Despite the chemical and biochemical analysis of the therapeutic properties of herbs, the molecular evidence for detecting the mode of action of natural drugs has been a novel trend of researchtoday. Hemigraphisalternata, the tropical herb ofAcanthaceae family has gained sufficient therapeutic attention as a potent phytomedicine for wound healing in folklore medicines. Transforming Growth Factor Beta 1 (TGF β1) is one such growth factor which plays effective role in wound healing. In the present study an attempt was made to revealthe molecular mechanism ofthe wound healing effectof Hemigraphis alternata leaf extract (HALE) fractions separated by silica based column chromatography. It was obvious from the data that the fractions 8 and 9 showed higher wound closure. The absolute quantification data of TGF β1 gene expression ofHALE fraction 9 showed higher level expression in HEK 293 treated cells. Thus the molecular data clearly confirms the role of TGF β1gene I in wound healing by the effect of HALE. To the best of our knowledge this is the first time report showing the molecular evidence on the wound healing effect of Hemigraphisalternatain human cell lines.