Effect of Atractylenolide Ⅲ on OGD/R-induced neuronal damage by regulating the Shh/Gli1 signaling pathway
HAN Mei
DANG Xiaohong
LI Yiqun
Abstract：Objective To investigate the effect of Atractylenolide Ⅲ(ATL Ⅲ)on neuronal damage induced by oxygen glucose deprivation and reoxygenation(OGD/R)via regulating the sonic hedgehog(Shh)/glioma-associated oncogene homolog 1(Gli1)signaling pathway.Methods The hippocampal neuron cell line HT22 was induced with blank control,OGD/R,OGD/R+low-dose,medium-dose and high-dose ATL Ⅲ(15,30,and 60 μmol/L ATL Ⅲ,respectively)and OGD/R+the Shh/Gli1 signaling pathway activator PM.The cell counting kit-8(CCK-8)assay was applied to detect cell viability.Flow cytometry was applied to detect cell apoptosis.Enzyme-linked immunosorbent assay(ELISA)was applied to measure the levels of inflammatory factors(interleukin-1beta[IL-1β],tumor necrosis factor alpha[TNF-α],transforming growth factor-beta[TGF-β],IL-10)and oxidative stress indicators(malondialdehyde[MDA],superoxide dismutase[SOD],reactive oxygen species[ROS])in HT22 cells.The colorimetric method was applied to detect Fe2+level.Western blot was applied to detect the protein expressions of apoptosis-related proteins(B-cell lymphoma 2-associated x protein[Bax],B-cell lymphoma 2[Bel-2]),Shh,and Gli1 in HT22 cells.Results Compared with those of blank control,OGD/R induction significantly decreased cell survival,relative levels of TGF-β and IL-10,SOD content,and protein level of Bcl-2,but significantly increased apoptotic rate,relative levels of IL-1β and TNF-α,MDA content,ROS,Fe2+,and protein levels of Shh,Gli1,and Bax(P＜0.05).Low-dose,medium-dose and high-dose ATL Ⅲ treatment in OGD/R-induced HT22 cells significantly increased cell survival,relative levels of TGF-β and IL-10,SOD content,and protein level of Bcl-2,but significantly decreased apoptotic rate,relative levels of IL-1β and TNF-α,MDA content,ROS,Fe2+,and protein levels of Shh,Gli1,and Bax in a dose-dependent manner(P＜0.05).Treatment of PM in OGD/R-induced HT22 cells resulted in significantly lower cell survival,relative levels of TGF-β and IL-10,SOD content,and protein level of Bcl-2,but significantly higher apoptotic rate,relative levels of IL-1β and TNF-α,MDA content,ROS,Fe2+,and protein levels of Shh,Gli1,and Bax than those induced with low-dose,medium-dose and high-dose ATL Ⅲ treatment(P＜0.05).Conclusion ATL Ⅲ can alleviate OGD/R-induced neuronal damage by inhibiting the Shh/Gli1 signaling pathway.
Keywords：Atractylenolide Ⅲsonic hedgehogglioma-associated oncogene homolog 1oxygen glucose deprivation and reoxygenationneuron damage
Publication Date：2025-06-26
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 885-889 )