Anisodamine delayed the progression of intracranial aneurysm by inhibiting ATF6 pathway mediated endoplasmic reticulum stress and vascular endothelial cell apoptosis
Objective To investigate the effects and molecular mechanisms of anisodamine on delaying the progression of intracranial aneurysms(IA).Methods In vivo experiment,30 mice were randomly divided into 3 groups:control group,sham group and model group,with 10 mice each.IA mouse model was established as model group.In vitro cell experiment,primary human umbilical vein endothelial cells(HUVECs)were divided into 3 groups:control group,lipopolysaccharide(LPS)group and LPS+anisodamine group.Western blot assay was used to detect the expression levels of endoplasmic reticulum stress related protein factors and apoptosis-related protein factors in mice Willis ring aneurysms and HUVECs cells.ELISA was used to detect the levels of TNF-α and IL-1[3 in the above tissues and cell supernatants.Results In vivo,compared with sham group,in model group the levels of endoplasmic reticulum stress-related protein factor activating transcription factor-6(ATF6(p50)),phosphorylated AKT serine/threonine kinase(p-AKT),nuclear factor kappa B(NF-K B(p-p65))ubunits in the vascular tissues of Willis ring aneurysms were up-regulated(P＜0.05),ATF6(p90)levels was down-regulated(P＜0.05).Apoptosis-related protein factor BCL2 associated X(Bax)and cleaved Caspase-3 levels were up-regulated(P＜0.05),the level of B cell leukemia/lymphoma 2(Bcl-2)was down-regulated(P＜0.05).There were no significant difference in AKT and NF-K B(p65)subunit levels between the two groups(P＞0.05).The levels of tumour necrosis factor alpha(TNF-α)and interleukin 1 beta(IL-1[3)were up-regulated(P＜0.05).Compared with the control group,there was no significant difference in the above detection indexes in the sham group(P＞0.05).In vitro HUVECs cell experiments,compared with control group,in LPS group cells ATF6(p50),p-AKT,NF-κ B(p-p65)ubunits were up-regulated(P＜0.05),ATF6(p90)levels was down-regulated(P＜0.05).Bax and cleaved Caspase-3 levels were up-regulated(P＜0.05),Bcl-2 was down-regulated(P＜0.05).There were no significant difference in AKT and NF-K B(p65)subunit levels between the two groups(P＞0.05).The levels of TNF-α and IL-1[3 in cell supernatants were up-regulated(P＜0.05).Compared with LPS group,LPS+anisodamine group reversed the levels of the above indexes(P＜0.05).Conclusion Anisodamine may delay the progression of intracranial aneurysm by inhibiting ATF6 pathway mediated endoplasmic reticulum stress and vascular endothelial cell apoptosis.
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