Exposure of rampant higher glucose level in diabetic condition could lead earlier and late diabetic complication eventually caused malign effect in body organs. The aim of the present work was to analyze the anti-diabetic potential of chloroform fraction of ethanol extract of Andrographis paniculata (AP) and diabetes laden gene expression alteration. In streptozotocin (60 mg/kg bw, i.p.) induced Type 2 diabetes albino mice, two weeks consecutive repeated dose treatment of chloroform fraction of AP at dose of 200 mg/kg, orally was evaluated for antidiabetic screening. Fasting blood glucose, oral glucose tolerance, serum lipid profile, tissues glycogen content, glucose 6-phosphatase and hexokinase enzymes level in liver, in vitro and in vivo insulin estimation were measured on last day of treatment. Subsequently presence of responsible phytoconstituents for respective activity was observed by HPLC. In diabetic mice, the fraction reduced the level of harmful cholesterol, while increased the favorable cholesterol significantly. Biochemical enzymes like glycogen, glucose-6-phosphatase and hexokinase were evaluated in body tissues. Apart from this in vitro, in vivo insulin estimation and diabetogenic gene expression analysis like GK, PEPCK, G-6pase, Glut-4, AR, PPAR-α,γ and TNF-α were evaluated using RT-PCR technique. Antidiabetic screening of fraction of AP at molecular level revealed significant antidiabetic activity.