【Animal modeling】-6-Hydroxydopa model rat salsolinol nitrogen methyltransferase activity detection

  Objective: To establish a rat model of Parkinson's disease with 6-hydroxydopa, and to determine the activity of salsolinol nitrogen methyltransferase (SNMT) in peripheral blood lymphocytes of rats.

  Methods: The rat striatum was damaged by unilateral and two-point injection of 6-OHDA, and the PD model was constructed and the model was evaluated for behavior; SNMT crude enzyme solution was extracted from the peripheral blood lymphocytes of the model rats, and the enzyme reaction product N- A high-performance liquid chromatography tandem triple quadrupole mass spectrometry method for methyl-salsolinol (NMSal) was used to characterize the activity of SNMT by the amount of NMSal generated.

  RESULTS: A total of 7 of the 18 6-OHDA-injected rats showed constant rotation to the uninjured side (>7 r/min) after induction with apomorphine, and the modeling success rate was 38.9%. A method for the characterization of SNMT activity with high selectivity, high sensitivity and high reproducibility based on multiple reaction monitoring technology was established. The detection limit and quantification limit of NMSal in this method reached 49 pmol/L and 98 pmol/L, respectively, and the intra-day and inter-day precisions were both below 6.0%. The test results showed that the activity of SNMT in the peripheral blood lymphocytes of the PD group was significantly di