CN106963844B - 番茄水溶性皂苷提取物在制备治疗高尿酸血症药物或保健品中的应用 - Google Patents
番茄水溶性皂苷提取物在制备治疗高尿酸血症药物或保健品中的应用 Download PDFInfo
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Abstract
本发明公开了番茄水溶性皂苷提取物在制备治疗高尿酸血症药物或保健品中的应用。申请人通过大量实验发现，来自番茄的番茄水溶性皂苷提取物可有效降低血清中的尿酸水平，可用于制备治疗高尿酸血症及相关疾病的药物或保健品。
Description
技术领域
本发明涉及番茄水溶性皂苷提取物的应用，具体涉及番茄水溶性皂苷提取物在制备治疗高尿酸血症药物或保健品中的应用。
背景技术
尿酸是人体嘌呤代谢的产物。人体嘌呤来源有两种，内源性依赖自身合成或核酸降解(大约600mg/d)，约占体内总尿酸量的80％；外源性为摄入的嘌呤饮食(大约100mg/d)，约占体内总尿酸量的20％。正常情况下，人体每天尿酸的产生和***基本上保持动态平衡，凡是影响血尿酸生成和***的因素均可以导致血尿酸水平增高，从而引起高尿酸血症。近年来，一些大规模、前瞻性研究表明，血尿酸升高与肾脏疾病、动脉粥样硬化、原发性高血压、脑卒中、心血管事件的发生和死亡等呈独立正相关。高尿酸血症是21世纪全球重要的公共健康问题之一，亦是我国愈来愈重要的代谢性疾病问题，随着我国经济社会的发展及饮食结构和生活方式的改变，在今后几十年，高尿酸血症的患病率预计将成倍增加，这些患者将给社会及家庭造成极大的压力，因此，研究有效方法预防和治疗高尿酸血症的发生具有重要意义。
番茄是我们日常食用的蔬菜，其中含有较高含量的水溶性皂苷类化合物。 2004年日本熊本大学药学部学者藤原章雄，野原稔弘等首先从日本产的一种称为桃太郎的番茄中分到得到两个结构复杂的甾体生物碱皂苷，命名为番茄皂苷A(Esculeoside A)和番茄皂苷B(Esculeoside B)(Tetrahedron 2004, 60:4915～4920)。番茄皂苷A(Esculeoside A)是番茄中主要的皂苷成分，在原料中的含量为番茄红素的3倍。通过进一步药理活性研究他们发现番茄皂苷A及其苷元(Esculeogenin A)体外可抑制泡沫细胞的形成，为一种较好的酰基辅酶A胆固醇酰基转移酶(acyl-coA:cholesterol acyltransferase,ACAT)抑制剂，是有前景的抗动脉粥样硬化症化合物 (Arterioscler Thromb Vasc Biol.2007,27:2400～2406)。但目前尚未见有番茄水溶性皂苷提取物可以应用于高尿酸血症的治疗或预防的相关报道。
发明内容
本发明要解决的技术问题是提供番茄水溶性皂苷提取物在制备治疗高尿酸血症药物或保健品中的应用。
本发明的技术方案为：番茄水溶性皂苷提取物在制备治疗高尿酸血症药物或保健品中的应用。
上述技术方案中，所述番茄水溶性皂苷提取物可采用现有常规技术进行制备，优选本申请中的番茄水溶性皂苷提取物的制备方法包括：将新鲜番茄打浆、酶解，酶解液过滤，收集滤液，干燥，制得番茄水溶性皂苷提取物粗品。在酶解时，所用的酶通常为果胶酶，其用量为原料重量的0.04～0.06％。
上述制得的是番茄水溶性皂苷提取物粗品，在其用量较大的情况下才会具有较好的治疗高尿酸血症的效果，为了减少在治疗时番茄水溶性皂苷提取物的用量，优选在番茄水溶性皂苷提取物的制备方法中进一步包括纯化步骤。所述的纯化步骤为现有技术中常规使用的可以纯化皂苷类物质的方法，如树脂柱层析或硅胶柱层析等操作，本申请中，具体的纯化步骤为：将番茄水溶性皂苷提取物粗品经大孔树脂柱或硅胶柱层析，用体积浓度为80～100％的乙醇洗脱，回收溶剂，干燥，得到纯化后的番茄水溶性皂苷提取物。其中，大孔树脂的型号具体可以是D101型、AB-8型或Diaion HP20型。由前述纯化步骤所得的纯化后的番茄水溶性皂苷提取物中，番茄皂苷A的含量通常都大于或等于17％。
进一步地，本发明还提供一种以番茄水溶性皂苷提取物为有效成分制备的治疗高尿酸血症的药物或保健品。所述药物或保健品的剂型可以是药学上可接受的剂型，如胶囊剂、片剂或颗粒剂等常规剂型。
申请人通过大量实验发现，来自番茄的番茄水溶性皂苷提取物可有效降低血清中的尿酸水平，可用于制备治疗高尿酸血症及相关疾病的药物或保健品。
具体实施方式
下面结合实验进一步说明番茄水溶性皂苷提取物在降低血清中尿酸水平的作用。
一、药物的制备与实验方案
1.番茄水溶性皂苷提取物的制备
1)精选成熟、无腐烂的新鲜樱桃小番茄果实1000g，洗净，用搅碎机打碎成浆汁；加入相当于鲜果重量0.05％的果胶酶，搅拌均匀，60℃保温酶解澄清2小时；
2)酶解液先用100目滤布粗滤，粗滤液经离心机离心10min，转速为 4000r/min，分离完毕后倾出上清液；
3)上清液上D101型大孔型吸附树脂(树脂用量为原料量的0.05倍，吸附流速为3BV/h)，然后用去离子水洗树脂柱直至流出液澄清，再用5倍柱体积、100％的乙醇洗脱(流速为9BV/h)，收集洗脱液；
4)将洗脱液置于旋转蒸发仪中，在0.05Mpa、60℃条件下减压回收乙醇，并进一步在0.07Mpa条件下减压浓缩至浸膏状，浸膏转入真空干燥箱，在真空度为0.09Mpa，温度为60℃条件下真空干燥，粉碎，得到番茄总皂苷2.7g，得率为0.27％。
经蒸发光散射－高效液相色法(色谱条件：色谱柱ZORBAX SB-C18柱；柱温30℃；流动相为甲醇-水梯度洗脱；检测时间20min；流速0.8mL/min；进样量10μL；蒸发光散射检测器参数为漂移管温度100℃；空气流速： 2.7L/min)检测番茄水溶性皂苷提取物中皂苷单体番茄皂苷A的含量为 23.83％。
2.番茄水溶性皂苷提取物对正常小鼠血尿酸的作用
取SPF级昆明种小鼠48只，按照体重随机分为4组，每组12只，即正常对照组(蒸馏水)、番茄水溶性皂苷提取物低剂量组(100mg/kg)、番茄水溶性皂苷提取物中剂量组(200mg/kg)、番茄水溶性皂苷提取物高剂量组 (400mg/kg)。小鼠适应性喂养1周后，连续给药14天。各组每天上午灌胃给药1次，给药体积均为0.2ml/10g。末次给药后，采血分离血清，测定血尿酸值。
3.番茄水溶性皂苷提取物对氧嗪酸钾和次黄嘌呤诱导的高尿酸血症小鼠血尿酸的作用
取SPF级昆明种小鼠60只，按照体重随机分为5组，每组12只，即正常对照组(蒸馏水)、高尿酸血症模型组(蒸馏水)、番茄水溶性皂苷提取物低剂量组(100mg/kg)、番茄水溶性皂苷提取物中剂量组(200mg/kg)、番茄水溶性皂苷提取物高剂量组(400mg/kg)。小鼠适应性喂养1周后，连续给药 14天。各组每天上午灌胃给药1次，给药体积均为0.2ml/10g。在14d时，除正常对照组外，其余各组用氧嗪酸钾(100mg/kg)皮下注射和次黄嘌呤 (200mg/kg)腹腔注射造模，2h后采血分离血清，测定血尿酸值。
4.番茄水溶性皂苷提取物对氧嗪酸钾和尿酸诱导的高尿酸血症小鼠血尿酸的作用
取SPF级昆明种小鼠60只，按照体重随机分为5组，每组12只，即正常对照组(蒸馏水)、高尿酸血症模型组(蒸馏水)、番茄水溶性皂苷提取物低剂量组(100mg/kg)、番茄水溶性皂苷提取物中剂量组(200mg/kg)、番茄水溶性皂苷提取物高剂量组(400mg/kg)。小鼠适应性喂养1周后，连续给药 14天。各组每天上午灌胃给药1次，给药体积均为0.2ml/10g。在14d时，除正常对照组外，其余各组用氧嗪酸钾(100mg/kg)皮下注射和尿酸(300mg/kg) 腹腔注射造模，2h后采血分离血清，测定血尿酸值。
二、实验结果
1.番茄水溶性皂苷提取物对正常小鼠血尿酸的作用
如表1所示，与正常对照组比较，番茄水溶性皂苷提取物低、中、高剂量组对血尿酸都没有显著性差异，说明番茄水溶性皂苷提取物对正常小鼠血尿酸没有影响。
表1番茄水溶性皂苷提取物对正常小鼠血尿酸的作用(
n＝12)
注：与正常对照组比较，#p<0.05，##p<0.01。
2.番茄水溶性皂苷提取物对氧嗪酸钾和次黄嘌呤诱导的高尿酸血症小鼠血尿酸的作用
如表2所示，与正常对照组比较，模型组血尿酸水平显著升高，有统计学意义(P<0.01)，说明高尿酸血症动物模型造模成功。与高尿酸血症模型组比较，番茄水溶性皂苷提取物低、中剂量组血尿酸降低，但都没有统计学意义；番茄水溶性皂苷提取物高剂量组血尿酸降低，有统计学意义(P<0.01)，说明番茄水溶性皂苷提取物对氧嗪酸钾和次黄嘌呤联合诱导高尿酸血症模型小鼠具有降尿酸作用。
表2番茄水溶性皂苷提取物对氧嗪酸钾和次黄嘌呤诱导的高尿酸血症小鼠血尿酸的作用(
n＝12)
注：与正常对照组比较，#p<0.05，##p<0.01；
与模型组比较，*P＜0.05，**P＜0.01。
3.番茄水溶性皂苷提取物对氧嗪酸钾和尿酸诱导的高尿酸血症小鼠血尿酸的作用
如表3所示，与正常对照组比较，模型组血尿酸水平显著升高，有统计学意义(P<0.01)，说明高尿酸血症动物模型造模成功。与高尿酸血症模型组比较，番茄水溶性皂苷提取物中、高剂量组血尿酸降低，都具有统计学意义 (P<0.01或P<0.05)，说明番茄水溶性皂苷提取物对氧嗪酸钾和尿酸联合诱导高尿酸血症模型小鼠具有降尿酸作用。
表3番茄水溶性皂苷提取物对氧嗪酸钾和尿酸诱导的高尿酸血症小鼠血尿酸的作用 (
n＝12)
注：与正常对照组比较，#p<0.05，##p<0.01；
与模型组比较，*P＜0.05，**P＜0.01。
Claims (2)
1.番茄水溶性皂苷提取物在制备治疗高尿酸血症药物中的应用；其中，
番茄水溶性皂苷提取物的制备方法包括：将新鲜番茄打浆、果胶酶酶解，酶解液过滤，收集滤液，干燥，制得番茄水溶性皂苷提取物粗品；将番茄水溶性皂苷提取物粗品经大孔树脂柱层析，用体积浓度为80～100％的乙醇洗脱，回收溶剂，干燥，得到纯化后的番茄水溶性皂苷提取物；
所述番茄水溶性皂苷提取物的给药剂量为400mg/Kg。
2.根据权利要求1所述的应用，其特征在于：所述纯化后的番茄水溶性皂苷提取物中，番茄皂苷A的含量≥17％。
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