| ObjectiveIn this experiment,SAMP8 AD model mice were used as the research object,and Shh signaling pathway was used as the entry point to observe the effects of early acupuncture intervention on the behavior and Shh signaling pathway of SAMP8 AD model mice.Discuss the possible mechanism of early acupuncture intervention on AD prevention and treatment,and provide experimental support for clinical acupuncture and best treatment time.MethodsThere are 18 SPF-grade SAMP8 mice aged 3 months and 10 months each,weighing 20-25 g.SAMP8 mice aged 3 months were divided into model group and acupuncture group according to the random number table method,and SAMR1 mice of the same month age were regarded as normal group,each group was 6 mice.The 10-month group is the same as the 3-month group.In the acupuncture group,acupuncture GV20 and bilateral HT7,moxibustion on both BL23,once a day,6d as a course of treatment,a total of 4 courses,1d interval between treatments.The normal group and the model group were bred routinely and restrained by the same method at the same time without acupuncture treatment.24 hours after the intervention,the Morris water maze was used to detect the spatial learning and memory abilities of each group of mice,the open field test was used to detect their depression,and the new object recognition test was used to detect their object recognition memory.After the behavioral test,Western-blot method and q PCR method were used to detect the expression of Shh,Smo,Ptch1,Gli1 protein and the level of protein transcription in the hippocampus.Results1.Behavioral changes of mice in each group:(1)The results of the Morris water maze experiment showed that in the positioning navigation experiment,the escape latency of the 3 and 10 months old model group was significantly longer than that of the normal group of the same month(P<0.01,P<0.05);On the first day,there was no significant difference in the escape latency between the three-month-old groups.Starting from the second day,the escape latency of the 3-month-old acupuncture group was significantly shorter than that of the same-month-old model group(P <0.01),and the 10-month-old acupuncture group’s escape latency The incubation period was shorter than that of the same month-old model group,but it was not statistically significant.In the space exploration experiment,compared with the normal group of the same month,the 3 and 10 months old model group had fewer times to cross the platform(P <0.01,P <0.05);the 3 month old acupuncture group had more times of crossing the platform than the same month old model group(P <0.05).The 10-month-old acupuncture group had more times of crossing the platform than the same-month-old model group,but there was no significant statistical significance.(2)The results of the opening experiment showed that the activity time in the central area was shorter in the 3 and 10 month-old model group compared with the normal group of the same month age(P<0.05);the activity time in the central area of ??the 3-month-old acupuncture group was better than that in the same month-old model group(P<0.05).The activity time in the central area of ??the10-month-old acupuncture group was longer than that of the same-month-old model group,but there was no significant statistical significance.(3)The results of the new object recognition experiment showed that: Compared with the normal group of the same month,the new object recognition coefficient of the3,10-month-old model group(P <0.01,P <0.05);the new object recognition coefficient of the 3-month-old acupuncture group was higher than that of the same-month-old model Group(P <0.05).The new object recognition coefficient in the 10-month-old acupuncture group was higher than that in the same-month-old model group,but there was no significant statistical significance.2.Changes in the protein expression of Shh,Ptch1,Smo,and Gli1 in the brain tissue of mice in each group: Compared with the normal group of the same month,the relative expression of Shh,Smo,and Gli1 in the brain tissue of the model group of 3 and 10 months was reduced(P< 0.01,P<0.05),the relative expression of Ptch1increased(P<0.01,P<0.05);the relative expression of Shh,Smo,and Gli1 in the brain tissue of mice in the 3-month-old acupuncture group was higher than that in the same-month-old model group(P<0.01)),the relative expression of Shh,Smo,and Gli1 in the brain tissue of mice in the 10-month-old acupuncture and moxibustion group was somewhat higher than that in the same-month-old model group,but there was no significant statistical significance;the relative expression of Ptch1 in the brain tissue of the 3-month-old acupuncture and moxibustion group Lower than the model group of the same month(P<0.01),the relative expression of Ptch1 in the brain tissue of mice in the 10-month-old acupuncture moxibustion group was somewhat lower than that of the model group of the same month,but there was no significant statistical significance.3.Changes in the transcription levels of Shh,Ptch1,Smo,and Gli1 proteins in the brain tissue of mice in each group: Compared with the normal group of the same month,the transcription levels of Shh,Smo,and Gli1 genes in the brain tissue of the 3 and 10month-old model groups were down-regulated(P <0.01,P<0.05),Ptch1 gene transcription level was up-regulated(P<0.01);the brain tissue Shh,Smo and Gli1 gene transcription levels of mice in the 3-month-old acupuncture group were higher than those in the same-month-old model group(P<0.01,P<0.05)),the transcription levels of Shh,Smo,and Gli1 genes in the brain tissue of mice in the 10-month-old acupuncture group were higher than those in the same-month-old model group,but there was no significant statistical significance.ConclusionEarly acupuncture intervention can significantly improve the spatial learning and memory impairment,object recognition memory impairment and depression in SAMP8 AD model mice.It is speculated that it may be related to the regulation of Shh signaling pathway and promote neurogenesis. |
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