Cytoprotective and Anti-inflammatory Effects of Derma Genie™-H002 against Paraphenylenediamine-induced Cytotoxicity and Inflammation in RAW 264.7 Cells

Abstract

Purpose: This study aimed to establish an <i>in vitro</i> model to examine paraphenylenediamine (PPD)-induced cytotoxicity and inflammation within RAW 264.7 cells. Additionally, we investigated the cytoprotective and anti-inflammatory effects of Derma Genie™-H002 derived from extracts of five medicinal herbs. Methods: Various biochemical analyses, including water-soluble tetrazolium salt, crystal violet staining, and lactate dehydrogenase leakage assays, were employed to evaluate the impact of Derma Genie™-H002 on PPD-induced cytotoxicity and inflammation. Furthermore, apoptosis- and inflammation-related gene expression was analyzed using reverse transcription-polymerase chain reaction and Western blotting. Results: Derma Genie™-H002 exhibited no significant cytotoxicity in RAW 264.7 cells, whereas PPD induced notable cytotoxicity at concentrations >25 μM. Cotreatment with PPD and Derma Genie™-H002 not only improved cell proliferation rates but also alleviated cell viability in PPD-induced RAW 264.7 cells. Furthermore, findings demonstrated that Derma Genie™-H002 mitigates the cytotoxicity of PPD-induced RAW 264.7 cells by reducing the expression of the proapoptotic protein Bax and increasing the expression of the antiapoptotic protein Bcl-2. Moreover, cotreatment with PPD and Derma Genie™-H002 downregulated the mRNA expression of inflammasome-related genes, inflammatory cytokines, and allergic contact dermatitis-related genes, all elevated by PPD. The increased expression of nuclear factor-κB protein induced by PPD was also reduced upon cotreatment with Derma Genie™-H002, providing compelling evidence of its anti-inflammatory efficacy in the PPD-induced inflammation model. Conclusion: This study demonstrated that Derma Genie™-H002 effectively alleviates PPD-induced cytotoxicity and inflammation in RAW 264.7 cells. Therefore, the potential application of Derma Genie™-H002 as a naturally derived functional anti-inflammatory material has been validated.