CN108158972B - 组合物及含有该组合物的抑制痤疮炎症的化妆品 - Google Patents
组合物及含有该组合物的抑制痤疮炎症的化妆品 Download PDFInfo
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- CN108158972B CN108158972B CN201810264465.6A CN201810264465A CN108158972B CN 108158972 B CN108158972 B CN 108158972B CN 201810264465 A CN201810264465 A CN 201810264465A CN 108158972 B CN108158972 B CN 108158972B
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Abstract
本发明涉及化妆品领域，特别涉及组合物及含有该组合物的抑制痤疮炎症的化妆品。该组合物包括黄藥树皮提取物、印度楝树叶提取物、辛酰基甘氨酸、苦参根提取物、丹参提取物、硅烷二醇水杨酸酯、槲皮素、积雪草提取物和L‑抗坏血酸磷酸酯镁。该组合物及含有该组合物的化妆品具有抑制痤疮炎症的功效。
Description
技术领域
本发明涉及化妆品领域，特别涉及组合物及含有该组合物的抑制痤疮炎症的化妆品。
背景技术
痤疮是一种与内分泌功能失调有关的毛囊，皮脂腺慢性炎症性皮肤病，是皮肤科常见的病和多发病，其临床表现为颜面部白头或黑头粉刺，丘疹，脓包，结节，囊肿，凹陷或增生性瘢痕等，严重影响患者的生活质量。痤疮的发病机制可能与皮脂腺分泌增多，毛囊口上皮角化过度，痤疮丙酸杆菌活跃或体内雄性激素水平增高有关。痤疮临床发病于青春期，她将影响到个人外貌及自尊，甚至引发心里疾病，导致生活亚健康状态。痤疮发病率高，研发发现其普遍发生在16-19岁男孩和14-17岁女孩中，在不同种族12-24岁青少年发病率高达85％，虽然痤疮影响大多数是青少年，但它在成人中仍普遍存在，比如20-30岁范围内占40-50％，甚至报道10-20％的40岁以上人亦患有痤疮。
由于痤疮患者以青春期男女较多，常可对容貌产生影响，而该人群更注重自己的外貌形象，故痤疮对患者的心理及生活质量产生较为严重的负面影响，造成患者正常生活，学习，工作，婚姻，社会交际，情绪，自信心等受到严重影响。国内外研究结果表明，痤疮会使患者产生情绪低落，焦虑，抑郁，精神紧张等不良的情绪，并因此产生严重的社会心理问题。最常见的是自尊心，自信心受损，产生尴尬，抑郁，紧张的心理，导致患者社交障碍和影响学习和工作，严重时甚至会导致患者产生自杀倾向。
随着临床医学模式从单纯的生物医学模式向生物－社会－心理医学模式转变，痤疮不仅仅是单纯生理赏的疾病，它已称为一种严重影响个人心理健康的疾病，能影响患者的容貌，情感，社交，人际关系，就业等方面，引起患者意志力减弱，产生自杀心理，焦虑抑郁，精神恍惚等症状。
目前针对痤疮的外用药物护理主要以下：
2.1维A酸
维A酸类药物外用治疗痤疮，可以消除皮脂腺堵塞，同事减少痤疮丙酸杆菌的菌群数量。但局部使用维A酸可引起不良反应，最常见的副作用对皮肤的刺激，包括干燥，红斑，刺痛，瘙痒等。
2.1过氧化苯甲酰
过氧化苯甲酰是一种特异性的微生物抑制剂，尤其对痤疮丙酸杆菌和金黄色葡萄球菌具有公安度选择性，可用于抑制毛囊内皮脂腺内的微生物并杀灭丙酸痤疮杆菌，从而改善炎症和非炎性皮损的星狂。但由于此成分对皮肤刺激性大，限制其应用。
2.3光动力治疗
光动力学是一种结合光，光敏剂和组织内分子氧，通过光动力学反应生成活性氧并选择性损伤靶向组织的治疗方法。
人体对痤疮丙酸杆菌产生卟啉光吸收的最高峰为蓝光(415nm)，其组织穿透力较弱，因此蓝光主要对于轻中度具有炎性皮损的痤疮患者有较好的治疗效果。红光(633nm)具有更强组织穿透力，不仅能刺激巨噬细胞释放细胞因子，达到一定的抗炎作用，还可以活化细胞中的纤维母细胞产生生长因子，诱导表达新胶原，促进对损伤组织的修复，因此，红光对深层次的皮损治疗效果较好。
强脉冲光
IPL所用原理是选择性光热解，常用的波长530-750nm，其靶组织主要是黑色素和血红蛋白，他们在吸收光子后被加热，破坏，最终被免疫细胞清除。IPL在临床应用于瘢痕，炎症，色素沉着的治疗，并具有创伤小，恢复快的有点。
采用光动力学治疗痤疮，主要针对细菌消灭，炎症消除，存在不足，单次治疗成本高，治疗周期长，且需要每月到医院或专业机构进行为其4-6月治疗。
2.4抗生素
红霉素，氯霉素，克林霉素，可用酒精或丙二醇配制成适宜浓度的外用药，但注意抗生素的耐药性问题，应慎重使用抗生素。外用抗生素与外用维A酸一样，可以引起皮肤局部红肿，干燥脱皮和瘙痒症状。
2.5雌激素
女性痤疮患者可采用抗雄性技术治疗，其治疗药物包括避孕药，螺内酯，西咪替丁。避孕药由乙炔雌二醇和环丙孕酮组成，是抗雄激素最常见的药物。研究显示，避孕药联用降血糖药二甲双胍治疗多卵性综合症，结果表面治疗方案可改善雄激素过多症状，缩减卵巢体积，但长期使用避孕药会影响机体糖代谢，引发高胰岛素血症。
螺内酯可选择性第破坏性腺和肾上腺的细胞色素P450酶系统，抑制雄激素生成酶活性，减少雄激素的产生。但螺内酯会产生高钾血症，肾功能损害和胃肠道反应等不良反应。
西咪替丁具有较弱的抗雄激素作用，能竞争性阻断DHT与受体结合，但不影响血清雄激素水平。但资料痤疮并不明显，且伴随精神紊乱，咽喉疼痛发热等不良反应。
2.5护理产品
市面上大多数针对青春痘护理的产品，绝大多数停留在清洁控油，化学剥离角质的应用层面，针对痤疮丙酸杆菌诱发炎症，细胞释放细胞因子，诱导免疫应答角度抑制痤疮导致的组织损伤与破坏。市面劣质化妆品，在配方中添加激素成分，严重破坏消费者皮肤屏障，导致激素脸的问题肌肤。
因此，提供一种抑制青春痘炎症的组合物及化妆品具有重要的现实意义。
发明内容
有鉴于此，本发明提供一种组合物及含有该组合物的抑制痤疮炎症的化妆品。该组合物及含有该组合物的化妆品具有抑制痤疮炎症的功效。
为了实现上述发明目的，本发明提供以下技术方案：
本发明提供了一种组合物，包括黄檗树皮提取物、印度楝树叶提取物、辛酰基甘氨酸、苦参根提取物、丹参提取物、硅烷二醇水杨酸酯、槲皮素、积雪草提取物和L-抗坏血酸磷酸酯镁。
在本发明的一些具体实施方案中，以质量份计，所述组合物包括：
在本发明的一些具体实施方案中，所述还包括甘油、丙二醇、丁二醇、寡聚透明质酸、β-葡聚糖、甜菜碱、尿囊素、烟酰胺、辛酰基甘氨酸、精氨酸、羟基乙酸、PEG-60氢化蓖麻油、乙基己基甘油、甘油辛酸酯、乳酸杆菌/梨汁发酵产物滤液、o-伞花烃-5-醇、中国地黄根提取物、黄芩根提取物、鱼腥草提取物、野大豆提取物中的一种或两者以上的混合物。
在本发明的一些具体实施方案中，以质量份计，所述组合物包括：
其中，以质量分数计，所述痤疮克星的组成为：
本发明还提供了所述的组合物在制备消除氧自由基，抑制丙酸痤疮杆菌和金黄色葡萄球菌、抑制丙酸痤疮杆菌胞外酶活性或抑制炎症因子释放的药物和/或化妆品中的应用。
本发明还提供了所述的组合物在制备治疗和/或预防痤疮中的应用。
本发明还提供了一种化妆品，包括所述的组合物以及化妆品中可接受的辅料。
在本发明的一些具体实施方案中，以质量分数计，所述化妆品包括：
其中，以质量分数计，所述痤疮克星的组成为：
在本发明的一些具体实施方案中，以质量分数计，所述化妆品包括：
在本发明的一些具体实施方案中，以质量分数计，所述化妆品包括：
在本发明的一些具体实施方案中，以质量分数计，所述化妆品包括：
在本发明的一些具体实施方案中，所述化妆品为祛痘洁面乳、祛痘水、祛痘精华、祛痘膏或祛痘面膜。凡是化妆品中可接受的剂型均在本发明的保护范围之内，本发明在此不做限定。
本发明还提供了所述的化妆品的制备方法，包括如下步骤：
步骤1：称量去离子水；
步骤2：分别称量甘油、丙二醇、丁二醇、甜菜碱、尿囊素、β-葡聚糖、寡聚透明质酸、烟酰胺、硅烷二醇水杨酸酯、辛酰基甘氨酸、精氨酸，加热至75～80℃，以150～200r/min，搅拌30min，以6000～7000r/min均质3min；
步骤3：分别称取L-抗坏血酸磷酸酯镁，黄原胶，降温至60～75℃加入，以200～250r/min，搅拌30min，以8000～9000r/min均质3min；
步骤4：降低搅拌速率至150～200r/min，温度降至45～55℃，分别称取苦参根提取物、槲皮素、痤疮克星、丹参提取物和积雪草提取物，搅拌20～30min；
步骤5：温度降低45～50℃，称量PHL，搅拌10min冷却。
本发明还提供了所述的化妆品或如所述的制备方法制得的化妆品在制备消除氧自由基，抑制丙酸痤疮杆菌和金黄色葡萄球菌、抑制丙酸痤疮杆菌胞外酶活性或抑制炎症因子释放的药物和/或化妆品中的应用。
本发明还提供了所述的化妆品或如所述的制备方法制得的化妆品在制备治疗和/或预防痤疮中的应用。
本发明针对痤疮诱发的炎症阶段产生的靶点进行有效控制，通过抑制痤疮丙酸杆菌胞外酶，包括脂肪酶，蛋白酶，透明质酸酶；通过抑制痤疮丙酸杆菌和金黄色葡萄球菌，通过消除氧自由基和炎症因子引起免疫应答。
本发明采用槲皮素和苦参根提取物有效抑制痤疮丙酸杆菌分泌脂肪水解酶；本发明通过L-抗坏血酸磷酸酯镁协同抑制痤疮丙酸杆菌分泌卟啉，从而有效抑制脂氧化；黄柏树皮提取物有效抑制透明质酸酶的活性；本发明采用辛酰基甘氨酸和苦参根提取物协同抑制丙酸痤疮杆菌和金黄色葡萄球菌增殖；
本发明采用槲皮素、硅烷二醇水杨酸酯有效清除氧自由基、采用L-抗坏血酸磷酸酯镁消除羟基自由基，有效避免自由基引起的组织损伤。
本发明采用积雪草提取物抑制炎症因子白介素1L-1α，采用槲皮素有效抑制组胺和白三烯的释放；采用硅烷二醇水杨酸酯有效抑制TNF-α和1L-8的释放，采用印度楝树叶提取物有效抑制花生四烯酸的活化，通过对炎症因子释放的有效抑制，避免机体免疫应答导致的组织损伤。
本发明采用槲皮素和苦参根提取物有效抑制痤疮丙酸杆菌分泌脂肪水解酶；本发明通过L-抗坏血酸磷酸酯镁协同抑制痤疮丙酸杆菌分泌卟啉，从而有效抑制脂氧化；黄柏树皮提取物有效抑制透明质酸酶的活性；本发明采用辛酰基甘氨酸和苦参根提取物协同抑制丙酸痤疮杆菌和金黄色葡萄球菌增殖；
本发明采用槲皮素、硅烷二醇水杨酸酯有效清除氧自由基、采用L-抗坏血酸磷酸酯镁消除羟基自由基，有效避免自由基引起的组织损伤。
本发明采用积雪草提取物抑制炎症因子白介素1L-1α，采用槲皮素有效抑制组胺和白三烯的释放；采用硅烷二醇水杨酸酯有效抑制TNF-α和1L-8的释放，采用印度楝树叶提取物有效抑制花生四烯酸的活化，通过对炎症因子释放的有效抑制，避免机体免疫应答导致的组织损伤。
本发明广泛应用于肌肤护理相关的产品中，如祛痘洁面乳，祛痘水，祛痘精华，祛痘膏，祛痘面膜等产品中。
本发明针对痤疮诱发的炎症阶段产生的靶点进行有效控制，通过抑制痤疮丙酸杆菌胞外酶，包括脂肪酶，蛋白酶，透明质酸酶；通过抑制痤疮丙酸杆菌和金黄色葡萄球菌，通过消除氧自由基和炎症因子引起免疫应答。
本发明提供的组合物及含有该组合物的化妆品能够消除氧自由基，抑制丙酸痤疮杆菌和金黄色葡萄球菌、抑制丙酸痤疮杆菌胞外酶活性，抑制炎症因子释放。
本发明广泛应用于肌肤护理相关的产品中，如祛痘洁面乳，祛痘水，祛痘精华，祛痘膏，祛痘面膜等产品中。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案，下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍。
图1示皮肤油脂含量的检测结果；
图2示一氧化氮释放量的检测结果。
具体实施方式
本发明公开了一种组合物及含有该组合物的抑制痤疮炎症的化妆品，本领域技术人员可以借鉴本文内容，适当改进工艺参数实现。特别需要指出的是，所有类似的替换和改动对本领域技术人员来说是显而易见的，它们都被视为包括在本发明。本发明的方法及应用已经通过较佳实施例进行了描述，相关人员明显能在不脱离本发明内容、精神和范围内对本文所述的方法和应用进行改动或适当变更与组合，来实现和应用本发明技术。
本发明提供的组合物及含有该组合物的抑制痤疮炎症化妆品中所用原料、辅料及试剂均可由市场购得。
其中，痤疮克星购自一丸公司；
本发明中所述的痤疮克星成分组成：
下面结合实施例，进一步阐述本发明：
实施例1
添加黄原胶0.2％，PHL为1％，剩余添加去离子水至100％,配制祛痘水组分进行功效评价，其配方工艺如下：
1)按照工艺配比称量其去离子水；
2)分别称量甘油、丙二醇、丁二醇、甜菜碱、尿囊素、β-葡聚糖、寡聚透明质酸、硅烷二醇水杨酸酯、辛酰基甘氨酸和精氨酸，加热至75℃，以180r/min，搅拌30min，以6000r/min均质3min；
3)分别称取L-抗坏血酸磷酸酯镁，黄原胶，降温至68℃加入，以230r/min，搅拌30min，以8500r/min均质3min；
4)降低搅拌速率至150r/min，温度降至45℃，分别称取苦参根提取物，槲皮素，痤疮克星、丹参提取物和积雪草提取物，搅拌20min；
5)温度降低48℃，称量防腐剂PHL，搅拌10min冷却室温即可。
实施例2
添加黄原胶0.2％，PHL为1％，剩余添加去离子水至100％,配制祛痘水组分进行功效评价，其配方工艺如下：
1)按照工艺配比称量其去离子水；
2)分别称量甘油、丙二醇、丁二醇、甜菜碱、尿囊素、β-葡聚糖、寡聚透明质酸，硅烷二醇水杨酸酯，辛酰基甘氨酸和精氨酸，加热至80℃，以200r/min，搅拌30min，以7000r/min均质3min；
3)分别称取L-抗坏血酸磷酸酯镁，黄原胶，降温至75℃加入，以250r/min，搅拌30min，以9000r/min均质3min；
4)降低搅拌速率至200r/min，温度降至50℃，分别称取苦参根提取物，槲皮素，痤疮克星，积雪草提取物和丹参提取物，搅拌30min；
5)温度降低50℃，称量防腐剂PHL，搅拌10min冷却室温即可。
实施例3
添加黄原胶0.2％，PHL为1％，剩余添加去离子水至100％,配制祛痘水组分进行功效评价，其配方工艺如下：
1)按照工艺配比称量其去离子水；
2)分别称量甘油、丙二醇、丁二醇、甜菜碱、尿囊素、β-葡聚糖、寡聚透明质酸，硅烷二醇水杨酸酯，辛酰基甘氨酸和精氨酸，加热至80℃，以200r/min，搅拌30min，以7000r/min均质3min；
3)分别称取L-抗坏血酸磷酸酯镁，黄原胶，降温至60℃加入，以200r/min，搅拌30min，以8000r/min均质3min；
4)降低搅拌速率至180r/min，温度降至55℃，分别称取苦参根提取物，槲皮素，痤疮克星，积雪草提取物和丹参提取物，搅拌25min；
5)温度降低45℃，称量防腐剂PHL，搅拌10min冷却室温即可。
对比例1
| 甘油 | 2％ |
| 丙二醇 | 3％ |
| 丁二醇 | 3％ |
| 寡聚透明质酸 | 0.01％ |
| β-葡聚糖 | 0.1％ |
| 甜菜碱 | 0.5％ |
| 尿囊素 | 0.15％ |
1)按照工艺配比称量其去离子水；
2)分别称量甘油、丙二醇、丁二醇、甜菜碱、尿囊素、β-葡聚糖、寡聚透明质酸，加热至75～80℃，以150～200r/min，搅拌30min，以6000～7000r/min均质3min；
3)称取黄原胶，降温至60～75℃加入，以200～250r/min,搅拌30min,以8000～9000r/min均质3min；
4)温度降低45～50℃，称量防腐剂PHL，搅拌10min冷却室温即可。
对比例2
| 甘油 | 2％ |
| 丙二醇 | 3％ |
| 丁二醇 | 3％ |
| 寡聚透明质酸 | 0.01％ |
| β-葡聚糖 | 0.1％ |
| 甜菜碱 | 0.5％ |
| 尿囊素 | 0.15％ |
| 辛酰基甘氨酸 | 0.5％ |
| 精氨酸 | 0.5％ |
| 痤疮克星 | 0.5％ |
1)按照工艺配比称量其去离子水；
2)分别称量甘油、丙二醇、丁二醇、甜菜碱、尿囊素、β-葡聚糖、寡聚透明质酸，辛酰基甘氨酸和精氨酸，加热至75～80℃，以150～200r/min，搅拌30min，以6000～7000r/min均质3min；
3)称取黄原胶，降温至60～75℃加入，以200～250r/min，搅拌30min，以8000～9000r/min均质3min；
4)降低搅拌速率至150～200r/min，温度降至45～55℃，称取痤疮克星，搅拌20～30min；
5)温度降低45～50℃，称量PHL，搅拌10min冷却。
对比例3
1)按照工艺配比称量其去离子水；
2)分别称量甘油、丙二醇、丁二醇、甜菜碱、尿囊素、β-葡聚糖、寡聚透明质酸，辛酰基甘氨酸和精氨酸，加热至75～80℃，以150～200r/min，搅拌30min，以6000～7000r/min均质3min；
3)称取黄原胶，降温至60～75℃加入，以200～250r/min，搅拌30min，以8000～9000r/min均质3min；
4)降低搅拌速率至150～200r/min，温度降至45-55℃，称取苦参根提取物和痤疮克星搅拌20-30min；
5)温度降低45～50℃，称量防腐剂PHL，搅拌10min冷却室温即可。
对比例4
| 甘油 | 2％ |
| 丙二醇 | 3％ |
| 丁二醇 | 3％ |
| 寡聚透明质酸 | 0.01％ |
| β-葡聚糖 | 0.1％ |
| 甜菜碱 | 0.5％ |
| 尿囊素 | 0.15％ |
| 辛酰基甘氨酸 | 0.5％ |
| 精氨酸 | 0.5％ |
| 痤疮克星 | 0.5％ |
| 苦参根提取物 | 0.1％ |
| 槲皮素 | 0.01％ |
| 积雪草提取物 | 0.3％ |
| 丹参提取物 | 0.2％ |
1)按照工艺配比称量其去离子水；
2)分别称量甘油、丙二醇、丁二醇、甜菜碱、尿囊素、β-葡聚糖、寡聚透明质酸，辛酰基甘氨酸和精氨酸，加热至75～80℃，以150～200r/min，搅拌30min，以6000～7000r/min均质3min；
3)称取黄原胶，降温至60～75℃加入，以200～250r/min，搅拌30min，以8000～9000r/min均质3min；
4)降低搅拌速率至150～200r/min，温度降至45～55℃，分别称取苦参根提取物，槲皮素，痤疮克星，丹参提取物和积雪草提取物，搅拌20～30min；
5)温度降低45～50℃，称量防腐剂PHL，搅拌10min冷却室温即可。
对比例5
| 甘油 | 6％ |
| 丙二醇 | 0.5％ |
| 丁二醇 | 1％ |
| 寡聚透明质酸 | 0.5％ |
| β-葡聚糖 | 0.001％ |
| 甜菜碱 | 0.01％ |
| 尿囊素 | 0.3％ |
| 辛酰基甘氨酸 | 0.01％ |
| 精氨酸 | 0.01％ |
| 痤疮克星 | 0.5％ |
| 苦参根提取物 | 0.1％ |
| 槲皮素 | 0.05％ |
| 积雪草提取物 | 0.001％ |
| 丹参提取物 | 0.001％ |
| L-抗坏血酸磷酸酯镁 | 3.2％ |
| 硅烷二醇水杨酸酯 | 1.2％ |
1)按照工艺配比称量其去离子水；
2)分别称量甘油、丙二醇、丁二醇、甜菜碱、尿囊素、β-葡聚糖、寡聚透明质酸，硅烷二醇水杨酸酯，辛酰基甘氨酸和精氨酸，加热至75～80℃，以150～200r/min，搅拌30min，以6000～7000r/min均质3min；
3)分别称取L-抗坏血酸磷酸酯镁，黄原胶，降温至60～75℃加入，以200～250r/min，搅拌30min，以8000～9000r/min均质3min；
4)降低搅拌速率至150～200r/min，温度降至45～55℃，分别称取苦参根提取物，槲皮素，痤疮克星，丹参提取物和积雪草提取物，搅拌20～30min；
5)温度降低45～50℃，称量防腐剂PHL，搅拌10min冷却室温即可。
实施例4 控油功效评价
测试原理：
本发明采用德国CK的皮肤油脂测试仪Sebumeter SM815进行吸油纸使用效果的功效评定。油份测试采用的是世界公认的SEBUMETER法，它是基于光度计原理，一种0.1mm厚的特殊消光胶带吸收人体皮肤上的油脂后，就会变成一种半透明的胶带，它的透光量就会发生变化，吸收的油脂越多，透光量就会越大，这样就可以测量出皮肤油脂的含量。最大的优点是测试探头体积小，使用方便，可测试皮肤的任何部位。这是一种油脂腺分泌物的间接测量法，结果可以用来区分不同的皮肤类型，使准确地了解由内部和外部原因而引起的油脂变化成为可能。
测试方法：
本发明此次测试选择21名测试对象，3人一组分成7组，分别在额头画出3个尺寸为3cm*3cm区域，分别针对空白样(不擦拭样品)、对照样和实施例样进行皮肤油脂含量评测，每个对照样取三人平均值计算油脂含量增量百分比进行相关评判。
(1)选择的测试对象肌肤健康，年龄18-30岁，经测试对象知情，同意，受试对象洁面15分钟后，测定其额头油脂含量作为初始值。
(2)在受试者额头空白区不涂抹样品，对照样区、发明样区称取0.5ml精华液以3cm*3cm无纺布膜布浸润覆盖肌肤20min后，取下无纺布，静待10分钟，分别针对三个区域进行油脂含量测定，每30min测试一次，持续评测2h；
(3)测试环境：测试环境温度T＝25℃，湿度H＝50；
(4)测试时间：在测试前进行洁面，15分钟测试初始油脂含量，分别测试30min,60min,90min,120min测试油脂含量数据；
(5)测试部位：额头标注空白，对照样，实施例样的测试区域。
功效评测结果如表1、图1所示：
表1
结论，本发明通过抑制痤疮丙酸杆菌胞外酶，蛋白酶，脂肪酶，透明质酸水解酶的作用，有效抑制油脂分泌，避免皮脂腺导管堵塞，从而降低痤疮丙酸杆菌增殖的几率。本发明具有较好的抑制油脂分泌作用。
实施例5 LPS诱导RAW264.7细胞释放NO抑制作用
实验原理：
LPS(脂多糖)：
位于革兰氏阴性细菌细胞壁最外层的一层较厚(8-10nm)的类脂多糖类物质，由类脂A、核心多糖和O-特异侧链3部分组成。脂多糖是内毒素和重要群特异性抗原(O抗原)。脂多糖由三部分组成。脂质A(LipidA)为构成内毒素活性的糖脂，以共价键联结到杂多糖链，有两部分：一是核心多糖，在有关的株内是恒定的；另一O特异性链(O-specific chain)是高度可变的。大肠杆菌的脂多糖在实验室免疫学中是常用的B细胞促分裂因子即多克隆活化因子(polyclonal activator)。本实验利用LPS诱导RAW264.7细胞产生过敏反应，释放NO(一氧化氮)。
Griess assay–NO含量测试方法
NO在体内或水溶液中极易氧化成NO2，在酸性条件下，NO与重氮盐磺胺发生重氮反应，并生成重氮化合物，后者进一步与萘基乙烯基二胺发生耦合反应，该反应生成的产物浓度与NO浓度具有线性关系，在540nm处有最大吸收峰。
实验材料和方法：
材料：DMEM，FBS，LPS，MTT，Griess reagent
实验方法：
RAW 264.7细胞培养
将RAW264.7细胞用移液枪吸取DMEM培基吹散分散细胞，利用Hemacytometer来计数细胞，然后利用DMEM来稀释细胞，稀释到浓度为5×104cells/ml。
稀释后的细胞溶液分别接种到96well中，每一孔为100ul，即5×103cells/well。
在37℃，5％CO2的培养箱中培养24小时。
待测样品和空白样准备：待测样品用DMEM培基稀释，稀释后的浓度分别为：1％(该浓度通过前面的MTT测试，结果为无毒)
名称Control Sample
样品组成LPS 10ppm LPS 10ppm+1％sample
细胞培养24小时后，观察细胞是否完全贴壁生长，如果细胞完全贴壁生长的话，将原培基移除，用DPBS洗涤。
将DPBS移除之后，分别加入前面准备好的样品。样品的溶度根据毒性测试的结果选择安全浓度1％(20ul/well)。
样品加入之后，放入37℃，5％CO2的培养箱中培养20小时。
20小时后，移取50ul的培养基到新的96Well上，加入相同体积的1％Griessreagent，充分混合后，于暗处反应10分钟。
利用ELISA reader在540nm处测试样品的吸光光度值。
NO抑制作用
NO Inhibition％＝((〖OD〗_Control-〖OD〗_Sample)/〖OD〗_Control)*100％
其中：〖OD〗_Control-空白样的吸光光度值
〖OD〗_Sample-样品的吸光光度值
抑制炎症结果如表2、图2所示：
表2
结论：本专利具有很好的抑制炎症介质ON释放，实例样抑制率可达36.37％-41.61％，对照样仅15.22％，其抑制率提升大于200％，具有显著差异(P＜0.05)。表明本发明提供的化妆品具有很好的抑制炎症因子表达，从而有效抑制痤疮引起炎症因子对皮肤组织的损伤。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。
Claims (3)
1.一种治疗和/或预防痤疮的化妆品，其特征在于，以质量分数计，由如下组分组成：
其中，以质量分数计，所述痤疮克星的组成为：
所述的化妆品的制备方法，包括如下步骤：
步骤1：称量去离子水；
步骤2：分别称量甘油、丙二醇、丁二醇、甜菜碱、尿囊素、β-葡聚糖、寡聚透明质酸、烟酰胺、硅烷二醇水杨酸酯、辛酰基甘氨酸、精氨酸，加热至75~80℃，以150~200r/min，搅拌30min，以6000~7000r/min 均质3min；
步骤3：分别称取L-抗坏血酸磷酸酯镁，黄原胶，降温至60~75℃加入，以200~250r/min，搅拌30min，以8000~9000r/min 均质3min；
步骤4：降低搅拌速率至150~200r/min，温度降至45~55℃，分别称取苦参根提取物、槲皮素、痤疮克星、丹参提取物和积雪草提取物，搅拌20~30min；
步骤5：温度降低45~50℃，称量PHL，搅拌10min 冷却。
2.根据权利要求1所述的化妆品在制备消除氧自由基，抑制丙酸痤疮杆菌和金黄色葡萄球菌、抑制丙酸痤疮杆菌胞外酶活性或抑制炎症因子释放的化妆品中的应用。
3.根据权利要求1所述的化妆品在制备治疗和/或预防痤疮的化妆品中的应用。
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