Cytoprotective effects of the medicinal herb Astragalus membranaceus on lipopolysaccharide-exposed cells

Astragalus membranaceus (AM) is a traditional Chinese medicinal herb, whose cytoprotective effects remain largely unknown. Here, the bacterial endotoxin lipopolysaccharide (LPS) was applied to a human pulmonary type II-like epithelial lung adenocarcinoma cell line, a human umbilical vein endothelial cell line, and a human bladder carcinoma cell line to construct in vitro models of intracellular oxidative stress. The authors assayed the cellular and mitochondrial cytoprotective effects of varying doses of AM root extract upon these cell lines. The cell lines were cultured as follows: LPS-only group, four LPS+AM groups treated with various AM concentrations plus LPS, and an untreated control group. Flow cytometry was used to assess apoptosis and cell cycle progression. A 2',7'-dichlorofluorescein-diacetate assay was used to quantitate reactive oxygen species (ROS) generation. Mitochondrial membrane potential (Delta Psi(mit)) was analyzed by Rhodamine 123 assay. Western blotting was performed to detect cleaved caspase-3, p53, and B cell lymphoma (Bcl)-2 levels. Across all cell lines, LPS significantly elevated apoptosis rates, shifted cells to S/G2 phase, increased ROS production, reduced Delta Psi(mit), upregulated cleaved caspase-3, upregulated p53, and downregulated Bcl-2 relative to controls (all P<0.05). As a general trend, increasing AM concentrations produced progressively greater reductions in the apoptosis rate, greater reductions in S/G2 phase %, greater reductions in ROS production, greater increases in Delta Psi(mit), greater reductions in cleaved caspase-3 and p53 expression, and greater increases in Bcl-2 expression. AM treatment protects human pulmonary and bladder epithelial cells, in addition to human endothelial cells, from LPS-induced apoptosis, in a dose-dependent manner.