Barleria prionitis is a well-known medicinal plant, traditionally used for the treatment of various inflammatory diseases because of its anti-inflammatory activity. But the effect of the extract on inflammatory mediators and cell membrane in response to toxic chemicals was not studied before. Here, we evaluated the membrane stabilization and mast cell protection activity of hydroalcoholic extract of B. prionitis whole plant. The hydroalcoholic extract significantly inhibited the hypo-saline induced erythrocyte membrane hemolysis and the compound 48/80 induced mast cells degranulation in a dose dependent manner. The extract at dose concentration of 10 µg/ml, reduced the rat mesenteric mast cells degranulation up to 64.91% and prevented hypotonic solution induced hemolysis of rat erythrocytes by 27.10%. These findings clearly validate the anti-inflammatory activity of B. prionitis whole plant extract and provide support for traditional usage for inflammatory disorders.