| Objective: Based on the theory of "kidney-brain combination",this study added Chinese medicine Bushen Huoxue decoction to acupuncture "Baihui",bilateral "Shenshu" and bilateral "Sanyinjiao" of rapidly aging SAMP8 mice Stomach,observe the intervention effect of Bushen Huoxue acupuncture and simultaneous therapy on SAMP8 mice.By observing the effect of acupuncture and medicine combination therapy on Aβ and p-tau protein in the cerebral cortex of SAMP8 mice,the effect of acupuncture and medicine combination therapy on Alzheimer’s disease(AD)was explored from the perspective of neuropathology AD provides evidence support.Materials and methods: Thirty clean seven-month-old male SAM germline mice were selected,including 24 rapidly aging SAMP8 mice and 6 anti-rapid aging SAMR1 mice.Twenty-four SAMP8 mice were randomly divided into a model group,an acupuncture group,a traditional Chinese medicine group,and an acupuncture group,with 6 mice in each group.Six SAMR1 mice of the same month age were used as the control group.Acupuncture group acupuncture "Baihui",bilateral "Shenshu",bilateral "Sanyinjiao",electric acupuncture selects continuous wave,frequency is 2Hz,current is 0.6m A,lasts 20 min,once per day;Chinese medicine group Bushen Huoxue Decoction Gavage,0.2ml/time,1 time/d;the acupuncture group was given acupuncture and traditional Chinese medicine at the same time.The control group and the model group were given the same degree of grasping and fixation and given distilled water by gavage,0.2ml/time,once/day for 4 weeks of continuous treatment.After the treatment,Morris water maze test and nesting test were used to detect the behavioral changes of mice.HE staining was used to observe the morphological changes of neurons in the cerebral cortex.Immunohistochemistry and ELISA were used to detect the expression of Aβprotein.Immunohistochemistry was used.Method and western blot method to detect tau protein expression.Results:1.Morris water maze experiment results show that: in the visual platform experiment,there is no significant difference in the escape latency and swimming speed of the mice in each group(P>0.05).With the increase of training time in the hidden platform experiment,the training incubation period of mice decreased.Compared with the control group,the escape latency of the model group was significantly increased(P<0.05);compared with the model group,the escape latency of the acupuncture group,Chinese medicine group,and acupuncture group was significantly reduced(P<0.05);and the needle Compared with the acupuncture group and the Chinese medicine group,the escape latency time of the acupuncture and medicine group had a decreasing trend,but there was no statistical significance(P>0.05).In the space exploration experiment,compared with the control group,the number of mice crossing the platform in the model group was significantly reduced(P<0.05);compared with the model group,the number of crossing the platform in the acupuncture group,Chinese medicine group,and acupuncture group was significantly increased(P<0.05);there was no statistically significant difference between the acupuncture group and the acupuncture group and the traditional Chinese medicine group(P>0.05).The nesting experiment results showed that: compared with the control group,the model group’s nesting experiment score was significantly reduced(P<0.05);compared with the model group,the traditional Chinese medicine group score was significantly increased(P<0.05).Compared with the model group,the acupuncture group had an increasing trend,but it was not statistically significant(P>0.05).2.HE staining results of mouse cerebral cortex showed that: the neurons in the control cerebral cortex of the mice in the control group were intact,arranged neatly,and the nuclear staining was clear and uniform;the number of neurons in the cerebral cortex of the model group decreased,and the arrangement was disordered and irregular.Nuclear shrinkage phenomenon;the number of neurons in the cerebral cortex of the acupuncture group,Chinese medicine group and acupuncture group increased,the arrangement was regular and neat,and the structure was clear.It shows that the acupuncture group,the traditional Chinese medicine group and the acupuncture group can alleviate the damage and nuclear shrinkage of cerebral cortex neurons in SAMP8 mice.3.Immunohistochemical method and ELISA method were used to detect the expression of Aβin the cerebral cortex of mice in each group.The results of Aβ immunohistochemistry showed that the neurons in the cerebral cortex of the control group SAMR1 mice were intact and arranged in an orderly manner without obvious Aβ positive staining;the number of Aβpositive staining cells in the cerebral cortex of the model group SAMP8 mice was significantly increased compared with the control group.The expression in the nucleus of the nucleus was brown(P<0.05);the number of Aβ positive staining cells in the cerebral cortex of SAMP8 mice in the acupuncture group,the Chinese medicine group and the acupuncture group was reduced to varying degrees compared with the model group,and the staining was lighter(P< 0.05);There was no significant difference in the number of Aβ positive staining cells in the cerebral cortex of SAMP8 mice between the acupuncture group,the traditional Chinese medicine group and the acupuncture group(P>0.05).The expression of Aβ protein was detected by ELISA.The results showed that compared with the control group,the Aβcontent in the cerebral cortex of the model group SAMP8 mice was significantly increased(P<0.05);compared with the model group mice,the brains of the three treatment groups SAMP8 mice The level of Aβ in the cortex has decreased,and the difference is statistically significant(P<0.05);but there is no significant difference in the content of Aβ in the cerebral cortex of the three groups of SAMP8 mice between the acupuncture group,the traditional Chinese medicine group and the acupuncture group(P>0.05).4.The expression of p-tau in the cerebral cortex of mice in each group was detected by immunohistochemistry and western blot.The results of p-tau immunohistochemistry showed that the cytoplasm of p-tau-positive cells was large,mostly brown or brown,and the expression mainly appeared in the nucleus of neurons.Compared with SAMR1 mice in the control group,there were fewer neurons in the cerebral cortex that showed positive expression of p-tau;compared with the SAMR1 mice in the control group,the number of p-tau positive staining cells in the model group SAMP8 mice was significantly increased,and the p-tau staining of positive cells was more Deep(P<0.05);compared with the model group SAMP8 mice,the number of p-tau positive staining cells and the staining intensity of SAMP8 mice in the acupuncture group,the traditional Chinese medicine group and the acupuncture group all decreased to varying degrees,close to the control group(P<0.05).Western blot method was used to detect the expression of p-tau protein in the cerebral cortex of SAMP8 mice.The results showed that compared with the control group SAMR1 mice,the p-tau protein content of the model group mice was significantly increased(P<0.05);compared with the model group SAMP8 Compared with mice,the expression levels of p-tau in acupuncture group,Chinese medicine group and acupuncture group were significantly lower(P<0.05);there was no significant difference between acupuncture group,Chinese medicine group and acupuncture group,no statistical significance(P>0.05).Conclusion:1 Acupuncture combined with medicine therapy can effectively improve the learning and memory function of SAMP8 mice;2 Combined acupuncture and medicine therapy can effectively reduce the expression of Aβ in the cerebral cortex of SAMP8 mice;3 Combined acupuncture and medicine can effectively reduce the expression of p-tau protein in the cerebral cortex of SAMP8 mice;4 Acupuncture and medicine combined therapy can inhibit the neuropathological changes of SAMP8 mice,but there is no significant difference compared with the acupuncture group and the traditional Chinese medicine group. |
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