To Explore The Mechanism Of "Xingnao Kaiqiao" Acupuncture On Vascular Dementia Based On Inflammatory Reaction

Objective: To observe the clinical efficacy of Xingnao Kaiqiao acupuncture in the treatment of vascular dementia patients,and to further observe the behavioral changes and the expression of TNF-α,IL-1β,IL-6,IL-10 mrna in the hippocampus of the model rats treated with xingnao Kaiqiao acupuncture on the basis of clinical experiment.To explore the mechanism of xingnao Kaiqiao acupuncture in improving VD cognitive impairment from the perspective of inflammation.Methods:1.Clinical experimental study: this study included 90 patients with vascular dementia who met the inclusion criteria,all of whom were inpatients and outpatients from the department of Neurology and rehabilitation medicine of Liuzhou Hospital of Traditional Chinese Medicine.Ninety patients with vascular dementia were randomly divided into western medicine group,ordinary acupuncture group and Xingnao Kaiqiao acupuncture group with 30 cases in each group by random number table method.The western medicine group was given Donepezil hydrochloride(5mg/qn,4 weeks).In xingnao Kaiqiao acupuncture group,neiguan,Shenmen,Shuigou,Baihui,Sishencong,Sanyinjiao,and Yongquan(except renren,Baihui,and Sishencong,all points were selected on both sides).Syndrome differentiation and acupoints: liver and kidney insufficiency,add Ganshu,Shenshu;If qi and blood are insufficient,add Zusanli,Qi Sea and Blood sea;Phlegm turbidity closed orifices,plus Fenglong,Zhongwan;Yu blood obstruction collaterals,plus diaphragm Shu.The main points in the ordinary acupuncture group were Baihui,Shenting,Yintang,Taixi,Xuanzhong and Si Shencong,and the points for syndrome differentiation were the same as those in the Xingnao Kaiqiao acupuncture group.Acupuncture treatment once a day,6 days a week for 4weeks.Before and after treatment,mental state Scale(MMSE),Clock drawing test(CDT),daily Living scale(ADL)were evaluated.SPSS 26.0 and Graph Pad Prism 8.0 were used for statistical analysis and plotting.2.Animal experimental study: In this study,healthy male SD rats were selected as the research object,and VD animal model was prepared by repeated ischemia reperfusion combined with intraperitoneal injection of sodium nitroprusside.In the sham operation group(12 rats),only bilateral common carotid arteries were separated without vessel clamping and intraperitoneal injection of sodium nitroprusside.The model was evaluated based on the results of the Morris Water Maze(MWM)experiment.After successful modeling,they were randomly divided into model group,Western medicine group,ordinary acupuncture group and Xingnao Kaiqiao acupuncture group(12 each).Western medicine group was given donepezil hydrochloride solution 0.45mg/(kg·d),once a day,for 4 weeks.The rats in the normal acupuncture group and the Xingnao Kaiqiao acupuncture group were needled according to the main points in the clinical experiment.For the bilateral acupoints,the right acupoint was needled twice a day and the left acupoint was needled one day in the animal experiment.The method of needling and the time of retention of the needle were referred to the clinical experiment.After 4 weeks of treatment,MWM(including positioning navigation experiment,spatial search experiment and visual platform experiment)was used again for behavioral analysis of rats in each group.After MWM,the rats in each group were decapitated,and the m RNA expressions of TNF-α,IL-1β,IL-6 and IL-10 in the hippocampus of rats in each group were detected by real-time PCR.SPSS 26.0 and Graph Pad Prism8.0 were used for statistical analysis and plotting.Results:1.Clinical trial: After treatment,MMSE,CDT and ADL scale scores of patients in the three groups were improved compared with those before treatment,with statistical significance(P<0.05).There were no significant differences in MMSE,CDT and ADL scores between western medicine group and ordinary acupuncture group(P>0.05).Compared with western medicine group and ordinary acupuncture group,MMSE,CDT and ADL scale scores of Xingnao Kaiqiao acupuncture group were significantly increased(P<0.05).2.Animal experiment:(1)MWM behavior test results:(1)Positioning navigation experiment: compared with sham operation group,the latency of escape in model group was significantly longer(P<0.01);Compared with model group,the latency period of escape of western medicine group,ordinary acupuncture group and Xingnao Kaiqiao acupuncture group was shortened in different degrees(P<0.05);There was no significant difference in escape latency between western medicine group and ordinary acupuncture group(P>0.05).Compared with western medicine group and ordinary acupuncture group,the escape latency time of xingnao Kaiqiao acupuncture group was significantly shortened(P<0.05).(2)Spatial search experiment:Compared with sham operation group,the first time to reach the platform in model group was significantly longer,The Times of crossing the original platform and the time to stay in the original platform quadrant were significantly reduced(P<0.01);Compared with the model group,there were no significant differences in the first time to reach the platform,The Times of crossing the original platform and the time to stay in the original platform quadrant between the western medicine group and the ordinary acupuncture group(P>0.05).Compared with model group,Western medicine group and ordinary acupuncture group,the time of reaching the platform for the first time in xingnao Kaiqiao acupuncture group was significantly shortened,The Times of crossing the original platform and the time of staying in the original platform quadrant were significantly increased(P<0.01);(3)Visual platform experiment: there was no difference in platform arrival time and swimming speed among the five groups(P>0.05).(2)RT-PCR results: Compared with sham operation group,m RNA expression levels of TNF-α,IL-1β and IL-6 in hippocampus of model group were significantly increased(P<0.01),and m RNA expression levels of IL-10 were significantly decreased(P<0.01);Compared with model group,the m RNA expression levels of TNF-α,IL-1β and IL-6 in hippocampus of western medicine group,ordinary acupuncture group and Xingnao-Kaiqiao acupuncture group were decreased(P<0.01),while the m RNA expression levels of IL-10 were increased(P<0.01).The m RNA expression levels of TNF-α,IL-1β,IL-6 and IL-10 in hippocampus of western medicine group and ordinary acupuncture group were not different(P>0.05).Compared with the Xingnao Kaiqiao acupuncture group,the m RNA expression levels of TNF-α,IL-1β and IL-6 in hippocampus of western medicine group and ordinary acupuncture group were higher,while the m RNA expression levels of IL-10 were lower(P<0.01).Conclusions:(1)Clinical trial study: Donepezil,ordinary acupuncture and xingnao Kaiqiao acupuncture can improve the cognitive function and daily living ability of VD patients to varying degrees,among which donepezil and ordinary acupuncture have a balanced therapeutic effect,while Xingnao Kaiqiao acupuncture has the best curative effect.(2)Animal experiments:(1)Donepezil and acupuncture could positively regulate inflammatory factors and spatial learning ability in hippocampus of VD rats,but did not significantly improve spatial memory ability of VD rats;(2)Xingnao Kaiqiao acupuncture can effectively improve spatial learning and memory ability of VD rats;(3)The mechanism of xingnao-Kaiqiao acupuncture to improve the cognitive function of VD rats may be through up-regulating the m RNA expression of inflammatory factor IL-10 in the hippocampus,down-regulating the m RNA expression of TNF-α,IL-1β and IL-6,inhibiting the inflammatory response in the hippocampus and playing a protective role in the brain.