Lipopolysaccharide (LPS) is a component of gram negative bacterial (E. coli) outer membrane. Previous studies showed that LPS present in plasma and follicular fluid of postpartum uterine infected animals and inducing the inflammatory response in bovine granulosa cells. Conjugated linoleic acid (CLA) is a dietary fatty acid that has unique properties like antiinflammatory, anti-cancer, anti-atherogenic and anti-diabetic effects. In present study we studied the effects of LPS on proinflammatory gene expression and CLA as anti-inflammatory agent to attenuate LPS induced inflammatory gene expression in cultured buffalo granulosa cells under serum free media. Granulosa cells were cultured and treated with LPS (1.0 μg/ml) for 6 h at different time intervals and pro-inflammatory cytokines (TNFα, IL-1β and IL-6) were analyzed using qRT- PCR. Result shown that all the three pro-inflammatory cytokines were significantly (P≤0.01) up regulated within 2h and then decreased sharply and reached to almost normal level within 6 h. The Conjugated linoleic acid (CLA), ligand of PPARγ, has been shown to be a potent anti-inflammatory agent. To test, whether CLA can prevent the LPS induced pro-inflammatory cytokines expression in granulosa cells, cells were cultured 48 hrs for basal growth, after basal growth cells were co-treated with CLA (10 μM) and LPS (1.0 μg/ml) for 2 h followed by analysis of expression of pro-inflammatory cytokines. Surprisingly, CLA had no effect on pro-inflammatory cytokine expression. Further, cells were pre-treated with CLA for 24hr after basal growth and then cells were exposed to LPS for 2 h. Result showed that cells pre-treated with CLA showed significantly (P≤0.01) less expression of pro inflammatory cytokines when comparison to LPS alone. In conclusion, the results of the present study showed that CLA pre-treatment can prevent the LPS induced pro-inflammatory cytokines expression in granulosa cells in vitro. (Work supported by Department of Biotechnology (DBT), Govt. of India.).