Research On Therapeutic Effects Of SSTF For MPTP Parkinson Mice

Objectives:Establish the Parkinson mice model through intraperitoneal injection of MPTP,observe and contrast ethological changes and degrees of brain neuronal apoptosis of micein each group, discuss protective effects and probable mechanisms of scutellariabaicalensis stem-leaf total flavonoid (SSTF) for nigral dopaminergic neuron damagecaused by MPTP, and evaluate therapeutic effects of SSTF with expectations to lay certaintheoretical basis for clinical application of SSTF into treatment of Parkinson’s disease.Methods:I: grouping and establishment of the MPTP Parkinson mice model; experimentalanimals are32healthy C57BL/6J male mice aged8-10weeks, which are randomlydivided into4groups.(1) Group A: the blank control group, free from any specialtreatment each day;(2) Group B: the MPTP group; subject to intraperitoneal injection of25mg/kg MPTP each day from the fourth day of the experiment, and intraperitonealinjection of normal saline in the equivalent volume for1time2hours later for5consecutive days;(3) Group C: the SSTF pretreatment group; from the first day of theexperiment, subject to intraperitoneal injection of SSTF once per day for3consecutivedays; from the fourth day, mice are subject to intraperitoneal injection of MPTP for1timetwo hours after intraperitoneal injection of SSTF, for5consecutive days;(4) Group D: theSSTF treatment group; from the fourth day of the experiment, mice are subject tointraperitoneal injection of MPTP, followed by intraperitoneal injection of SSTF once perday2hours later, for5consecutive days; II: ethological inspection for mice; observeethological changes of mice every day,and score and record shaking palsy of mice,suspension test, and climbing test according to relevant standards; III: preparation forexperimental specimen; narcotized mice are subject to heart perfusion; cut their heads to take their brains after cleaning and fixing, take midbrain tissues to prepare paraffin slices,and then store for backup; IV: after toasting, dewaxing, hydration, and antigen retrieval oftissue slices, perform immumohistochemical staining of TH, Bcl-2, caspase-3and GFAPaccording to relevant steps in the specifications, observe cellular morphology of eachgroup under an inverted microscope, and count positive immune cells.Results:I: within3-5minutes after injection of MPTP, all mice in Group B (the MPTP modelgroup) have ethological changes, such as body trembling, bradykinesia, roachback,increased step width, Instability of gait, tail erecting, and piloerection. After lasting for30minutes, these symptoms begin to alleviate;24hours later, the mice basically recover to anormal state. When compared with Group B, mice in Group C (the SSTF pretreatmentgroup) and Group D (the SSTF treatment group) are found to alleviate in different degrees,and the lasting period is shortened respectively. II:(1) according to test results of tremblingscores, suspension capability, and climbing capability, obvious improvements are made tomotor coordination capabilities of mice in Group C and Group D, with statisticalsignificance with Group B (the MPTP model group) in terms of difference value. Whencompared with Group A (the blank control group), however, there is still statisticaldifference. There is no statistical difference (P>0.05) between Group C and Group D. Withincreased times of injection, difference value in each day has no statistical significance(P>0.05). III:(1) according to TH immune chemical staining results, considerable THpositive nerve cells are seen in Group A, with obvious axon and normal form. In Group B,the number of TH positive nerve cells remarkably decreases. In Group C and Group D, thenumber of TH positive nerve cells is more than that in Group B and less than that in GroupA, all with statistical significance (P<0.05) in terms of difference value. In Group B,obvious changes are found in cellular morphology, with swelling soma and unclear outline.Moreover, axon decreases or disappears.(2) According to Bcl-2staining results, Group Aenjoys the largest number of positive cells, with relatively dark brownish yellow staining.The number of positive cells in Group B obviously decreases, still with differences andstatistical difference (P<0.05) in terms of difference value when compared with the normalgroup.(3) according to caspase-3immune chemical staining results, the number of positivecells in Group B (the model group) is remarkably more than that in Group A, withstatistical significance (P<0.05) in terms of difference value. The number of positive cellsin Group C and Group D obviously declines when compared with that in Group B, with statistical significance (P<0.05) in terms of difference value. When compared with that inGroup A, the number of positive cells in Group C and Group D increases, with statisticalsignificance (P<0.05) in terms of difference value.(4) according to GFAP immunechemical staining results, the number of positive cells in Group B, Group C and Group Dincreases in different degrees, with statistical significance (P<0.05) when compared withGroup A. Group B has the largest number of GFAP positive cells, with statisticalsignificance (P<0.05) when compared with difference values of Group C and Group D.Changes in cellular morphology are found in Group B, Group C and Group D, and stainingis deepened in different degrees when compared with Group A.Conclusion:(1) SSTF can obviously improve symptoms of MPTP Parkinson mice.(2) SSTF canobviously inhibit midbrain nigral neuronal apoptosis of MPTP Parkinson mice.(3) Whencompared with mice subject to later treatment, MPTP Parkinson mice under preventive useof SSTF have no obvious improvement to their symptoms.(4) When compared with micesubject to later treatment, preventive use of SSTF fails to effectively inhibit midbrainnigral neuronal apoptosis of MPTP Parkinson mice.(5) SSTF has wide research prospectsin terms of its application in treatment for Parkinson’s disease.