Effects of Longhu Xingnao Granules on mitochondrial dynamics in an oxygen sugar deprivation reperfusion injury model of SH-SY5Y cells

Fifteen rats were randomly assigned to three groups: blank serum, LHXN-containing serum, and Ginaton-containing serum. The corresponding drug-containing serum was collected after seven days of gavage administration. SH-SY5Y cells were subjected to in vitro OGD/R. Successfully treated cells were then randomly divided into five groups: a control group, a model group, an LHXN-containing serum group, a Ginaton-containing serum group, and an LHXN plus mitochondrial division inhibitor (FCCP) group. The LHXN- and Ginaton-containing serum groups were selected based on the CCK8 assay following six hours of treatment. The optimal concentrations were determined through this assay. Flow cytometry was used to assess apoptosis rates, mitochondrial superoxide levels (using dihydroethidium, DHE), and mitochondrial membrane potential. Transmission electron microscopy (TEM) was employed to examine ultrastructural changes, including mitochondrial swelling, cristae rupture, and vacuolization induced by ischemia/reperfusion. qRT-PCR and Western blotting were used to analyze the relationship between mitochondrial dynamics and the mitochondrial fragmentation inhibitor, along with the expression of relevant proteins (Fis1, Mfn2, Opa1, Drp1) and key factors of the Nurr1/YAP-INF2 pathway (Nurr1, YAP1, INF2).