血红素加氧酶-1对脂肪干细胞的保护性研究

Lentiviral Mediated HO-1 Gene transfer Can Enhance the Survival of Transfected ADSCs in Vitro Under Ischemic Environment

目的探讨慢病毒介导的血红素加氧酶-1(Heme oxygenase-1,HO-1)对脂肪干细胞(ADSCs)在低氧无血清的条件下的保护作用。方法分离培养SD大鼠ADSCs,取第三代ADSCs分为三组:对照组(Control);低氧无血清(serum—free and oxygen deprivation,SOD)组;慢病毒介导的HO-1组。采用DAPI染色检测细胞凋亡率;蛋白印迹(Western-blot)法检测三组HO-1的表达量;ELISA法测定三组细胞培养上清液中TNF-а,IL-1β,IL-10含量。结果 (1)携带HO-1基因的慢病毒转染ADSCs可明显增加ADSCs中HO-1的表达;(2)对照组凋亡率对照组(4.21±2.56)%;SOD组(26.69±1.29)%;HO-1组(12.32±1.28)%。SOD组与对照组相比,凋亡率有显著升高(P<0.01),HO-1组与SOD组相比凋亡率显著降低(P<0.05);(3)与对照组比较,SOD组TNF-а,IL-1β浓度显著升高(P<0.01);IL-10浓度显著降低(P<0.01)。与SOD组比较,HO-1组TNF-а,IL-1β浓度显著降低(P<0.05);IL-10浓度显著升高(P<0.05)。结论提高ADSCs中HO-1的表达可以显著降低ADSCs在低氧无血清条件下的凋亡,其机制与降低炎症因子TNF-а,IL-1β分泌,升高抗炎因子IL-10表达有关。

Objective To analyze the protective effects of viral mediated Heme oxygenase-1（HO-1） to Adipose-deprived stem cells（ADSCs） under the circumstance with serum free and oxygen deprivation. Methods Fractional cultivate the ADSCs of SD rats, divided the third generation of ADSCs into three groups ： control group, serum free and oxygen deprivation group（SOD group）, viral mediated HO-1group. The cell apoptotic rates were detected by using DAPI staining;Western blot was used to identify the protein expressions of HO-1 in three groups; The concentrations of serum TNF-а, IL-1β, IL-10 were determined by using ELISA. Results（1）The viral mediated ADSCs with HO-1gene can significantly increase the expression of HO-1;（2）The cell apoptotic rates of the three groups were（4.21±2.56）%; SOD（26.69±1.29）%; HO-1（12.32±1.28）%. Compared with the control group, the cell apoptotic rates of SOD group was significantly higher（P〈0.01）. Compared with the SOD group, the cell apoptotic rates of HO-1 group was significantly lower（P〈0.05）.（3）In SOD group, the concentrations of serum TNF-а, IL-1βwere significantly higher than that of control group（P〈0.01）, the concentrations of serum IL-10 was significantly lower than that of control group（P〈0.01）. The concentrations of serum TNF-а, IL-1βin HO-1group were significantly lower than that of SOD group（P0.05）, The concentrations of serum IL-10 was significantly higher than that of SOD group（P〈0.05）. Conclusions The apoptotic of ADSCs under the circumstance with serum free and oxygen deprivation can be significantly cut down by eveulating the expression of HO-1 in ADSCs. The mechanisms are associated with the lower excretion of inflammatory factors： TNF-а, IL-1β; and the higher secretion of anti-inflammatory factor-IL-10.