| Purpose: To study the expression characteristics of GFAP in astrocytes and the expression of connexin CX43 in three courses of treatment of VD rats with Huayu Tongluo moxibustion,and to analyze the effect of Huayu Tongluo moxibustion on astrocytes and its correlation with connexin.Methods: Experiment 1: The SPF rats were removed from the water maze and rotated in situ,floating and stationary.Then the experimental rats were divided into two groups: sham operation group and model group by random list method.In the sham operation group,only the common carotid artery was separated and no other treatment was done.The rat model was established by 2-VO method modified by the experimental group.Then the rats identified by the water maze experiment and successfully modeled were randomly divided into three groups: model group,moxibustion group and western medicine group.Each group of rats was fed for 7 days and then began different treatment.Rats in the model group and the sham-operation group continued to be fed conventionally without any treatment.Moxibustion group: suspension moxibustion was performed on rats "Dazhui","Baihui" and "Shenting" for 20 minutes at each point and one course of treatment every 7 days,with a total of 3 courses of moxibustion.Western medicine group: western medicine piracetam was given to rats in western medicine group at the same time period as moxibustion group,and the dosage was determined according to the table of equivalent dose ratio between human and animal.After the treatment was completed,the water maze test results of each group were evaluated to analyze the effect of Huayu Tongluo moxibustion on the spatial memory ability of VD rats.Experiment 2: VD rats were randomly divided into sham operation group,model group,western medicine group and moxibustion group.Each group was further divided into three subgroups according to the course of treatment,namely,one course of treatment was 7 days,two courses of treatment were 14 days,and three courses of treatment were 21 days.In the same way as in experiment 1,the sham-operation group was randomly selected as the control group,and the other VD rat models were reproduced as above.After the model was successful,different treatment methods were adopted according to different groups.The method was the same as that in experiment 1.Rat brain tissue was obtained 24 hours after treatment.The pathological changes of VD rats were observed by HE staining.Immunohistochemistry and Western blot were used to detect the expression of GFAP protein in hippocampal astrocytes of VD rats.To explore whether the mechanism of Huayu Tongluo moxibustion on VD rats is related to astrocytes.Experiment 3: Grouping and sampling are the same as experiment 2.Western blot was used to detect the expression of CX43 in gap junction protein of VD rats.Statistical data analysis of GFAP and CX43 expression correlation study.To explore the effect of Huayu Tongluo moxibustion on astrocytes in VD rats and the correlation between gap junction.SPSS23.0 was used for data statistical analysis in this experimental study,and the measurement data were expressed by mean (?)tandard deviation.For data satisfying normal distribution and variance homogeneity test,variance analysis is used for comparison between the two groups,and SNK and LSD analysis are used for multiple comparison.Data that do not conform to normal distribution and variance homogeneity are analyzed by rank sum test.Pearson multivariate correlation was used to analyze the correlation,and the correlation coefficient was r.Setting p < 0.05 is statistically significant,p<0.01 is statistically significant,p>0.05 is not statistically significant.Results:(1)Effect of Huayu Tongluo Moxibustion on Spatial Memory Ability of VD Rats: On the 5th day of water maze experiment,the incubation period and times of crossing the platform were counted.Compared with the sham operation group,the model group takes longer and less time.Compared with the model group,the rats in moxibustion group and western medicine group have less time and more times.Moxibustion group has more obvious effect than western medicine group.Compared with before treatment,the time of rats in moxibustion group and western medicine group decreased more and the number of times increased.The difference was statistically significant(P<0.05).(2)Effect of Huayu Tongluo Moxibustion on GFAP Expression in VD Rats: The expression in model group is higher than that in sham operation group;After treatment,the expression of GFAP in hippocampus of rats in western medicine group and moxibustion group was lower than that in model group,and that in moxibustion group was lower than that in western medicine group.After three courses of treatment,the overall expression of GFAP showed a downward trend.The difference was statistically significant(P<0.05).(3)Study on the expression of connexin CX43 in hippocampus of VD rats and its correlation with GFAP by Huayu Tongluo moxibustion: the expression in model group is higher than that in sham operation group;After treatment,the expression of CX43 in western medicine and moxibustion group was lower than that in VD model group,and that in moxibustion group was lower than that in western medicine group.The overall expression of CX43 also showed a downward trend in the three courses after treatment.Statistical software was used to compare the protein expression results of GFAP and CX43,and correlation analysis was conducted,showing significant positive correlation.The difference was statistically significant(P<0.05).Conclusion:(1)Huayu Tongluo moxibustion can effectively improve the spatial memory of VD rats.(2)Huayu Tongluo moxibustion can promote the repair of cerebral ischemia injury by down-regulating the expression of astrocytes in the CA1 region of VD rats.(3)The effect of Huayu Tongluo moxibustion on the expression of astrocytes in VD rats may be related to the expression of gap junction CX43. |
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