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Aim To explore the neuroprotective mechanism of thin recipe of Buyang Huanwu decoction on CDK5 and apoptotic factors in an oxidative stress cell model. Methods A PC 12 cell model of oxidative stress damage was established using hydrogen peroxide (H202) at the concentration of 0. 3 mmol • L"1 co-cultured for 2 h, PC 12 cells were divided into four groups, namely normal control, model control, Buyang Huanwu decoction and its thin recipe groups. We evaluated the content of extracellular L-LDH and intracellular GSH by ELISA, and the extracellular ROS levels by FCM. In addition, the expressions of CDK5 and Tau were further analyzed to facilitate the understanding of neuroprotective mechanism by IF, and the mR-NA levels of apoptotic factors such as caspase-3 and the ratio of Bax/Bcl-2 mRNA were determined by qPCR. Results The thin recipe of Buyang Huanwu decoction increased GSH activity, decreased L-LDH and ROS levels compared with model cells at the concentration of 1.0 g • L"1 (P < 0. 05). Furthermore, we observed lower expression of CDK5 and Tau, as well as less mRNA of caspase-3 and the ratio of Bax/Bcl-2 in the thin recipe of Buyang Huanwu decoction groups than that of model groups (P < 0. 05). Conclusions The thin recipe of Buyang Huanwu decoction could play the neuroprotective effects by CDK5 and Tau signal sites to inhibit neuronal apoptosis after oxi-dative stress damage of PC12, which illuminate the partial mechanism and the reasonability of this new recipe simplified from Buyang Huanwu decoction in molecular level.
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Aim To evaluate the regulation of thin recipe of Buyang Huanwu decoction on cyclin-dependent kinase 5(Cdk5)expressions in hippocampus tissue of rats after cerebral ischemia.Methods Male SD rats were divided into sham-operation group,MCAO group,Buyang Huanwu decoction group(ig.3.15 g·kg-1)and its thin recipe composition group(ig.2.41 g·kg-1).Each group was then divided into five subgroups based on the time after administration for 1,3,7,14,28 d respectively.Cdk5 protein and mRNA levels in each group were examined by using immunohistochemistry,Western blot and real-time PCR respectively.Results The up-regulation of Cdk5 was observed in model rat hippocampus after cerebral ischemia 1 day,and kept increasing with the aggravation of ischemia injury,the peaked expression was observed after 7～14 d,while the downtrend was observed after 28 days compared with the corresponding sham-operation groups(P0.05).Conclusion The thin recipe of Buyang Huanwu decoction could exert the protective effect by regulating Cdk5 after cerebral ischemia.
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Aim To observe the effects of thin recipe of Buyang Huanwu Decoction on angiogenesis and the signal pathway of Nrf2 / HO-1 after cerebral ischemic injury in rats. Methods Totally 120 SD rats were randomly divided into four groups: sham operation group,model group,Buyang Huanwu Decoction group and thin recipe of Buyang Huanwu Decoction group. The focal cerebral ischemia rat model was established by middle cerebral arterial occlusion. Each group was treated with corresponding treatment. Each group was detected after cerebral ischemia for day 1,day 3 and day 7, respectively. Immunohistochemical method was used to detect the plasma levels of factor VIII related antigen( vWF), determination of microvessel density (MVD). The expression of Nrf2,HO-1 gene and pro-tein in brain tissues was detected by Real-Time PCR (RT-PCR) and Western blot. Results ① Compared with sham-operation group, the expression of vWF in the model group was significantly increased on day 3(P< 0. 05). Compared with model group, the expression levels increased differently in each drug group on day 7 (P < 0. 05). ② The expression of Nrf2, HO-1 gene and protein in sham operation group showed a small a-mount of gamma expression. Compared with sham op-eration group at the same time point, the expression of Nrf2 protein was significantly increased on day 3(P <0. 01). Compared with model group at the same time point, the Nrf2mRNA and protein expression was up-regulated in each drug group. The Nrf2mRNA on day 1,the Nrf2 protein on day 1 and day 7 were significant-ly increased( P < 0. 01). Compared with sham opera-tion group at the same time point, the expression of HO-1mRNA and protein in the model group was signif-icantly increased on day 7(P < 0. 05). Compared with model group at the same time point, the HO-1mRNA on day 3, the HO-1 protein on day 3 and day 7 in each drug group were significantly increased (P < 0. 05,P <0. 01). Conclusions The thin recipe of Buyang Hua-nwu Decoction promotes brain angiogenesis after ische-mia. The effect may be related wih the expression of Nrf2 / HO-1 signal pathway.