| [Object]To provide clinical data for correlative research of CCCI and define the improvement of the symptomes, cognition and cerebral blood flow perfusion, we compare the differences between before and after treatment of them. Furthermore, to explore the pathological changes of CCCI and the effect and mechanism of Huoluo Yinao prescriptionm, Fructus broussonetiae and Tribulus terrestris, we study the cognition, neural network, neuroplasticity and the polarization of microglial cell in CCCI rats and drug treatment group rats.[Methods]In clinical research, study of 10 patients which visited encephalopathy department outpatient service and inpatient ward of Dongfang hospital affiliated to Beijing University of Chinese Medicine in September 2014 - December 2015, they all conformed to the included standard and has chief complaint for dizziness and woozy. Three of the patients have CTP comparison before and after treatment. The personal data, scales of symptoms, the multi-field cognitive function and CTP data were record before treatment. After 3 months of treatment by Huoluo Yinao prescription, we evaluate the scales of symptoms and the multi-field cognitive function again, meanwhile select 3 patients to obtain CTP data. Using SPSS 19.0 to analysis of the data before and after treatment, and definite the efficacy of Huoluo Yinao prescription. In experimental research, a model of CCCI was induced by 2-VO. Rats randomly divided into 5 groups:Sham, Model, Fufang, CSZ and XNST. Each group was intragastric administration one time every day from modeling day to drawing materials day for 42 days. The Morris water maze test were executed one week before drawing materials day to evaluating the memory of rats. Western blotting and immunofluorescence was used to detect the expression levels of MBP, NF200, MAP2, synaptophysin, Ibal, iNOS, CD16, CDllb, Argl, CD206, p-AKT(Thr 308), p-AKT(Ser 473), AKT, p-Erk, Erk, p-AMPK and AMPK, and a comparison was performed among the groups.[Results]Clinical observation showed:(1) Symptom observation:European vertigo scale, DHI vertigo disorder scale showed the symptoms of 4 patients improved significantly in 7 days, and 2 patients in 14 days whose primary symptom is vertigo, while all of the patients with dizziness were improved significantly in 7 days. The TCM evaluation of primary symptoms clear that both vertigo and dizziness could be improved in 7 days. After treatment for 3 months, vertigo and dizziness were improved significantly, but the improvement of secondary symptoms and other symptoms were not obvious, while more patients show the dark tongue and string pulse before and after treatment. (2) Multi-field cognitive function score:The comparison of scores of MES-M, MES-T, short delay memory of California verbal learning test and the logical memory test have remarkable difference before and after treatment, the others difference are not obvious. Categorizing patients according to the affected blood vessels around the side:the score increased of MES-M, MES-T, short delay memory of California verbal learning test and the logical memory test mainly related to the patients with left side blood vessels stenosis. Remarkly, the correct words within 15s of language fluency test and positive order recite of digit span test were improved obviously in patients with left side blood vessels stenosis. In the patients with right side blood vessels stenosis, the number reached within 1 min of trail making test and digital model test were improved obviously. The others difference are not obvious. (3) CTP:The CBF was significantly decreased, rMTT rise obviously, but CBV and TTP did not have significantly difference comparison with normal side of the brain. The rCBF and rCBV of whole brain and anterior circulation area were rise, both of the mean value were greater than 1, but there was no statistically significant difference between pretherapy and post-treatment. Meanwhile, the rMTT of whole brain and anterior circulation area were decline. Experiment results indicated: (1) Morris water maze:Navigation results showed that each group of the relationship between escape latency and training time is negative correlation. The escape latency of Model were significantly longer than Sham from 2d to 5d. Nevertheless, the escape latency of Fufang, CSZ and XNST were significantly shorter than Model and close to Sham. Track diagram shows the strategy of searching platform was random of all groups, but at 5d, the strategy of Sham and treatment groups was straight, and Model group still presents random or rounding. The results of space exploration show the frequency of platform crossing was reduced of Model, it has a tendency to increase of Fufang, CSZ and XNST, but there was no significant difference between them. The target quadrant occupancy and swimming speed did not show remarkable difference between five groups. (2) Western blot and immunofluorescence: CCCI did not cause obvious change of MBP expression, CSZ and XNST have a tendency to raise its expression. NF200 of Model group increased significantly compared with Sham group, each treatment group can down-regulate its expression, and XNST is the most apparent. No obvious difference was found between the expression of MAP2 in the rats of five groups. CCCI destroyed the orderly expression of MAP2, and all of treatment groups have certain improvement effect. The level of synaptophysin of Model group was significantly less than Sham group. All of treatment groups cloud enhance the expression of it, and effect of XNST is most obvious. For Ibal expression, the Model group was obviously increased, all of the drug groups have a tendency to reduce it. The body of Ibal cell appears as dot, and around cell body there were a small amount of slender branches. The green fluorescent points in Model significant increase, cell body become larger and branches increase. The treatment groups reduce green fluorescent points and branches. The expression of iNOS, CD 16 and CD11b were increased in Model, iNOS and CD 16 were more apparent, each drug could reduce their expression. Argl expression of the Model group was obviously reduced, and all of the drug groups could increase the expression of Arg1. XNST can obviously increase the expression of P-AKT(Thr 308), and the others groups have no effect on p-AKT(Thr 308), p-AKT(Ser 473), p-Erk and p-AMPK.[Conclusions]Huoluo Yinao prescription could improve symptoms such as dizziness, enhance the memory and increase the cerebral blood flow perfusion of injured side of patients with CCCI. Huoluo Yinao prescriptionm, Fructus broussonetiae and Xinnao Shutong capsule may play improvement effect for cognition by promoting neural network and the neural plasticity. And this protective effect may be associated with the reduced microglia activation, inhibition of M1 polarization direction and promotion of M2 polarization, and then the microglia cell tends to guide the protection effect. |
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