic mechanism of 11-keto-β-boswellic acid and Z-Guggulsterone on ischemic stroke revealed by single-cell transcriptomics | |
Ravelli et al. | NADPH oxidase contributes to streptozotocin-induced neurodegeneration | |
CN109276711B (en) | Application of manganese type high-stability superoxide dismutase in improving autism and product | |
Yang et al. | Comprehensive analysis of lncRNA expression profiles in rats with cerebral ischemia-reperfusion injury after treatment with 20 (R)-ginsenoside Rg3 | |
Rasouli et al. | Evaluation of associated genes with traumatic pain: a systematic review | |
Zhao et al. | Argatroban promotes recovery of spinal cord injury by inhibiting the PAR1/JAK2/STAT3 signaling pathway | |
Teles et al. | Differential TNFα mRNA regulation detected in the epidermis of leprosy patients | |
CN109589329B (en) | Pharmaceutical composition for reducing brain injury caused by stroke by applying rosmarinic acid | |
Shao et al. | Higenamine improves DSS-induced ulcerative colitis in mice through the Galectin-3/TLR4/NF-κB pathway | |
CN111012765A (en) | Application of pseudopterogorgia-hancei alcohol as active substance in preparing medicine for treating cerebral apoplexy | |
CN115317538B (en) | Externally-applied traditional Chinese medicine composition for treating psoriasis and preparation method and application thereof | |
Liu et al. | Altered expression of neuronal CCR6 during pilocarpine induced status epilepticus in mice |
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 | ||
RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200417 |
|
RJ01 | Rejection of invention patent application after publication |