| Objective1To study the effects of treating cognitive impairment after stroke by electroacupuncture at "Shenting" and "Baihui" in the cognitive function and activity of daily living, for the further promotion and application providing the scientific clinical and practice basis.2Using a rat model with focal cerebral ischemia/reperfusion (I/R) injury, in the present study we evaluated the therapeutic efficacy of electroacupuncture against post-stroke cognitive impairment and explored the molecular mechanisms from the ethology/histopathology/molecular boilogy and other kinds of modern experiment technology, for providing the scientific theory basis.Methods:1Clinlal ResearchThere are84patients who matched the inclusion criteria were randomly divided into two groups with every42cases to the clinical observations, the therapeutic group who were used electroacupuncture treatment and the control group who were used foundation treatment only. The conventional therapy was given to patients in two groups, incluing the reducing intracranial pressure, improving the brain circulation, good limb position, cognitive function rehabilitation and so on. The electroacupuncture group was treated with electroacupuncture at "Shenting" and "Baihui" on the basis of the treatment for the control group, once a day,6days every week, total4weeks. Useing the Montreal cognitive function assessment scale (MoCA)、the Loewenste inoccupational therapy cognition assessment (LOTCA) and improved Bathel index (BI) to evaluate the effects of electroacupuncture at "Shenting" and "Baihui" on the neuropsychology/cognition function and the influence of daily living skills after stroke in the before treatment, treatment two weeks and at the end of the treatment.2Experiments Researeh120male SD rats were randomly diveded into the sham operation control group (SC), ischemia control group (IC) and electroacupuncture group (EA),40rats in each group. Using a rat model with focal cerebral ischemia/reperfusion (I/R) injury, in the present study we evaluated the therapeutic efficacy of electroacupuncture against post-stroke cognitive impairment with the Morris water maze and the step down test, TTC staining was used to observe the effect of the cerebral infarct volume, HE staining was used to observe the pathomorphism of the brain, Tunel staining was used to observe the effect of electroacupuncture on apoptotic cell induced by focal cerevbral ischemia in rats, NF-κB-p65activaton-nuclear transfer was observed by laser scanning confocal Microscopy, the activity of translation and transcription NF-κB-p65/IκB/p-IκB/Bax/Fas was also tested by western bloting and RT-PCR in the10d with electroacupuncture at "Shenting" and "Baihui" after MCAO. Then we analysis the relationship between the animal behavior changing and the gene/proterin regulation.Results1Clinical ResultAt the end of the test, we found1patients withdrew in the EA group and3patients in the control group. There were not statistically significant (P>0.05) between the treatment group and the control group in the eassessmen of MoCA, LOCAT and BI index before treatment, the baseline of both groups were in order and the comparability was good. Significant differences in the eassessmen of MoCA, LOCAT and BI index were found between two groups electroacupuncture2weeks and4weeks (P<0.05, compaire with before treatment), which indicated electroacupuncture at "Baihui" and "Shenting" can significantly improve the cognitive function and the ability of daily living. Compaired with electroacupuncture2weeks, the socres of MoCA, LOCAT and BI index were also significance improved in electroacupuncture4weeks (P<0.05), which demonstrated, to some extent, the curative effect of electroacupuncture has a time-dependent. There was a significant difference in MoCA, the orientation/perception/the visual motion perception in LOCAT and BI between electroacupuncture group and the control group (P<0.05), however, there are no achieve statistical signigicance in the thinking operation/the attention and concentration in LOCAT (P>0.05) in electroacupuncture2weeks. There was a significant difference in MoCA/LOCAT and BI index between electroacupuncture group and the control group in electroacupuncture4weeks (P<0.05), which indicated the improvment of the attention lag behind from other cognitive functions.2Experimental Result(1) Result of neurological deficits and the cognitive impairment in cerebral I/R injured rats indicates that:The neuroprotective effect of electroacupuncture at "Shenting" and "Baihui" acupoints was evaluated by examining the neurological deficit scores. As expected, rats in sham operation group (SC) did not show any manifestation of neurological deficits, whereas all rats in both ischemia control (IC) and electroacupuncture (EA) groups displayed obvious signs of cerebral injury(P<0.05). To evaluate the effect of electroacupuncture on cognitive function, we performed Morris water maze test from the fourth to the ninth day after MCAO surgery. In the Morris water maze test the latency and route for IC group rats to reach the hidden platform markedly increased, whereas the times that rats crossed the location of the platform were significantly decreased in the9th day after MCAO(P<0.05versus SC group), indicating that cerebral I/R injury resulted in cognitive impairment. However, electroacupuncture significantly decreased the latency and route and increased the times of crossing the area which the platform located in Morris water maze test (P<0.05, versus IC group). Collectively, these data suggested that electroacupuncture at "Shenting" and "Baihui" could ameliorate cognitive impairment in cerebral I/R injured rats. Compaire with the ischemia control group, eletroacupuncture could reduce the times which the cerebral I/R injured rats was shocked by electric current (P<0.05) in3min and increase the average latency jumping from the safe plat in the step down test in the ninth and the tenth day (P<0.05).(2) TTC staining shows that:Electroacupuncture treatment profoundly reduced cerebral infarct volumes in I/R injured rats (P<0.05, EA group versus IC group). Taken together, it was suggested that electroacupuncture at "Shenting" and "Baihui" acupoints had therapeutic efficacy against cerebral I/R injury.(3) HE staining shows that:In the IC group, the number of the nerve cells detected, arranged loosely, lysis, the cellular structure fuzzy and deformation and has a few nerve cell dyeing deepen. However, in the EA group, the lysis was ligher and the tightly and the structure was relatively completely. There was no pathological changes in the sham group.(4) TUNEL staining shows that:The percentage of TUNEL-positive cells in SC or IC group was1.2±0.3%or46.7±9.7%(P<0.05), indicating that I/R injury profoundly promoted cerebral cell apoptosis. However, electroacupuncture significantly inhibited I/R-induced apoptosis of neural cells as the percentage of TUNEL-positive cells in EA group was18.9±4.5%(P<0.05, versus IC group), suggesting that electroacupuncture at "Shenting" and "Baihui" inhibited ischemia-mediated cerebral cell apoptosis.(5) Immunofluorescence staining to examine the nuclear translocation of NF-κB shows that:The cerebral I/R injury profoundly induced in the nuclear translocation of NF-κB p65 subunit, which however was inhibited by electro acupuncture.(6) The activity of translation and transcription of NF-κB-p65/p-IκB/IκB/Bax/Fas by western bloting and RT-PCR in the10d after MCAO. To further investigate the mechanism of NF-κB signal pathway of activity of electroacupuncture, we examined the mRNA level and protein expression of NF-κB-p65/IκB/Bax/Fas in ischemic cerebral tissues using RT-PCR and Western Blotting, respectively. The results show:cerebral I/R injury remarkably enhanced NF-κB-p65/Bax/Fas expression at both transcriptional and translational levels and weakened the IκB/p-IκB expression, which, however, was significantly neutralized by electroacupuncture (n=6, P<0.05).Conclusion(1) Electroacupuncture at "Shenting" and "Baihui" can improve patient,s overall cognitive function and the activities of daily living, however, the improve of daily living lag in the cognitive function. Close relationships were found between the disease course and the curative, the sooner of the intervention, the more obvious of the improvment in the cognitive function and the daily living.(2) Electroacupuncture at "Shenting" and "Baihui" can improve the cognitive function and has protective effect in cerebral ischemia-reperfusion injured rats. The inhibition of NF-κB-mediated neuronal cell apoptosis might be one of the mechanisms whereby electroacupuncture at "Shenting" and "Baihui" exerts therapeutic effect on post-stroke cognitive impairment. |
PDF Full Text Request