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Aim To study the neuroprotective effects of Herba siegesbeckiae extract on cerebral ischemia/ reperfusion rats and its mechanism. Methods Sixty SD rats were randomly divided into model group, low, middle and high dose groups of Herba siegesbeckiae, and Sham operation group, and the drug was given continuously for seven days. The degree of neurologic impairment was evaluated by mNSS, and the infarct volume was measured by MRI. The number of Nissl-posi- tive cells was detected by Nissl staining, and the apop- tosis was accessed by Tunel staining. Furthermore, the expression of Bax, Bcl-2 and NeuN was observed by Western blot, and the expression of NeuN was detected by immunofluorescence staining. The expression of IL- 1β, TNF-α and IL-6 mRNA was performed by RT- qPCR. Results The mNSS score and the volume of ischemic cerebral infarction in the model group were significantly increased, and Herba siegesbeckiae extract treatment significantly decreased the mNSS score and infarct volume (P<0.05, P<0.01). Herba siegesbeckiae extract could increase the number of Nissl-pos- itive cells and the expression of NeuN (P<0.01), and reduce the number of Tunel-positive cells (P<0.01). Western blot showed that Herba siegesbeckiae extract inhibited the expression of Bax, increased Bcl-2 and NeuN in ischemic brain tissue (P<0.01). RT-qPCR showed that Herba siegesbeckiae extract inhibited the expression of IL-1 β, TNF-α and IL-6 mRNA in the is-chemic brain tissue (P<0.01). Conclusions Herba siegesbeckiae extract can reduce the cerebral infarction volume, improve the neurological function damage, inhibit the apoptosis of nerve cells and the expression of inflammatory factors and promote the expression of NeuN, there by exerting protective effects on MCAO rats.
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Aim To explore the mechanism of Buyang huanwu decoction attenuating cerebral ischemia-reper- fusion injury in rats by regulating autophagv through AMPK/mTOR/ULKl signaling pathway.Methods Left eerebral ischemia model in rats was established by modified thread ocelusion method, then the rats in each group were given medieine onee every 24 hours for 3 times.After 72 hours of reperfusion, the nerve injury and the changes of cerebral infarction volume were observed; the morphology, number and apoptosis of nerve cells were observed by Nissl staining and TUNEL staining; the expression of autophagy protein and AMPK/mTOR/LJLKl autophagy signaling pathway related proteins were detected by Western blot.Results Buyang huanwu decoction could improve the neurological deficit of rats, reduce the volume of cere bral infarction and neuronal apoptosis, reduce the pathological injury of brain tissues, inhibit the phosphorylation activation of AMPK, relieve the inhibition of AMPK on downstream mTOR and LJLK1 , promote the phosphorylation activation of both, and inhibit autophagy.AMPK agonist metformin increased the level of autophagy and reversed the protective effect of Buyang huanwu decoction on cerebral ischemia-reperfusion injury in rats.Conclusion Buyang huanwu decoction mediates AMPK/mTOR/ULKl autophagy signaling pathway to play a neuroprotective effect on cerebral is- chemia-reperfusion injury in rats.
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Aim To explore the protective effect of ac-tive components of Xiaoxuming decoction ( XXMD ) on focal cerebral ischemia/reperfusion injury in rats dur-ing early recovery period .Methods The ischemia and reperfusion of middle cerebral artery model rats were established by nylon wire with 2 hours ischemic time on healthy male SD rats .The models of MCAO were eval-uated by Zea-Longa′s standard score .The model rats were randomly divided into sham operation group , the ischemia/reperfusion model group , the active compo-nents group of Xiaoxuming decoction and the positive group (extract of ginkgo biloba leaves EGb 761).Rats were orally administrated with different drugs 24 h after operation for up to 14 days, once a day.The effect of active components of Xiaoxuming decoction on behavior changes of MCAO rats in different recovery period was evaluated by a series of behavioral assessent methods such as modified neurological severity scores and cor-ner test.The infarct volume was observed by TTC stai-ning.Moreover, the contents of MDA and the activities of GSH-Px, SOD and NOS in the penumbra and core tissues of rat brain were detected by spectrophotometric method.Results Compared with the I/R model group, the active components group of XXMD could significantly alleviate the neurological deficit scores with prolonged administration . The motion function tended to be normal , stayed longer on the balance beam, sensory function gradually restored sensitivity , reduced the radio of turning to the right .Among them , compared with model group , the active components of XXMD could effectively improve the neurological dys-function after five days of administration ( P<0.05, P<0.01).Meanwhile, the active components of Xiaox-uming decoction could significantly reduce the infarct volume percentage in the cerebral tissue on post opera-tion day 5 and 14(P<0.01).In addition, the active components of XXMD could reduce the content of MDA and the activity of NOS , increase the activity of SOD and GSH-Px.Conclusion The active components of XXMD can generate neuroprotective effect in early stage of recovery and may play a major role in regula-ting the level of oxidative stress in rat brain .
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Aim To discuss the application of tissue spontane-ous fluorescence for draw focal cerebral ischemia reperfusion in-jury in rats based on specific fluorescence detection technology. Methods The change of spontaneous fluorescence WAS COM-PARED between the brains of normal rats and those of rats with draw focal cerebral ischemia-reperfusion injury and an quantita-tive analysis was then made. Result The results showed that spontaneous fluorescence of brain tissue for focal cerebral ische-mia reperfusion injury changed significantly. Spontaneous fluo-rescence signal of injury considerably enhanced. The fluores-cence signal which was quantified by IVIS had significant statisti-cal significance compared with normal brain tissue, P <0. 05. Conclusion Our research shows that spontaneous fluorescence of brain tissue enhances obviously after focal cerebral ischemia-reperfusion injury. Our research provides a method for the re-search and evaluation of focal cerebral ischemia-reperfusion inju-ry model in rats.