摘要
背景:枸杞多糖具有多种生物活性,研究发现其具有促进创面愈合的潜在效用。目的:探讨枸杞多糖在肿瘤坏死因子α介导角质细胞凋亡中的作用以及对烧伤创面愈合的作用。方法:①体外实验:将角质细胞分4组培养,正常组加入含体积分数15%胎牛血清及1%谷氨酰胺的α-MEM培养基(完全培养基),阳性对照组加入含枸杞多糖的完全培养基,模型组加入含肿瘤坏死因子α的完全培养基,实验组加入含枸杞多糖与肿瘤坏死因子α的完全培养基。培养24 h后,采用CCK-8法检测细胞增殖,流式细胞仪检测细胞凋亡,Western blot检测凋亡蛋白Cleaved caspase-8、TNF R1、FADD及自噬蛋白LC3的表达。在正常组、模型组、实验组的基础上,再增加自噬抑制剂组(加入含3-甲基腺嘌呤的完全培养基)、自噬抑制剂+枸杞多糖组(加入含枸杞多糖、肿瘤坏死因子α与3-甲基腺嘌呤的完全培养基),培养24 h后,采用流式细胞术检测角质细胞凋亡情况。②体内实验:采用随机数字表法将6只SD大鼠分为对照组与实验组,每组3只。在每只大鼠背部制作4个直径2 cm的深Ⅱ度烧伤创面模型,造模24 h后,对照组与实验组小鼠创面分别涂抹生理盐水、枸杞多糖溶液,1次/d。治疗后定期观察创面愈合情况,治疗后28 d取材,观察创面病理形态。结果与结论:①体外实验:单独加入枸杞多糖不影响角质细胞的增殖、凋亡与凋亡及自噬蛋白的表达;加入肿瘤坏死因子α后,角质细胞增殖受到抑制、凋亡率增加、凋亡及自噬蛋白的表达升高,枸杞多糖可拮抗肿瘤坏死因子α介导的上述作用;枸杞多糖联合自噬抑制剂组可进一步降低角质细胞的凋亡率;②体内实验:实验组大鼠治疗后12,16,20,24,28 d的创面愈合率均高于对照组(P<0.05,P<0.01);治疗后28 d的苏木精-伊红染色显示,与对照组相比,实验组大鼠创面表皮完整、清晰;③结果表明:枸杞多糖通过抑制角�
BACKGROUND:Lycium barbarum polysaccharide has many biological activities and has been found to have potential effects on promoting wound healing.OBJECTIVE:To investigate the mechanism of lycium barbarum polysaccharide in tumor necrosis factor-α-mediated keratinocyte apoptosis and its effect on the healing of burn wounds.METHODS:(1)In vitro experiment:Keratinocytes were divided into four groups:cells were cultured in theα-MEM medium(complete medium)containing 15%fetal bovine serum and 1%glutamine in normal group,cultured in the complete medium containing lycium barbarum polysaccharide in positive control group,cultured in the complete medium containing tumor necrosis factor-αin model group,and cultured in the complete medium containing lycium barbarum polysaccharide and tumor necrosis factor-αin experimental group.After 24 hours of culture,cell proliferation was detected using cell counting kit-8 assay;Cleaved caspase-8,TNF R1,FADD,and LC3 were detected using western blot.Then an autophagy inhibitor group(the complete medium containing 3-methyladenine)and an autophagy inhibitor+lycium barbarum polysaccharide group(the complete medium containing lycium barbarum polysaccharide,tumor necrosis factor-α,and 3-methyladenine)were set up.After 24 hours of culture,keratinocyte apoptosis was detected by flow cytometry.(2)In vivo experiment:Six Sprague-Dawley rats were randomly divided into a control group and an experimental group,with three rats in each group.Four deep II degree burn wounds with a diameter of 2 cm were made on the back of each rat.At 24 hours after modeling,mice in the control and experimental groups were coated with normal saline and lycium barbarum polysaccharide solution,respectively,once a day.Wound healing was observed regularly after treatment.Samples were taken 28 days after treatment and the pathologic pattern of the wound was observed.RESULTS AND CONCLUSION:(1)In vitro experiment:Addition of lycium barbarum polysaccharide alone did not affect cell proliferation and apoptosis and the expressio
作者
朱永朝
房超
赵芳
张庆
赵丹
Zhu Yongzhao;Fang Chao;Zhao Fang;Zhang Qing;Zhao Dan(General Hospital of Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China)
出处
《中国组织工程研究》
CAS
北大核心
2024年第23期3686-3691,共6页
Chinese Journal of Tissue Engineering Research
基金
宁夏回族自治区自然科学基金项目(2020AAC03412)
项目负责人:赵丹。
关键词
角质细胞
肿瘤坏死因子Α
枸杞多糖
凋亡
自噬
创面愈合
keratinocyte
tumor necrosis factor-α
lycium barbarum polysaccharide
apoptosis
autophagy
wound healing
