| Objective: To observe the effect of "Tiao Shen Changzhi" acupuncture on the behavior of LID model rats,the expression of PKC and PP-1 protein and the concentration of Ca2+ in striatum,and to explore the possible mechanism of acupuncture treatment of Parkinson’s LID by "Tiao Shen Changzhi" acupuncture method.Methods: A total of 90 healthy male SD rats were selected,and 24 rats were randomly divided into normal group and sham operation group.The remaining 66 rats were injected into the 6-OHDA striatum by double-target injection to establish the rat model of partial Parkinson’s disease(PD).The successful PD model rats were evaluated by apmorphine(APO)induced rotation test after surgery,and the rats were intraperitoneally injected with levodopa methyl methyl(10mg/kg)combined with benserazid(2.5mg/kg)to prepare the levodopainduced disorder(LID)rat model required for this study.During administration,the rats were subjected to behavioral evaluation of abnormal involuntary movement(AIM).4 weeks later,successful LID rats were randomly divided into LID model group,Western medicine group,ordinary acupuncture group and Tiaoshenchangzhi group,with 12 rats in each group.AIM behavioral score was used to observe the behavior of rats in each group after corresponding intervention.The expression of PKC and pp-1 protein in striatum of rats were detected by immunohistochemistry and Western Blot.Fura-2/AM fluorescence probe was used to detect Ca2+ concentration in striatum tissues.To explore the possible mechanism of "Tiaoshenchangzhi" acupuncture in treating Parkinson’s LID from the perspective of PKC,PP-1 and Ca2+ concentration.Results:1.Behavioral results: after PD model rats were induced by L-DOPA injection,the AIM score score increased the fastest in the first 14 days,and the AIM score tended to be stable after 14 days.The LID rat model was successfully prepared.After 4 weeks of intervention treatment,the AIM score in the LID model group was significantly lower than that in the western medicine group,common acupuncture group and Tiaoshen Changzhi group after intervention treatment(P < 0.01).From the comparison table of AIM scores in rats,it can be seen that the intervention effect of Tiaoshen Changzhi group is lower than that of western medicine group(P<0.05),but it is significantly better than that of ordinary acupuncture group in the same acupuncture intervention(P<0.01).2.Immunohistochemical results: Compared with the normal group,the expression of PKC protein in the sham operation group was similarly low(P>0.05),and the positive rate of PKC protein in the LID model group was significantly higher(P < 0.01).Compared with THE LID model group,the expression of PKC protein in the WESTERN medicine group,Tiaoshenchangzhi group and ordinary acupuncture group was significantly reduced under therapeutic intervention;According to the positive area of PKC in cells,compared with the Tiaoshenchangzhi group,the intervention effect of western medicine group was significantly lower than that of the Tiao Shenchangzhi group(P<0.01).Similarly,the down-regulation of Shenchangzhi group by acupuncture was better than that of the ordinary acupuncture group(P<0.01).Protein expression of STRIatum PP-1 was also lower in the sham operation group than in the normal group,and the difference was not significant.The positive rate of PP-1 in the LID model group was significantly higher(P<0.01).Compared with LID model group,the expression of PP-1 protein in the WESTERN medicine group,Tiaoshenchangzhi group and ordinary acupuncture group was significantly decreased under therapeutic intervention(P<0.01);Compared with the Western medicine group,the expression of PP-1 protein in the Tiaoshen Changzhi group and the ordinary acupuncture group was higher(P<0.01).Compared with the Tiaoshen Changzhi group,the positive expression of PP-1 cells showed that the intervention effect of Tiao Shen Changzhi group was better than that of the ordinary acupuncture group(P<0.01).3.Western Blot results: compared with the normal group,the expression of PKC protein in the sham operation group was also lower,the difference was not obvious,and the gray level of PKC in the LID model group was significantly increased(P < 0.01).Compared with LID model group,the gray value of PKC in western medicine group,Tiaoshenchangzhi group and ordinary acupuncture group was significantly reduced under therapeutic intervention.As can be seen from the gray level of PKC cell protein expression,compared with the Tiaoshenchangzhi group,the intervention of Western medicine group was superior to the Tiao Shen Changzhi group to a certain extent(P < 0.05),and the gray level of PKC in tiaoshen Changzhi group was significantly superior to the ordinary acupuncture group(P <0.01).Experimental data showed that the gray level of PP-1 in the sham operation group was lower than that in the normal group,and the difference was not significant(P>0.05),while the expression of PP-1 in the LID model group was significantly increased(P <0.01).Compared with LID model group,PP-1 gray value of LID rats in western medicine group,Tiaoshenchangzhi group and ordinary acupuncture group was significantly reduced under therapeutic intervention.Compared with the tiaoshenchangzhi group,the intervention effect of western medicine group was better than that of tiao Shen Changzhi group to some extent(P < 0.05).Similarly,acupuncture intervention to reduce Shenchangzhi group was better than that of ordinary acupuncture group(P < 0.01).4.The concentration of Ca2+ was detected by Fura-2/AM fluorescence probe method:compared with the normal group,the concentration of ca2+ in the sham operation group was also lower,the difference was not statistically significant(P >0.05),and the concentration of ca2+ in the lid model group was significantly higher(P < 0.01).Compared with lid model group,western medicine group,tiaoshengchang Zhi group and ordinary acupuncture group,the concentration of ca2+ decreased significantly under therapeutic intervention(P < 0.01);It can be seen from the concentration value of ca2+ that the intervention effect of Western medicine group is better than that of regulating mind and free will group to a certain extent P< 0.05).Similarly,the acupuncture intervention of regulating mind and free will group is significantly better than that of the ordinary acupuncture group(P < 0.01).Conclusion:1."Tiaoshen Changzhi" acupuncture could significantly inhibit calcium ion concentration in striatum and abnormal expression and aggregation of ca2 + related regulating enzymes PKC and PP-1 in RATS with Parkinson’s Disease MODEL.2."Tiaoshenchangzhi" acupuncture method can significantly improve the abnormal involuntary movement of the mouth and face,upper limbs and neck body cadres of LID model rats.3."Tiaoshen Changzhi" acupuncture treatment can improve LID,and one of its mechanisms may be through regulating calcium ion concentration and the content of CALCIUM ion-related regulating enzymes PKC and PP-1,so as to improve the abnormal involuntary movement of LID model rats. |
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