DEVELOPMENT, OPTIMIZATION AND PHARMACOLOGICAL EVALUATION OF OLIVE OIL BASED CURCUMIN NANOEMULSION WITH ARQ-E-MAKO AND ARQ-E-KASNI AGAINST DIETHYLNITROSAMINE INDUCED HEPATOCELLULAR CARCINOMA IN RATS

Abstract

OBJECTIVE:  Development and optimization of the olive oil-based curcumin nanoemulsion and its evaluation with Ark-e-mako and Ark-e-kasni against diethylnitrosamine induced hepatocellular carcinoma in rats. MATERIAL AND METHODS: The nanoemulsion was constructed via an aqueous titration method. Distilled water served as the aqueous phase, and olive oil, tween 80, and ethanol served as the oil phase, surfactant, and co surfactant. Nanoemulsion was characterized on the basis of particle size and stability study. Further, in vivo pharmacological study, the rats were divided into 7 groups (each 6). The hepatocellular carcinoma (HCC) was induced in all groups except in group I. The HCC was induced by administering diethynitrosamine (DEN) (200 mg/kg, i.p.) single dose and carbon tetrachloride (CCl4 3 ml/kg, i.p.) weekly for 6 weeks. At the end of the six week, blood was collected from each group through retro orbital puncture and subjected to the alpha fetoprotein (AFP) and carcinoembryonic antigen (CEA) test. After confirmation of HCC in rats, all the treated groups I, II, III, IV, V, VI and VII were administered orally with Vehicle (Normal saline), Methotrexate, Curcumin nanoemulsion, Arq-e-mako, Arq-e-kasni and combination of curcumin nanoemulsion, Arq-e-mako, Arq-e-kasni respectively, onec a day for 8 weeks. After the completion of experimental period the rats were anesthetized (Ketamine 60 mg/kg, i.p.) and blood sample was collected through cardiac puncture for estimation of HCC markers such Alpha feto protein and Carcinoembryonic antigen and other biochemical parameters (SGOT, SGPT, ALP, Total Protein etc). Further rats were euthanized; liver were carefully removed and put in a 10% neutral buffered formalin solution for biochemical and histological examination. RESULTS:  The refined formulation exhibited an average particle size of 200 nm and a zeta potential of -13.76. The stability of the formulation under stressful condition was found to be stable. The impact on percentage body weight variation across several treatment groups revealed a notable rise in body weight relative to the disease control. The treated groups had a notable reduction in biochemical parameters of SGPT, SGOT, and ALP when contrasted with the disease control group. The treated groups also exhibit a notable reduction in alpha feto protein and carcinoembryonic antigen levels compared to the disease control groups. Histopathological observations indicate enhancement in cellular architecture within the treatment group relative to the disease control groups. CONCLUSION: The overall results indicate that the optimised formulation when administered along with Arq e-Mako and Arq-e-Kasni has promising hepatoprotective and anticancer potential by improving body weight, liver function biomarkers, tumour related indicators and histological changes.