CN113082192A - 胶原蛋白与脂肪干细胞的组合物在制备药物或者化妆品中的用途 - Google Patents
胶原蛋白与脂肪干细胞的组合物在制备药物或者化妆品中的用途 Download PDFInfo
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- CN113082192A CN113082192A CN202110470742.0A CN202110470742A CN113082192A CN 113082192 A CN113082192 A CN 113082192A CN 202110470742 A CN202110470742 A CN 202110470742A CN 113082192 A CN113082192 A CN 113082192A
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Abstract
本发明涉及胶原蛋白与脂肪干细胞的组合物在制备药物或者化妆品中的用途。本发明制备了新的胶原蛋白肽,所述胶原蛋白肽具有较好的自由基清除能力。同时本发明还通过使用多肽来培养脂肪干细胞后能够有效的防止脂肪干细胞的衰老及促进脂肪干细胞的增殖,将胶原蛋白肽和多肽培养的脂肪干细胞以及维生素C一起使用能够有效的预防鼠衰老及美白,具有很好的应用价值。
Description
技术领域
本申请涉及药物或者化妆品领域,具体的,涉及胶原蛋白与脂肪干细胞的组合物在制备药物或者化妆品中的用途。
背景技术
ADSCs因来源广泛、取材方便、组织损伤小、扩增迅速等优点已成为细胞治疗和组织工程应用中的理想工具。众多研究发现,ADSCs在体内损伤修复、抗衰老等方面均可发挥较为显著的作用;其生物学作用的实现一部分通过多向分化能力,ADSCs可分化为脂肪、骨、软骨、肌肉、肌腱等多种细胞,补充替代老化、缺损的组织细胞;另一部分通过旁分泌功能,ADSCs可合成及分泌多种细胞因子、多肽类、气体分子等活性成分,提高受损细胞的生命力和抗凋亡能力。ADSCs通过分泌一系列活性成分在抑制皮肤老化、美白肌肤、辅助脂肪移植、促进毛发再生等诸多抗衰老方面均有较显著的效果。
众多实验证实,ADSCs分泌的多种细胞因子具有增强细胞增殖和迁移能力,促血管形成及抗细胞凋亡等作用,促进组织功能恢复正常,减少损害,在抗衰老领域有较显著的应用效果。研究表明,ADSCs所分泌的胰岛素样生长因子、表皮生长因子、转化生长因子、白细胞介素和肿瘤坏死因子,可改善中波紫外线引起的皮肤成纤维细胞损伤,加强成纤维细胞增殖及合成胶原蛋白的能力,其中转化生长因子在体外实验中刺激成纤维细胞的作用最强。将ADSCs注射于光老化裸鼠模型,可观察到注射ADSCs部位真皮层厚度和成纤维细胞数量显著增加,成纤维细胞合成胶原蛋白增多,真皮血管生成增加,波紫外线照射导致的皮肤老化外观明显改善。将ADSCs条件培养基应用于患者面部皮肤除皱,注射治疗后2个月观察到显著的皮肤质地改善、真皮厚度增加及皱纹淡化。对手术联合ADSCs注射进行面部年轻化治疗的患者进行随访,发现接受联合治疗患者的皮肤较单纯手术治疗的更好。
胶原蛋白作为一种常用的化妆品原料,近年来应用发展很快。它可以滋润肌肤,赋予其平滑感觉,对头发也有很好的调理作用。研究表明,护肤品中加入很少的胶原蛋白纯溶液就有良好的抗各种辐射的作用,且能形成很好的保水层,全面提供皮肤所需要的水分。胶原蛋白可以进入皮肤深层,给予皮肤所必需的营养成分(氨基酸),维持胶原纤维结构的稳定性和完整性,增强皮肤中胶原的活性,使皮肤细胞的生存环境得到改善,并促进组织的新陈代谢,达到营养滋润、美容养发的目的。胶原蛋白与皮肤角质层结构的相似性决定了它与皮肤良好的相容性、良好亲和力和渗透性,能渗透入皮肤表皮层,被皮肤充分吸收,并在皮肤表面形成一层极薄的膜层,从而使皮肤丰满、皱纹舒展,同时提高皮肤密度,产生张力,具有抗皱作用。胶原蛋白中的酪氨酸残基与皮肤中的酪氨酸竞争,与酪氨酸酶的活性中心结合,从而抑制酪氨酸酶催化皮肤中的酪氨酸转化为多巴,阻止皮肤中黑色素的形成,达到美白的作用。
目前,将干细胞与胶原蛋白一起使用用于皮肤美白或者治疗的情形还不多,而且相应的改进也研究较少。
发明内容
本发明克服现有技术的缺陷,提供一种具有美白、保水、抗皱的化妆品或者药物。
具体的,本发明提供一种胶原蛋白肽,所述胶原蛋白肽采用如下方法制备获得:
用自来水清洗黑鱼皮并用刀切成小块。将鱼皮浸泡在NaOH溶液中。碱处理的鱼皮用自来水洗至中性,然后用HCl(v/v)浸泡1h,然后用自来水漂洗至中性。将无菌水加入清洁过的鱼皮中,并将混合物在恒温下温育。以8000r/min离心20min,得到的上清液为胶原蛋白,冷冻干燥后使用。用复合蛋白酶(木瓜蛋白酶,广西庞博生物制剂有限公司,酶活2900U/mg;胰蛋白酶,上海南宁生化试剂有限公司,酶活3600U/mg;碱性蛋白酶,山东隆大生物工程有限公司,酶活2000U/mg;)酶解鱼皮胶原蛋白,在pH8.0和40℃下酶解,酶解后沸水中加热灭酶,冷却至室温,酶解液离心,取上清液浓缩并冷冻干燥得到鱼皮胶原蛋白肽。
本发明进一步的,通过前期鉴定分离筛选,获得了能够有效促进脂肪间充质干细胞增殖及防止干细胞本身衰老的多肽,所述多肽的序列如SEQ ID NO:1所示。
进一步的,本发明提供一种延缓衰老、美白的药物组合物或者化妆品组合物,所述组合物含有鱼皮胶原蛋白肽以及采用SEQ ID NO:1所示多肽处理的脂肪间充质干细胞和维生素C。
在实施方案中,该组合物或组合包括药学上可接受的载体。在实施方案中,药学上可接受的载体包括乳液。在实施方案中,乳液是水包油乳液或油包水乳液。在实施方案中,组合或组合包括霜(cream)、露(lotion)、悬浮液或水性溶液或以霜、露、悬浮液或水性溶液的形式存在。
在实施方案中,将组合物配制成用于局部施用于皮肤(即,该组合物是局部的(topical)组合物)。在实施方案中,该组合物是药物组合物。
所述组合物还包括药学上可接受的载体。
在实施方案中,将药物组合物配制成用于局部施用于皮肤。在实施方案中,药学上可接受的载体包括乳液。在实施方案中,乳液是水包油乳液或油包水乳液。在实施方案中,药物组合物为霜、露、悬浮液或水性溶液的形式。所述组合物可以用于皮下注射或者表面涂抹。
所述组合物能够下调酪氨酸酶活性减少酪氨酸酶相关蛋白的表达,最终抑制黑色素的合成,起到美白皮肤作用。而且,所述组合物还能够导致黑素细胞数量更少、美白皮肤的作用。
本发明还涉及治疗或美容处理方法,该方法包括给予药用组合物或化妆品组合物,所述组合物作为一种抑制黑色素合成的组合物。在一种具体模式中,本发明还涉及治疗或美容方法,该方法包括给予含所述干细胞的药用组合物或化妆品组合物,用于治疗色素病症。
进一步的,该组合物还可以选自所有以下所述的化合物及其功能等价物:
该其它化合物可以特别地选自皮肤病学或化妆品中常规使用的活性剂,例如润肤剂、保湿活性剂、角蛋白合成活化剂、角质调节剂、角质剥脱剂、重建皮肤屏障的试剂(皮脂合成活化剂)、过氧化物酶体增殖剂-激活受体(PPAR)激动剂、RXR或LXR激动剂、皮脂调节剂、抗刺激剂、舒缓剂、抗炎剂、抗氧化剂和抗老化剂、脱色剂或低脱色剂、着色剂、脂解剂或脂肪生成抑制剂或抗橘皮组织或还原剂、无机或有机滤光剂和防晒剂、抗真菌化合物、防腐剂、抗细菌剂、益生元和益生菌、抗生素、免疫调节剂。
更特别地,在组合中能够使用的用于愈合和/或重建皮肤屏障的试剂有利地为泛醇(维生素B5)、阿拉伯半乳聚糖、氧化锌、神经酰胺、胆固醇、角鲨烷和磷脂。
在组合中能够使用的皮脂调节剂有利地选自5-α-还原酶抑制剂。锌(和锌衍生物例如其葡糖酸盐、水杨酸盐和焦谷氨酸盐)和螺内酯也具有皮脂抑制活性。其它作用于皮脂质量的脂质源的皮脂调节剂,例如亚油酸也是感兴趣的。
所述抗炎和/或抗刺激和/或舒缓剂可以是阿拉伯半乳聚糖。
在组合中能够使用的防晒活性剂有利地为UVB和/或UVA滤光剂和防晒剂,例如本领域技术人员已知的无机和/或有机防晒剂或滤光剂,本领域技术人员能够根据所要求的保护程度调整它们的选择和浓度。
在组合中能够使用的防腐剂例如为在化妆品中通常使用的那些,具有抗菌活性的分子(伪防腐剂)例如辛酸衍生物,如,例如辛酰基甘氨酸和辛酸甘油酯;己二醇、乙酰丙酸纳、以及铜和锌衍生物(葡糖酸盐和PCA)。
有益效果
本发明制备了新的胶原蛋白肽,所述胶原蛋白肽具有较好的自由基清除能力。同时本发明还通过使用多肽来培养脂肪干细胞后能够有效的防止脂肪干细胞的衰老及促进脂肪干细胞的增殖,将胶原蛋白肽和多肽培养的脂肪干细胞以及维生素C一起使用能够有效的预防鼠衰老及美白,具有很好的应用价值。
附图说明
图1酶添加量对胶原蛋白得率的影响图
图2鱼皮胶原蛋白多肽对DPPH自由基的清除率结果图
图3脂肪间充质干细胞衰老结果图
具体实施方式
为了更进一步的说明本发明的目的、技术方案和优点,我们结合以下具体实施例来阐述本发明,这些实施例仅为了更好的说明本发明专利,而不用于限制本发明范围。基于本发明中的实施例,本领域的技术人员在没有做出创造性的情况下所得到的其他所有实施方式均属于本发明所保护的范围。
实施例1胶原蛋白肽的制备
用自来水清洗黑鱼皮并用刀切成0.5cm×0.5cm的小块。将鱼皮浸泡在1.0mmol/L的NaOH溶液中2h(鱼皮:溶液=1:10,w/v)。碱处理的鱼皮用自来水洗至中性,然后用0.5%HCl(v/v)浸泡1h(鱼皮:溶液=1:10,w/v),然后用自来水漂洗至中性。将无菌水加入清洁过的鱼皮中,并将混合物在恒温下温育。以8000r/min离心20min,得到的上清液为胶原蛋白,冷冻干燥后使用。用复合蛋白酶(木瓜蛋白酶,广西庞博生物制剂有限公司,酶活2900U/mg;胰蛋白酶,上海南宁生化试剂有限公司,酶活3600U/mg;碱性蛋白酶,山东隆大生物工程有限公司,酶活2000U/mg;)酶解鱼皮胶原蛋白,加酶量设置0.5%、1%、2%、3%、4%、5%共五个梯度,在pH8.0和40℃下酶解4h,酶解后沸水中加热15min灭酶,冷却至室温,酶解液8000r/min离心20min,取上清液浓缩并冷冻干燥得到鱼皮胶原蛋白肽。
胶原蛋白在经过酶解后分解为氨基酸,用氯胺T氧化而形成含有吡咯环的氧化物,过氯酸终止氧化后用对二甲氨基苯甲醛作显色剂,生成红色的化合物,在550nm波长下进行比色测定。所测定羟脯氨酸含量经过计算得出胶原蛋白含量。结果如图1所示。
从图1可以看出,酶添加量对胶原蛋白得率的影响为得率的提高并不是随着比例增大而一直升高,在酶添加量为2%时是酶发挥活力的最佳比例,此时酶活最高,更高的添加量可能会起到障碍的作用,阻拦酶解反应的进行。因此,以2%酶添加量制备得到胶原蛋白肽粉末用于后续实验。
实施例2 DPPH自由基清除能力的测定
将实施例1制备得到的胶原蛋白肽制备成不同浓度的样品溶液和0.1mmol/L DPPH乙醇溶液。精确吸取待测不同浓度的稀释样品0.4mL于5mL的离心管中,加入2.0mL DPPH乙醇溶液,混匀,暗处放置30min,不断震荡,517nm测定吸光值。DPPH清除率(%)=((A对照-(A样品-A样品对照))/A对照)×100式中:A对照为0.4mL去离子水+2.0mLDPPH溶液;A样品对照为0.4mL样品+2.0mL乙醇;A样品为0.4mL待测稀释样品+2.0mLDPPH溶液。结果如图2所示。
DPPH法是评估天然抗氧化剂清除自由基活性的快速可行的方法。由图2可知,本发明制备的鱼皮胶原蛋白多肽对DPPH自由基的清除率都随着样品浓度的增加而显著增大,在50μg/mL浓度下即可达到80%以上的清除率,具有较好的效果。
实施例3脂肪干细胞的制备
无菌条件下,获得脂肪生理盐水混合物50mL,离心、PBS清洗两遍去除血细胞,获得纯度较高的脂肪颗粒。0.075%Ⅰ型胶原酶37℃恒温摇床消化60min,1500r/min,离心10min,去上层未消化的脂肪组织及油脂,沉淀重悬200目筛网过滤,再次离心,红细胞裂解液裂解红细胞5min、磷酸盐缓冲液洗涤两遍,10%胎牛血清高糖DMEM重新悬,以5×104/cm2接种至10cm2培养板中。以第1次接种的细胞为0代,记作P0,细胞90%融合后0.25%胰酶消化,1∶4传代接种,取第3代细胞进行实验。
第3代的hADSCs胰蛋白酶消化后制成单细胞悬液。1×106细胞分别加入抗人CD34PE、CD73PE、CD90FITC、CD105PE、CD45FITC单抗各1μL,PE-IgG1、FITC-IgG1为阴性对照。4℃避光孵育30min,流式细胞仪检测。结果如下表1所示:
表1细胞表型鉴定结果
细胞表型鉴定 | 表达率 |
CD105 | (96.2±2.6)% |
CD90 | (95.8±1.9)% |
CD34 | (4.2±1.2)% |
CD45 | (3.1±0.80)% |
从表1可以看出,经检测CD105表达率(96.2±2.6)%,CD90表达率(95.8±1.9)%,CD34表达率(4.2±1.2)%,CD45表达率(3.1±0.80)%。本发明分离的脂肪干细胞表现出明显的脂肪干细胞的生物学特性,高表达CD90、CD105,低表达CD34、CD45等造血系分子。
实施例4脂肪干细胞抗衰老实验
将实施例3分离的脂肪干细胞进行消化、制成细胞悬液,按细胞密度1×105个/mL接种,分组处理:对照组采用10%胎牛血清的DMEM培养基传代培养48h;衰老组:应用D-半乳糖8g/L+10%胎牛血清的DMEM培养基培养48h;衰老拯救组:应用D-半乳糖8g/L+SEQ ID NO:1抗衰老肽1mg/mL+10%胎牛血清的DMEM培养基培养48h;刺激组:应用SEQ ID NO:1抗衰老肽1mg/mL+10%胎牛血清的DMEM培养基培养48h;随后,按β-半乳糖苷酶检测试剂盒进行操作,吸除细胞培养基,PBS洗涤1次,加入适量SA-β-gal染色固定液,室温固定15min,吸除细胞固定液,加入PBS洗涤3次,每次3min,吸除PBS,加入适量细胞染色工作液,37℃孵育过夜,在光学显微镜下观察衰老细胞呈淡蓝至深蓝色,标本阳性率以阳性标本占总标本数的百分率表示。结果如图3所示。
从图3可以看出,SA-β-gal染色结果显示,脂肪间充质干细胞在1mg/mL的多肽的作用下,能够显著的降低脂肪间充质干细胞本身的衰老速度,多肽培养的间充质干细胞培养基中蓝绿色的衰老细胞较少,与对照组比较差异有显著性意义(P<0.01),说明多肽处理后的脂肪干细胞具有显著抗细胞衰老作用。
此外,根据培养后的细胞计数结果可以看出,所述多肽在抑制细胞衰老的同时,还能够促进干细胞本申请的增殖,与对照组相比,增殖效果可以达到(23.1±2.5)%。
实施例5胶原蛋白肽与脂肪干细胞对豚鼠酪氨酸酶和皮肤黑色素的影响
选取花色豚鼠,雌雄兼用,体重250g左右;将豚鼠随机分成5组,即正常对照组(皮下注射生理盐水),阳性对照组:(为市售胶原蛋白肽100mg/kg);蛋白肽处理组(皮下注射实施例1制备的胶原蛋白肽100mg/kg),脂肪干细胞处理组(皮下注射实施例3制备的干细胞2*109个/mL/0.1mL/kg),胶原蛋白肽+脂肪干细胞处理组(皮下注射实施例1制备的胶原蛋白肽50mg/kg+实施例3制备的干细胞1*109个/mL/0.1mL/kg),各组给药频率3d/次。
实验前各组豚鼠用电动剃刀剪去黑灰色皮肤的硬毛备用。连续给药30d后,各组豚鼠于末次给药后,用20%乌拉坦麻醉,分离颈总动脉采血,2500r/min离心10min,取血浆测定酪氨酸酶的含量;在剃毛区取2cm×2cm皮肤,用4%甲醛固定,常规石蜡包埋切片,用HE染色,另用硫酸亚铁Lillie法进行黑色素染色,随意观察50个毛囊,计算其中有黑色素的毛囊数。进行统计学处理并比较组间差异。结果如表2所示。
表2各组对豚鼠的酪氨酸酶活性和皮肤黑色素的影响
由表1可见,阳性对照组、胶原蛋白肽处理组、脂肪干细胞处理组、胶原蛋白肽+脂肪干细胞处理组均能够导致豚鼠血浆中酪氨酸酶活性水平明显降低,与同期正常对照组相比,具有显著性差异(*P<0.05),特别是采用胶原蛋白肽和干细胞一起处理能够更显著降低豚鼠血清酪氨酸酶,显示出了明显的协同效果。此外,阳性对照组、胶原蛋白肽处理组、脂肪干细胞处理组、胶原蛋白肽+脂肪干细胞处理组均能够导致豚鼠皮肤毛囊含黑色素颗粒细胞数明显减少,与同期正常对照组相比,有显著性差异(P<0.05)。由此提示特别是胶原蛋白肽+脂肪干细胞处理组具有较好的美白作用,能够减少豚鼠皮肤中黑色素颗粒物的含量。
实施例6对D-半乳糖衰老模型小鼠皮肤羟脯氨酸和水分含量测定
采用腹腔注射D-半乳糖致小鼠亚急性衰老模型:各组小鼠每日腹腔注射D-半乳糖100mg/kg,连续注射6d,正常对照组腹腔注射等量的无菌生理盐水。6d后,开始给药,正常对照组(皮下注射生理盐水),模型对照组(皮下注射生理盐水),阳性对照组:(为市售胶原蛋白肽100mg/kg);胶原蛋白肽+脂肪干细胞处理组(皮下注射实施例1制备的胶原蛋白肽50mg/kg+实施例3制备的干细胞1*109个/mL/0.1mL/kg),胶原蛋白肽+维生素C+脂肪干细胞处理组(皮下注射实施例1制备的胶原蛋白肽50mg/kg+维生素C10 mg/kg+实施例3制备的干细胞1*109个/mL/0.1mL/kg),各组给药频率3d/次,给药处理5次。取几个组处理的小鼠的一块皮肤,称湿重,然后60℃烘至恒重后称重,计算两者差值即为皮肤的水分含量。另取小鼠皮肤0.5g,在冰冷的生理盐水中漂洗,滤纸拭干,放入5ml的小烧杯内。用移液管量取预冷0.86%生理盐水2ml,用眼科剪尽快剪碎组织块。将剪碎的组织倒入玻璃匀浆管中,加0.9ml的冷生理盐水冲洗残留在烧杯中的组织块,一起倒入匀浆管中进行匀浆,左手持匀浆管将下端插入盛有冰水混合物的器皿中,右手将捣杆垂直插入套管中,上下转动研磨数10次,充分研碎,使组织匀浆化。将制备好的10%匀浆用普通离心机2 000r/min离心15min,将离心好的匀浆取上清0.1ml加0.4ml生理盐水,稀释成2%的组织匀浆进行羟脯氨酸检测,进行统计学处理并比较组间差异。结果如表3所示。
表3各组对豚鼠皮肤羟脯氨酸和水分含量的影响
由表3可见,胶原蛋白肽+维生素C+脂肪干细胞处理组能够明显地增加衰老小鼠皮肤的羟脯氨酸含量,与模型对照组相比具有显著性差异(P<0.05)。同时胶原蛋白肽+维生素C+脂肪干细胞处理组均能够明显地增加衰老小鼠皮肤的水分含量,与模型对照组相比具有显著性差异(P<0.05)。具有较好的应用价值。
以上结合具体实施方式和范例性实例对本发明进行了详细说明,不过这些说明并不能理解为对本发明的限制。本领域技术人员理解,在不偏离本发明精神和范围的情况下,可以对本发明技术方案及其实施方式进行多种等价替换、修饰或改进,这些均落入本发明的范围内。本发明的保护范围以所附权利要求为准。
序列表
<110> 北京瀚梅生物科技有限公司
<120> 胶原蛋白与脂肪干细胞的组合物在制备药物或者化妆品中的用途
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 45
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 1
Ser Phe His His Met Asn Gln Lys Thr Arg Pro Arg Lys Trp Ile Pro
1 5 10 15
Ser Pro Ile Ile Thr Arg Gln Val Thr Ala Trp Gly Leu Pro Gln Lys
20 25 30
His Pro His Lys Gln Ser Arg Gln Trp Val Pro Trp Met
35 40 45
Claims (6)
1.胶原蛋白肽、微生素C以及具有抗衰老作用的脂肪干细胞在制备抗衰老、美白、保湿的药物组合物中的用途;其中所述胶原蛋白肽采用如下方法制备获得:
用自来水清洗黑鱼皮并用刀切成0.5cm×0.5cm的小块,将鱼皮浸泡在1.0mmol/L的NaOH溶液中2h,其中鱼皮∶溶液=1∶10,w/v;碱处理的鱼皮用自来水洗至中性,然后用0.5%HCl(v/v)浸泡1h,其中鱼皮∶溶液=1∶10,w/v,然后用自来水漂洗至中性;将无菌水加入清洁过的鱼皮中,并将混合物在恒温下温育;以8000r/min离心20min,得到的上清液为胶原蛋白,冷冻干燥后使用;用复合蛋白酶:木瓜蛋白酶,酶活2900U/mg;胰蛋白酶,酶活3600U/mg;碱性蛋白酶,酶活2000U/mg酶解鱼皮胶原蛋白,加酶量2%,在pH8.0和40℃下酶解4h,酶解后沸水中加热15min灭酶,冷却至室温,酶解液8000r/min离心20min,取上清液浓缩并冷冻干燥得到鱼皮胶原蛋白肽;
所述脂肪干细胞是将分离制备的脂肪干细胞进行消化、制成细胞悬液,按细胞密度1*105个/mL接种,采用添加了1mg/mL的SEQ ID NO:1多肽的10%胎牛血清的DMEM培养基传代培养后获得。
2.胶原蛋白肽、微生素C以及具有抗衰老作用的脂肪干细胞在制备抗衰老、美白、保湿的化妆品中的用途;其中所述胶原蛋白肽采用如下方法制备获得:
用自来水清洗黑鱼皮并用刀切成0.5cm×0.5cm的小块,将鱼皮浸泡在1.0mmol/L的NaOH溶液中2h,其中鱼皮∶溶液=1∶10,w/v;碱处理的鱼皮用自来水洗至中性,然后用0.5%HCl(v/v)浸泡1h,其中鱼皮∶溶液=1∶10,w/v,然后用自来水漂洗至中性;将无菌水加入清洁过的鱼皮中,并将混合物在恒温下温育;以8000r/min离心20min,得到的上清液为胶原蛋白,冷冻干燥后使用;用复合蛋白酶:木瓜蛋白酶,酶活2900U/mg;胰蛋白酶,酶活3600U/mg;碱性蛋白酶,酶活2000U/mg酶解鱼皮胶原蛋白,加酶量2%,在pH8.0和40℃下酶解4h,酶解后沸水中加热15min灭酶,冷却至室温,酶解液8000r/min离心20min,取上清液浓缩并冷冻干燥得到鱼皮胶原蛋白肽;
所述脂肪干细胞是将分离制备的脂肪干细胞进行消化、制成细胞悬液,按细胞密度1*105个/mL接种,采用添加了1mg/mL的SEQ ID NO:1多肽的10%胎牛血清的DMEM培养基传代培养后获得。
3.如权利要求1所述的用途,其特征在于该药物包括药学上可接受的载体。
4.如权利要求2所述的用途,其特征在于该化妆品包括药学上可接受的载体,。
5.如权利要求4所述的用途,药学上可接受的载体包括乳液。
6.如权利要求5所述的用途,乳液是水包油乳液或油包水乳液。
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