| Objective:In recent years,the proportion of the elderly population in China has been rising,and the aging of the population has become a trend.The number of patients with degenerative diseases of the nervous system has also increased year by year.Parkinson’s disease(PD),as a neurodegenerative disease that seriously endangers the health and quality of life of elderly patients,has become a research hotspot in the field of neurology in recent years.Traditional Chinese medicine has its unique advantages and curative effects in the treatment of PD,but its mechanism of action is still unclear.In recent years,with the advancement of traditional Chinese medicine theory and traditional Chinese medicine in the field of autophagy,more and more studies have shown that the intervention of autophagy-lysosomal pathway to regulate autophagy in Chinese medicine may explain the treatment of PD by Chinese medicine.The mechanism of action of neurodegenerative diseases.In this study,a PD rat model prepared by single-point injection of 6-hydroxydopamine(6-OHDA)was used as the research object,and the autophagy-associated protein Beclin1 in the substantia nigra of the PD rat model was observed.The effects of P62 and LC3 II expression and the behavioral changes of rats in each group were explored whether the "Blood Kidney Method"(Ma Ling Paan Pill)can delay the progression of PD by repairing and dredging the damaged nodes of autophagy-lysosomal degradation pathway.To clarify the autophagy mechanism of autophagy may be an important material basis for the treatment of PD,and provide a new method and idea for clinical PD treatment.Methods:SPF grades weighing 180-220 g and 90 male SD rats were used for one week after routine feeding.The single-point method was used to drill the right mesencephalic substantia nigra parsing point of the SD rat from the anterior iliac crest center-5.0 mm to the anteriorcondyle center-1.7 mm,and the meningeal surface depth-7.6 mm to drill and inject 24 μg 6-OHDA To establish a rat model of PD,behavioral testing of model rats was performed by intraperitoneal injection of apomorphine hydrochloride(APO)1 week after surgery.A randomized PD model of 50 models was randomly selected and weighed according to the random number table method.The rats were divided into high-,medium-,and low-dose groups of Chinese medicine,Medoga group and model group,with 10 rats in each group and10 normal rats.Rats were only treated with skin preparation,scalp incision,skull drilling,scalp suture,etc.,and were treated with 6-OHDA solution instead of 6-OHDA solution.Drug administration was grouped according to the established protocol.The general records and behavioral tests were performed on each group before and 30 days after administration.After 30 days of gavage,HE staining was used to observe the damage of mesangial tissue in rats after immunotherapy.Immunohistochemistry The method was used to detect the expression of Beclin1,P62 and LC3 II proteins in the substantia nigra pars compacta of rats.Statistical methods: If the results are normal distribution and the variance is uniform,single factor analysis of variance is used;if not,multiple independent samples are used for nonparametric test.Kruskal-Wallis test.Results:1.The single point method was used to make the right mesencephalic substantia nigra parsing point of the SD rat-5.0 mm from the anterior iliac crest center,-1.7 mm from the anterior condyle center,and 7.6 mm from the meningeal surface depth and inject 24 μg 6-OHDA,a rat model of PD was created,and 56 rats were successfully modeled with a success rate of 70%.2.In general,the mental,activity,hair,diet and other conditions of the Chinese herbal medicines in the high,middle,low and Midota groups were improved compared with the model group.The PD symptoms gradually decreased with the prolonged administration time.3.The LSD test method was used to test the difference ratio(K)of rat rotation circle.The three doses(K)of high,medium and low doses in Chinese medicine group were higher(P<0.01),which proved that the symptoms of PD rats were improved and the effect of Chinese medicine group was improved.More obvious.4.HE staining showed that the number of neurons in the sham-operated group was dense,arranged neatly,and the morphology was normal,with no obvious damage.The neuron cells in the model group showed obvious damage.The neuron injury after 30 days of administration in the Chinese medicine group was compared with the model.The groupimproved.Among them,the middle dose group of Chinese medicine had the largest number of neurons,and the improvement was the most obvious.The Meiduoba group also improved compared with the model group,but the improvement was not as good as the middle dose group of traditional Chinese medicine.5.The expression of Beclin1 in the high,middle and low dose groups of Chinese medicine was higher than that in the model group(P<0.01).Compared with the Chinese medicine group,the expression of Beclin1 was the highest in the middle dose group and the lowest in the low dose group.Compared with the low dose group but less than the medium dose group,the difference was significant(P<0.01),indicating that the Chinese medicine Guiling Paan Pill has the effect of increasing the expression of Beclin1 in the PD rat model,and its effect is obvious,which can improve the expression of Beclin1 in PD rats.The effect is more pronounced with medium dose(1.8 g/Kg/d).6.The expression of P62 protein in the high,middle and low dose groups of Chinese medicine was lower than that in the model group(P<0.01).Compared with the traditional Chinese medicine group,the expression of P62 was the highest in the low dose group and the lowest in the middle dose group.The difference was higher in the middle dose group but not in the low dose group(P<0.01).It indicated that Chinese herbal medicine Guiling Paan Pill has the effect of reducing the accumulation of P62 protein in PD rat model,and the effect is remarkable.The effect is related to the dosage,and the effect of medium dose(1.8g/Kg/d)is more obvious.7.The expression of LC3 II in the high,middle and low dose groups of Chinese medicine was significantly higher than that in the model group(P<0.01).The middle dose group was the highest in the middle dose group and the lowest in the low dose group.The lower dose group in the high dose group was higher than the medium dose group.In the dose group,the difference was significant(P<0.01),indicating that the Chinese medicine Guiling Paan Pill has the effect of increasing the expression of LC3 II in the PD rat model,ie increasing the number of autophagosomes.The effect is obvious,and the effect is related to the dosage,medium dose(1.8 g)./Kg/d)The effect is more obvious.Conclusions:A single-point injection of 6-OHDA in the substantia nigra pars compacta was used to establish a PD rat model,which was convenient,efficient,and has a high success rate."Blood Kidney Method"(Ma Ling Paan Pill)can effectively improve the symptoms of PD rats,the curative effect is exact,and the improvement of medium dose administration ismore obvious.Intervention of PD model in the rat model of tonifying kidney can effectively increase the expression of Beclin1 and LC3 II in the substantia nigra of PD rats and decrease the accumulation of P62."Blood Kidney Method" is effective in treating the mechanism of PD and clearing autophagy-lysosomal degradation pathway,increasing autophagy activity and improving cell homeostasis. |
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