美洲大蠊药渣中多糖制备工艺优化及促创面愈合的综合利用研究
|
摘要
目的采用响应面法优化美洲大蠊药渣中多糖提取工艺,并考察其促创面愈合的作用,为美洲大蠊药渣综合利用提供思路。方法在单因素实验基础上,采用Box-Behnken设计最佳美洲大蠊多糖提取工艺,并分析其单糖组成。采用0.1、0.05、0.025 g/mL美洲大蠊多糖作用于创面损伤模型小鼠,计算创面愈合率,用苏木精-伊红和天狼猩红染色评价创面愈合程度。结果美洲大蠊多糖的最佳提取工艺为碱液浓度0.02mol/L,提取时间1.65h,提取温度62℃,该条件所得多糖转移率为13.68%,且由甘露糖、鼠李糖、葡萄糖醛酸、半乳糖醛酸、葡萄糖、半乳糖、岩藻糖7种单糖组成。创面愈合实验表明,第3、6、9、14天多糖给药组创面愈合率均高于模型组,且组织切片染色表明该多糖组分能促进胶原和肉芽组织生成,从而促进创面愈合。结论响应面法优化的提取工艺切实可靠,提取率高;美洲大蠊药渣提取多糖能促进小鼠创面愈合,提示美洲大蠊药渣中多糖具有开发为创面用相关产品的应用前景。
Objective To optimize the extraction of polysaccharide from the residue of Periplaneta americana by response surface method and study the effect of P. americana polysaccharide(PAP) on wound healing, so as to provide ideas for the comprehensive utilization of P. americana residues. Methods On the basis of single factor experiment, Box-Behnken response surface method was used to design the best extraction process. And the monosaccharide composition of polysaccharide was analyzed. Polysaccharide concentrations of 0.1, 0.05, and 0.025 g/mL were applied to the wound injury model mice. The wound healing rate was calculated, and hematoxylin-eosin(HE) and Sirius Red staining were used to evaluate the wound healing degree. Results The optimum extraction technology of PAP were as follows: The concentration of NaOH was 0.02 mol/L; And the extraction time was 1.65 h; The extraction temperature was 62 ℃. The transfer rate of polysaccharide extracted by this process was 13.68%. The polysaccharide of the P.americana consisted of seven monosaccharides of mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose, and fucose. Wound healing experiment showed that wound healing rate of the polysaccharide group on day 3, 6, 9, and 14 was higher than that of the model group, and HE and Sirius Red staining showed that the polysaccharide could promote the formation of collagen and granulation tissue, thus promoting the wound healing. Conclusion The optimized extraction process of PAP from P. americana residues by response surface method was reliable, with high extraction rate. And PAP could promote wound healing in mice, suggesting that the polysaccharide from P. americana residues could be used as wound related products.
Objective To optimize the extraction of polysaccharide from the residue of Periplaneta americana by response surface method and study the effect of P. americana polysaccharide(PAP) on wound healing, so as to provide ideas for the comprehensive utilization of P. americana residues. Methods On the basis of single factor experiment, Box-Behnken response surface method was used to design the best extraction process. And the monosaccharide composition of polysaccharide was analyzed. Polysaccharide concentrations of 0.1, 0.05, and 0.025 g/mL were applied to the wound injury model mice. The wound healing rate was calculated, and hematoxylin-eosin(HE) and Sirius Red staining were used to evaluate the wound healing degree. Results The optimum extraction technology of PAP were as follows: The concentration of NaOH was 0.02 mol/L; And the extraction time was 1.65 h; The extraction temperature was 62 ℃. The transfer rate of polysaccharide extracted by this process was 13.68%. The polysaccharide of the P.americana consisted of seven monosaccharides of mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose, and fucose. Wound healing experiment showed that wound healing rate of the polysaccharide group on day 3, 6, 9, and 14 was higher than that of the model group, and HE and Sirius Red staining showed that the polysaccharide could promote the formation of collagen and granulation tissue, thus promoting the wound healing. Conclusion The optimized extraction process of PAP from P. americana residues by response surface method was reliable, with high extraction rate. And PAP could promote wound healing in mice, suggesting that the polysaccharide from P. americana residues could be used as wound related products.
引文
[1]孙星衍.神农本草经[M].北京:商务印书馆,1955.
[2]高洁,沈咏梅,岳碧松.美洲大蠊药理作用及其临床疗效的研究进展[J].中药药理与临床,2018,34(4):203-208.
[3]张臻,傅超美,胡慧玲,等.D-最优混料设计优化美洲大蠊乳膏的基质配方研究[J].中草药,2013,44(12):1574-1578.
[4]甘平,张旭强,何旭,等.美洲大蠊醇提物对小鼠急性肝损伤的保护作用[J].现代药物与临床,2011,26(2):123-128.
[5]蒋永新,王熙才,金从国,等.美洲大蠊提取物对小鼠3LL肺癌的抑制作用及其机制探讨[J].中国肺癌杂志,2006,9(6):488-491.
[6]李晓庆.美洲大蠊多糖提取工艺优化与抗肿瘤作用研究[D].佳木斯:佳木斯大学,2016.
[7]李武,段丽芳,何贵清,等.美洲大蠊提取物对实验性肝纤维化的影响[J].时珍国医国药,2010,21(5):1137-1138.
[8]林喆,李全学,徐静,等.美洲大蠊化学成分和生物活性研究进展[J].中草药,2017,48(17):3645-3649.
[9]罗廷顺,高孟婷,马芳芳,等.美洲大蠊药理作用及临床应用研究进展[J].安徽农业科学,2012,40(10):5933-5935.
[10]中药大品种科技竞争力报告(2017版)[J].中国现代中药,2018,20(1):6-13.
[11]张汉超,耿福能,沈咏梅,等.康复新液药理作用及临床应用的研究进展[J].中国民族民间医药,2017,26(3):57-60.
[12]黄丫丫,杨杭,林家莉,等.载美洲大蠊提取物纳米粒的表征及体外释放研究[J].中草药,2016,47(19):3407-3414.
[13]Predel R,Kellner R,Rapus J,et al.Isolation and structural elucidation of eight kinins from the retrocerebral complex of the American cockroach,Periplaneta americana[J].Regul Pept,1997,71(3):199-205.
[14]Zhu J J,Yao S,Guo X,et al.Bioactivity-guided screening of wound-healing active constituents from American cockroach(Periplaneta americana)[J].Molecules,2018,23(1):101-112.
[15]Neupert S,Fusca D,Schachtner J,et al.Toward a singlecell-based analysis of neuropeptide expression in Periplaneta americana antennal lobe neurons[J].J Comp Neurol,2012,520(4):694-716.
[16]He Z C,Liu G M,Wang X Y,et al.Research advance on neuropeptides from Periplaneta americana[J].Nat Prod Res Dev,2008,20(1):180-186.
[17]廖芳,朱伟,周洁,等.大孔吸附树脂纯化美洲大蠊多肽的工艺研究[J].中草药,2016,47(19):3420-3425.
[18]张锐,刘建群,舒积成,等.美洲大蠊药渣制备壳聚糖工艺的优化[J].中成药,2017,39(12):2625-2627.
[19]李玉寒,卞君醒,狄斌,等.壳聚糖复合创面敷料研究进展[J].药物生物技术,2017,24(4):90-93.
[20]莫开菊,赵娜,朱照武,等.葛仙米多糖的单糖组成分析[J].食品科学,2015,36(18):89-92.
[21]李晓庆,许龙,富尧,等.盐提法提取美洲大蠊多糖工艺优化的研究[J].安徽农学通报,2016,22(10):38-39.
[22]李晓庆,许龙,陈光道,等.稀碱法提取美洲大蠊多糖工艺条件的研究[J].安徽农业科学,2016,44(15):125-126.
[23]李晨晨,贾成友,郭华,等.正交试验法优化美洲大蠊糖蛋白的提取工艺[J].成都中医药大学学报,2014,37(3):4-7.
[24]李晓庆,富尧,陈光道,等.木瓜蛋白酶提取蜚蠊多糖最佳工艺研究[J].安徽农业科学,2015,43(34):81-82.
[2]高洁,沈咏梅,岳碧松.美洲大蠊药理作用及其临床疗效的研究进展[J].中药药理与临床,2018,34(4):203-208.
[3]张臻,傅超美,胡慧玲,等.D-最优混料设计优化美洲大蠊乳膏的基质配方研究[J].中草药,2013,44(12):1574-1578.
[4]甘平,张旭强,何旭,等.美洲大蠊醇提物对小鼠急性肝损伤的保护作用[J].现代药物与临床,2011,26(2):123-128.
[5]蒋永新,王熙才,金从国,等.美洲大蠊提取物对小鼠3LL肺癌的抑制作用及其机制探讨[J].中国肺癌杂志,2006,9(6):488-491.
[6]李晓庆.美洲大蠊多糖提取工艺优化与抗肿瘤作用研究[D].佳木斯:佳木斯大学,2016.
[7]李武,段丽芳,何贵清,等.美洲大蠊提取物对实验性肝纤维化的影响[J].时珍国医国药,2010,21(5):1137-1138.
[8]林喆,李全学,徐静,等.美洲大蠊化学成分和生物活性研究进展[J].中草药,2017,48(17):3645-3649.
[9]罗廷顺,高孟婷,马芳芳,等.美洲大蠊药理作用及临床应用研究进展[J].安徽农业科学,2012,40(10):5933-5935.
[10]中药大品种科技竞争力报告(2017版)[J].中国现代中药,2018,20(1):6-13.
[11]张汉超,耿福能,沈咏梅,等.康复新液药理作用及临床应用的研究进展[J].中国民族民间医药,2017,26(3):57-60.
[12]黄丫丫,杨杭,林家莉,等.载美洲大蠊提取物纳米粒的表征及体外释放研究[J].中草药,2016,47(19):3407-3414.
[13]Predel R,Kellner R,Rapus J,et al.Isolation and structural elucidation of eight kinins from the retrocerebral complex of the American cockroach,Periplaneta americana[J].Regul Pept,1997,71(3):199-205.
[14]Zhu J J,Yao S,Guo X,et al.Bioactivity-guided screening of wound-healing active constituents from American cockroach(Periplaneta americana)[J].Molecules,2018,23(1):101-112.
[15]Neupert S,Fusca D,Schachtner J,et al.Toward a singlecell-based analysis of neuropeptide expression in Periplaneta americana antennal lobe neurons[J].J Comp Neurol,2012,520(4):694-716.
[16]He Z C,Liu G M,Wang X Y,et al.Research advance on neuropeptides from Periplaneta americana[J].Nat Prod Res Dev,2008,20(1):180-186.
[17]廖芳,朱伟,周洁,等.大孔吸附树脂纯化美洲大蠊多肽的工艺研究[J].中草药,2016,47(19):3420-3425.
[18]张锐,刘建群,舒积成,等.美洲大蠊药渣制备壳聚糖工艺的优化[J].中成药,2017,39(12):2625-2627.
[19]李玉寒,卞君醒,狄斌,等.壳聚糖复合创面敷料研究进展[J].药物生物技术,2017,24(4):90-93.
[20]莫开菊,赵娜,朱照武,等.葛仙米多糖的单糖组成分析[J].食品科学,2015,36(18):89-92.
[21]李晓庆,许龙,富尧,等.盐提法提取美洲大蠊多糖工艺优化的研究[J].安徽农学通报,2016,22(10):38-39.
[22]李晓庆,许龙,陈光道,等.稀碱法提取美洲大蠊多糖工艺条件的研究[J].安徽农业科学,2016,44(15):125-126.
[23]李晨晨,贾成友,郭华,等.正交试验法优化美洲大蠊糖蛋白的提取工艺[J].成都中医药大学学报,2014,37(3):4-7.
[24]李晓庆,富尧,陈光道,等.木瓜蛋白酶提取蜚蠊多糖最佳工艺研究[J].安徽农业科学,2015,43(34):81-82.