Sepsis　induces　critical　myocardial　dysfunction,　resulting　in　an　increased　mortality.　Gracillin　(GRA)　is　a　natural　steroidal　saponin,　showing　strong　capacities　of　anti-inflammation,　but　its　pharmacological　effects　on　lipopolysaccharide　(LPS)-induced　acute　cardiac　injury　still　remain　unclear.　In　this　study,　we　attempted　to　explore　if　GRA　was　effective　to　attenuate　cardiac　injury　in　LPS-challenged　mice　and　the　underlying　mechanisms.　First,　we　found　that　GRA　treatments　markedly　up-regulated　the　expression　of　miR-29a　in　cardiomyocytes.　LPS-induced　cytotoxicity　in　cardiomyocytes　was　significantly　alleviated　by　GRA　treatment,　as　evidenced　by　the　improved　cell　viability　and　reduced　lactate　dehydrogenase　(LDH)　release.　In　addition,　LPS-triggered　apoptotic　cell　death　was　clearly　ameliorated　in　cardiomyocytes　co-treated　with　GRA.　Notably,　LPS-exposed　cells　showed　significantly　reduced　expression　of　miR-29a,　while　being　rescued　by　GRA　treatment.　In vivo,　LPS　apparently　impaired　cardiac　function　in　mice,　which　was,　however,　alleviated　by　GRA　administration.　In　addition,　GRA　markedly　attenuated　apoptosis　in　hearts　of　LPS-challenged　mice　by　decreasing　the　expression　of　cleaved　Caspase-3.　LPS-triggered　inflammatory　response　in　cardiac　tissues　was　also　suppressed　by　GRA　through　blocking　nuclear　factor　κB　(NF-κB)　signaling　pathway.　We　also　found　that　miR-29a　expression　was　highly　reduced　in　hearts　of　LPS-treated　mice　but　was　rescued　by　GRA　pretreatment.　Besides,　miR-29a　mimic　alleviated　LPS-induced　apoptosis　and　inflammation　in　cardiomyocytes;　however,　LPS-caused　effects　were　further　accelerated　by　miR-29a.　Of　note,　the　protective　effects　of　GRA　on　LPS-injured　cardiac　tissues　were　significantly　abrogated　by　miR-29a　suppression.　In　conclusion,　our　findings　demonstrated　that　GRA　exerted　an　effective　role　against　LPS-induced　acute　cardiac　injury　through　impeding　apoptosis　and　inflammation　regulated　by　miR-29a.