| Objectives To observe the effects of Selegiline on tyrosine hydroxylase(TH) and neuronal nitric oxide synthase(n NOS) and α-Synuclein(α-Syn) in the gastric antrum and colon of Parkinson’s disease model rats, so as to investigate the therapeutic effect and mechanism of selegiline for treatment of gastric and colon disfunction in PD.Methods 1 Experimental animal grouping. A total of 72 healthy male SD trial rats were randomly divided into normal control group(control group), PD model group(model group) and selegiline treatment group(selegiline group). Each group was randomly divided into 4-day subgroup and 8-day subgroup, there were 12 rats in each group. 2 Parkinson’s disease model rats preparation We dissolved the rotenone in the sunflower oil, made them mixed fully,made them mixed fully, injectied it in the subcutaneous tissue of each rat daily by 2mg/kg in the two groups. Ready to observe and record the behavior changes after the rote-none was given, we terminated the drug once occurred the behavioral scoring as soon as possible. We carried on behavior study examination according to the Parkinson animal model behavior study grading standard, then rats who got 2 to 6 points were selected as PD model rats. 3 Method of administration. Control group: Daily subcutaneous injection sunflower oil 2mg/kg, Rats were daily subcutaneous injected sunflower oil 2mg/kg, and after successful model preparation we stopped injection and fed to an equal volume of saline until the point time. model group: Rats were daily injected the rotenone sunflower oil emulsion(2mg/kg) in hypodermic tisse every day, after the success of the model preparation, we stopped subcutaneous injection and filled the same volume of saline into stomach until the time point. Selegiline group: after the success of model preparation, rats in treatment group were given Selegiline 0.5mg·kg-1 intragastrically every day. We continuous administrated for 4 d or 8 d respectively according to the subgroup grouping. 4 The amount of solid food residue in gastric and 1 h fecal excretion and water content. At the time point of 4 d and 8 d after therapy, solid food residue in gastric and 1 h fecal excretion and water content were detected. 5 We detected the expression of TH, n NOS and α-Syn in gastric and colon in each group by Immunohistochemistry and Western Blot. 6 We detected the expression of TH, n NOS and α-Syn in gastric and colon in each group by Immunohistochemistry and Western Blot. 7 We observed tissue slices with microscope camera OLYMPUS(400×), then analysis them by Motic Med 6.0 digital medical image analysis system. The results of Western Blot were analyzed with Image J. 7 SPSS 19.0 statistic software was used for data analysis. Measurement data were presented as mean±SD, and the Normality W test and the Homogeneity of variance Levene test wereused to test the data, and then we use randomized block design analysis of varianc, P<0.05 for the difference which was statistically significant.Results 1 Compared with control group, model group at each time point solid food residue in gastric was increased(all P<0.01); compared with model group, treatment group at each time point solid food residue in gastric was decreased( P<0.05 or P<0.01); and compared with the treatment 4d group, the treatment 8d group solid food residue was decreased(P<0.05). 2 Compared with control group, model group had a lower 1 h fecal excretion and water content at each time point(all P<0.01); compared with model group, 1 h fecal excretion and water content was all increased in therapy group at each time point(P<0.05); and compared with treatment 4d subgroup,treatment 8 d subgroup 1 h fecal excretion and water content was higher(P<0.05). 3 Compared with control group, model group at each time point TH in gastric and colon was significantly lower,n NOS and α-Syn was significantly increased, the difference was statistically significant(all P<0.01); Compared with the model group, the expression of TH was significantly increased, n NOS and α-Syn was significantly reduced in the treatment group at each time point, the difference was statistically significant(P<0.05 or P<0.01); Compared with the treatment 4d group, the expression of TH in gastric and colon in the treatment 8d group time point was all increased(P<0.05), and there were less expression(P<0.05) of n NOS and α-Syn at 8d group time point.Conclusions 1 The expression of TH in gastric and colon of Parkinson’s disease rat model was all reduced,but n NOS and α-Syn was increased, TH, n NOS and α-Syn involved in Parkinson’s disease pathogenesis of gastrointestinal dysfunction. 2 Selegiline can improve gastric and colon dysfunction in PD rats, and its mechanism may be related to reduce PD model rats gastric and colon dopaminergic neuronal injury and inhibition of n NOS and α-Syn expression in gastric and colon of PD model rats, And with time, the effect is more pronounced. |
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