Glaucoma, the collection of syndromes, is the 2nd leading basis of vision loss in the world. Primary open angle glaucoma (POAG), the mostly occurring form is described by optic nerve head excavation, defective fields of vision & increased or regular intraocular pressure (IOP), with no sign of closed angle on Gonioscopic examination. Though the actual reason of POAG is unclear yet but inherited factors may perform key function in understanding actual mechanism of the disease. This study was conducted for linkage analysis between POAG and two genes (MYOC & CYP1B1) related with POAG in the people of Sindh province of Pakistan and elaborate the protecting properties of antioxidants (Resveratrol & Ascorbate) on cultured HTMC subjected to reactive oxygen species. For this purpose, twenty-six families having multiple affected members diagnosed with primary open angle glaucoma (POAG) were enrolled through hospitals from different cities of Sindh, Pakistan and were numbered from POAG-1 to POAG-26. After genomic DNA isolation, exclusion studies were carried out to analyze enrolled families for linkage to MYOC and CYP1B1 genes. Out of twenty-six families, two families were found linked with the MYOCgene and one family was found linked with CYP1B1 gene. In family POAG-26, a novel mutation for MYOC gene, c.1130 C>G resulting in p.T377R was identified where as in family POAG-06, one reported mutation for MYOC in Indian population, c.144G>T resulting in p.Q48H was found. In another family POAG-02, a novel mutation for CYP1B1 gene, c.650 G>A resulting in p.D217N was detected. For exploration of potential effects of antioxidants (Resveratrol & Ascorbic acid) on cultured Human Trabecular meshwork cell line, Cells were purchased from The ScienCell Research Laboratories, Canada. Cultured HTM cells were co-incubated with varying concentrations of H2O2 in growth media for twenty-four hours. Effects of different concentrations of H2O2 on TM cell metabolism were determined by observing the cells under light microscope and detecting mitochondrial enzymes functional activity with the help of MTT assay. It was proved that Resveratrol has more protecting properties than the Ascorbic acid, even at 4.0 mM H2O2, the TM cellular movement was 76%. Underneath co-treatment circumstances, the 1mM Resveratrol maintained TM cell metabolism. Related to H2O2- treated TM cells, resveratrol enhanced role of mitochondria up to 4.0 mM H2O2 (76%). For evaluation of protective effects of co-incubation of Ascorbate and Resveratrol was because of metabolic alteration of Trabecular Meshwork cells, which were pre-treated by Ascorbate and Resveratrol for 24 hours followed by H2O2 treatment termed as “pretreat-only’’. Highly significant differences were noticed for the 1.0 mM H2O2 concentration (Chi Square test, p=0.0001). A considerable decrease in metabolism of Trabecular Meshwork cells was noticed estimating sixty one percent at 1.0 mM of hydrogen peroxide associated to Ascorbic acid – ninety nine percent and Resveratrol ninety nine percent (p=0.0001).