| Vascular dementia(VD)is the acquired intelligent damage syndrome caused vasogenic factor.The clinical manifestation is a series of abnormal changes,including memory,cognition,emotion and behavior.In recent years,the incidence of VD increases rapidly.The specific pathogenesis of VD in modern medicine is still unclear,so there is no explicit treatment.Traditional Chinese Medicine(TCM)has obvious advantages in the prevention and treatment of VD.In this study,to elucidate the therapeutic effect of Wu Zang Wen Yang Hua Yu Decoction on VD by use of morris water maze、HE staining、 WB、PCR and proteomics were used.The study was divided into two parts:1.Theoretical research of Chinese medicine This paper mainly expounds the etiology,pathogenesis,diagnosis and treatment of vascular dementia in TCM,the study of Professor Tang Nong’s academic thought and the relationship between brain function and the five viscera;the interpretation of the pathogenesis and treatment of vascular dementia based on the theory of yin and yang ontological structure of inner yang external yin.2.Experimental researchObjectives: To establish the model of vascular dementia rats,observe the effects of Wu Zang Wen Yang Hua Yu Decoction on learning and memory of vascular dementia rats,and to study the effects of Wu Zang Wen Yang Hua Yu Decoction on the signal pathway P I 3 K / A k t / m T O R and the apoptosis protein of BCL-2、BCL-XL、Caspase-9 and Caspase-3 apoptosisprotein.To study the effect of Wu Zang Wen Yang Hua Yu Decoction on the expression of related proteins,to identify the differential protein expression,and to explore the effect mechanism of Wu Zang Wen Yang Hua Yu Decoction on VD.Methods: The healthy male SD rats of SPF grade were purchased.Then the experimental rats were randomly divided into blank group and model group.Then the model of VD rat was established by repeated double-sided carotid reperfusion method.Then the rats were randomly divided into five groups,including blank group、 model group、 high-dose Chinese medicine group、 low-dose Chinese medicine group and western medicine group.Continuous for 14 days.The protein expression of PI3K、Akt、 m TOR、BCL-2、BCL-XL 、Caspase3、 Caspase9 were detected by WB and PCR.To study the proteomics and identify the differential proteins and determine the effect of Wu Zang Wen Yang Hua Yu Decoction on the related proteins of VD rats by use of i TRAQ/TMT labeling and liquid chromatography-mass spectrometry(LC-MS).Results:(1)The analysis of Water maze test demonstrated that the comparison between VD model group and sham operation group has significantly different in decreasing the ability learning and memory.This two kinds of ability were recovered in all treatment groups comparing with the VD model group,and the differences were statistically significant(P<0.05).(2)HE staining of hippocampal tissue showed that: compared with the model group,the pathological damage of hippocampus was improved in the treatment group of Wu Zang Wen Yang Hua Yu Decoction,the therapeutic effect of high-dose Chinese medicine group was significant.(3)The analysis of RT-PCR and WB showed:Compared with control group,protein expression of PI3K、Akt、m TOR、BCl-2、BCL-XL、Caspase-9、Caspase-3 in hippocampal CA1 area of rats of model group,the difference between both groups was statistically significant(P<0.05).Compared with model group,protein expression of PI3K、Akt、m TOR、BCl-2、BCL-XL in hippocampal CA1 area of rats of in high-doses and western medicine group up-regulated,and the protein expression of Caspase3 and Caspase9 were decreased in high-doses and western medicine group,the difference between both groups was statistically significant(p<0.05).Compared with western medicine group,protein expression of PI3K、Akt、m TOR、BCl-2、BCL-XL、Caspase-9、Caspase-3 in the low-doses showed statistic difference(p<0.05);protein expression of PI3K、Akt、m TOR、BCl-2、BCL-XL、Caspase-9、Caspase-3 in the high-doses showed no statistic difference(P>0.05).Compared with low-doses group,protein expression of PI3K、Akt、m TOR、BCl-2、BCL-XL、Caspase-9、Caspase-3 in high-doses group showed statistic difference(p<0.05).The difference of treating effect between high-doses group and western medicine groups was no statistically significant(P>0.05).(4)The results of proteomics showed that : In the VD model,23 species were up-regulated and 17 were down-regulated among the more than 2 times the differential protein(P<0.05)Compared with the sham-operation group.16 kinds of proteins were up-regulated and 10 were down-regulated among the more than 2 times the differential protein(P<0.05)in the Chinese medicine group compared with the model group.Differential protein expression level was clustered to show:In the control group,the expression of the cluster1 region was down-regulated,the cluster2 and cluste4 region were up-regulated.In the model group,the expression of the related protein in cluster1 region was up-regulated,and the expression of the related protein in the cluste4 region were down-regulated.In the treatment group,the expression of the related protein in cluster1 region was down-regulated,the expression of the related protein in the cluster2 and cluste4 region were up-regulated.Conclusions: The results showed that the Wu Zang Wen Yang Hua Yu Decoction significantly improved the learning and memory ability of VD rats and inhibite the neuronal apoptosis in hippocampus.Wu Zang Wen Yang Hua Yu Decoction might play a neuroprotective role by activating the signaling pathway P I 3 K / A k t / m T O R,s t r e n g t h e n i n g t h e e x p r e s s i o n o f BCL-2and BCL-XL,reducing the e x p r e s s i o n o f Caspase-9 and Caspase-3,and effectively inhibiting the apoptosis of nerve cells.The therapeutic effect on vascular dementia rats may be achieved by restoring the expression of related abnormal proteins. |
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