Ameliorative effect of helichrysum cymosum extracts and biosynthesized zinc oxide nanoparticles on c3a hepatocytes, l6 myocytes and tm4 sertoli cells.

Abstract

The current study investigated the antidiabetic, antioxidant, anti-inflammatory, cytotoxicity, and reproductive effect of Helichrysum cymosum aqueous, ethanol and zinc oxide nanoparticles using In vitro assays. The shoots of H. cymosum were extracted using two solvents; aqueous by infusion and ethanol by extraction. Meanwhile zinc oxide nanoparticle was obtained by green synthesis from the aqueous extract. Bioactive compounds were identified on the aqueous and ethanol extracts using High-Performance Liquid Chromatography and UHPLC-ESI-MS. Cytotoxicity of all tested sample was carried out using MTT (colorimetric assay) on C3A hepatocytes, L6 myocytes and TM4 Sertoli cell, to establish a non-toxic concentration. Antidiabetic effect of aqueous and ethanol, and zinc oxide nanoparticles were tested by enzymatic inhibitory assays; alpha glucosidase, alpha amylase and pancreatic lipase inhibition. Additionally glucose uptake and utilization were assessed on C3A and L6 cell lines. The mouse macrophage cell line, RAW 264.7 was used to assess anti-inflammatory activity of aqueous, ethanol and nanoparticles, by measuring the levels of nitrite produced. The ferric reducing antioxidant power (FRAP), trolox equivalent antioxidant capacity (TEAC) and 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay were used to examine the antioxidant activity of aqueous and ethanol extracts, while average cellular CellROX® Orange fluorescent intensity was used to determine the intracellular antioxidant activity of both extracts and nanoparticles on C3A and L6 myocytes cells. The antioxidant enzyme activity, superoxide dismutase, glutathione and other non-enzymatic assays like total antioxidant capacity, reactive oxygen species and morphological changes were performed on TM4 Sertoli cell lines to evaluate the antioxidant activity on reproductive cells. The findings reveal that the extracts and zinc oxide-mediated nanoparticles demonstrated potent hypoglycemic activity and antioxidant activities, mitigating the effect of oxidative stress induced complications associated with diabetes. The extracts also showed enhanced antioxidants such as glutathione and superoxide dismutase and positively interfered in reactive oxygen species production in reproductive cells indicating that the extracts contain important bioactive constituents which can alleviates oxidative stress induced-infertility complications. The current study suggest that H.cymosum extracts and zinc oxide mediated nanoparticles have prospect for future development of antidiabetic and infertility drug. However, additional research in vivo and clinical trials is necessitated.