Effect of Zhenxin Shengshui Yizhi Fang on Damage of Human Brain Microvascular Endothelial Cells Induced by Aβ25-35 / 中国实验方剂学杂志

ABSTRACT

Objective:To observe the neuroprotective effect and potential mechanism of Zhenxin Shengshui Yizhi Fang(XSF) aqueous extract on human brain microvascular endothelial cells (HBMEC) injury induced by amyloid-β protein(Aβ)25-35. Method:HBMEC cells damage induced by Aβ25-35 was used as Alzheimer' s disease(AD) cell model. The study included control group, Aβ25-35 group, and low, medium and high-dose XSF aqueous extract groups (125, 250, 500 mg·L-1). After treatment, the cytotoxicity of different concentrations of drugs and Aβ25-35 was determined by methyl thiazolyl tetrazolium(MTT) colorimetry. Apoptosis was observed by Hoechst-33258 staining. The activity of Caspase-3 was detected by colorimetry. Western blot was used to detect the expression levels of the receptor of advanced glycation end products (RAGE) and low-density lipoprotein receptor-related protein (LRP1). Result:Compared with the control group, the cell viability of Aβ25-35 group was significantly decreased (P<0.01). Hoechst-33258 staining showed bright blue fluorescence, chromatin condensation, dense staining or fragmentation dense staining, whitening in color, and significant increase of the percentage of apoptotic cells (P<0.01). Caspase-3 activity increased significantly (P<0.01). Western blot showed that RAGE protein expression increased significantly (P<0.01), while low-density lipoprotein receptor-related protein(LRP1), glucose transporter 1(GLUT1) and GLUT3 protein expressions decreased significantly (P<0.01). Compared with the Aβ25-35 group, the cell viability of XSF aqueous extract groups was significantly increased in a dose-dependent manner. The XSF aqueous extract had a more significant protective effect of than the other groups (P<0.05). The XSF aqueous extract group (500 mg·L-1) significantly inhibited the number of apoptotic cells (P<0.01), but significantly reduced the Caspase-3 activity (P<0.01). RAGE protein expression was not significantly decreased in XSF aqueous extract group (125 mg·L-1), but significantly decreased in XSF aqueous extract group (250, 500 mg·L-1, P<0.01), while LRP1, GLUT1 and GLUT3 protein expression significantly increased (P<0.05, P<0.01) in a dose-dependent manner. Conclusion:XSF aqueous extract can attenuate the cytotoxicity of HBMEC induced by Aβ25-35 oligomer, inhibit apoptosis, decrease caspase-3 activity and RAGE protein expression, increase LRP1, GLUT1 and GLUT3 protein expressions, and reduce the abnormal accumulation and deposition of Aβ in the brain, which may be its mechanisms for prevention and treatment of AD.