Identification of Quercetin, Kaempferol and Luteolin from Methanolic Extract of Corchorus depressus and its Pre-Clinical Wound Healing

Abstract

Corchorus depressus Linn. (Tiliaceae) has been extensively utilized in Odisha, India, and is considered sacred and religious in addition to being used in Pakistan, Baluchistan, and Sind for ethnopharmacological purposes. Ethno-pharmacological claims and lack of sufficient scientific data with respect to the wound-healing activity of the plant encouraged us to the preparation of herbal formulation and its evaluation against wound-healing models with the identification of the phytoconstituents present in the plant extract. The physiochemical properties of the plant like ash value, the preliminary phytochemical screening, the HPLC, HPTLC, FTIR analysis, and the dermal toxicity of the methanolic extract were carried out using standard methods. A stability study was conducted on the prepared ointment formulations. With the help of in vivo excision, incision, dead-space wound, mice-burn model, and along with in vitro stimulation of Interleukin-10 models, the wound healing potential was evaluated. The granulation tissue was collected for nitric oxide estimation and histopathological analysis. The tensile strength of the granuloma tissue and hydroxyproline content were estimated using standard methods. The methanolic extract ointment resulted in a steady contraction of wounds with time and caused significant inhibition in the level of nitric oxide in the scar tissue and aggregation of macrophages in histological examination. The incision model showed a significant (381.33±4.58, p<0.05) increase in the tensile strength of granuloma tissue when compared to the control (156.5±5.75). The dead space wound model demonstrated that the plant extract significantly increased (75.43±2.97, p<0.001) hydroxyproline content against the control. Methanolic extract ointment caused significant wound healing and reduced epithelialization time in the mice burn model from the 12th day till the 24th day as compared to the negative control. Observations of hematological and serum biochemical changes in the dermal toxicity study suggested that the herbal extracts are safe for use topically. In the accelerated stability study, there were no marks of physical instability in the prepared ointment. IR spectral analysis was successfully used to detect flavanones like Luteolin and flavonol such as Quercetin and Kaempferol in methanolic extract. The present findings provide scientific evidence that the plant Corchorus depressus Linn. (Tiliaceae) displayed wound healing activity, corroborating its traditional use by the Tribals of Odisha and the Indian system of medicine.