BackgroundPalmatine is a protoberberine alkaloid which identified to be the bioactive compound present in stem extract of Cosininum Fenestratum plant. Our previous research have reported the plasma glucose lowering activity of Palmatine and its effect on liver, kidney and antioxidant enzymes parameters in STZ induced diabetic rat model. We also reported that the molecular mechanism of action of palmatine insulin release is through activation insulin‐dependent pathway in vivo and in vitro.ObjectiveThe focus of this study was to determine the effect of Palmatine on insulin dependent signaling pathway in insulin resistance rat skeletal muscle ex‐vivo.MethodNormal Male SD rats (180–200g) were sacrificed and skeletal muscles were harvested. The smaller size skeletal muscles from rats were mounted in the Krebs‐Henseleit buffer (KHB) buffer to maintain the resting position. The tissues were then incubated at 37°C under 95% O2‐ 5%CO2 in Glucose free KHB containing 01% BSA and 1mM pyruvate for 1hr, and KHB in 500nmol/L insulin and 125nmol/L glucose for 24hrs. After incubation with insulin/glucose KHB, tissues were transferred to KHB buffer with 250mmol/L glucose for 5hrs, and then KHB with insulin for 30 mints. After induction tissue were replaced with fresh KHB drug was administered by pipetting drug into the organ bath and was allowed to incubate for 24hrs. RNA was extracted and converted to cDNA according manufacturer protocol and the expression of IRS‐1, PI3K, PKC, AKT2 and GLUT4 genes involved in insulin dependent signaling pathway were evaluated using qPCR.ResultPalmatine showed an increase in fold expression of PI3K, PKC, AKT2 and GLUT 4 compared to negative control. The result was similar to standard drug metformin. The increase in fold expression of the insulin dependent pathway genes shows they were revived as a result of Palmatine treatment.ConclusionPalmatine were able to rejuvenate PI3K, PKC, AKT2 and GLUT 4 in Palmatine treated group, which is usual impaired in insulin resistance skeletal muscle. The result is in consonance with the previous finding of in vitro and in vivo. Currently work is on‐going to investigate the effect of palmatine on the insulin‐independent pathway.Support or Funding InformationUCSI University