流化床气流超微粉碎协同超高压法提取赶黄草黄酮工艺优化及其保肝作用研究

Optimization of extraction process of Penthorum chinense flavonoids by fluidized bed jet milling assisted ultrahigh pressure extraction method and its liver protecting effect

目的优化流化床气流超微粉碎协同超高压法提取赶黄草黄酮的工艺,并对其保肝作用进行研究。方法采用流化床气流超微粉碎协同超高压法提取赶黄草黄酮,通过单因素和响应面实验优化提取工艺参数。同时采用随机法把小鼠分为正常对照组、CCl4模型组、联苯双酯组和赶黄草黄酮低、中、高剂量组。除正常对照组外,其余5组注射0.1%的CCl4构建肝损伤小鼠模型。采用全自动生化仪测定小鼠血清中谷草转氨酶(aspartate aminotransferase,AST)、谷丙转氨酶(alanine aminotransferase,ALT)、乳酸脱氢酶(lactate dehydrogenase,LDH)值和丙二醛(malondialdehyde,MDA)含量,肝组织中谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)和超氧化物歧化酶(superoxide dismutase,SOD)活性。结果最佳提取工艺条件为:溶剂倍数25、压力400 MPa、保压时间13 min,在此条件下,赶黄草黄酮得率为3.98%,此法的提取效果优于其他3种单一的提取方式。体外保肝实验结果表明:赶黄草黄酮能显著降低CCl4所致肝损伤小鼠血清中AST、ALT、LDH的值和MDA含量,提高小鼠肝组织中GSH-Px和SOD活性,表明赶黄草黄酮对CCl4所致肝损伤具有较好的保护作用。结论优化后的提取条件能够有效提取赶黄草黄酮,且得到的赶黄草黄酮具有明显的保肝作用。

Objective To optimize the extraction process of Penthorum chinense flavonoids by fluidized bed jet milling assisted ultrahigh pressure extraction method,and study its liver protecting effect.Methods The fluidized bed jet milling assisted ultrahigh pressure extraction method was used to extract Penthorum chinense flavonoids.And the extraction process parameters were optimized through single factor and response surface tests.At the same time,mice were randomly divided into normal control group,CCl4 model group,biphenyl diester group,and low,medium,and high dose groups of Penthorum chinense flavonoids.Except for the normal control group,the remaining 5 groups were injected with 0.1%CCl4 to construct a liver injury mouse model.The aspartate aminotransferase(AST),alanine aminotransferase(ALT),lactate dehydrogenase(LDH)values and malondialdehyde(MDA)content in mouse serum,as well as the activities of glutathione peroxidase(GSH-Px)and superoxide dismutase(SOD)in liver tissue were measured by using a fully automated biochemical analyzer.Results The optimum extraction conditions were:Solvent ratio of 25,pressure of 400 MPa,and pressure holding time of 13 minutes.Under these conditions,the yield of Penthorum chinense flavonoids was 3.98%,and the extraction effect of this method was superior to the other 3 kinds of single extraction methods.The results of in vitro liver protection test showed that Penthorum chinense flavonoids could significantly reduce the values of AST,ALT,LDH and MDA content in serum of CCl4 induced liver injury mice,and increased the activities of GSH-Px and SOD in liver tissue of mice.This indicated that Penthorum chinense flavonoids had a good protective effect on CCl4 induced liver injury.Conclusion The optimized extraction conditions can effectively extract Penthorum chinense flavonoids,and the obtained Penthorum chinense flavonoids has a significant liver protective effect.