Geniposide-mediated Sirt3 molecules ameliorate nerve injury in rat subarachnoid haemorrhage
Objective To study the mitochondrial mechanism of improvement of subarachnoid haemorrhage by geniposide and the molecular relevance of Sirt3.Methods A total of 40 SD rats were randomly divided into the sham-operation group,the model group,the nicotinamide ribose group and the geniposide group,with 10 rats in each group.In the sham-operation group,only the middle of the neck was incised and sutured;in other groups,a subarachnoid haemorrhage model was constructed by the thread bolus method;in the sham-operation group and the model group,equal amounts of saline were given;in the nicotinamide-ribose group,nicotinamide-ribose was given at 20 mg/(kg·d),and in the geniposide group,geniposide was given at 30 mg/(kg·d);Neurological severity score(NSS)were used to evaluate the cognitive function of rats;absent field experiment was used to evaluate the cognitive function;water maze experiment was used to evaluate the learning and memory ability;hematoxylin-eosin(HE)staining was used to observe the neuronal cell damage in the brain tissue of rats;ELISA kit was used to evaluate the level of inflammation by detecting tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ),transforming growth factor-β(TGF-β);and the damage to the mitochondrial structure and function was weighed by electron microscopy and ATPase activity.Finally,the alteration of Sirt3 expression in rat brain tissue was determined by Western blot and immunohistochemistry.Results Compared with the sham-operated group,the NSS score of rats in the model group was elevated(t=23.760),the distance travelled and the number of centre traversals were reduced in the open field experiments(t=14.250,10.720),the evasion latency was increased and the number of traversals across the platforms was decreased in the water maze experiments(t=4.618,9.238);HE staining results showed that the neuronal cell damage was aggravated(t=4.959);ELISA results suggested that the expression of pro-inflammatory factors TNF-α and IFN-γwas elevated,and the expression of anti-inflammatory factor TGF-β was decreased(t=4.355,6.716,5.317);electron microscopy and ATP kit results suggested that the mitochondrial damage was aggravated,and the activity of ATPase was decreased(t=9.882);Western blot and immunohistochemistry results suggested that the content of Sirt3 was reduced(t=9.145,7.236),and all of the above differences were statistically significant(all P＜0.05).Compared with the model group,NSS scores were reduced in the nicotinamide ribose and geniposide groups(t=6.325,3.678),the distance travelled and the number of centre traversals were increased in the wilderness field experiments(t=4.782,7.324;6.430,4.191),and the evasion latency was reduced and the number of traversals across the platform was increased in the water maze experiments(t=3.247,6.248;5.145,4.293),fewer damaged neuronal cells(t=4.272,3.184),lower expression of TNF-α,IFN-γ,and higher expression of TGF-β(t=6.824,3.284,5.149;7.145,6.248,4.135),and improvement of mitochondrial structure,increased ATPase activity(t=3.241,6.214)and the amount of Sirt3(Western blot and immunohistochemistry)was elevated in rat brain tissues(t=5.324,2.241;2.356,4.128),and all of the above differences were statistically significant(all P＜0.05).Conclusion Geniposide might alleviate mitochondrial inflammatory damage and ameliorate the complications of subarachnoid haemorrhage and behavioral dysfunction by activating Sirt3 molecules.
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