The relationship of Eucommia ulmoides promoting the therapeutic efficacy of bone marrow stromal cells transplantation for traumatic brain injury
Objective To investigate the effects of Eucommia ulmoides(EU)on the biological features of bone marrow stromal cells(BMSCs)and the therapeutic efficacy of grafted BMSCs for traumatic brain injury.Methods In vitro,the C57BL/6 mice-derived BMSCs were divided into EU and Control groups(8 in each group),which were treated with EU extract(100 ng/ml)and the dissol-vent of the equal volume respectively.Phase contrast microscopy and CCK-8 assay were used to detect cell morphology and prolifera-tion.Immunofluorescence(IF)staining and quantitative reverse transcription polymerase chain reaction(qRT-PCR)were used to de-tect the expression of genes related to neural differentiation[Oct4,neurofibrilar light chain(Nefl),neuro differentiation factor(Neurod1),and nerve growth factor(Ngf)].In vivo,thirty-two 3-month old male C57BL/6 mice were divided randomly into Sham surgery,TBI,TBI +BMSCs and TBI +EU_BMSCs groups(8 in each group).The TBI group,TBI +BMSCs group and TBI +EU_BM-SCs group were treated with 100 μl PBS 7 days after operation,and the TBI +BMSCs group was injected with GFP transgenic murine BMSCs without special treatment via intraorbital vein.GFP transgenic mouse derived BMSCs in TBI +EU_BMSCs group were pretrea-ted with the same amount of EU extract by intraorbital vein for 14 days.Mice in the Sham surgery group underwent all procedures ex-cept TBI surgery and cell transplantation.The locomotor of mice was monitored by Rotarod and Beam walking tests.The histopathologi-cal analyses were performed by IF staining.Results In vitro experiments,after 14 days of EU,phase contrast microscope image analy-sis and CCK-8 test showed that the proliferation activity of EU group was higher than that of Control group(P<0.001).The expres-sion levels of Neurod1 and Ngf mRNA in EU group were higher than those in Control group by qRT-PCR(P<0.001).After treated with EU extract for 70 days,the cells in EU group showed neuron-like morphology.IF staining showed that Nefl and MAP2 were ex-pressed in a higher proportion of cells in EU group,but not in Control group(P<0.001).qRT-PCR analysis showed that the expres-sion of Oct4 mRNA in EU group was lower than that in Control group,while the transcription levels of Nefl,Neurod1 and Ngf were higher than that in Control group(P<0.001).In vivo experiment,the rotating rod fatigue meter and balance beam walking tests showed that the recovery of motor function in TBI +BMSCs group was significantly better than that in TBI group(P<0.001),and the improvement of motor function in TBI +EU_BMSCs group was further improved than that in TBI +BMSCs group(P<0.001).IF stai-ning of brain tissue showed that the expression level of MAP2 in the lesion of TBI +BMSCs group was higher than that of TBI group(P<0.001),and the expression level of MAP2 in TBI +EU_BMSCs group was further recovered(P<0.001).The content of GFP + transplanted cells and the proportion of GFP + transplanted cells differentiated into MAP2 + cells in TBI +EU_BMSCs group were sig-nificantly higher than those in TBI +BMSCs group(P<0.001).Conclusion EU extract promoted the differentiation of BMSCs into neurons and enhanced the therapeutic efficacy of grafted BMSCs for TBI.
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维普
