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  • 吴 晓,李宇飞,金佳慧,等.缺氧预处理的脂肪间充质干细胞对小鼠硬皮病模型的治疗作用[J].同济大学学报(医学版),2020,41(4):411-417.    [点击复制]
  • WU Xiao,LI Yu-fei,JIN Jia-hui,et al.Therapeutic effect of hypoxia-pretreated adipose tissue-derived stromal cells on mice with scleroderma[J].同济大学学报(医学版),2020,41(4):411-417.   [点击复制]
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缺氧预处理的脂肪间充质干细胞对小鼠硬皮病模型的治疗作用
吴晓,李宇飞,金佳慧,沈亮亮,BhavanaRajbanshi,赵敬军
0
(同济大学医学院,上海 200092; 同济大学附属同济医院皮肤科,上海 200065)
摘要:
目的 观察脂肪间充质干细胞(adipose tissue-derived stromal cells, ADSCs)及缺氧预处理的ADSCs(hypoxia-pretreated ADSCs, hpADSCs)对硬皮病小鼠皮损组织的治疗作用。方法 分离提取小鼠的脂肪间充质干细胞,体外培养至第3代后,部分细胞进行缺氧预处理。使用博来霉素(bleomycin, BLM)对小鼠进行造模,根据处理方式分为PBS处理的空白对照组、BLM模型组、ADSCs治疗组、hpADSCs治疗组。各组皮损样本石蜡切片分别进行H-E染色、Masson染色,检测真皮厚度、羟脯氨酸评估胶原水平,检测H2O2评估氧化应激水平,使用Western印迹法和免疫组化评估α-SMA及TGF-β的表达水平。结果 与空白对照组相比,BLM模型组真皮厚度和组织内H2O2水平明显增加(P<0.05),α-SMA和TGF-β表达水平也有一定程度的上调。虽然ADSCs治疗能够一定程度上降低真皮厚度,但是差异无统计学意义(P>0.05),而hpADSCs治疗能够显著降低小鼠真皮厚度(P<0.05)。ADSCs与hpADSCs治疗均能够明显降低组织内H2O2水平(P<0.05)。相比ADSCs治疗,hpADSCs治疗能够更好地降低真皮内TGF-β和α-SMA的表达水平。结论 ADSCs能够降低硬皮病小鼠的胶原沉积并下调氧化应激水平,hpADSCs有更好的治疗效果。
关键词:  硬皮病  脂肪间充质干细胞  预缺氧  氧化应激
DOI:10.16118/j.1008-0392.2020.04.002
投稿时间:2020-01-07
基金项目:国家自然科学基金(81874240)
Therapeutic effect of hypoxia-pretreated adipose tissue-derived stromal cells on mice with scleroderma
WU Xiao,LI Yu-fei,JIN Jia-hui,SHEN Liang-liang,Bhavana Rajbanshi,ZHAO Jing-jun
(Tongji University School of Medicine, Shanghai 200092, China; Dept. of Dermatology, Tongji Hospital, Tongji University School of Medicine, Shanghai 200065, China)
Abstract:
Objective To investigate the therapeutic effects of adipose tissue-derived stromal cells(ADSCs) and hypoxia-pretreated ADSCs(hpADSCs) on bleomycin-induced scleroderma in mice. Methods ADSCs were isolated from mouse adipose tissue and cultured to 3th passage. Hypoxia-pretreated ADSCs were obtained by culturing ADSCs in low-oxygen incubator for 2 days. Bleomycin(BLM) was used to induce scleroderma in mice. Mice were randomly divided into four groups: PBS-treated blank group, BLM-treated group, ADSCs-treated group and hpADSCs-treated group. The skin tissue specimens were pathologically examined with H-E staining. Dermal thickness was measured by Masson staining. Hydroxyproline level in skin was measured to evaluate the collagen level. The level of H2O2 was measured to evaluate the oxidative stress. The expression of α-SMA and TGF-β was detected by Western blotting and immunochemistry. Results In BLM-treated group, the collagen deposition and oxidative stress were significantly increased(P<0.05), and the expression of α-SMA and TGF-β was also increased. The treatment of hpADSCs significantly reduced the thickness of derma(P<0.05). ADSCs and hpADSCs therapy both attenuated oxidative stress(P<0.05). Compared with ADSCs, hpADSCs had more marked effect on decreasing the expression of TGF-β in dermis. Conclusion ADSCs can attenuate BLM-induced scleroderma by reducing the skin oxidative stress level and abnormal collagen deposition, while hpADSCs have better therapeutic effect on scleroderma in mice than ADSCs.
Key words:  scleroderma  adipose tissue-derived stromal cells  hypoxia pretreatment  oxidative stress

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