| Purpose:To explore the effects of acupuncture combined with thorns and abundance on inflammatory injury and apoptosis in autistic ratsMethod:Autism rat model was established according to the method of Roullet.A sham operation group,a model group,a head acupuncture group,a rich environment group,a head acupuncture group and a rich environment group were established.The sham operation group and the model group were not treated.The head acupuncture group was treated with acupuncture,and the enriched environment group was given rich environmental treatment.The head acupuncture combined with the enriched environment group was given acupuncture combined with rich environmental treatment.Field experiments assessed anxiety,buried bead experiments evaluated repeated stereotypes,three boxes of experiments assessed social abilities and social preferences,and Morris water maze assessed spatial learning and memory abilities.HE and Nissl staining were used to detect the inflammatory lesions of the frontal lobe and the morphology and quantity of neurons.The serum proinflammatory factors IL-1?,TNF-a,anti-inflammatory factors IL-1Ra and TGF-?1 were detected by ELISA.Immunohistochemical detection was used.The expression of proinflammatory cytokines IL-1?,TNF-a,inflammatory factors IL-1Ra and TGF-?1 were detected by TUNEL method.The frontal lobe Ras and Mek1/2were detected by Western Blot.p-ERK1/2,Bcl-2,Bax protein expression.Result:1.Behavioral assessment:(1)Market experimentCompared with the sham operation group,the spontaneous activity distance and time of the model group were prolonged(P<0.05).Compared with the model group,the head acupuncture group,the enriched environment group,the head acupuncture group combined with the enriched environment group spontaneous activity distance and time The difference was significant(P<0.05).Compared with the head acupuncture group and the enrichedenvironment group,the spontaneous activity distance and time of the head acupuncture combined with the enriched environment group were the shortest(P<0.05).(2)Buried beads experimentCompared with the sham operation group,the number of beads in the model group was more than that of the model group(P<0.05).Compared with the model group,the head acupuncture group,the enriched environment group,the head acupuncture group and the enriched environment group had fewer beads.(P<0.05);compared with the enriched environment group and the head acupuncture group,the number of buried beads in the head-to-acupoint cluster-enriched environment group was less(P<0.05).(3)Three boxes of experiments3.1 Compared with the sham operation group,the social index and social preference index of the model group were both small(P<0.05);compared with the model group,the head acupuncture group,the rich environment group,the head acupuncture group combined with the rich environment group socialization The index and social preference index increased(P<0.05).Compared with the head acupuncture group and the rich environment group,the social index and social preference index of the head cavity acupuncture combined with the rich environment group were the largest(P<0.05).3.2 Compared with the sham operation group,the social Sniff and social preference Sniff of the model group were less(P<0.05).Compared with the model group,the head acupuncture group,the rich environment group,the head acupuncture group and the rich environment group socialized.Sniff and social preference Sniff were both increased(P<0.05).Compared with the head acupuncture group and the enriched environment group,the social Sniff and social preference Sniff of the head cavity cluster combined with the rich environment group were the largest(P<0.05).(4)Morris water maze4.1 Positioning cruise experiment: Compared with the sham operationgroup,the number of exploration platforms in the model group was small(P<0.05);compared with the model group,the head acupuncture group,the rich environment group,the head acupuncture group combined with the enriched environment group exploration platform The number of times was higher(P<0.05).Compared with the head acupuncture group and the enriched environment group,the number of times of the head acupuncture combined with the enriched environment group was more(P<0.05).4.2 Space exploration experiment: Compared with the sham operation group,the number of crossing the platform of the model group was less(P<0.05);compared with the model group,the head acupuncture group,the rich environment group,the head acupuncture group combined with the rich environment group crossing the platform times The increase was(P<0.05).Compared with the acupuncture group and the enriched environment group,the head acupuncture combined with the enriched environment group crossed the platform more frequently(P<0.05).2.HE stainingIn the sham operation group,the frontal lobe cells were arranged evenly and tightly,with a clear outline.The nucleus was stained blue,and it was oval.The nucleolus was seen.No degeneration and necrosis of nerve cells and infiltration of inflammatory cells were observed.In the model group,the cells were arranged disorderly,the cell gap became large,the cell structure was blurred or disappeared,and there were neuronal cell degeneration,necrosis,disintegration,nuclear pyknosis,and even softening of the foci.At the same time,obvious inflammatory cell infiltration,cell edema,and glue were observed.Proliferating cells.The pathological injury was alleviated in the three intervention groups after treatment,and the head group acupuncture combined with enriched environment group changed most obviously.3.Nissl stainingCompared with the sham operation group,the survival of the frontal lobe neurons in the model group was reduced(P<0.05).Compared with the modelgroup,the number of frontal acupuncture groups,the enriched environment group,and the head acupuncture group combined with the enriched environment group.The increase was(P<0.05).Compared with the head acupuncture group and the enriched environment group,the number of frontal neurons in the head acupuncture combined with enriched environment group was the highest(P<0.05).The number of neurons in the sham operation group was the highest,the structure of the frontal lobe cells was intact,the cell body was not atrophy,the number of Nissl bodies was large,there was no dissolution phenomenon,and the cytoplasm was deeply stained;the model group had the smallest number of nerve cells,the cell structure was destroyed,and the normal neuron cells were rare.The cell body atrophy and the number of Nissl bodies decreased significantly.After three kinds of interventions,the neuronal damage was alleviated,and the head group acupuncture combined with enriched environment group was the most obvious.4.ELISA testProinflammatory cytokines IL-1? and TNF-a: Compared with the sham operation group,the expression of serum IL-1? and TNF-a in the model group was increased(P<0.05).Compared with the model group,the head acupuncture group and the environment were rich.The expression of group and head acupuncture combined with enriched environment group decreased(P<0.05);compared with head acupuncture group and rich environment group,head group acupuncture combined with enriched environment group had the lowest expression(P<0.05).Inflammatory factors IL-1Ra and TGF-?1: Compared with the sham operation group,the serum IL-1Ra and TGF-?1 in the model group were significantly lower(P<0.05);compared with the model group,the head acupuncture group and the rich environment group The expression of the head acupuncture combined with the enriched environment group was increased(P<0.05).Compared with the head acupuncture group and the enrichedenvironment group,the expression of the head acupuncture combined with the enriched environment group was the highest(P<0.05).5.ImmunohistochemistryProinflammatory cytokines IL-1? and TNF-a: Compared with the sham operation group,the expression of IL-1? and TNF-a in the frontal lobe of the model group increased(P<0.05);compared with the model group,the head acupuncture group and the environment The expression of group and head acupuncture combined with enriched environment group decreased(P<0.05).Compared with head acupuncture group and rich environment group,head acupuncture combined with enriched environment had the lowest expression(P<0.05).Inflammatory factors IL-1Ra and TGF-?1: Compared with the sham operation group,the expression of IL-1Ra and TGF-?1 in the frontal lobe of the model group was decreased(P<0.05).Compared with the model group,the head acupuncture group and the environment were rich.The expression of group and head acupuncture combined with enriched environment increased(P<0.05).Compared with head acupuncture and rich environment,the head acupuncture combined with enriched environment group had the most expression(P<0.05).6.TUNEL method detectionCompared with the sham operation group,the TUNEL positive number in the frontal lobe of the model group increased(P<0.05).Compared with the model group,the number of TUNEL positive cells in the head acupuncture group,the enriched environment group,and the head acupuncture combined with the environmental group decreased.(P<0.05);Compared with the head acupuncture group and the enriched environment group,the number of TUNEL positive cells in the frontal lobe of the rats with head acupuncture combined with enriched environment was the least,and the difference was significant(P<0.05).7.Western Blot testApoptosis-promoting Bax: Compared with the sham-operated group,the expression of Bax protein in the frontal lobe of the model group increased significantly(P<0.05).Compared with the model group,the head-point cluster,the enriched environment group,and the head-cup cluster combined with abundant environment.The expression of Bax protein in the frontal lobe was significantly decreased(P<0.05).Compared with the acupuncture group and the enriched environment group,the expression of Bax protein in the acupuncture group was the least(P<0.05).Inhibition of Bcl-2: Compared with the sham operation group,the expression of Bcl-2 protein in the frontal lobe of the model group was significantly reduced(P<0.05).Compared with the model group,the head acupuncture group,the enriched environment group,and the cephalic point The expression of Bcl-2 protein in the frontal lobe of the burr-enriched environment group was significantly increased(P<0.05).Compared with the acupuncture group and the enriched environment group,the head group acupuncture combined with the environmentally rich Bcl-2 expression.The most significant difference was significant(P < 0.05).Bcl-2/Bax: Among the five groups,the sham operation group was the largest,and the model group was reduced.Compared with the model group,the Bcl-2/Bax ratio was significantly increased after the head cavity acupuncture,enriched environment,and head acupuncture combined with environmental treatment.High;close to 1,in which the Bcl-2/Bax ratio is highest in the acupuncture combined with enriched environment.Ras/ERK1/2 pathway: compared with the sham operation group,the model group frontal lobe Ras,Mek1/2 and P-ERK1/2 was increased(P<0.05).Compared with the model group,the expression of the head acupuncture group,the enriched environment group,the head acupuncture group and the rich environment group decreased(P<0.05).Compared with the group and the rich environment group,the expression of the head acupuncture combined with the enriched environment group was the least(P<0.05).In conclusion:1.The combination of acupuncture and acupuncture combined with rich environment can improve the anxiety,stereotypes,social interaction,cognitive learning and memory ability of selevar-induced autistic rats,which is better than single head acupuncture group and rich environment group.2.The combination of acupuncture and thorns can reduce the inflammatory response induced by sodium valproate in rats with autism,inhibit the pro-inflammatory factors TNF-a,IL-1? and enhance the anti-inflammatory factor TGF in serum and frontal lobe brain tissue.TGF-?1,IL-1Ra expression,reduce inflammatory lesions in the frontal lobe cells,and exert neuroprotective effects.3.The combination of acupuncture and acupuncture combined with rich environment can reduce the number of apoptotic cells in the frontal lobe induced by sodium valproate,promote the expression of Bcl-2 in the frontal lobe,and inhibit Bax,Ras,MEK1/2,P-ERK1/2.Expression reduces apoptosis. |
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