Three-way Cell-based Screening of Antistress Compounds: Identification, Validation, and Relevance to Old-age Related Pathologies.

A variety of environmental stress stimuli have been linked to poor quality of life, tissue dysfunctions and ailments including metabolic disorders, cognitive impairment, and accelerated aging. Oxidative, metal and hypoxia stresses are largely associated with these phenotypes. Whereas drug development and disease therapeutics have advanced remarkably in last three decades, there are still limited options for stress management. Since the latter can effectively decrease the disease burden, we performed cell-based screening of antistress compounds by recruiting three chemical models of oxidative (paraquat), metal (cadmium nitrate) or hypoxia (cobalt chloride) stresses. The screening of 70 compounds for their ability to offer protection against oxidative, metal and hypoxia stresses resulted in selection of 5 compounds: Withaferin-A (Wi-A), methoxy Withaferin-A (mWi-A), Withanone (Wi-N), triethylene glycol (TEG), and Ashwagandha (Withania somnifera) leaf extract (M2DM). Molecular assays revealed that whereas stress caused increase in (i) apoptosis, (ii) ROS accumulation coupled with mitochondrial depolarization, (iii) DNA double-strand breaks, (iv) protein aggregation, low nontoxic doses of the selected compounds caused considerable protection. Furthermore, Wi-N, TEG and their mixture treated normal human fibroblasts (at young, mature and senescent stages representing progressively increasing accumulation of stress) showed increase in proliferation. Taken together, these results suggested three-way (oxidative, metal and hypoxia) antistress potential of Wi-N and TEG that may be useful for management of environmental and old-age related pathologies.