ABSTRACT
To study the protective effect of Ershiwuwei Zhenzhu Pills on ischemic stroke rats. Ninety 4-weeks-old SPF male SD rats were randomly divided into 6 groups(n=15):sham operation group, model group, nimodipine group(12 mg·kg~(-1)), Ershiwuwei Zhenzhu Pills high-dose group(400 mg·kg~(-1)), Ershiwuwei Zhenzhu Pills medium-dose group(200 mg·kg~(-1)), Ershiwuwei Zhenzhu Pills low-dose group(100 mg·kg~(-1)).The permanent middle cerebral artery occlusion model(PMCAO) was established in the model group, nimodipine group, and Ershiwuwei Zhenzhu Pills groups by the improved thread plug method, while the sham operation group did not insert the thread plug.Nimodipine group and Ershiwuwei Zhenzhu Pills groups were given intragastric administration once a day for 24 days before the modeling operation, and once 1 hour before the modeling operation, while sham operation group and model group were given equal volumes of distilled water.The neuroethology of the surviving rats was measured; The volume of cerebral infarction in rats was measured by TTC method; The histopathology of rat brain was observed by HE method; The expression levels of tumor necrosis factor α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),malondialdehyde(MDA),superoxide dismutase(SOD) and catalase(CAT) in serum were detected by ELISA;The mRNA expressions of Notch 1,Jagged 1,Hes 1 and Bcl-2 in rat brain were detected by RT-PCR;Western blot was used to detect the expression levels of caspase-3 protein in rat brain; the expression levels of vascular endothelial growth factor(VEGF) and CD34 positive cells in rat brain were detected by immunofluorescence.The low, medium and high dose groups of Ershiwuwei Zhenzhu Pills and nimodipine group could significantly reduce the neurobehavioral score and cerebral infarction volume of rats with permanent middle cerebral artery occlusion, reduce the morphological changes of nerve cells, decrease the expression of TNF-α,IL-1β and IL-6 in rat serum, increase the activity of SOD and CAT,and reduce the level of MDA.Furthermore, the expression levels of Notch l, Jagged l, Hes l and Bcl-2 mRNA were significantly increased, and the expression level of caspase-3 protein was decreased.Meanwhile, the number of VEGF and CD34 positive cells increased in the treatment group.The differences were statistically significant. Ershiwuwei Zhenzhu Pills has a protective effect on ischemic stroke rats, and its mechanism may be related to anti-inflammation, anti-oxidation, promotion of nerve cell proliferation, inhibition nerve cell apoptosis and promotion of angiogenesis.
Subject(s)
Animals , Male , Rats , Caspase 3/metabolism , Drugs, Chinese Herbal/pharmacology , Infarction, Middle Cerebral Artery/drug therapy , Interleukin-6/metabolism , Ischemic Stroke/drug therapy , Nimodipine/pharmacology , Proto-Oncogene Proteins c-bcl-2/metabolism , RNA, Messenger , Rats, Sprague-Dawley , Superoxide Dismutase/metabolism , Tumor Necrosis Factor-alpha/metabolism , Vascular Endothelial Growth Factor A/metabolismABSTRACT
This study was aimed to observe the influence of Catalpol on real-time gait analysis of early recovery after permanent middle cerebral artery occlusion (pMCAO) in rats to evaluate its effect on the improvement after cerebral ischemia. Rats were randomly divided into 6 groups, which were the normal control group, model group, Tianbaoning group, Catalpol 15, 30, 60 mg·kg-1 group. Rats were trained on the gait instrument for 7 days before pMCAO, 3 times/day. After the training, pMCAO model was made. And continuous infusion was performed from the 3rd to the 14th day after the operation. Then, the real-time gait behavior was detected on the 14th day. The re-sults showed that 14 days after the surgery, compared with the normal control group, the models had a significant extending in the duty cycle (P < 0.01), and obvious increasing of the average body angle of absolute value (P <0.05), the shortening of the two feet supporting time (P < 0.05), and extending of three feet supporting time (P <0.01), and increasing of the coordination index (P < 0.01). Compared with the model group, the Catalpol of 30 mg·kg-1 and 60 mg·kg-1 group has obviously decreased duty cycle and average body angle of absolute value (P <0.05, or P < 0.01). The Catalpol of 30 mg·kg-1 can obviously reduce the coordination index of the right front foot relative to the other three feet (P < 0.05), which improved the coordination of cerebral ischemia animals. It was concluded that Catalpol can improve the real-time gait behavior changes of cerebral ischemia model rats. There-fore, Catalpol have a neural protective effect on cerebral ischemia.