Intervention and mechanism of Huoxue Rongluo Formula on BMEC after OGD/R based on JAK2/STAT3 pathway
Objective To investigate the intervention and mechanism of Huoxue Rongluo Formula(HXRLF)on brain microvascular endothelial cell(BMEC)after oxygen-glucose deprivation/reoxygenation(OGD/R)by activating Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3)pathway.Methods Rat BMEC(bEnd.3 cell)was cultivated and an OGD/R model was established.The cells were randomized into normal(10%blank serum),model(10%blank serum),HXRLF(10%HXRLF containing serum),butylph-thalide(10%butylphthalide containing serum),Stattic(10%blank serum+10 μmol/mL Stattic),and combination(10%HXRLF serum+ 10%μmol/mL Stattic)groups,and were given corresponding drug interventions for 24 h,respectively.CCK-8 assay was used to determine the effects of HXRLF on the activity of bEnd.3 cells after OGD/R.Wound healing assay and Transwell migration assay were used to test the effects of HXRLF on scratch healing and cell migration ability of bEnd.3 cells after OGD/R;the protein expression levels of JAK2,p-JAK2,STAT3,p-STAT3,and VEGFA in bEnd.3 cells were determined by Western-blot.Results Compared with the normal group,the survival rate,scratch healing rate,and cell migration number of bEnd.3 cells in the model group were significantly lower(P<0.01),while the expression levels of JAK2,p-JAK2,STAT3,p-STAT3,and VEGFA were higher(P<0.05 or P<0.01).Compared with the model group,the survival rate,scratch healing rate,and migration number of bEnd.3 cells in the HXRLF group were significantly higher(P<0.01),and the expression levels of JAK2,p-JAK2,p-STAT3,STAT3,and VEGFA were significantly higher(P<0.05 or P<0.01).Compared with the Stattic group,the survival rate,scratch healing rate,and cell migration number of bEnd.3 cells in the combination group were significantly higher(P<0.05 or P<0.01),and the expression levels of STAT3 and VEGFA were higher(P<0.05).Conclusion HXRLF may activate the JAK2/STAT3 signaling pathway,induce endothelial cell proliferation,migration,and survival,activate VEGFA expression,and alleviate cerebral ischemia-reperfusion injury,thus exerting neuroprotective effects.
ischemic strokeHuoxue Rongluo FormulaJanus kinase 2/signal transducer and activator of transcription 3 path-wayoxygen-glucose deprivation/reoxygenationbrain microvascular endothelial cell
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维普
