| Object:Through this study,we further elucidated the efficacy and mechanism of Tong Du Tiao Shen acupuncture treatment of type 1 diabetic cognitive impairment,and provided a scientific theoretical basis for the further promotion and application of Tong Du Tiao Shen acupuncture in clinical practice.method:in male C57BL/6J mice as the research object,type 1 diabetes mod el induced by intraperitoneal injection of high dose of STZ,in CA 1 L PS injection induced local inflammation,200 male C57BL/6J mice(6~8weeks old),were randomly divided into 8 groups,25 rats in each grou p:(1)normal glucose injection+CA1 normal saline+sham acupuncture grou p;(2)normal glucose injection+CA1 normal saline+electroacupuncture gr oup;(3)normal glucose injection+CA1 LPS+sham acupuncture group;(4)nor mal glucose injection+CA1 LPS+group;(5)high glucose+CA1 injection of saline+sham acupuncture group;(6)high glucose+CA1 injection ofphysiol ogical saline+electroacupuncture group;(7)high glucose+CA1 injection in LPS+sham acupuncture group;(8)high glucose injection+CA1 LPS+group.Electroacupuncture acupoints “Baihui” and “Dazhui”,ioppi depth of 2 mm,the frequency of electroacupuncture was 2 Hz,the voltage is2 V,once every other day,20 min each time,for 2 consecutive months,sham acupuncture “Baihui”and “Dazhui” by 1 mm non acupoints,not connected to electrical stimulation instrument.After treatment,the cognitive function of each group was analyzed by new object recogniti on test and water maze test.Golgi staining was used to analyze the m orphology of neurons.Western blotting was used to analyze the level of tau protein phosphorylation in each group.ELISA was used to analy ze the expression of IL-1,TNF-? and IL-6 in plasma and hippocampus of each group.Result:1.The body weight of normal blood glucose mice gradually increased(P<0.05),and the normal blood glucose mice were injected with LPS in the hippocampal CA1 area(P<0.05).Either EA(P<0.05)or sham(P<0.05)body weight increase.The body weight of hyperglycemia mice was gradually reduced(P < 0.05);body weights were reduced in both groups after electro-acupuncture(P<0.05)or sham(P<0.05)after injection of LPS(P<0.05)in the hippocampal CA 1 area of hyperglycemic mice.After acute high-dose intraperitoneal injection of STZ,blood glucose levels in the hippocampal CA1 injection LPS group and saline group increased(P<0.01,P<0.01).2.New object recognition experiment results show that in the experime ntal stage,the new object recognition index in the hyperglycemia gro up is lower than that in the normal blood glucose group(P<0.05).The index of identification of new objects in the group of injection of LPS in the hippocampal CA1 region of normal blood glucose mice was lo wer than that of the Saline group(P<0.05).Compared with Saline inje ction group,the index of new object recognition in hyperlipidemic mo use hippocampal CA1 injection group was significantly lower(P<0.05).After the LPS was injected into the hippocampal CA1 region of normal blood glucose mice,the new object recognition index of the electroa cupuncture group was higher than that of the pseudoneedle group(P<0.05).In the hyperglycemia mouse hippocampal CA1 zone,the new object r ecognition index was higher in the electroacupuncture group than in t he pseudoneedle group after injection of LPS(P<0.05).3.The result of water maze showed that during the navigation experime nt,the escape latency of hyperglycemic group was higher than that of normal blood glucose group(P<0.01).The escape latency of the group of normal blood glucose mice in the hippocampal CA1 injection group was higher than that of the Saline group(P<0.05),and the escape la tency of the hyperglycemic group in the hippocampal CA 1 injection gr oup was higher than that in the Saline group(P<0.05).The evasion l atency of the electroacupuncture group after the injection of LPS in the hippocampal CA1 region of mice with normal blood glucose was lowe r than that of the sham group(P<0.05).Compared with the sham group,the latency of the hyperglycemic mice in the hippocampal CA1 area rec eiving LPS after electrotherapy was lower than that of the sham group(P<0.05).In the space exploration experiment period,the time spent in the tar get quadrant and the number of platform crossings in the hyperglycemi a group were lower than those in the normal blood glucose group(P<0.01,P<0.05).The time of the injection of LPS in the hippocampal CA 1region of the mice with normoglycemia was lower than that of the Sal ine group in the target quadrant(P<0.05,P<0.05),and the LPS of th e hyperglycemic mice in the hippocampal CA1 injection group.The time spent in the target quadrant of the injection Saline group and the n umber of crossing platforms decreased(P<0.05,P<0.05).Compared wit h the pseudoneedle group,the number of time spent in the target quad rant and the number of traversing platforms increased after injection of LPS in the hippocampal CA1 region of normal blood glucose mice(P<0.05,P<0.05).The time that the electroacupuncture group was trea ted with injection of LPS in hippocampal CA1 area of hyperglycemic mi ce was higher than that of the pseudoneedle group in the target quadr ant(P<0.05,P<0.05).4.Golgi staining results showed that the total dendritic spine densit y and mushroom-like dendritic spine density in the hyperglycemia grou p were lower than those in the normoglycemic group(P<0.01,P<0.05);and the hyperglycemic mice were injected with LPS in the hippocampal CA1 area.The total dendritic spine density and mushroom-shaped dend ritic spine density were decreased in the Saline injection group(P<0.05,P<0.05).The total dendritic spine density and mushroom-like den dritic spine density in hyperglycemic mice receiving EA group were hi gher than those in sham group(P<0.05,P<0.05);the hyperglycemic mic e received LPS after CA1 injection in hippocampal CA1 area.The total dendritic spine density and mushroom-like dendrite spine density in t he EA group were higher than those in the sham group(P<0.05,P<0.05).5.Western blotting results showed that there was no significant diffe rence in total tau protein expression among the groups(P>0.05).In t he hyperglycemia group,p-Tau(Ser396)and p-Tau(Ser404)increased in the hippocampus of the normal blood glucose group(P<0.05,P<0.05). Compared with the Saline group,p-Tau(Ser396)and p-Tau(Ser404)inc reased in the hyperglycemic mouse hippocampal CA1 injection group(P<0.05,P<0.05).In hyperglycemic mice receiving electroacupuncture,p-Tau(Ser396)and p-Tau(Ser404)decreased compared with the sham gro up(P<0.05,P<0.05).In the hyperglycemic mouse hippocampal CA1 after injection of LPS received electro-acupuncture group than the pseudo-n eedle group p-Tau(Ser396),p-Tau(Ser404)were reduced(P<0.05,P<0.05).6.ELISA results show:(1)Comparison of the expression of IL-1?,TNF-? and IL-6 in hypergl ycemia group and normoglycemic group Compared with hyperglycemic group and normoglycemic group,there was no significant difference in levels of IL-1?,TNF-?,and IL-6 in plas ma of normal saline injected in hippocampal CA1 region for 12 h(P>0.05,P>0.05,P>0.05);The levels of IL-1?,TNF-?and IL-6 in plasma of hyperglycemia group were higher than those of normoglycemia group(P<0.05,P<0.05,P<0.05).There was no significant difference in levels of IL-1?,TNF-?,and IL-6 in the hippocampus of normal hippocampal CA1 region after 12 h i njection of normal saline(P>0.05,P>0.05,P>0.05),60 day hyperglyc emic group had higher levels of IL-1?,TNF-?,and IL-6 in the hippoca mpus than the normoglycemic group(P<0.05,P<0.05,P<0.05).(2)Comparison of IL-1?,TNF-?and IL-6 expression in lipopolysacchari de group and normal saline group The levels of IL-1?,TNF-?,and IL-6 in plasma were increased in hype rlipidemic mice injected with LPS at hippocampal CA1 area compared wi th Saline group at 12 h(P<0.01,P<0.01,P<0.01).There was no signif icant difference in plasma levels of IL-1?,TNF-?and IL-6 between th e injection of LPS in hippocampal CA1 region and the injection of Sal ine(P>0.05,P>0.05,P>0.05).The levels of IL-1?,TNF-?and IL-6 in the hippocampus of the hypergl ycemic mice injected with LPS in the hippocampal CA1 area were signif icantly higher than those in the Saline group(P>0.01,P>0.01,P>0.01).There was a significant difference in TNF-?between the injection of LPS and Saline in the CA1 hippocampus of hyperglycemic mice(P<0.05).There was no significant difference in IL-1? and IL-6 between the LPS group and the hippocampal CA1 region(P>0.05,P>0.05).(3)Comparison of IL-1?,TNF-?,IL-6 expression in electroacupuncture group and sham group There was no significant difference in plasma levels of IL-1?,TNF-?and IL-6 between the electroacupuncture group and the sham group at 12 h(P>0.05,P>0.05,P>0.05).On the 60 th day,the levels of IL-1?,TN F-?,and IL-6 in the EA group were lower than those in the sham group(P<0.05,P<0.05,P<0.05).Compared with the sham group,the levels of IL-1?,TNF-?and IL-6 in the hippocampus of the hyperglycemic mice in the hippocampal CA1 region after injection of LPS were significantly lower than those in the sham group(P<0.05,P<0.05).On the 60 th day,the levels of IL-1?,TNF-?,and IL-6 in hippocampus of electroacupunc ture group were lower than those of sham group(P<0.05,P<0.05,P<0.05).conclusion:1.Acute high-dose intraperitoneal injection of STZ can cause chr onic hyperglycemia.Electroacupuncture “Baihui”and “Dazhui” have no effect on blood glucose and insulin in STZ-induced type 1 diabet ic mice.2.Hyperglycemia and LPS can cause cognitive impairment in mice.Injection of LPS into the hippocampal CA1 region aggravates cognitiv e impairment in hyperglycemic mice.Electroacupuncture“Baihui”and“Dazhui”can effectively relieve hyperglycemia and LPS induced reco gnition.3.Hyperglycemia and injection of LPS in hippocampal CA1 area wil l result in decreased hippocampal dendritic spine density.Injection of LPS in hippocampal CA1 area will increase dendritic spine density in hyperglycemic mice.Electroacupuncture “Baihui”and“Dazhui”ca n be performed.Improved STZ induced type 1 diabetes and hippocampal CA1 injection of LPS caused by dendritic spine defects.4.Hyperglycemia and injection of LPS into hippocampal CA1 area w ill increase the phosphorylation of hippocampal Tau.Injection of LPS in hippocampal CA1 area will increase the phosphorylation level of hippocampal Tau in hyperglycemic mice.Electroacupuncture“Baihui”an d“Dazhui”can improve.STZ-induced type 1 diabetes and TPA hyperphos phorylation of hippocampal CA1 injection of LPS.5.The levels of IL-1?,TNF-?and IL-6 in plasma and hippocampus of STZ-induced type 1 diabetic mice showed an increasing trend.Injec ting LPS into hippocampal CA1 area exacerbated the plasma and hippoc ampal formation in hyperglycemic mice.Levels of IL-1?,TNF-?and IL-6 and electro-acupuncture“Baihui”and“Dazhui”can reduce the level s of IL-1?,TNF-?and IL-6 induced by STZ-induced type 1 diabetes an d LPS injection in hippocampal CA1 area. |
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