机构地区:[1]徐州医科大学附属医院内分泌科,徐州221000 [2]邳州中医院内科,徐州221300 [3]徐州医科大学附属医院整形美容外科,徐州221000 [4]徐州市中心医院骨脊柱外科,徐州221000 [5]徐州医科大学附属医院风湿免疫科,徐州221000 [6]首都医科大学附属北京同仁医院内分泌科,北京100010
出 处:《中华内分泌代谢杂志》2023年第9期790-796,共7页Chinese Journal of Endocrinology and Metabolism
摘 要:目的研究氧化石墨烯(graphene oxide, GO)调控脂肪源间充质干细胞(adipose-derived mesenchymal stem cells, Ad-MSCs)促进糖尿病皮肤创面修复的作用。方法将Ad-MSCs与浓度分别为100、200、400 μg/mL GO在25 mmol/L葡萄糖浓度的环境下共同培养48 h, 用流式细胞仪和Western印迹法检测Ad-MSCs的凋亡情况。选用18只BALB/c无胸腺雌性裸鼠, 建立糖尿病裸鼠皮肤缺损模型后, 将磷酸盐缓冲液、Ad-MSCs、GO+Ad-MSCs以皮内注射方式注入裸鼠创缘皮肤进行治疗, 即为对照组、Ad-MSCs组、GO+Ad-MSCs组, 每组6只。观察创面细胞存活情况和裸鼠皮肤愈合情况, HE染色及Masson染色明确创面愈合情况, 酶联免疫吸附试验(ELISA)测定表皮生长因子(EGF)、血管内皮生长因子(VEGF)、肿瘤坏死因子-α(TNF-α)、白细胞介素6(IL-6)水平。结果与未进行GO共培养的高糖组(对照组)相比, Ad-MSCs与GO共培养中Ad-MSCs的凋亡率明显降低(P<0.05), 且随GO的浓度增加, 细胞凋亡率也随之降低, Bax蛋白表达水平下降, Bcl-2蛋白表达水平增高(均P<0.05)。糖尿病裸鼠皮肤缺损模型显示, GO+Ad-MSCs组中Ad-MSCs存活率、创面愈合度明显增高(均P<0.05), 且上皮化程度和胶原再生的厚度均优于其他组。ELISA结果表明, GO+Ad-MSCs组中EGF、VEGF表达明显增加(均P<0.05), TNF-α、IL-6表达明显降低(均P<0.05)。结论 GO在体外条件下可以抑制Ad-MSCs凋亡;在体内情况下, 使用经GO处理后Ad-MSCs比单纯应用Ad-MSCs存活率更高, 创面愈合速度更快, 效果更佳。Objective To study the effect of graphene oxide(GO)on adipose-derived mesenchymal stem cells(Ad-MSCs)to promote diabetic skin wound healing.Methods Ad-MSCs were cultured with GO at concentration of 100,200,and 400μg/mL and with 25 mmol/L of glucose for 48 h.The apoptosis of Ad-MSCs was detected by flow cytometry and Western blotting.Eighteen BALB/c thymus free female nude mice were selected to establish the skin defect model of diabetic nude mice.Then phosphate-buffered saline,Ad-MSCs,and GO+Ad-MSCs were injected into the damaged skin of nude mice by intradermal injection for treatment,thereby created the control group,Ad-MSCs group,and GO+Ad-MSCs group,6 mice each group.The survival of wound cells and skin healing of mice were observed after surgery,and the wound healing was determined by HE staining and Masson staining.The level of epidermal growth factor(EGF),vascular endothelial growth factor(VEGF),tumor necrosis factor-α(TNF-α),and interleukin 6(IL-6)were determined by enzyme-linked immunosorbent assay(ELISA).Results Compared with the high glucose group without GO(control group),the apoptosis rate of Ad-MSCs co-culture with GO was significantly reduced(P<0.05),and the apoptosis rate decreased with the increase of GO concentration.The expression level of Bax protein decreased,and the expression level of Bcl-2 protein increased(P<0.05).Diabetic skin defect model showed that the survival rate of Ad-MSCs and wound healing degree in GO+Ad-MSCs group were significantly increased(P<0.05),and the degree of epithelialization and thickness of collagen regeneration were better than those in other groups.ELISA results showed that the expressions of EGF and VEGF were significantly increased in GO+Ad-MSCs group(P<0.05),while the expressions of TNF-αand IL-6 were significantly decreased(P<0.05).Conclusion GO can inhibit the apoptosis of Ad-MSCs in vitro.In vivo,Ad-MSCs treated with GO have higher survival rate,faster wound healing,and better effect than Ad-MSCs treated alone.
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