广地龙纯化蛋白促进创伤修复作用机制研究  

Mechanism of Purified Protein in Pheretima aspergillum in Promoting Wound Repair

作　　者：田林华 姚琳 冯丽娜 高辛 王伟明[1] 董坤 TIAN Linhua;YAO Lin;FENG Lina;GAO Xin;WANG Weiming;DONG Kun(Southern Medical University,Guangzhou 510515,China;Heilongjiang Academy of Chinese Medical Sciences,Harbin 150010,China)

机构地区：[1]南方医科大学,广州510515 [2]黑龙江省中医药科学院,哈尔滨150010

出　　处：《世界中医药》2023年第9期1236-1239,共4页World Chinese Medicine

基　　金：国家中药材产业技术体系专项项目(CARS-21);黑龙江省自然科学基金项目(LH2019H091)。

摘　　要：目的:研究广地龙纯化蛋白促进创伤修复的作用机制。方法:采用细胞计数试剂(CCK-8)法,检测给药后对人永生化角质形成细胞(HaCaT)和小鼠成纤维细胞(NIH3T3)增殖的影响;通过划痕实验,检测HaCaT细胞和NIH3T3细胞迁移的影响;利用羟脯氨酸检测试剂盒和酶联免疫吸附试验法,检测NIH3T3细胞培养上清中羟脯氨酸、胶原蛋白-Ⅰ和基质金属蛋白酶-1水平。结果:在一定范围内,地龙蛋白能促进HaCaT细胞和NIH3T3细胞增殖和迁移,分别在0.313 mg/mL和1.250 mg/mL时,细胞增殖活性最强,划痕愈合率最高;给药后NIH3T3细胞培养上清中羟脯氨酸、胶原蛋白-Ⅰ水平明显升高,基质金属蛋白酶-1水平显著降低,在1.250 mg/mL时,与对照组比较最显著。结论:广地龙纯化蛋白组分通过促进HaCaT细胞和NIH3T3细胞增殖和迁移,促进NIH3T3细胞胶原蛋白合成与分泌,抑制胶原蛋白降解,从而促进创伤修复。Objective:To study the mechanism of purified protein in Pheretima aspergillum in promoting wound repair.Methods:The Cell Counting Kit-8(CCK-8)assay was used to detect the effect of P.aspergillum purified protein on the proliferation of immortalized human keratinocytes(HaCaT cells)and mouse fibroblasts(NIH3T3 cells).The cell scratch test was used to investigate the effect on cell migration of HaCaT and NIH3T3 cells.The hydroxyproline assay kit and the enzyme-linked immunosorbent assay(ELISA)were used to detect the levels of hydroxyproline(HYP),collagen-Ⅰ(Col-Ⅰ),and matrix metalloproteinase-1(MMP-1)in the culture supernatant of NIH3T3 cells.Results:Within a certain concentration range,P.aspergillum purified protein could promote the proliferation and migration of HaCaT and NIH3T3 cells,with the strongest cell proliferation activity and the highest scratch healing rate at 0.313 mg/mL and 1.250 mg/mL,respectively.After administration of P.aspergillum purified protein,the levels of HYP and Col-Ⅰin NIH3T3 cell culture supernatant increased significantly,while the level of MMP-1 decreased significantly.The administration of P.aspergillum purified protein at the concentration of 1.250 mg/mL showed the most significant results compared with the control group.Conclusion:P.aspergillum purified protein can promote the proliferation and migration of HaCaT and NIH3T3 cells,improve the synthesis and secretion of collagen in NIH3T3 cells,and inhibit the degradation of collagen,thereby promoting wound healing.

关 键 词：广地龙 纯化蛋白 创伤修复 NIH3T3细胞 HACAT细胞 胶原蛋白 细胞增殖 细胞迁移 

分 类 号：R284[医药卫生—中药学]