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携带血管紧张素转换酶2基因的间充质干细胞影响博来霉素诱导的急性肺损伤的凋亡途径。

MSCs with ACE II gene affect apoptosis pathway of acute lung injury induced by bleomycin.

作者信息

Zhang Xiaomiao, Gao Fengying, Li Qian, Dong Zhixia, Sun Bo, Hou Lili, Li Zhuozhe, Liu Zhenwei

机构信息

1Department of Thoracic Surgery, First People's Hospital Affiliated to Shanghai Jiaotong University , Shanghai , PR China.

出版信息

Exp Lung Res. 2015 Feb;41(1):32-43. doi: 10.3109/01902148.2014.963901. Epub 2014 Nov 24.

Abstract

PURPOSE

The aim of this study was to evaluate the effect and related mechanisms of Mesenchymal stem cells (MSCs) and Angiotensin converting enzyme II (ACE II) on acute lung injury (ALI).

METHODS

MSCs were separated from umbilical cord cells, and the changes of phenotype before and after ACE II silence were observed using Flow Cytometer. ALI model was induced by 10 mg/mL bleomycin in 60 Balb/c mice, and the rest 8 mice were regarded as the baseline group. The mice were randomly divided into four groups (n = 15): control, ACE II, stem, and stem + ACE II. The apoptotic index (AI) was calculated using TUNEL, and the detection of protein and mRNA of Bax, Bak and p53, Bcl-2, Grp78, CHOP and Caspase 12 were used by western-blot and RT-PCR, respectively.

RESULTS

The umbilical cord cells differentiated into stable MSCs about 14 days, and ACE II transfection reached a peak at the 5th day after transfection. ACE II silence did not affect the phenotype of MSCs. All the proteins and mRNAs expression except Bcl-2 in the stem and stem + ACE II were significantly lower than those in control from 8 h (p < 0.05, p < 0.01), while Bcl-2 exhibited an opposite trend. Stem + ACE II performed a better effect than single stem in most indexes, including AI (p < 0.05, p < 0.01).

CONCLUSIONS

The co-administration of MSCs and ACE II can significantly suppress apoptosis in ALI mice, and may be an effective clinical treatment for ALI.

摘要

目的

本研究旨在评估间充质干细胞(MSCs)和血管紧张素转换酶II(ACE II)对急性肺损伤(ALI)的影响及相关机制。

方法

从脐带细胞中分离出MSCs,使用流式细胞仪观察ACE II沉默前后的表型变化。用10 mg/mL博来霉素诱导60只Balb/c小鼠建立ALI模型,其余8只小鼠作为基线组。将小鼠随机分为四组(n = 15):对照组、ACE II组、干细胞组和干细胞+ACE II组。使用TUNEL法计算凋亡指数(AI),分别通过蛋白质免疫印迹法和逆转录聚合酶链反应检测Bax、Bak和p53、Bcl-2、Grp78、CHOP及半胱天冬酶12的蛋白质和mRNA表达。

结果

脐带细胞约14天分化为稳定的MSCs,ACE II转染在转染后第5天达到峰值。ACE II沉默不影响MSCs的表型。从8小时起,干细胞组和干细胞+ACE II组中除Bcl-2外的所有蛋白质和mRNA表达均显著低于对照组(p < 0.05,p < 0.01),而Bcl-2呈现相反趋势。在大多数指标中,干细胞+ACE II组比单一干细胞组效果更好,包括AI(p < 0.05,p < 0.01)。

结论

MSCs与ACE II联合应用可显著抑制ALI小鼠的细胞凋亡,可能是ALI有效的临床治疗方法。

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