苦瓜伤流液降血糖功效及其作用机制研究
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摘要
苦瓜是一种药食同源的植物,在亚洲民间广泛用于治疗糖尿病。已有报道仅限于对苦瓜果实、种子等部位功能成分提取及其降血糖等生物活性的研究,而对苦瓜伤流液的研究甚少。苦瓜伤流液(Momordica charantia L.bleeding sap)是指苦瓜收获结束后,将其主茎在一定高度剪断,从切口处收集到的液体。因其不影响苦瓜收成,是对苦瓜资源充分利用的极好途径。
本文以苦瓜伤流液与苦瓜汁为原料,采用冷冻干燥技术制备苦瓜伤流液与苦瓜汁干粉,对苦瓜伤流液的组成、抗氧化活性、降血糖活性及降糖机制等进行研究。论文的主要研究内容与结果如下：
(1)通过化学分析法对苦瓜伤流液的成分进行分析,并通过电感耦合等离子体原子发射光谱法检测伤流液中部分矿质元素的含量。结果显示,苦瓜伤流液的总固形物含量为0.19～0.22%,其中包含蛋白质、多糖、酚类等有机成分和K、Ca等常量元素及Mg、Cu、Zn、Fe、Cr、Se等微量元素。
(2)采用体外抗氧化模型,研究苦瓜伤流液与苦瓜汁的抗氧化活性。结果表明,苦瓜伤流液与苦瓜汁浓度分别为20 mg/mL、1.6 mg/mL时,对DPPH-的清除率与0.02 mg/mL的Vc相当。苦瓜伤流液浓度为10 mg/mL时,对Fe2+的螯合力与0.05 mg/mL的EDTA-Na2效果相当,而苦瓜汁对Fe2+的螯合力无明显作用。苦瓜伤流液与苦瓜汁浓度分别为20 mg/mL、15 mg/mL时,抑制AGEs能力分别为44.55%与49.07%。
(3)建立STZ糖尿病小鼠模型,并对苦瓜伤流液与苦瓜汁的降血糖活性进行比较研究。试验结果表明,苦瓜伤流液与苦瓜汁对糖尿病小鼠均有一定的降血糖作用,且两者作用效果无显著差异；苦瓜伤流液450 mg/kg剂量降糖效果优于900mg/kg。
(4)采用自建的STZ糖尿病小鼠模型,研究苦瓜伤流液的降血糖作用机制。结果显示苦瓜伤流液能提高STZ糖尿病小鼠的肝糖原含量、血清胰岛素水平及HDL含量,降低STZ糖尿病小鼠血清中TC、TG含量；胰腺切片显示,苦瓜伤流液能够改善胰岛组织形态,促进胰岛细胞的修复。结果表明,苦瓜伤流液的降糖机制与促进肝糖原合成、调节脂代谢、改善胰岛组织功能、促进胰岛p细胞修复及增加胰岛素分泌相关。
(5)采用体外胰岛p细胞模型,研究苦瓜伤流液对胰岛细胞的修复作用,验证苦瓜伤流液的降糖机制。结果显示,1.5 mmol/L STZ损伤胰岛p细胞后,模型组细胞存活率为75.89%；受损细胞经苦瓜伤流液修复后,存活率明显上升,苦瓜伤流液浓度为0.112 mg/mL,细胞存活率为95.20%,且其胰岛素分泌量也与正常对照组相当。此结果进一步表明,苦瓜伤流液的降糖机制与修复受损胰岛细胞、刺激胰岛素分泌相关。
Momordica charantia L. has the medical and edible value, which is widely used in the treatment of diabetes in Asia. The previous researches on the bioactivities of Momordica charantia L. were only focus on the extraction and bioactivity such as Momordica charantia L. fruit and seed, while neglecting the research of Momordica charantia L.bleeding sap. Momordica charantia L. bleeding sap refers to the sap collected from the cut stem of Momordica charantia L.. Since it is collected after the harvest of Momordica charantia L., it can be an excellent way to make full use of the resources.
Based on Momordica charantia L. bleeding sap and Momordica charantia L. fruit juice as raw materials, the research adopts freeze-drying technology to make Momordica charantia L. bleeding sap and Momordica charantia L. fruit juice into powder. Then the composition of Momordica charantia L. bleeding sap and its antioxidant activity, hypoglycemic activity, and hypoglycemic mechanism are studied. The main contents and results are presented as follows:
(1) The chemical analysis is carried out to analyze the composition of Momordica charantia L. bleeding sap, and ICP-AES is applied to the determination of some mineral element content. The results show that in the bleeding sap, soluble solid concentration is 1.9～2.2%o, protein, sugar, phenols are detected; major elements such as K, Ca and trace elements such as Mg, Cu, Zn, Fe, Cr, Se are also detected.
(2) Use antioxidant model in vitro to make a research to the antioxidant activity of Momordica charantia L. bleeding sap and Momordica charantia L. fruit juice. The results show that:at the respective concentration of 20 mg/mL and 1.6 mg/mL of Momordica charantia L. bleeding sap and Momordica charantia L. fruit juice, the ability to scavenging the DPPH-is similar to Vc of 0.02 mg/mL; the chelating capacity for ferrous irons is similar to 0.05 mg/mL EDTA-Na2 when the Momordica charantia L. bleeding sap concentration at 10 mg/mL, but Momordica charantia L. fruit juice has no significant effect on the system; at the respective concentration of 20 mg/mL and 15 mg/mL of Momordica charantia L. bleeding sap and Momordica charantia L. fruit juice, inhibition capacity to AGEs are 44.55% and 49.07%.
(3) Establish STZ diabetic mice model and make a comparison research on the the hypoglycemic activity between bleeding sap and fruit juice. The dates demonstrate that both Momordica charantia L. bleeding sap and Momordica charantia L.fruit juice could reduce the levels of blood glucose in diabetic mice, and they have no significant difference between each other. The effect of Momordica charantia L. bleeding sap at 450 mg/kg doses of hypoglycemic is better than that of 900 mg/kg.
(4) Adopt STZ diabetic mice to study the hypoglycemic mechanism of Momordica charantia L. bleeding sap. The results show that bleeding sap could significantly reduce the levels of blood-fat in diabetic mice, with an increase in hepatic glycogen and insulin content. The pathological sections show Momordica charantia L. bleeding sap could improve the function of pancreas islet tissues, and accelerate the renovation of pancreaticβcells in diabetic mice. The results indicate that bleeding sap can accelerate the repair of pancreatic inletβcells injured by streptozotocin, stimulate the secretion of insulin, increase the synthesis of liver glycogen and regulate blood lipid metabolism so as to reduce blood glucose level of the diabetic mice.
(5) Use insulinβcells to study the repairing effect of bleeding sap and test the hypoglycemic mechanism of bleeding sap. The results show that:when 1.5 mmol/L STZ acting inβcells, cells security rate is 75.89%; when the damaged cells are repaired by bleeding sap, the security rate rose significantly; at the concentration of 0.112 mg/mL, the security rate is 95.20%, and the insulin secretion is equivalent to normal group. This result further indicates the hypoglycemic mechanism of bleeding sap is related to the renovation of insulinβcells and the stimulation of insulin secretion.
本文以苦瓜伤流液与苦瓜汁为原料,采用冷冻干燥技术制备苦瓜伤流液与苦瓜汁干粉,对苦瓜伤流液的组成、抗氧化活性、降血糖活性及降糖机制等进行研究。论文的主要研究内容与结果如下：
(1)通过化学分析法对苦瓜伤流液的成分进行分析,并通过电感耦合等离子体原子发射光谱法检测伤流液中部分矿质元素的含量。结果显示,苦瓜伤流液的总固形物含量为0.19～0.22%,其中包含蛋白质、多糖、酚类等有机成分和K、Ca等常量元素及Mg、Cu、Zn、Fe、Cr、Se等微量元素。
(2)采用体外抗氧化模型,研究苦瓜伤流液与苦瓜汁的抗氧化活性。结果表明,苦瓜伤流液与苦瓜汁浓度分别为20 mg/mL、1.6 mg/mL时,对DPPH-的清除率与0.02 mg/mL的Vc相当。苦瓜伤流液浓度为10 mg/mL时,对Fe2+的螯合力与0.05 mg/mL的EDTA-Na2效果相当,而苦瓜汁对Fe2+的螯合力无明显作用。苦瓜伤流液与苦瓜汁浓度分别为20 mg/mL、15 mg/mL时,抑制AGEs能力分别为44.55%与49.07%。
(3)建立STZ糖尿病小鼠模型,并对苦瓜伤流液与苦瓜汁的降血糖活性进行比较研究。试验结果表明,苦瓜伤流液与苦瓜汁对糖尿病小鼠均有一定的降血糖作用,且两者作用效果无显著差异；苦瓜伤流液450 mg/kg剂量降糖效果优于900mg/kg。
(4)采用自建的STZ糖尿病小鼠模型,研究苦瓜伤流液的降血糖作用机制。结果显示苦瓜伤流液能提高STZ糖尿病小鼠的肝糖原含量、血清胰岛素水平及HDL含量,降低STZ糖尿病小鼠血清中TC、TG含量；胰腺切片显示,苦瓜伤流液能够改善胰岛组织形态,促进胰岛细胞的修复。结果表明,苦瓜伤流液的降糖机制与促进肝糖原合成、调节脂代谢、改善胰岛组织功能、促进胰岛p细胞修复及增加胰岛素分泌相关。
(5)采用体外胰岛p细胞模型,研究苦瓜伤流液对胰岛细胞的修复作用,验证苦瓜伤流液的降糖机制。结果显示,1.5 mmol/L STZ损伤胰岛p细胞后,模型组细胞存活率为75.89%；受损细胞经苦瓜伤流液修复后,存活率明显上升,苦瓜伤流液浓度为0.112 mg/mL,细胞存活率为95.20%,且其胰岛素分泌量也与正常对照组相当。此结果进一步表明,苦瓜伤流液的降糖机制与修复受损胰岛细胞、刺激胰岛素分泌相关。
Momordica charantia L. has the medical and edible value, which is widely used in the treatment of diabetes in Asia. The previous researches on the bioactivities of Momordica charantia L. were only focus on the extraction and bioactivity such as Momordica charantia L. fruit and seed, while neglecting the research of Momordica charantia L.bleeding sap. Momordica charantia L. bleeding sap refers to the sap collected from the cut stem of Momordica charantia L.. Since it is collected after the harvest of Momordica charantia L., it can be an excellent way to make full use of the resources.
Based on Momordica charantia L. bleeding sap and Momordica charantia L. fruit juice as raw materials, the research adopts freeze-drying technology to make Momordica charantia L. bleeding sap and Momordica charantia L. fruit juice into powder. Then the composition of Momordica charantia L. bleeding sap and its antioxidant activity, hypoglycemic activity, and hypoglycemic mechanism are studied. The main contents and results are presented as follows:
(1) The chemical analysis is carried out to analyze the composition of Momordica charantia L. bleeding sap, and ICP-AES is applied to the determination of some mineral element content. The results show that in the bleeding sap, soluble solid concentration is 1.9～2.2%o, protein, sugar, phenols are detected; major elements such as K, Ca and trace elements such as Mg, Cu, Zn, Fe, Cr, Se are also detected.
(2) Use antioxidant model in vitro to make a research to the antioxidant activity of Momordica charantia L. bleeding sap and Momordica charantia L. fruit juice. The results show that:at the respective concentration of 20 mg/mL and 1.6 mg/mL of Momordica charantia L. bleeding sap and Momordica charantia L. fruit juice, the ability to scavenging the DPPH-is similar to Vc of 0.02 mg/mL; the chelating capacity for ferrous irons is similar to 0.05 mg/mL EDTA-Na2 when the Momordica charantia L. bleeding sap concentration at 10 mg/mL, but Momordica charantia L. fruit juice has no significant effect on the system; at the respective concentration of 20 mg/mL and 15 mg/mL of Momordica charantia L. bleeding sap and Momordica charantia L. fruit juice, inhibition capacity to AGEs are 44.55% and 49.07%.
(3) Establish STZ diabetic mice model and make a comparison research on the the hypoglycemic activity between bleeding sap and fruit juice. The dates demonstrate that both Momordica charantia L. bleeding sap and Momordica charantia L.fruit juice could reduce the levels of blood glucose in diabetic mice, and they have no significant difference between each other. The effect of Momordica charantia L. bleeding sap at 450 mg/kg doses of hypoglycemic is better than that of 900 mg/kg.
(4) Adopt STZ diabetic mice to study the hypoglycemic mechanism of Momordica charantia L. bleeding sap. The results show that bleeding sap could significantly reduce the levels of blood-fat in diabetic mice, with an increase in hepatic glycogen and insulin content. The pathological sections show Momordica charantia L. bleeding sap could improve the function of pancreas islet tissues, and accelerate the renovation of pancreaticβcells in diabetic mice. The results indicate that bleeding sap can accelerate the repair of pancreatic inletβcells injured by streptozotocin, stimulate the secretion of insulin, increase the synthesis of liver glycogen and regulate blood lipid metabolism so as to reduce blood glucose level of the diabetic mice.
(5) Use insulinβcells to study the repairing effect of bleeding sap and test the hypoglycemic mechanism of bleeding sap. The results show that:when 1.5 mmol/L STZ acting inβcells, cells security rate is 75.89%; when the damaged cells are repaired by bleeding sap, the security rate rose significantly; at the concentration of 0.112 mg/mL, the security rate is 95.20%, and the insulin secretion is equivalent to normal group. This result further indicates the hypoglycemic mechanism of bleeding sap is related to the renovation of insulinβcells and the stimulation of insulin secretion.
引文
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