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Objective To investigate the protective effect of Buyang Huanwu Decoction ( BYHWD) on PC12 cell apoptosis induced by oxygen-glucose deprivation ( OGD) and to screen out the main ingredients of BYHWD on protecting PC12 cell from injury induced by OGD. Methods Seven herbs of BYHWD were selected as the influencing factors, and they were Radix Astragali, Radix Paeoniae Rubra, the carda part of Radix Angelicae Sinensis, Pheretima Asiatica, Rhizoma Chuanxiong, Semen Persicae, and Flos carthami. Two levels were designed for each factor, and the experiment was arranged according to L8 ( 27) orthogonal test table. Apoptosis of PC12 cells induced by OGD was detected by serum pharmacology method and flow cytometry techniques. With apoptotic rate as the evaluation index, visual analysis and variance analysis were used for the comparison of apoptosis of PC12 cells in the blank control group ( oxygen + sugary medium + blank serum) , model group (oxygen-glucose-free medium + blank serum) and drug groups (oxygen-glucose-free medium +serum containing BYHWD or the separate recipe of BYHWD ) . Results Compared with the blank control group, apoptotic rate was increased in PC12 cells of the model group, the difference was statistically significant ( P<0.01) . Compared with the model group, the apoptotic rate was decreased in BYHWD group, and the differences was statistically significant (P<0.01) . The results of orthogonal analysis showed that Semen Persicae, Flos carthami and Radix Paeoniae Rubra were the main active ingredients of BYHWD on depressing apoptosis of PC12 cell induced by OGD ( P<0.01). The apoptotic rate of serum containing the prescription with Radix Paeoniae Rubra was lower than that of the prescription without Radix Paeoniae Rubra. Conclusion BYHWD could inhibit OGD-induced PC12 cell apoptosis, and Radix Paeoniae Rubra is the main active herb on depressing apoptosis in BYHWD.
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Objective To observe the influence of Buyang Huanwu Decoction ( BYHWD) on the inhibition of rat myocardial H9C2 cell activity and SH2-containing tyrosine phosphatase-1 ( SHP-1) activity induced by trastuzumab, and to explore the possible regulatory mechanism after observing the intervention of BYHWD on rat myocardial H9C2 cell transfected with SHP-1 or SHPC/S-1 gene. Methods The eukaryotic expression vectors pcDNA3.1 (+)- SHP-1 and pcDNA3.1 (+) -SHPC/S–1 were constructed and then were transfected to rat myocardial H9C2 cells using the method of liposome transfection. The cells with positive clones were screened out with G418, and then were cultured with trastuzumab for maintaining growth. Using quantitative RT-PCR, we detected the expression of SHP-1 gene and SHPC/S - 1 gene in rat myocardial H9C2 cells. The phosphatase activity analysis was used for observing the regulatory effect of BYHWD on SHP-1 in myocardial cells. Furthermore, we observed the apoptosis of rat myocardial H9C2 cells by methyl thiazolyl tetrazolium (MTT) assay after treatment with BYHWD. Results Sequencing results indicated the successful construction of eukaryotic expression vectors, which had stable expression in myocardial H9C2 cells even under the intervention of trastuzumab. The results of phosphatase analysis showed that H9C2-SHP-1 had the highest activities of phosphatase, but the activities were decreased after the intervention with BYHWD ( P<0.05) . The results of MTT assay also showed the apoptotic rate of H9C2-SHP-1 cells was decreased after treatment with BYHWD ( P <0.05) . Conclusion BYHWD can promote the proliferation of myocardial H9C2 cells inhibited by trastuzumab, and can regulate the expression of SHP-1 in myocardial cells, which will supply reference to the further study of treatment of trastuzumab-induced cardiac toxicity.
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【Objective】 The effect of pretreatment with Buyang Huanwu Decoction(BHD) on L-type calcium channels of cortical neurons in rats with global cerebral ischemia was observed,thus to explore the molecular therapeutic mechanism of BHD for cerebral ischemia as well as the pathogenic mechanism of Ca2+ message in ischemic injury of neurons.【Methods】SD rats were randomized into pseudo-operation group,model groups and BHD groups.And the model groups and BHD groups were divided into six groups according to the time points after reperfusion(2nd,12th,24th,48th and 72nd hour after reperfusion respectively).BHD groups received 0.64g/kg BHD by gastric infusion,bid,for 5 continuous days.Except the rats in pseudo-operation groups,the rats in other groups were induced global cerebral ischemia by modified Pulsinelli's four-vessel occlusion.Cerebral cortical neurons in rats were isolated promptly in 2nd,12th,24th,48th and 72nd hour after reperfusion.The recorded single-channel current was amplified by the EPC-9 patch-clamping amplifier,and then input into the computer by Pulse+Pulsefit.The analytical software TAC was used to detect the opening time and opening probability of the L-type Ca2+ channels in rats at different time points after reperfusion.【Results】In the model groups,the opening time of the L-type Ca2+ channels was prolonged after reperfusion as compared with that in the pseudo-operation group,and the opening probability of the L-type Ca2+ channels arrived at the peaks in 2nd and 24th hour after reperfusion.In BHD groups,the opening time in 72nd hour after reperfusion was decreased(P
ABSTRACT
[Objective] To study the effects of Buyang Huanwu Decoction and Radix Astragali on astrocytes in gerbils with cerebral ischemia and reperfusion injury. [Methods] Gerbils model of cerebral ischemia was set up by occlusion of bilateral common carotid arteries. The dynamic expression of glial fibrillary acidic protein (GFAP) were determined by immunohistochemical method in reperfusion for 24 and 48 hours after 15 minutes of cerebral ischemia. [Results] Positive expression of GFAP reached a peak in reperfusion for 24 hours and was decreased by Buyang Huanwu Decoction and Radix Astragali. Positive GFAP expression was attenuated in reperfusion for 48 hours and enhanced by Buyang Huanwu Decoction and Radix Astragali increased the expression. [ Conclusion ] The regulatory effect of Buyang Huanwu Decoction and Radix Astragali on astrocytes may be one of its mechanisms in repairing nervous function after cerebral ischemia.