ABSTRACT
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.
ABSTRACT
Objective To screen the optimized Buyang Huanwu Decoction (BYHWD);To verify it. Methods H2O2 was used to induce PC12 cell oxidative stress models. MTT method was used to determine the prevention effects of BYHWD at different concentrations (0.1, 0.2, 0.5, 1.0, 2.0, 3.5 mg/mL) on in vitro oxidative stress cell models to define the optimized concentration. Orthogonal design was used to divide BYHWD single medicine into decomposed BYHWD groups, control group (only with DMEM), normal group (without H2O2 and medicine processing), and model group, to investigate the protective effects on PC12 cells. Optimized BYHWD was screened to decide the compatibility ratio of each medicine. MTT was used to detect the cell survival rate in each group. Middle cerebral artery occlusion was used to replicate MACO rat models. SD rats were randomly divided into sham-operation group, model group, BYHWD group and optimized BYHWD high-, medium-and low-dose groups. Each medication group was given relevant medicine for gavage. The screened results were verified. Results Compared with other decomposed BYHWD groups, the protective effects of the compatibility of Astragali Radix+Chuanxiong Rhizoma+Pheretima on PC12 cells was the best (P<0.05), which was nearly equaled to BYHWD. Compared with the model group, BYHWD and the optimized one could evidently reduce cerebral cortex infarction area and improve the impaired brain edema (P<0.05), and the medium-dose group was the best. Conclusion The optimized BYHWD ratio is:Astragali Radix:Chuanxiong Rhizoma:Pheretima=10:3:1.
ABSTRACT
Objective To investigate the preventive effects of Buyang Huanwu Decoction (BYHWD) and its recipe composition (BYJJF) in focal ischemic brain injury condition in vivo/in vitro. Methods In vivo studies, SD rats were divided into sham-operation group, MCAO group, BYHWD group and BYJJF group based on rat weight, 10 rats in each group. The body weight, infarct area and brain water contents were determined. In vitro studies, H2O2 was used to damage PC12 cells, and the vitro oxidative stress cell model was established. PC12 cells were divided into normal group, blank control group, BYHWD and BYJJF groups with different concentrations (0.1, 0.2, 0.5, 1.0, 2.0, 3.5 mg/mL). MTT method was employed to determine the protective effects of BYHWD and BYJJF on model cells.Results Vivo studies showed that after 7 days of treatment with BYHWD and BYJJF, those determinated quotas were all significantly improved compared with MCAO model rats (P0.05).Vtiro studies showed that the protective effects of BYHWD and BYJJF took place 2 hours later, and it was obvious in oxidative stress injury caused by H2O2, with statistical differences with model group (P0.05).Conclusion The research confirmed that BYJJF plays a significant role in improving the cerebral ischemia injury, which is the same performance as BYHWD, and BYJJF can save TCM resources under the precondition of TCM efficacy.