The endometriosis treatment was critical issue due to complications associated with current drug delivery system. Present investigation compared the curative effect of Vitamin D3 (VTD3) and Omega–3 (OG3) with Diphereline during the treatment of endometriosis. Endometriosis was induced in different groups containing 60 adult female rats. The rat model is categorized into 6 groups untreated and treated (Olive Oil (solvent), VTD3 (42 mcg/kg/day), OG3 (450 mg/kg/day), VTD3+OG3, Diphereline (3 mg/kg/day)). The suspension containing combination of Diphereline and supplements was injected and treated for 4 weeks to analyses the effect of supplements. The interleukin -6 (IL-6) and Tumor necrosis factor – alpha (TNFα) inflammatory responses were measured from the serum samples while endometrial implants was dissected and histopathological investigation was done. At the end of four weeks pathologic score was decreases significantly with simultaneous measurement of inflammation score of endometriotic lesion, size of implant area, IL-6, TNFα response and compared with untreated female rat. No significant different was observed in groups undergoing treatment of VTD3, OG3 and Diphereline. The combined effect of VTD3+OG3 has similar responses with Diphereline treated endometrial implants. In conclusion, treatment of VTD3 deficiency and making a change in dietary habits of high-risk population for endometriosis from adolescence may also play a preventative role in adulthood.