基于响应曲面法优化壳聚糖猴头菇多糖PLGA纳米颗粒的制备及其免疫效应的研究

Optimization of preparation and immune effect of chitosan Hericium erinaceus polysaccharide PLGA nanoparticles based on response surface method

为探究壳聚糖猴头菇多糖PLGA纳米颗粒(CS-PLGA-HEP)对免疫活性调节的影响,通过制备CS-PLGA-HEP纳米颗粒,采用响应曲面法优化制备工艺,傅里叶红外光谱检测、Zeta电位检测等方法对CS-PLGA-HEP纳米颗粒的结构进行表征检测,并通过鸡新城疫疫苗免疫佐剂注射试验来探究其免疫效果。结果显示,CS-PLGA-HEP的最优条件:F68浓度为0.38%,有机相(O)与内水相(W_(1))之比为5.75,外水相(W_(2))与初乳(PE)之比为7.81,在此条件下其包封率的实测值为(91.32±0.13)%,接近预测值91.60%。纳米颗粒均匀,表面光滑且状态稳定。当CS-PLGA-HEP组浓度为20 mg/kg时,与猴头菇多糖(HEP)组相比更能有效提高鸡血清新城疫特异性抗体浓度,促进IL-4和IFN-γ的分泌水平,增强鸡脾淋巴细胞的成熟分化,提升机体免疫调节能力。结果表明,采用响应曲面法优化壳聚糖猴头菇多糖PLGA纳米颗粒制备条件合理,且纳米修饰可以增强HEP的免疫调节活性,为免疫增强剂的设计与应用提供了参考。

The aim of this study was to investigate the effect of chitosan Hericium erinaceus polysaccharide PLGA nanoparticles(CS-PLGA-HEP)on the modulation of immune activity.The structure of CSPLGA-HEP nanoparticles was characterized through the preparation of CS-PLGA-HEP nanoparticles,optimization of preparation process using response surface methodology(RSM).Fourier infrared spectroscopy and Zeta potential detection.The immunological effect was explored through a chicken Newcastle disease vaccine immune adjuvant injection test.The result show that the optimal condition for CS-PLGA-HEP was F68 concentration of 0.38%,the ratio of organic phase(O)to internal aqueous phase(W_(1))of 5.75,the ratio of external aqueous phase(W_(2))to colostrum(PE)of 7.81,and the measured value of its encapsulation rate under these conditions was(91.32±0.13)%,which closely matched the predicted value of 91.60%.The nanoparticles were uniform,with a smooth surface and stable state.When the concentration of CS-PLGAHEP group was 20 mg/kg,it was more effective in increasing the concentration of chicken serum Newcastle disease-specific antibody,promoting the secretion level of IL-4 and IFN-γ,enhancing the maturation and differentiation of chicken splenic lymphocytes,and improving the immunomodulatory ability of the organism as compared with that of HEP group.The study found that the response surface method was utilised to optimize the preparation conditions of chitosan Hericium erinaceus polysaccharide PLGA nanoparticles(CS-PLGA-HEP).The nano-modification was shown to enhance the immunomodulatory activity of HEP,making it a potential reference for the design and application of immune-enhancing agents.