We aimed to develop and standardize a diet and rotenone-based animal model of Parkinson's disease. To achieve this, 21 days old male C57BL/6 mice were fed on a high-fat diet [HFD] (60%kcal) for 17 weeks; starting from week 11, animals were orally gavaged daily with 10mg/kg rotenone dose till the end of the 17th week. Further, they were subjected to behavioral tests (Rota-rod, Y maze, and fecal pellet output test). Fecal pellets were collected for 16S rRNA sequencing and animals were subsequently euthanized. Our data revealed that rotenone and its combination with HFD exhibited loss of motor functions and constipation-like effects in the fecal pellet output test. The combination of rotenone and HFD caused gut barrier disruption (ZO-1 and occludin levels). The combination also exhibited colonic inflammation signified by elevation of TRPV1, GFAP, MPO, inflammatory cytokine levels, and peripheral CRP levels. Alpha-synuclein pathology was detected in the colon of the combination group. Gut dysbiosis was also prominent, signified by an increased Firmicutes/Bacteroidetes ratio and an alteration at phylum, order, and family levels. The combination group elicited increased neuroinflammation signified by increased pro-inflammatory cytokine levels in the striatum, combination group also exhibited mild dopaminergic neuronal loss in nigra and striatum and gross neuronal loss in the striatum. Elevated levels of GFAP and TRPV1 were noted in the combination group brain. Thus, the study revealed holistic non-motor and motor behavioral deficits along with gut dysbiosis and a gut inflammation state, thus pathology initiated in the gut and anterogradely traveled in the brain.