WO2022047779A1 - 烟酰胺单核苷酸在制备提高损伤皮肤肌膜中Na+-K+-ATP酶水平试剂中的应用 - Google Patents

烟酰胺单核苷酸在制备提高损伤皮肤肌膜中Na+-K+-ATP酶水平试剂中的应用 Download PDF

Info

Publication number
WO2022047779A1
WO2022047779A1 PCT/CN2020/113751 CN2020113751W WO2022047779A1 WO 2022047779 A1 WO2022047779 A1 WO 2022047779A1 CN 2020113751 W CN2020113751 W CN 2020113751W WO 2022047779 A1 WO2022047779 A1 WO 2022047779A1
Authority
WO
WIPO (PCT)
Prior art keywords
skin
sarcolemma
reagent
level
atpase
Prior art date
Application number
PCT/CN2020/113751
Other languages
English (en)
French (fr)
Inventor
赵欣
周先容
余建军
徐钦源
黄浩
Original Assignee
音芙医药科技(上海)有限公司
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by 音芙医药科技(上海)有限公司 filed Critical 音芙医药科技(上海)有限公司
Priority to PCT/CN2020/113751 priority Critical patent/WO2022047779A1/zh
Publication of WO2022047779A1 publication Critical patent/WO2022047779A1/zh

Links

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
    • A61K31/70—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
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/18—Antioxidants, e.g. antiradicals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings

Definitions

  • the invention belongs to the field of biomedicine, in particular to the application of nicotinamide mononucleotide in the preparation of medicines for repairing damaged skin sarcolemma Na + -K + -ATPase, health care products and cosmetics.
  • the skin is the largest and most complex organ of the human body. It is the first barrier to isolate the human body from the external environment. It plays many roles in protecting the body, regulating body temperature, feeling, secretion, excretion and immunity.
  • Skin damage includes physical damage and chemical damage. Physical damage mainly includes ultraviolet (UV), infrared, dust and haze, among which UV radiation is the most significant.
  • UV radiation belongs to the category of diseases, and its clinical symptoms mainly include redness, swelling and dryness of the cortex, epidermal damage, muscle relaxation, rough folds, abnormal local pigment deposition and leather-like appearance, etc., and may even induce skin cancer.
  • ultraviolet radiation is usually divided into long-wave ultraviolet UVA (320nm-400nm), medium-wave ultraviolet UVB (280nm-320nm), and short-wave ultraviolet UVC (200nm-280nm) according to the length of the wavelength.
  • Medium-wave ultraviolet UVB (280nm-320nm) is the most harmful wavelength band to organisms, so animal skin photodamage models are often modeled by UVB.
  • the skin tissue changes caused by this modeling method are extremely similar to human photodamaged skin. resemblance.
  • Na + -K + -ATPase also known as sodium-potassium pump
  • It is a membrane transport protein with dual activities of carrier and enzyme. It is embedded in the cell membrane and can transfer ATP is hydrolyzed into ADP and phosphoric acid, thereby releasing energy.
  • the level of ATPase activity can directly reflect the changes in mitochondrial function.
  • mitochondrial sodium pump function is dysfunctional, resulting in mitochondrial inner membrane Edema, the mitochondrial membrane fluidity also decreases. Free radicals are one of the important factors for the damage of Na + -K + -ATPase.
  • DAMPs damage-associated molecular patterns
  • ATP is not only the main energy substance to maintain the metabolic function of cells in physiological state, but also can be used as DAMPs molecule to regulate various cellular effects in stress state.
  • ATP metabolism and The level of purine signal is low, and exogenous ATP supplementation can accelerate the healing of diabetic wounds; it is suggested that the metabolism of extracellular ATP is related to the process of wound inflammation repair, and the amplification of extracellular ATP signal in the early stage of acute injury can promote the healing of diabetic wounds
  • the normal level of Na + -K + -ATPase is one of the important factors to maintain the normal shape of the skin or promote the skin's self - repair .
  • the study of ATPase is relatively rare.
  • Nicotinamide ribonucleotide is a biologically active nucleotide produced by the reaction of a phosphate group with a riboside containing ribose and nicotinamide. It usually has two forms, ⁇ and ⁇ , of which ⁇ -nicotinamide mononucleotide is the active form between the two, and its molecular weight is 334.221 g/mol.
  • NMN opens a new frontier for modern therapy, and the biomolecule has shown substantial beneficial pharmacological activity in several preclinical disease models, including myocardial and cerebral ischemia, neurodegenerative diseases such as Alzheimer's disease, and diabetes.
  • NAD + is a coenzyme for dehydrogenases, such as alcohol dehydrogenase (ADH), used to oxidize ethanol. It plays an irreplaceable role in glycolysis, gluconeogenesis, tricarboxylic acid cycle and respiratory chain.
  • ADH alcohol dehydrogenase
  • the intermediate product will pass off the hydrogen to NAD + , making it NADH, and NADH will be used as the carrier of hydrogen to synthesize ATP by chemical osmotic coupling in the respiratory chain.
  • ADH alcohol dehydrogenase
  • NMN nicotinamide mononucleotide
  • the purpose of the present invention is to provide a new medical application of nicotinamide mononucleotide (NMN).
  • NPN nicotinamide mononucleotide
  • the first aspect of the present invention provides the application of nicotinamide mononucleotide in preparing a reagent for increasing the level of Na + -K + -ATPase in damaged skin sarcolemma.
  • the damaged skin is skin damage caused by ultraviolet radiation
  • the skin damage caused by ultraviolet radiation is skin damage caused by medium-wave ultraviolet radiation.
  • the second aspect of the present invention provides a reagent for increasing the level of Na + -K + -ATPase in damaged skin sarcolemma, using nicotinamide mononucleotide as the only active ingredient.
  • the third aspect of the present invention provides the application of the above-mentioned reagent for increasing the level of Na + -K + -ATPase in damaged skin sarcolemma in the preparation of medicines, health products or skin care products for relieving skin oxidative stress damage.
  • the above-mentioned reagent for improving the level of Na + -K + -ATPase in the sarcolemma of damaged skin can be glue, ointment, emulsion, mixture, oral ampoule, tablet or capsule; above-mentioned medicines are glue, ointment, Emulsions, aerosols, injections, mixtures, oral ampoules, tablets or capsules; the health food is in oral forms such as capsules or tablets, powders, etc.
  • the cosmetic is in the form of external use such as lotion, spray, facial mask, eye cream and the like.
  • nicotinamide mononucleotide can maintain the normal sodium-potassium pump by increasing the level of Na + -K + -ATPase in the skin, and improve the related serum, skin and liver.
  • Oxidative stress indicators thereby alleviating the oxidative stress injury induced by UVB irradiation; further by inhibiting the pro-inflammatory cytokines TNF- ⁇ , IL-1 ⁇ and IL-6 and increasing the anti-inflammatory cytokines I ⁇ B- ⁇ , IL-4 and The activation of IL-10 can ultimately reduce the inflammatory damage of the body, and provide a basis for the development of NMN-related drugs and health products that improve skin aging and damage and improve the body's antioxidant capacity.
  • nicotinamide mononucleotide has been sold as a health care product, its pharmacological action is clear, the toxic and side effects are small, and the drug safety has been clinically recognized, and the new indication of nicotinamide mononucleotide provided by the present invention can be realized quickly Clinical translation or marketing. Therefore, nicotinamide mononucleotide has great potential in the protection of clinical acute skin damage or daily skin photodamage. The invention also provides a new clinical medicine for relieving and treating skin photodamage.
  • Figure 1 shows the mRNA expression levels of SOD1, SOD2, CAT and GSH in skin and liver tissues.
  • the data are presented as mean ⁇ standard deviation ( ⁇ SD), where * means p ⁇ 0.05, ** means p ⁇ 0.01, *** means p ⁇ 0.001;
  • Figure 2 shows the mRNA expression levels of NF- ⁇ B p65, I ⁇ B- ⁇ , TNF- ⁇ , IL-6 and IL-10 in skin and liver tissues, the data are presented as mean ⁇ standard deviation ( ⁇ SD), where * means p ⁇ 0.05, ** means p ⁇ 0.01, *** means p ⁇ 0.001;
  • Figure 3 shows the observation of the pathological morphology of the liver in different experimental groups
  • Figure 4 is the observation of skin pathological morphology in different experimental groups, (A) skin H&E staining; (B) skin Masson staining; (C) skin TB staining.
  • Uthever TM NMN was provided by Yinfu Pharmaceutical Technology (Shanghai) Co., Ltd. (Shanghai, China), and the purity of NMN was determined by high performance liquid chromatography, and the purity was >98.5%.
  • mice Forty 7-week-old female ICR mice were purchased from the Experimental Animal Center of Chongqing Medical University [Chongqing, China, SCXK(YU) 2018-0003]. Mice were kept under constant temperature and humidity conditions (temperature 25 ⁇ 2°C, relative humidity 50 ⁇ 5%), 12h light/dark cycle. Mice were allowed free access to standard rat chow and drinking water.
  • mice Forty mice were randomly divided into 4 groups with 10 mice in each group, namely normal group, UVB group, VC+UVB group, and nicotinamide mononucleotide (NMN)+UVB group. The whole experiment lasted for 4 weeks. From the 3rd week onwards, the skin damage model was established on the mice in the other groups except the normal group by using ultraviolet radiation equipment, irradiating for 2h every day.
  • Nicotinamide mononucleotide can improve oxidative stress injury including Na + - K + -ATPase in UVB mice
  • GSH glutathione
  • T-SOD, CAT, AGES and H 2 O 2 in mouse serum and skin tissue were determined according to the kit instructions, and the levels of Na + K + -ATP and NAD + in skin tissue were also determined.
  • the levels of oxidative stress indicators in mouse skin tissue are shown in Table 1, and the levels of oxidative stress indicators in mouse serum are shown in Table 2:
  • oral NMN can increase the level of NAD + in skin tissue, and the increased NAD + will further increase the level of Na + -K + -ATPase, which ultimately prevents skin damage caused by UVB irradiation. role of cancer.
  • the regulation of the activity of Na + K + -ATPase can endow the cells with the necessary vitality, and the decrease of its activity will lead to the disturbance of intracellular energy production and ion transport, thus affecting the cell function and accelerating the cell damage.
  • 1 ⁇ L of cDNA, 10 ⁇ L of TaqMan TM Multiplex Master Mix, 2 ⁇ L of 10 ⁇ M primer and 7 ⁇ L of ddH 2 O were mixed evenly, and amplification detection was performed on a real-time fluorescence quantitative PCR instrument.
  • the amplification conditions were: deformation at 95°C for 15s, annealing at 55°C for 30s, and extension at 72°C for 35s, for a total of 40 cycles.
  • the relative expression of each target gene was calculated by 2 - ⁇ CT , in which ⁇ -actin was used as the internal reference gene.
  • the primer sequences used are shown in Table 3.
  • Nicotinamide mononucleotide can reduce inflammatory damage in the body
  • NF- ⁇ B is an important transcription factor in the immune response process of the body. Under normal circumstances, it binds to its inhibitory protein I ⁇ B in an inactive manner. Studies have found that when the body is abnormally stimulated, it will accelerate the phosphorylation of I ⁇ B protein, making NF- ⁇ B protein.
  • the experimental method is the same as that in 3.2, and the primer sequences used are shown in Table 3. The results are shown in Figure 2.
  • UVB irradiation increased the mRNA expression level of NF- ⁇ Bp65 in the skin and liver of mice, decreased the expression of the inhibitory gene I ⁇ B- ⁇ of NF- ⁇ Bp65, and also down-regulated the expression of NF- ⁇ Bp65 inhibitory gene I ⁇ B- ⁇ .
  • the expression of IL-10 related to the NF- ⁇ Bp65 signaling pathway up-regulated the expression of TNF- ⁇ and IL-6, among which the UVB group had the most significant changes.
  • the mRNA expressions of I ⁇ B- ⁇ and IL-10 in the skin and liver of the VC+UVB group and the NMN+UVB group were increased to different degrees, while the mRNA expressions of NF- ⁇ Bp65, TNF- ⁇ and IL-6 were increased.
  • the expression was decreased, and the mRNA expression levels of the above indicators in the NMN+UVB group were close to those in the normal group. It indicated that NMN could inhibit the activation of NF ⁇ B-p65 signaling pathway by up-regulating the expression of I ⁇ B- ⁇ in skin and liver tissue.
  • NMN can down-regulate the expression of pro-inflammatory cytokines TNF- ⁇ and IL-6, and up-regulate the expression of anti-inflammatory cytokine IL-10 from the serum level, skin tissue level and mRNA level, respectively, thus reducing the effect of these inflammatory mediators on the skin. Degree of inflammatory damage in injured mice.
  • Nicotinamide mononucleotide can improve the pathological morphology of mice after UV irradiation
  • mice were dissected and the liver and back hairless skin of the mice were removed, and the liver and skin tissues of the size of soybean were cut and fixed in 4% paraformaldehyde solution. The remaining liver and skin tissues were stored in a -80°C refrigerator for later use.
  • the thickness of the dermis in the UVB group was significantly thinner, the number of collagen fiber bundles was significantly reduced, the subcutaneous tissue was disordered, and the boundary was not obvious; in addition, inflammatory cell infiltration around the adnexa was seen, indicating the occurrence of chronic inflammation in the skin, indicating that long-term UVB irradiation can induce skin photodamage.
  • the thickness of the skin dermis of the mice in the VC+UVB group increased, but the collagen fibers were loosely dispersed.
  • the thickness of the whole skin layer of the mice in the NMN+UVB group was significantly thicker than that in the UVB group, and the distribution of collagen fiber bundles was relatively uniform and orderly, and the overall structure was similar to that of the normal group.
  • Collagen fibers after Masson staining appeared blue-purple.
  • the number and arrangement of collagen fibers in the normal group were significantly higher than those in the UVB group, indicating that UVB irradiation would cause the degradation of collagen fibers.
  • the number of collagen fibers in the dermis of the mice in the VC+UVB group and the NMN+UVB group increased, and the collagen fibers in the NMN+UVB group were arranged more neatly and orderly, with almost no shrinkage. , breakage, etc.
  • the skin dermis of the UVB group mice became thinner, the number of collagen fibers decreased, the number of mast cells increased, and at the same time, the liver tissue also showed a certain degree of pathological changes, indicating that the UVB-induced skin damage model was successful.
  • the pathological morphology of the skin was greatly improved, and the liver of the mice also remained in a normal shape without obvious damage.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Medicinal Chemistry (AREA)
  • Epidemiology (AREA)
  • Organic Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Dermatology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Engineering & Computer Science (AREA)
  • Toxicology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Cosmetics (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

提供了烟酰胺单核苷酸在制备提高损伤皮肤肌膜中Na +-K +-ATP酶水平试剂中的应用。通过UVB诱导的小鼠皮肤损伤模型实验，烟酰胺单核苷酸能够通过增加皮肤中Na +-K +-ATP酶活力，进而维持损伤皮肤肌膜正常的钠钾泵；能够通过改善血清、皮肤和肝脏中相关氧化应激指标，进而缓解因UVB照射诱发的氧化应激损伤；同时能够通过抑制促炎细胞因子TNF-α、IL-1β和IL-6和增加抑炎细胞因子IκB-α、IL-4和IL-10的活化，最终降低机体的炎症损伤，从而为开发改善皮肤衰老与损伤、提高机体抗氧化能力的NMN相关药物、保健食品或化妆品等提供依据。

Description

烟酰胺单核苷酸在制备提高损伤皮肤肌膜中Na +-K +-ATP酶水平试剂中的应用 技术领域
本发明属于生物医药领域，具体涉及烟酰胺单核苷酸在制备修复损伤皮肤肌膜Na +-K +-ATP酶药物或保健品、化妆品中的应用。
背景技术
皮肤是人体最大、最复杂的器官，是人体与外界环境隔离的第一道屏障，起着保护身体、调节体温、感觉、分泌、排泄和免疫等诸多作用。皮肤损伤包括物理损伤和化学损伤，物理损伤主要包括紫外线(UV)、红外线、粉尘和雾霾等，其中以UV辐射最为显著。通常UV辐射引起的急性皮肤损伤属于疾病范畴，其临床症状主要表现为皮层红肿干燥、表皮破损、肌肉松弛、褶皱粗糙、局部色素异常沉积和皮革样外观等，甚至可能诱发皮肤癌。研究表明皮肤光损伤会导致皮肤的严重角质化，阻止物质和能量的传递质，主要表现为表皮层趋于扁平，表皮突起和乳头层消失，表皮毛细血管扭曲变形，免疫功能降低，黑色素瘤细胞炎症增生。皮肤组织病理特征多表现为ECM的胶原纤维的断离崩解、胶原蛋白的流失和炎症细胞的浸润。目前，皮肤光损伤日益受到人们的关注，它不仅影响人类的健康，同时也给人身心的带来压力，影响美观甚至就业。
近年来，皮肤光损伤硏究已成为皮肤科领域特别是美容医学的研究热点，鉴于此，建立切实可行的光损伤模型对于深入研究其发生发展机制及筛选抗光老化的药物显得尤为重要。目前国内外研究皮肤光损伤的模型主要包括细胞模型、体外人皮肤模型、人体志愿者模型和动物模型等四种，其中动物模型是迄今为止应用最为广泛的皮肤光损伤模型。动物模型研究皮肤光损伤时常用的造模方法是采用紫外线长期照射动物裸露的皮肤。在研究和应用中，紫外辐射按波长长短通常分为长波紫外线UVA(320nm-400nm)、中波紫外线UVB(280nm-320nm)、短波紫外线UVC(200nm-280nm)。中波紫外线UVB(280nm-320nm)是对生物体最具危害性的波段，所以动物皮肤光损伤模型也常采用UVB进行造模，该造模方法引起的皮肤组织改变与人光损伤的皮肤极其相似。
Na +-K +-ATP酶又称为钠钾泵，是1957年Jens C.Skou"21发现的，是一种膜转运蛋白，具有载体和酶的双重活性，镶嵌于细胞膜上，它能把ATP水解成ADP和磷酸，从而释放出能量。同时，ATP酶活性水平可以直接反映线粒体功能的改变。当Na +-K +-ATP酶活性 下降时，线粒体钠泵功能出现障碍，导致线粒体内膜水肿，线粒体膜流动性也随之下降。自由基是Na +-K +-ATP酶受损的重要因素之一，膜流动性的降低导致线粒体氧化磷酸化功能出现障碍，而ATP合成的不足，又可直接降低Na +-K +-ATP酶的活性。研究发现，当外源性皮肤组织受损后，损伤刺激因子将启动炎症-修复-重塑进程，最终使创面愈合。组织细胞受损后会释放多种具有免疫调节活性的内源性物质，即损伤相关模式分子(damage associated molecular patterns，DAMPs)，参与协同激活先天免疫系统及调节获得性免疫方向，对组织修复或异常重构进行调控。其中，ATP不仅是生理状态下维持细胞代谢功能的主要能量物质，应激状态下也可作为DAMPs分子实现对多种细胞效应的调控。在糖尿病创面愈合过程的炎症反应阶段，ATP代谢及嘌呤信号水平均呈低水平状态，外源性补充ATP可加速糖尿病创面的愈合；继而提示，胞外ATP代谢与创面炎症修复进程相关，且在急性损伤早期放大胞外ATP信号可促进糖尿病创面愈合。由此说明，在皮肤受到外界刺激时，正常的Na +-K +-ATP酶水平是维持皮肤正常形态或者促进皮肤自我修复的重要因素之一，而目前关于皮肤中Na +-K +-ATP酶的研究较为少见。
烟酰胺单核苷酸(nicotinamide ribonucleotide，NMN)是一种生物活性核苷酸，由磷酸基与含有核糖和烟酰胺的核苷反应生成。它通常有α和β两种形式，其中β-烟酰胺单核苷酸是两者之间的活性形式，其分子量为334.221g/mol。NMN为现代治疗开辟了一个新的领域，该生物分子在几种临床前疾病模型中显示出大量有益的药理活性，包括心肌和脑缺血、阿尔茨海默病等神经退行性疾病和糖尿病。NMN大部分药理作用是通过促进NAD +合成进行的，研究表明服用NMN可以使人体内NAD+得含量增加60％。NAD +是脱氢酶的辅酶，如乙醇脱氢酶(ADH)，用于氧化乙醇。它在糖酵解，糖异生，三羧酸循环和呼吸链中发挥着不可替代的作用。中间产物会将脱下的氢递给NAD +，使之成为NADH，而NADH则会作为氢的载体，在呼吸链中通过化学渗透偶联的方式，合成ATP。在小鼠模型中最新发现的抗衰老、延长寿命的特性，使NMN更有吸引力。研究表明补充烟酰胺单核苷酸(NMN)可以改善小鼠随着年龄增长的代谢和应激反应，所以NMN被认为是一种很有前途的治疗与年龄相关的生理功能障碍和疾病的方法。然而目前关于NMN对UVB诱导的皮肤光损伤的研究少有报道。
发明内容
本发明的目的在于提供一种烟酰胺单核苷酸(NMN)新的医药用途。
本发明第一方面，提供了烟酰胺单核苷酸在制备提高损伤皮肤肌膜中Na +-K +-ATP酶水平试剂中的应用。
优选的，损伤皮肤为紫外线辐射引起的皮肤损伤，进一步，紫外线辐射引起的皮肤损伤为中波紫外线辐射诱导的皮肤损伤。
本发明的第二方面，提供了一种提高损伤皮肤肌膜中Na +-K +-ATP酶水平的试剂，以烟酰胺单核苷酸作为唯一活性成分。
本发明的第三方面，提供了上述提高损伤皮肤肌膜中Na +-K +-ATP酶水平的试剂在制备缓解皮肤氧化应激损伤药物、保健品或护肤品中的应用。
本发明的第四方面，提供了上述提高损伤皮肤肌膜中Na +-K +-ATP酶水平的试剂在制备缓解皮肤光损伤药物、保健品或护肤品中的应用。
形式上，上述提高损伤皮肤肌膜中Na +-K +-ATP酶水平的试剂可以为胶剂、膏剂、乳剂、合剂、口服安瓿剂、片剂或胶囊剂；上述药物为胶剂、膏剂、乳剂、气雾剂、注射剂、合剂、口服安瓿剂、片剂或胶囊剂；所述保健食品为胶囊剂或片剂、粉剂等口服型式。所述化妆品为乳液、喷剂、面膜、眼霜等外用型式。
发明的作用与效果
通过UVB诱导的小鼠皮肤损伤模型实验，烟酰胺单核苷酸能够通过增加皮肤中Na +-K +-ATP酶的水平，进而维持正常的钠钾泵，并改善血清、皮肤和肝脏中相关氧化应激指标，进而缓解因UVB照射诱发的氧化应激损伤；进一步能够通过抑制促炎细胞因子TNF-α、IL-1β和IL-6和增加抑炎细胞因子IκB-α、IL-4和IL-10的活化，最终降低机体的炎症损伤，为开发改善皮肤衰老与损伤、提高机体抗氧化能力的NMN相关药物和保健品等提供依据。
此外，烟酰胺单核苷酸作为保健品已在出售，其药理作用明确、毒副作用小，药物安全性已得到临床认可，本发明提供的烟酰胺单核苷酸的新适应症可较快实现临床转化或市场推广。因此，烟酰胺单核苷酸在临床急性皮肤损伤或日常皮肤光损伤的防护应用上具有巨大潜力。本发明也为缓解和治疗皮肤光损伤提供了新的临床药物。
附图说明
图1为皮肤和肝脏组织中SOD1、SOD2、CAT和GSH的mRNA表达水平，数据呈现为平均值±标准偏差(±SD)，其中*表示p<0.05，**表示p<0.01，***表示p<0.001；
图2为皮肤和肝脏组织中NF-κBp65、IκB-α、TNF-α、IL-6和IL-10的mRNA表达 水平，数据呈现为平均值±标准偏差(±SD)，其中*表示p<0.05，**表示p<0.01，***表示p<0.001；
图3为不同实验组肝脏病理学形态观察；
图4为不同实验组皮肤病理学形态观察，(A)皮肤H&E染色；(B)皮肤Masson染色；(C)皮肤TB染色。
具体实施方式
下面结合实施例和附图对本发明进行详细描述。但下列实施例不应看作对本发明范围的限制。
实施例1：实验材料和方法
(1)NMN来源
Uthever TMNMN由音芙医药科技(上海)有限公司(上海，中国)提供，通过高效液相色谱仪测定NMN的纯度，纯度>98.5％。
(2)实验动物
40只7周龄雌性ICR小鼠购自重庆医科大学实验动物中心[重庆，中国，SCXK(YU)2018-0003]。小鼠饲养在恒定温度与湿度的条件下(温度25±2℃，相对湿度50±5％)，12h亮/暗循环。允许小鼠自由摄入标准鼠粮和饮用水。
(3)UVB诱导皮肤损伤
40只小鼠随机分为4组，每组10只，分别为正常组、UVB组、VC+UVB组、烟酰胺单核苷酸(NMN)+UVB组。整个实验周围为4周，从第3周起，采用紫外辐射设备对除正常组外的其余组小鼠进行皮肤损伤模型的建立，每天照射2h。
在进行UV照射前，采用电动剃须刀剃除小鼠背部约2cm 2的毛发。每组小鼠的具体处理方式如下：(1)正常组：小鼠整个实验期间自由饮食和饮水，不进行造模；(2)UVB组：小鼠整个实验期间自由饮食和饮水，但是在第3周和第4周每天接受2h的UV照射；(3)VC+UVB组：每天按照300mg/kg.bw的剂量给小鼠灌胃VC溶液，同时在第3周和第4周每天给予小鼠2h的UV照射；(4)NMN+UVB组：每天按照300mg/kg.bw的剂量给小鼠腹腔注射NMN，同时在第3周和第4周每天给予小鼠2h的UV照射。所有实验均经重庆市功能性食品协同创新中心批准。
实施例2：烟酰胺单核苷酸能够改善UVB小鼠包括Na +-K +-ATP酶在内的氧化应激损伤
机体氧化-抗氧化平衡障碍与氧化应激损伤的发生和发展密切相关，长期UVB照射会诱发剧烈的氧化应激反应，从而加速皮肤的损伤。研究表明UVB照射会引起皮肤组织中NAD +和Na +-K +-ATP酶的减少；超氧化物歧化酶(SOD)是生物机体内唯一能清除超氧阴离子自由基(O2·-)的抗氧化酶，其活性高低与衰老有着密切的联系，一般随年龄增加而下降。MDA是自由基诱发脂质过氧化链式反应产生的代谢终产物，也是一种极活泼的交联剂，能使真皮结构发生大分子交联，使皮肤变硬、皱缩并生成老年斑。自由基累积越多，MDA生成越多，对机体的损伤也越大。过氧化氢酶(CAT)属于氧化还原酶，大多数为血红素过氧化氢酶，它在生物合成、降解及防御方面发挥着重要的作用。过氧化氢(H 2O 2)是一种活性氧簇的中间产物，如果它在体内过度积累，就会导致抗氧化系统失衡，进而加大氧化应激反应，造成细胞的损伤和凋亡。谷胱甘肽(GSH)作为体内一种重要的抗氧化剂，能够清除掉人体内的自由基；由于GSH本身易受某些物质氧化，所以它在体内能够保护许多蛋白质和酶等分子中的巯基不被有害物质氧化，从而保证蛋白质和酶等分子生理功能的正常发挥。
2.1小鼠血清和皮肤组织中T-SOD、CAT、AGES和H 2O 2的水平
根据试剂盒说明分别测定小鼠血清和皮肤组织中T-SOD、CAT、AGES和H 2O 2的水平，此外还需测定皮肤组织中Na +K +-ATP和NAD +的水平。
小鼠皮肤组织中氧化应激指标水平如表1所示，小鼠血清中氧化应激指标水平如表2所示：
表1小鼠皮肤组织中氧化应激指标水平
Figure PCTCN2020113751-appb-000001
注：表中数据呈现为平均值±标准偏差
Figure PCTCN2020113751-appb-000002
同一列中的不同字母(a-d)表示组别之间存在显著差异(p<0.05)。
由表1可知，正常组小鼠皮肤组织中T-SOD、CAT、Na +K +-ATP和NAD +水平均显著高于UVB组，而H 2O 2和AGEs含量显著低于UVB组。与UVB组相比，VC+UVB组、NMN+UVB组小鼠血清中以上指标的水平都得到不同程度的改善，其中给予NMN处理后 的小鼠血清中的以上指标的水平与正常组之间不存在显著差异。
作为合成NAD +的前体物质，口服NMN能够增加皮肤组织中NAD +的水平，而增加的NAD +会进一步增加Na +-K +-ATP酶的水平，最终起到预防因UVB照射诱发的皮肤癌的作用。Na +K +-ATP酶的活性调节可赋予细胞必要的生命活力，其活性下降会导致细胞内能量生产和离子转运发生障碍，从而影响细胞功能，加速细胞损伤。本实验发现，UVB组小鼠皮肤组织中NAD +和Na +K +-ATP的水平显著低于正常组，说明UVB照射引起了皮肤能量代谢障碍。给予小鼠NMN处理后，皮肤中NAD +和Na +K +-ATP酶水平明显上升，且与正常组的水平接近。
表2小鼠血清中氧化应激指标水平
Figure PCTCN2020113751-appb-000003
注：表中数据呈现为平均值±标准偏差
Figure PCTCN2020113751-appb-000004
同一列中的不同字母(a-d)表示组别之间存在显著差异(p<0.05)。
如表2所示，正常组小鼠血清中T-SOD和CAT的酶活力是最高的，H 2O 2和AGES的含量是最低的，而以上指标在UVB组的水平呈现出完全相反的趋势，且正常组与UVB组之间存在显著差异(p＜0.05)。与UVB组相比，经过VC、NMN处理后，小鼠血清中的T-SOD、CAT酶活力显著升高，H 2O 2和AGES含量显著降低(p＜0.05)。值得注意的是，NMN+UVB组小鼠血清中的T-SOD、CAT、H 2O 2和AGES的水平均接近于正常组，其中T-SOD的酶活力更是显著高于正常组(p＜0.05)，说明NMN能极大提高小鼠血清抗氧化酶活力。
2.2皮肤和肝脏组织中SOD1、SOD2、CAT和GSH的mRNA表达水平
用Trizol试剂对肝脏和皮肤组织进行匀浆和总RNA提取，然后使用cDNA试剂盒将RNA逆转录成cDNA。接着将1μL cDNA、10μL TaqMan TMMultiplex Master Mix、2μL of 10μM primer和7μL ddH 2O混合均匀后，在实时荧光定量PCR仪上进行扩增检测。扩增 条件为：95℃变形15s，55℃退火30s，72℃延伸35s，总共40个循环。最后，通过2 -ΔΔCT计算各目的基因的相对表达量，其中β-actin作为内参基因。用到的引物序列见表3。
表3实验所用引物序列
Figure PCTCN2020113751-appb-000005
如图1可知，与正常组相比，模型小鼠皮肤和肝脏中的SOD1、SOD2、CAT和GSH的mRNA表达水平显著降低。与UVB组相比，VC+UVB组、NMN+UVB组皮肤和肝脏中的SOD1、SOD2、CAT和GSH的mRNA表达均有不同程度的升高，其中NMN+UVB组以上指标的mRNA表达水平接近于正常组。
NMN能够维持小鼠皮肤组织中的T-SOD、CAT、MDA和H 2O 2在一个相对正常的水平。更加值得注意的是，NMN还能极大的提高血清中T-SOD、CAT水平，其中T-SOD的酶活力甚至高于正常组。另外，NMN还能从基因层面上调皮肤和肝脏组织中SOD1、SOD2、CAT和GSH的mRNA表达。如此说明NMN能够通过调节机体的氧化应激水平从而改善小鼠因UVB导致的皮肤损伤，还能从血清、肝脏等层面提高机体的整体抗氧化水平。
实施例3：烟酰胺单核苷酸能够降低机体的炎症损伤
当皮肤长期暴露在紫外线下，Na +K +-ATP酶会出现降低，从而导致皮肤细胞内外渗透压出现异常，细胞膜的完整性和流动性被破坏，进而会出现干燥、瘙痒、红斑和水肿等炎症反应。NF-κB是机体免疫应答过程中重要的转录因子，正常情况下以无活性的方式和其抑制蛋白IκB结合，研究发现当机体受到非正常刺激时会加速IκB蛋白的磷酸化作用，使得NF-κB被激活，而活化的NF-κB会进一步增加促炎性细胞因子TNF-α、IL-6、IL-12、COX-2、iNOS等的释放。许多研究都表明NF-κB信号通路与机体的炎症反应密切相关，而且皮肤损伤过程中也通常会出现NF-κB信号功能障碍。IκB-α是NF-κB的阻遏蛋白，它可以掩蔽NF-κB的核定位信号，从而使NF-κB以非活性复合物形式存在于细胞内。研究表明几乎所有的NF-κB诱导物都会因为IκB-α的降解而迅速激活NF-κB，所以可以通过阻止IκB-α的磷酸化来防止NF-κB的活化。通过测定小鼠血清和皮肤中IL-4、IL-6、IL-10、TNF-α和IL-1β的水平，能够对机体的炎症损伤水平进行窥探。
3.1小鼠血清和皮肤中IL-4、IL-6、IL-10、TNF-α和IL-1β的水平
根据ELISA试剂盒说明书分别测定血清和皮肤组织中IL-4、IL-6、IL-10、TNF-α和IL-1β的水平，结果如表4所示：
表4小鼠血清与皮肤中炎症指标水平
Figure PCTCN2020113751-appb-000006
注：表中数据呈现为平均值±标准偏差
Figure PCTCN2020113751-appb-000007
同一列中的不同字母(a-d)表示组别之间存在显著差异(p<0.05)。
由表4可知，UVB组小鼠血清和皮肤中的促炎细胞因子TNF-α、IL-1β和IL-6的水平是最高的，而抑炎细胞因子IL-4和IL-10是最低的，且UVB组与正常组之间存在统计学差异(p＜0.05)。与UVB组比较，经过VC、NMN处理之后，小鼠血清和皮肤中的促炎细胞因子得到不同程度降低，而抑炎细胞因子有所提高，其中NMN+UVB组改善炎症指标的效果好于VC+UVB组，且与正常组的结果相近。
3.2皮肤和肝脏组织中NF-κBp65、IκB-α、TNF-α、IL-6和IL-10的mRNA表达水平
实验方法同3.2，所用引物序列参见表3。结果如图2所示，与正常组相比，UVB照射增加了小鼠皮肤和肝脏中NF-κBp65的mRNA表达水平，降低了NF-κBp65的抑制基因IκB-α的表达，此外还下调了与NF-κBp65信号通路相关的IL-10的表达，上调TNF-α和IL-6的表达，其中UVB组的变化最为显著。与UVB组比较，VC+UVB组、NMN+UVB组皮肤和肝脏中的IκB-α和IL-10的mRNA表达均有不同程度的升高，而NF-κBp65、TNF-α和IL-6的表达有所降低，其中NMN+UVB组以上指标的mRNA表达水平接近于正常组。说明NMN能够通过上调皮肤和肝脏组织中IκB-α的表达，从而较好地抑制NFκB-p65信号通路的活化。
经UVB照射后，小鼠皮肤和肝脏组织中NFκB-p65的mRNA表达显著升高，IκB-α的mRNA表达明显降低，说明紫外线照射引起了机体炎症的发生。NMN可以分别从血清水平、皮肤组织水平和mRNA水平下调促炎细胞因子TNF-α、IL-6的表达，上调抑炎细胞因子IL-10的表达，从而很好地减少了这些炎症介质对皮肤损伤小鼠的炎症损伤程度。
实施例4：烟酰胺单核苷酸能够改善小鼠紫外照射后组织的病理学形态
解剖小鼠取出小鼠的肝脏和背部无毛皮肤，剪取黄豆大小的肝脏和皮肤组织固定在4％的多聚甲醛溶液中，剩余的肝脏和皮肤组织保存在-80℃冰箱备用。
肝脏进行H&E染色，皮肤进行H&E、Masson’s trichrome和toluidine blue染色。最后采用正置显微镜观察组织的病理学形态，结果如图3和图4所示：
4.1肝脏病理学形态
如图3所示，正常组小鼠肝脏结构完整，肝细胞以卫星发射状整齐有序地排列在中央静脉周围，细胞核大而圆，不存在炎症细胞浸润的现象，呈现出正常肝脏该有的形态特征。与正常组相比，UVB组小鼠肝脏细胞排列比较紊乱，中央静脉周围的肝细胞出现部分坏死，并存在炎症细胞浸润的现象。与UVB组相比，VC+UVB组、NMN+UVB组的肝细胞结构 均有不同程度的改善，其中NMN+UVB组小鼠肝细胞形态与正常组接近。
4.2皮肤病理学形态
H&E染色主要用来观察皮肤的整体结构，Masson染色主要用来观察皮肤中的胶原纤维，TB染色则主要用于观察皮肤中的肥大细胞。由图4(A)可知，正常组小鼠皮肤结构完整，表皮层较薄，表皮与真皮连接处呈波浪状，无过度角质化的角质层；真皮层较厚，胶原束形态结构完整，排列有序，分布均匀。UVB组小鼠真皮层厚度明显变薄，胶原纤维束的数量显著减少，皮下组织排列紊乱，分界不明显；此外，可见附属器周围有炎症细胞浸润，表明皮肤有慢性炎症反应的发生，说明长期UVB照射可诱发皮肤光损伤。与UVB组比较，VC+UVB组小鼠皮肤真皮层厚度有所增加，但是胶原纤维分散疏松。NMN+UVB组小鼠皮肤整皮层厚度明显厚于UVB组，胶原纤维束分布相对均匀有序，整体结构与正常组接近。
Masson染色后的胶原纤维呈现蓝紫色。由图4(B)可知，正常组胶原纤维的数量和排列整齐度明显比UVB组高，说明UVB照射会引起胶原纤维的降解。与UVB组相比，VC+UVB组、NMN+UVB组小鼠皮肤真皮层中的胶原纤维数量均有所增加，其中NMN+UVB组的胶原纤维排列更为整齐有序，几乎不存在皱缩、断裂等现象。
由TB染色(图4(C))结果可知，与正常组比较，UVB组小鼠真皮层中的肥大细胞数量显著升高，说明UVB照射会诱发皮肤肥大细胞的产生，从而导致皮肤炎症。与UVB组相比，VC+UVB组小鼠皮肤中的肥大细胞有所减少，但数量仍旧明显多于正常组。给予小鼠NMN处理之后，NMN+UVB组小鼠皮肤中的肥大细胞数量得到明显减少，结果接近于正常组。
本发明中，UVB组小鼠皮肤真皮层变薄，胶原纤维数量减少，肥大细胞数量增多，同时肝脏组织也出现了一定程度的病理改变，说明UVB诱导的皮肤损伤模型成功。给予皮肤损伤小鼠NMN经口灌胃之后，皮肤病理学形态得到了极大改善，小鼠的肝脏也保持在正常的形态，未出现明显损伤。
以上显示和描述了本发明的基本原理、主要特征和本发明的优点。本行业的技术人员应该了解，本发明不受上述实施例的限制，上述实施例和说明书中描述的只是说明本发明的原理，在不脱离本发明精神和范围的前提下本发明还会有各种变化和改进，这些变化和改进都落入要求保护的本发明范围内。本发明要求保护范围由所附的权利要求书及其等同物界定。
Figure PCTCN2020113751-appb-000008
Figure PCTCN2020113751-appb-000009
Figure PCTCN2020113751-appb-000010
Figure PCTCN2020113751-appb-000011
Figure PCTCN2020113751-appb-000012

Claims (8)

  1. 烟酰胺单核苷酸在制备提高损伤皮肤肌膜中Na +-K +-ATP酶水平试剂中的应用。
  2. 根据权利要求1所述的烟酰胺单核苷酸在制备提高损伤皮肤肌膜中Na +-K +-ATP酶水平试剂中的应用，其特征在于：
    其中，所述损伤皮肤为紫外线辐射引起的皮肤损伤。
  3. 根据权利要求2所述的烟酰胺单核苷酸在制备提高损伤皮肤肌膜中Na +-K +-ATP酶水平试剂中的应用，其特征在于：
    其中，所述紫外线辐射引起的皮肤损伤为中波紫外线辐射诱导的皮肤损伤。
  4. 一种提高损伤皮肤肌膜中Na +-K +-ATP酶水平的试剂，其特征在于，以烟酰胺单核苷酸作为唯一活性成分。
  5. 权利要求4所述的提高损伤皮肤肌膜中Na +-K +-ATP酶水平的试剂在制备缓解皮肤氧化应激损伤药物、保健食品或化妆品中的应用。
  6. 权利要求4所述的提高损伤皮肤肌膜中Na +-K +-ATP酶水平的试剂在制备缓解皮肤光损伤药物、保健食品或化妆品中的应用。
  7. 根据权利要求4所述的提高损伤皮肤肌膜中Na +-K +-ATP酶水平的试剂，其特征在于：所述试剂为胶剂、膏剂、乳剂、合剂、口服安瓿剂、片剂或胶囊剂；
  8. 权利要求6所述的提高损伤皮肤肌膜中Na +-K +-ATP酶水平的试剂在制备缓解皮肤光损伤药物、保健食品或化妆品中的应用，其特征在于：
    其中，所述药物为胶剂、膏剂、乳剂、气雾剂、注射剂、合剂、口服安瓿剂、片剂或胶囊剂等；
    所述保健食品为胶囊剂或片剂、粉剂等口服型式。
    所述化妆品为乳液、喷剂、面膜或眼霜。
PCT/CN2020/113751 2020-09-07 2020-09-07 烟酰胺单核苷酸在制备提高损伤皮肤肌膜中Na+-K+-ATP酶水平试剂中的应用 WO2022047779A1 (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/CN2020/113751 WO2022047779A1 (zh) 2020-09-07 2020-09-07 烟酰胺单核苷酸在制备提高损伤皮肤肌膜中Na+-K+-ATP酶水平试剂中的应用

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2020/113751 WO2022047779A1 (zh) 2020-09-07 2020-09-07 烟酰胺单核苷酸在制备提高损伤皮肤肌膜中Na+-K+-ATP酶水平试剂中的应用

Publications (1)

Publication Number Publication Date
WO2022047779A1 true WO2022047779A1 (zh) 2022-03-10

Family

ID=80492462

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2020/113751 WO2022047779A1 (zh) 2020-09-07 2020-09-07 烟酰胺单核苷酸在制备提高损伤皮肤肌膜中Na+-K+-ATP酶水平试剂中的应用

Country Status (1)

Country Link
WO (1) WO2022047779A1 (zh)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116879283A (zh) * 2023-07-19 2023-10-13 南京峦创生命科技有限公司 用于测定atp的试剂盒

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015070280A1 (en) * 2013-11-14 2015-05-21 Newsouth Innovations Pty Limited Senescence and senescence associated secretory phenotype
CN108137639A (zh) * 2015-08-05 2018-06-08 麦德龙国际生物科技有限责任公司 烟酰胺单核苷酸衍生物及其用途
CN110461339A (zh) * 2017-02-08 2019-11-15 东方酵母工业株式会社 皮肤色素沉着抑制剂
CN110974781A (zh) * 2020-01-02 2020-04-10 崔立峰 一种用于皮肤创面修复的凝胶
CN111166760A (zh) * 2019-12-17 2020-05-19 浙江安赛新材料科技有限公司 β-烟酰胺单核苷酸或其前体的组合物及制备方法、应用

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015070280A1 (en) * 2013-11-14 2015-05-21 Newsouth Innovations Pty Limited Senescence and senescence associated secretory phenotype
CN108137639A (zh) * 2015-08-05 2018-06-08 麦德龙国际生物科技有限责任公司 烟酰胺单核苷酸衍生物及其用途
CN110461339A (zh) * 2017-02-08 2019-11-15 东方酵母工业株式会社 皮肤色素沉着抑制剂
CN111166760A (zh) * 2019-12-17 2020-05-19 浙江安赛新材料科技有限公司 β-烟酰胺单核苷酸或其前体的组合物及制备方法、应用
CN110974781A (zh) * 2020-01-02 2020-04-10 崔立峰 一种用于皮肤创面修复的凝胶

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
ANDREW C CHEN; DIONA L DAMIAN: "Nicotinamide and the skin", AUSTRALASIAN JOURNAL OF DERMATOLOGY, AUSTRALIAN COLLEGE OF DERMATOLOGISTS, SYDNEY,, AU, vol. 55, no. 3, 17 March 2014 (2014-03-17), AU , pages 169 - 175, XP071010115, ISSN: 0004-8380, DOI: 10.1111/ajd.12163 *
BRIGITTE PELZMANN; SETH HALLSTRöM; PETER SCHAFFER; PETRA LANG; KARL NADLINGER; GEORGE D BIRKMAYER; KAROLINE VRECKO; GILBERT R: "NADH supplementation decreases pinacidil‐primed IK(ATP) in ventricular cardiomyocytes by increasing intracellular ATP", BRITISH JOURNAL OF PHARMACOLOGY, WILEY-BLACKWELL, UK, vol. 139, no. 4, 29 January 2009 (2009-01-29), UK , pages 749 - 754, XP071079638, ISSN: 0007-1188, DOI: 10.1038/sj.bjp.0705300 *
HUANG, CHUNYUN : "Ultraviolet-induced oxidative damage to skin", INTERNATIONAL JOURNAL OF DERMATOLOGY AND VENEREOLOGY, vol. 35, no. 1, 31 January 2009 (2009-01-31), CN, pages 51 - 53, XP009535045, ISSN: 1673-4173, DOI: 10.3760/cma.j.issn.1673-4173.2009.01.024 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116879283A (zh) * 2023-07-19 2023-10-13 南京峦创生命科技有限公司 用于测定atp的试剂盒

Similar Documents

Publication Publication Date Title
CN109125107B (zh) 一种有效改善和修复面部激素依赖性皮炎的多肽组合物
US11191712B2 (en) Skin treatment compositions, masks, and related methods
CN1951410A (zh) 含药用人参的脑细胞或神经细胞保护剂
Zhou et al. Nicotinamide mononucleotide combined with lactobacillus fermentum TKSN041 reduces the photoaging damage in murine skin by activating AMPK signaling pathway
WO2022077276A1 (zh) 烟酰胺单核苷酸与发酵乳杆菌联合在制备缓解皮肤光老化制剂中的应用
Wikramanayake et al. Targeting mitochondria in dermatological therapy: beyond oxidative damage and skin aging
WO2022047779A1 (zh) 烟酰胺单核苷酸在制备提高损伤皮肤肌膜中Na+-K+-ATP酶水平试剂中的应用
Kuete et al. Toxicological evaluation of the hydroethanol extract of Tabernaemontana crassa (Apocynaceae) stem bark
Zhang et al. Glycine-Histidine-Lysine (GHK) alleviates astrocytes injury of intracerebral hemorrhage via the Akt/miR-146a-3p/AQP4 pathway
Wang et al. Oral administration of punicalagin attenuates imiquimod‐induced psoriasis by reducing ROS generation and inflammation via MAPK/ERK and NF‐κB signaling pathways
Fan et al. Ginsenoside Rh2 attenuates myocardial ischaemia‑reperfusion injury by regulating the Nrf2/HO‑1/NLRP3 signalling pathway
US20070042976A1 (en) Method of treating cosmetic and dermatologic conditions by a demethylating agent
CN117547524A (zh) 吉马酮化合物的新用途
CN110742818A (zh) 一种抗紫外线损伤的辅酶q10脂质体的制备方法
González et al. Eruptive vellus hair cyst with singular histologic finding
US20090068292A1 (en) Therapeutic herb cosmetic composition
CN111135124A (zh) 含有蔗糖、吲哚-3-乙酸以及玫瑰果提取物的混合物的皮肤外用剂组合物
KR100881445B1 (ko) 피부보호 및 피부질환 개선용 조성물 및 그 제조방법
Leem et al. Creatine supplementation with exercise reduces α-synuclein oligomerization and necroptosis in Parkinson's disease mouse model
TWI815349B (zh) 山竹果殼萃取物用於製備促進糖尿病傷口癒合的藥物的用途
Wang et al. Tanshinone IIA, a melanogenic ingredient basis of Salvia miltiorrhiza Bunge
Yang et al. Research on Mechanism of miR-106a Nanoparticles Carrying Dexmedetomidine in Regulating Recovery and Metabolism of Nerve Cells in Hypoxia-Reoxygenation Injury
Yang et al. Isoalantolactone Regulates the Activity of Mitogen-Activated Protein Kinases and Nuclear-Factor Erythroid 2-Related Factor 2 Pathways to Attenue Angiotensin II-Induced Cardiomyocyte Hypertrophy and Oxidative Stress.
CN113244288B (zh) 一种中药组合物及其应用
US20240287133A1 (en) Peptide having activity of preventing hair loss or promoting hair growth, and use thereof

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 20952023

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 20952023

Country of ref document: EP

Kind code of ref document: A1