CN113143899A - 辣椒素在制备治疗溃疡性结肠炎药物中的应用 - Google Patents
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Abstract
本发明公开了辣椒素在制备治疗溃疡性结肠炎(UC)中的应用，辣椒素通过肠黏膜下微动脉的舒张，恢复肠黏膜供血，加快肠黏膜损伤后的修复，维持肠黏膜屏障的功能，从而达到有效的治疗作用，不但拓宽了辣椒素的医药用途，还为UC的治疗提供新的有效药物，具有重要的临床意义。
Description
技术领域
本发明涉及生物医药领域，具体涉及辣椒素在制备治疗溃疡性结肠炎药物中的应用。
背景技术
溃疡性结肠(UC)与克罗恩病(CD)是炎性肠病(Inflammatory bowel disease,IBD)的两种主要类型，二者皆为全球流行性疾病。在我国，UC患者的数量及发病率均显著高于CD，并呈现逐年上升的趋势。UC明显影响我国人民的健康水平，而其引起的严重并发症可威胁患者的生命。现有治疗UC的药物主要分为氨基水杨酸类药物、糖皮质激素以及生物制剂等。这些治疗药物多用于控制疾病症状，需要长期甚至终身用药，且具有较大的药物副作用。UC的发病机制不清是当前缺乏有效治疗药物的重要原因。
当前临床上用于治疗UC的药物，主要通过作用于肠道内的微生物及炎症细胞以获得抗菌消炎的有限疗效。目前急需从不同的视野寻找新的药物靶标，研发更为有效的治疗药物。众所周知，肠黏膜下微血管有效的供血功能，对维持正常的黏膜屏障功能和组织学结构发挥关键作用；而在UC中，肠黏膜下微血管的供血功能障碍。例如，UC患者的肠黏膜下微血管对迷走神经递质乙酰胆碱的舒张反应受损，导致结肠慢性炎症区域内的血流供应减少。于是，一方面可能会导致对炎症介质清除能力的下降而造成其在局部的堆积，另一方面会导致慢性炎症区域内肠黏膜损伤后修复能力的下降。二者将进一步加重肠黏膜细胞的损伤及黏膜屏障功能的丧失，从而促进UC的发生及发展。因此，肠黏膜下微血管功能的障碍可能是UC难治性的重要基础，急需一种通过恢复黏膜下微血管功能的药物，为临床防治难治性UC提供新的治疗手段。
发明内容
有鉴于此，本发明的目的之一在于提供一种辣椒素在制备治疗溃疡性结肠炎药物中的应用。
为达到上述目的，本发明提供如下技术方案：
辣椒素在制备治疗溃疡性结肠炎药物中的应用，所述辣椒素的结构如式Ⅰ所示：
优选的，所述辣椒素通过舒张人体肠黏膜下微动脉治疗溃疡性结肠炎。
优选的，所述辣椒素通过降低过氧化物酶活性治疗溃疡性结肠炎。
优选的，所述辣椒素在制备恢复溃疡性结肠炎小鼠的体重的药物中的应用。
优选的，所述辣椒素在制备恢复溃疡性结肠炎结直肠长度的药物中的应用。
本发明中所述辣椒素的有效浓度为1mg/mL。
本发明的有益效果在于：本发明公开了辣椒素在制备治疗溃疡性结肠炎中的应用，主要通过对肠黏膜下微动脉的舒张，恢复肠黏膜供血，加快肠黏膜损伤后的修复，维持肠黏膜屏障的功能，从而达到有效的治疗作用，并为UC的治疗提供了新的思路，也提供了辣椒素治疗UC的药物的新用途，与目前临床上用于治疗UC的药物，主要通过作用于肠道内的微生物及炎症细胞以获得抗菌消炎的有限疗效。本发明从不同的视野寻找全新的药物靶标，不仅提供了为溃疡性结肠炎治疗寻找到新的治疗药物，也拓宽了辣椒素的药物用途。
附图说明
为了使本发明的目的、技术方案和有益效果更加清楚，本发明提供如下附图进行说明：
图1为辣椒素能够引起人体肠黏膜下微血管内皮依赖性的舒张；
图2为辣椒素能够引起小鼠肠系膜2级微动脉内皮依赖性的舒张(A：检测结果；B:统计结果)；
图3为辣椒素具有缓解小鼠UC的作用(A体重检测结果；B：结肠长度；C：粪便特征评分；D：髓过氧化物酶(MPO)活性)。
具体实施方式
下面结合附图和具体实施例对本发明作进一步说明，以使本领域的技术人员可以更好的理解本发明并能予以实施，但所举实施例不作为对本发明的限定。
本发明中辣椒素购自美国MCE公司(货号：HY-10448)；DSS购自美国MPbio公司；MPO检测试剂盒购自美国Abcam公司(货号：ab105136)。
实施例1
利用辣椒素体外舒张人体肠黏膜下微动脉的方法，具体步骤如下：
A.将手术切除的肠道及周围肠系膜组织放入K-H液中，Krebs-Henseleit溶液包含(mM)：118NaCl，4.7KCl，1.18MgSO4、25NaHCO3、1.2KH2PO4、1.6CaCl2和11.1D-葡萄糖。
B.在显微镜下用血管镊和血管剪仔细分离肠黏膜下动脉周围脂肪结缔组织，取出2段长度为1.5～2mm的肠黏膜下动脉。用两段钨丝线(直径40μm)将血管固定于丹麦微血管张力测定仪含K-H液浴槽中(体积为5mL)，其中一段钨丝线与张力换能器连接，另一段钨丝线与血管微调装置连接。实验过程中使浴槽温度始终维持在37℃左右，持续泵入混合气(含95％O2+5％CO2)，pH值的范围维持在7.35至7.45之间。血管在正常生理状态下，通过神经体液调节处于一定张力条件下，故对分离后的血管需进行标准化，使其初始张力等于在100mmHg下血管直径的0.9倍条件下的张力。固定好的血管在K-H液中稳定20min，完成标准化后进行后续实验。
C.微动脉安装后，使血管在归一化之前在零张力下稳定20分钟。平衡期过后，通过用5μM去甲肾上腺素(NE)预先最大程度地预收缩环，然后用卡巴胆碱(CCh，100μM)松弛来评估内皮的动脉完整性。冲洗后，加入浓度为0.5μM、1μM、2μM、3μM、4μM和5μM的辣椒素进行实验。
D.将检测结果通过Graphpad软件统计分析结果，结果如图1所示。统计结果显示，辣椒素能够剂量依赖性而且内皮依赖性舒张人体的肠黏膜下微动脉。
实施例2
利用辣椒素体外舒张小鼠2级肠系膜微动脉的方法，具体步骤如下：
A.颈椎脱位处死C57BL/6小鼠后，进行剖腹手术并切除肠系膜，立即浸入冰冷的Krebs-Henseleit溶液中；Krebs-Henseleit溶液包含(mM)：118NaCl，4.7KCl，1.18MgSO4、25NaHCO3、1.2KH2PO4、1.6CaCl2和11.1D-葡萄糖。
B.在显微镜下获取肠系膜微动脉(肠系膜上动脉第二级分支，长度为2mm，直径为100至150μm)。在Krebs-Henseleit溶液中将其从周围的脂肪和结缔组织中清除，然后安装在Mulvany式钢丝肌电图仪(520A型，DMT，丹麦奥胡斯，丹麦)中进行功能评估。Powerlab分析系统(美国科罗拉多州科罗拉多斯普林斯的AD仪器公司)记录等轴测张力的变化。两条钨丝(每根直径40μm)穿过微血管内腔并固定在肌电记录仪的钳口上。室温下，将Krebs-Henseleit溶液(5mL)于37℃下用95％O2+5％CO2的混合气体连续充气。
C.微动脉安装后，使血管在归一化之前在零张力下稳定20分钟。平衡期过后，用5μM去甲肾上腺素(NE)预收缩血管，冲洗后，加入不同浓度的辣椒素进行实验。
D.将检测结果如图2中A所示。通过Graphpad软件统计分析结果，结果如图2中B所示。结果显示，辣椒素能够剂量依赖性而且内皮依赖性舒张小鼠的肠系膜微动脉。
实施例3
利用辣椒素缓解小鼠UC的方法，具体步骤如下：
A.将6周龄的C57BL/6小鼠随机分为对照组和实验组。对照组小鼠接收辣椒素(10mg/kg/天，灌胃)，而实验组的小鼠则接受含2.5％DSS的饮用水或2.5％DSS加辣椒素的组合(10mg/kg/天，灌胃)。将所有小鼠治疗7天作为实验期，并在整个实验期内每天通过测量体重，直肠出血和水消耗进行监测。粪便特征评分如下：正常(得分0)，软软且形成良好的药丸(得分1)，非常柔软(得分2)，腹泻(得分3)和严重腹泻(得分4)。隐血的评分如下：无血液(得分0)，粪便中的隐血(得分1)，粪便中可见的痕迹血液(得分2)，大出血(得分3)和直肠出血(得分4)。将这些分数相加即可获得粪便分数(最高分数为8)。实验结束时，通过颈脱位法处死小鼠以测量结肠长度，并按照MPO分析试剂盒(ab111749)所述，检测髓过氧化物酶(MPO)活性。
B.将检测结果通过Graphpad软件统计分析结果，统计结果如图3中所示。结果显示，辣椒素能够恢复DSS-结肠炎小鼠的体重与结直肠长度，改善其粪便评分。同时，辣椒素还能够通过降低MPO的活性缓解小鼠结肠的炎症。
以上所述实施例仅是为充分说明本发明而所举的较佳的实施例，本发明的保护范围不限于此。本技术领域的技术人员在本发明基础上所作的等同替代或变换，均在本发明的保护范围之内。本发明的保护范围以权利要求书为准。
Claims (5)
1.辣椒素在制备治疗溃疡性结肠炎药物中的应用，其特征在于：所述辣椒素的结构如式Ⅰ所示：
2.根据权利要求1所述辣椒素在制备治疗溃疡性结肠炎药物中的应用，其特征在于：所述辣椒素通过舒张人体肠黏膜下微动脉治疗溃疡性结肠炎。
3.根据权利要求1所述辣椒素在制备治疗溃疡性结肠炎药物中的应用，其特征在于：所述辣椒素通过降低过氧化物酶活性治疗溃疡性结肠炎。
4.根据权利要求1所述辣椒素在制备治疗溃疡性结肠炎药物中的应用，其特征在于：所述辣椒素在制备恢复溃疡性结肠炎小鼠的体重的药物中的应用。
5.根据权利要求1所述辣椒素在制备治疗溃疡性结肠炎药物中的应用，其特征在于：所述辣椒素在制备恢复溃疡性结肠炎结直肠长度的药物中的应用。
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