Cistanche extracts ameliorates the neurotoxicity induced by hydrogen peroxide in new mutant DJ‐1‐transfected neuroblastoma cellular models

Abstract

Introduction DJ‐1 mutation is a causative reason for familial Parkinson's disease (PD). Leucine166Proline (L166P) and C106S are two important DJ‐1 mutations. In this study, we established hydrogen peroxide (H2O2) induced L166P and C106S DJ‐1‐transfected neuroblastoma (SH‐SY5Y) cellular models of PD and investigated the effects of Cistanche extracts and key bioactive compounds, including acteoside, echinacoside, caffeic acid, and Cistanche total glycosides on these two models. Methods After expressing FLAG‐tagged L166P and C106S DJ‐1 plasmids in Escherichia coli, the expressed plasmids were collected, treated with restriction enzyme, and identified using DNA electrophoresis. After purification, the L166P DJ‐1 and C106S DJ‐1 plasmids were separately transfected into SH‐SY5Y cells using liposomes. Transfected SH‐SY5Y cells were detected by western blotting and immunocytochemistry. Cell viability was determined using MTT assay. Results Both western blotting and immunocytochemistry showed that L166P and C106S DJ‐1 were highly expressed in the transfected SH‐SY5Y cells. MTT assays showed that transfection with L166P or C106S DJ‐1 reduced the viability of SH‐SY5Y cells exposed to H2O2, as compared to untransfected SH‐SY5Y cells. In addition, Cistanche extracts and key bioactive compounds, including acteoside, echinacoside, caffeic acid, and Cistanche total glycosides, significantly inhibited the decreases of cell viability caused by H2O2 in L166P and C106S DJ‐1‐transfected SH‐SY5Y cells. Conclusions These findings suggest that we successfully established sensitive and stable H2O2 induced L166P DJ‐1‐ and C106S DJ‐1‐transfected SH‐SY5Y cell models of PD and Cistanche extracts may thus be useful for treating PD.

Cistanches Herba, from an endangered species to a big brand of Chinese medicine

“…Second, CH owns therapeutic benefits toward PD. CH and its key bioactive components, including VB, ECH, caffeic acid and total glycoside extract (CTGs) significantly boosts cell viability against H 2 O 2 ‐induced SH‐SY5Y cells, 109 indicating a therapeutic potential for PD. Noteworthily, ECH shows neuroprotective effect on PD pathological mice via regulating p38 MAPK and nuclear factor κB (NF‐κB) p52 signals 110,111 .…”