CN116904392A - 烟酸在促进脂肪细胞分化中的用途 - Google Patents
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Abstract
本发明公开了烟酸在促进脂肪细胞分化中的用途。本发明通过在诱导3T3‑L1细胞向脂肪细胞分化过程中施加烟酸，发现3T3‑L1细胞向脂肪细胞分化的能力明显增强。该用途不仅扩展了烟酸作为必需维生素的功能，还为功能受损脂肪细胞的补充和修复提供了新的思路。
Description
技术领域
本发明属于药物研发技术领域，具体涉及烟酸在促进向脂肪细胞分化中的用途。
背景技术
脂肪组织功能障碍会引发一系列的代谢性疾病，例如，胰岛素抵抗、脂代谢异常以及高脂血症，严重威胁着人类的生命健康。然而，由于脂肪组织功能障碍是一种复杂的多因素疾病，其致病机制呈现出多样性，因此目前并没有很好的靶向治疗方案。近年来，有研究表明脂肪形成缺陷可能是导致脂肪组织功能障碍的影响因素，脂肪细胞分化异常会造成机体内脂代谢紊乱。3T3-L1细胞系是用于体外研究脂肪分化的经典细胞系，通过探索发现能够促进3T3-L1细胞向脂肪细胞分化的新药物，将会为修复脂肪细胞的异常分化增添一种新可能，有助于预防和缓解脂肪功能障碍。
烟酸(Nicotinic acid，NA)，又称作尼可丁酸，属于维生素B族维生素，化学式：C6H5NO2，分子量约为123.11，CAS号59-67-6，其结构式为如下所示：
在常温下NA纯品呈无色针状晶体，味微酸，可溶于水及酒精。烟酸理化性质稳定，结构简单，是人体所必须的13种维生素之一。NA以辅酶形式存在于食物中，经肝脏转化为烟酰胺腺嘌呤二核苷酸(辅酶Ⅰ，NAD)和烟酰胺腺嘌呤二核苷酸磷酸(辅酶Ⅱ，NADP)，在呼吸链中起到递氢体的作用。并且烟酸可通过cAMP/PKA通路抑制脂解，进而降低游离脂肪酸和甘油三酯水平，从而发挥治疗动脉硬化作用。但是，烟酸是否能够促进3T3-L1细胞向脂肪分化的能力目前尚未见报道。
发明内容
本发明的目的在于提供烟酸在促进向脂肪细胞分化中的用途。
烟酸在制备促进向脂肪细胞分化的培养基中的用途。
具体的，通过在培养基中添加烟酸，促进3T3-L1细胞向脂肪细胞分化。
烟酸在制备促进脂肪细胞分化的药物中的用途。
一种促进3T3-L1细胞向脂肪细胞分化的药用组合物，包括烟酸和其药学上可接受的盐。
优选的，还包括一种或多种药学上可接受的辅料或载体。
所述辅料或载体包括稀释剂、赋形剂、填充剂、粘合剂、湿润剂、崩解剂、吸收促进剂、表面活性剂、吸附载体或润滑剂。
所述药用组合物可制成的剂型为片剂、胶囊剂、泡腾片、颗粒剂、散剂、分散片、口服液、丸剂或注射剂。
本发明的有益效果：本发明发现烟酸能促进3T3-L1细胞向脂肪细胞的分化和成熟，进而补充和修复功能受损的脂肪细胞。该作用可应用于预防、治疗和改善体内脂肪发育不良所引起的胰岛素抵抗、血脂异常等代谢性疾病。
附图说明
图1是采用Western blot方法检测所诱导的脂肪细胞在烟酸处理后脂肪分化标志分子Pparγ和Adipsin的蛋白表达水平。
图2是采用油红O染色检测所诱导的脂肪细胞在烟酸处理后脂滴大小和含量的变化情况。
具体实施方式
为了便于理解本发明，下面将对本发明进行更全面的描述。但是，本发明可以以许多不同的形式来实现，并不限于本文所描述的实施例。相反地，提供这些实施例的目的是使对本发明的公开内容的理解更加透彻全面。
实施例1烟酸对3T3-L1细胞向脂肪细胞分化的作用
1.烟酸的配制
本发明实施例中所使用的烟酸购自Selleck公司，产品目录号：S1744，CAS号59-67-6，取50mg烟酸粉末，加入4.0614ml二甲基亚砜，震荡使其充分溶解，混匀，此储液终浓度为100mM。分装后保存于-20℃。
2.诱导3T3-L1细胞向脂肪细胞分化的方案
将3T3-L1细胞按照3x105/ml的密度铺于60mm培养皿中，待细胞长满，更换新鲜培养基继续培养48h使细胞接触抑制。按表1的培养基配方配制相应的分化培养基和维持培养基，细胞接触抑制后更换分化培养基培养48h，再更换维持培养基培养，之后每48h更换一次培养基。
表1
3.细胞的处理与实验分组
取诱导分化5天的3T3-L1细胞并按如下组别进行干预：
对照组：对照组为阴性对照，培养基中不添加烟酸；
烟酸干预组：在培养基中加入烟酸并使终浓度为200nM。
4.Western Blot实验
按照常规方法收集并裂解诱导分化6天的3T3-L1细胞，提取总蛋白后进行蛋白免疫印迹分析，检测脂肪分化标志分子Pparγ和Adipsin的表达变化。实验所用抗体如表2所示：
表2
5.油红O染色
3T3-L1细胞诱导分化6天后，缓慢吸去细胞培养液，用PBS洗涤1次，加入4％多聚甲醛固定10min，PBS漂洗2次。根据样品数量及每个样品所需的染色工作液的体积，按照油红O溶液和油红O稀释液3:2的比例配制油红O染色工作液，混匀后静置10min，然后用0.45μm针头滤器过滤，需在两小时内使用。加入适量染色洗涤液覆盖细胞20秒，吸除染色洗涤液，加入适量油红O染色工作液均匀覆盖细胞，染色10-20分钟。去除油红O染色工作液，加入适量染色洗涤液，静置30秒，然后再去除染色洗涤液，用PBS洗涤20秒。加入适量PBS均匀覆盖细胞，在光学显微镜下观察和拍照。
实验结果：
如图1所示，在3T3-L1细胞向脂肪细胞分化过程中施加烟酸，与DMSO对照组相比，烟酸显著促进了Pparγ和Adipsin蛋白的表达。同时，油红O染色的结果也显示，烟酸处理后细胞中脂滴的数目增多且脂质含量也显著升高(图2)。以上结果说明，烟酸促进了3T3-L1细胞向脂肪细胞的分化。
以上所述实施例仅表达了本发明的几种实施方式，其描述较为具体和详细，但并不能因此而理解为对发明专利范围的限制。应当指出的是，对于本领域的普通技术人员来说，在不脱离本发明构思的前提下，还可以做出若干变形和改进，这些都属于本发明的保护范围。因此，本发明专利的保护范围应以所附权利要求为准。
Claims (7)
1.烟酸在制备促进向脂肪细胞分化的培养基中的用途。
2.根据权利要求1所述烟酸在制备促进脂肪细胞分化的培养基中的用途，其特征在于，通过在培养基中添加烟酸，促进3T3-L1细胞向脂肪细胞分化。
3.烟酸在制备促进3T3-L1细胞向脂肪细胞分化的药物中的用途。
4.一种促进3T3-L1细胞向脂肪细胞分化的药用组合物，其特征在于，包括烟酸和其药学上可接受的盐。
5.根据权利要求4所述促进3T3-L1细胞向脂肪细胞分化的药用组合物，其特征在于，还包括一种或多种药学上可接受的辅料或载体。
6.根据权利要求5所述促进3T3-L1细胞向脂肪细胞分化的药用组合物，其特征在于，所述辅料或载体包括稀释剂、赋形剂、填充剂、粘合剂、湿润剂、崩解剂、吸收促进剂、表面活性剂、吸附载体或润滑剂。
7.根据权利要求4所述促进3T3-L1细胞向脂肪细胞分化的药用组合物，其特征在于，所述药用组合物可制成的剂型为片剂、胶囊剂、泡腾片、颗粒剂、散剂、分散片、口服液、丸剂或注射剂。
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