Experimental Study Of The ADT-OH’s Inhibition Of Neuroinflammation And Anti-tumor Pharmacological Activity

Aim: Recent studies suggest neuroinflammation and microglia activation play important roles in PD pathogenesis, may be a cause of the selective loss of dopaminergic neurons in the substantia nigra. 5-(4-hydroxyphenyl)-3H-1,2-dithiole-3-thione(ADT-OH) is a H2S-releasing compound, H2 S is part of a group of biologically active gases that are termed gasomediators. It has been shown that H2 S attenuates the expression of pro-inflammatory cytokines by lipopolysaccharide(LPS)-activated microglia. This study is thought to discuss wether ADT-OH has inhibitive effect on neuroinflammation and anti-tumor activity.Methods: Using LPS stimulate BV2 cells to establish the model of microglia over activation. The cell model was pretreated by dosing ADT-OH, according to the drug treatment of concentration gradient and gradient time experimental results to determine the best treatment time and the most effective concentration. Weston-blot was used to detect the protein level of pro-inflammatory cytokines iNOS and COX2 in the downstream, and the secretion of pro-inflammatory factors IL-6 and TNF-α in the supernatant were detected by ELISA. Detection of transcription levels of a series of inflammatory cytokines by RT-PCR and Q-PCR. In addition, Weston-blot and fluorescence analysis were applied to detect the kinase activity, protein degradation and translocation of NF-κB pathway. SH-SY5 Y cells were treated with condition medium of BV2, the viability and apoptosis of SH-SY5 Y were detected by MTT and flow cytometry after PI stain. Using MTT to detect ADT-OH is capable of inhibiting the activity and proliferation of tumor cell lines, the cell cycle of tumor cells was detected by flow cytometric.Results: The experiment results suggested that ADT-OH had great inhibition effect on microglia over activation. Pretreatment with ADT-OH significantly inhibited the transcription and protein levels of pro-inflammatory cytokines(e.g., iNOS, COX2, IL-6, TNF-α, etc.). ADT-OH can down-regulate the phosphorylation of IKK and p65, also the p65 translocation were inhibited. Moreover, ADT-OH showed neuroprotective effect through inhibiting the over activation of microglia. MTT and flow cytometric results showed that ADT-OH had anti-tumor activity by interfering with the cell cycle.Conclusion: ADT-OH has significantly anti-neuroinflammation and anti-tumor activity, this may be a worth considering therapeutic method of Parkinson’s disease.