Huntington’s disease (HD) is an autosomal neurodegenerative disease characterized by chorea, dystonia, motor ataxia, cognitive decline and psychiatric disorders with gradual loss of nerve cells and has no existing cure for the disease. In the present study, a mitochondrial toxin, 3-nitropropionic acid (3-NP) is used to induce HD like symptoms in rats. Tetramethylpyrazine is one of the active ingredients of Chuan Xiong which was reported to have neurotrophic and neuroprotective activities. The present study was designed to evaluate the role of TMP on 3-NP induced behavioral, biochemical, neurochemical, and histological alterations in the different regions of the brain. Animals were pretreated with normal saline/TMP for 7 days. From 8th day, the treatment groups were co-administered with 3-NP (10 mg/kg, i.p) and continued to the 21st day of the treatment protocol. At the end of the study, we found that the TMP improved all the behavioral performances of 3-NP induced neurotoxic rats, significantly. Further, oxidative stress parameters (lipid peroxidation, reduced glutathione, catalase, and superoxide dismutase), succinate dehydrogenase enzyme, and neurochemical (GABA and glutamate) estimations were done in the brain homogenate. In our study, the treatment with TMP ameliorated the 3-NP induced alterations, in the biochemical and neurochemical parameter in the brain homogenate, dose-dependently. The protective role of TMP further confirmed by measuring the lesion area with the 2,3,5-triphenyltetrazolium chloride staining of the brain slices and histopathological alteration in the hippocampus (CA1 and CA3) and striatal regions of the brain. Hence, the present findings suggest that the protective role of TMP against 3-NP induced behavioral, biochemical, neurochemical, and histological alterations in rats.