| Objective:From four aspects of Animal, Molecular Biology, spectrum-effect relationship and pharmacokinetics, study on the protective effect of cerebral ischemic injury.On the basis of the establishment of focal cerebral ischemia in rats, to study the protective effect of Naojian Tablet (the thin recipe of Buyanghuanwu), which is based on the method of benefiting vital energy, removing stasis and producing new, on cerebral ischemia injury in rat and the effect on the expression of hippocampal tissue cyclin, CyclinB1and CyclinB2. On this basis, combined with matlab software programming to analyze the spectrum-effect relationship between the effective component of Naojian Tablet and the protective effect on cerebral injury, as well as the pharmacokinetics,and to investigate the the material basis for efficacy and law of action of Buyanghuanwu recipe and Naojian Tablet, and to establish mathematical model fitting the spectrum-effect relationship and pharmacokinetics of Buyanghuanwu recipe and Naojian Tablet, providing scientific experiments basis for the effectiveness of clinical applications of Naojian Tablet.Methods:The rat model of focal cerebral ischemia was built using middle cerebral artery occlusion, and the animals were randomly divided into sham operation group, model group, Buyanghuanwu Decoction (BYHWD) group, the high, medium and low dose of Naojian Tablet groups. The rats of each group were weighed at1d to7d, and the neurological score of the rats of each group was evaluated at7d, which was used to calculate infarct size with TTC after processing. The brain tissue was taken at1d,3d,7d,14d,28d to run WB, RT-PCR experiments to test the expression of their hippocampus cyclin, Cyclin B1and Cyclin B2. The experimental results were analyzed with PCA and GRA method to obtain the relationship between the fingerprints of BYHWD and Naojian Tablet, the pharmacodynamic indexes such as neurological score, cerebral infarct size, and dry-wet ratio of brain, and the Cyclin B1and Cyclin B2, then the spectrum-effect relationship and pharmacokinetics mathematical model was established based on curve-fitting method.Results:(1) Compared with the model group, body weight of rats in Naojian Tablet group increased significantly (P<0.05), neurological score (P<0.05), infarct size (P<0.05) and brain dry-wet ratio (P<0.05) were significantly decreased, and the medium dose group of Naojian Tablet had the optimal therapeutic effect on cerebral ischemia, which efficacy is slightly better than BYHWD group (normal human dose).(2) The test results of WB and RT-PCR showed that, compared with the model group, the expression of Cyclin B1and Cyclin B2can be reduced effectively in Naojian Tablet group, the difference was statistically significant (P<0.05), the regulation of Naojian Tablet on cyclin was most obvious at7d (P<0.05); the expression of Cyclin B1and Cyclin B2had no significant difference between Naojian Tablet group and BYHWD group (P>0.05).(3) The study result of spectrum-effect relationship showed that, for the samples of Naojian Tablet and BYHWD, the ranking of common peaks was larger than their respective unique peaks, and the contributions of chemical components at the retention time tR=20.649,6.474and26.404min on neurological score, infarct size, brain dry-wet ratio as well as Cyclin B1and Cyclin B2were of the top five.(4) The results of evaluation of pharmacokinetics model showed that there was no significant difference (P>0.05) between Naojian Tablet and BYHWD on the absorption, distribution and elimination of ferulic acid and ligustilide, but the senkyunolide in BYHWD was more easily absorbed、distributed and eliminated in body.Conclusion:(1) Naojian Tablet can improve neurological symptoms, reduce infarct size, relieve cerebral edema effectively, confirming that Naojian Tablet has protective effect on cerebral ischemia injury, and its effect is equivalent to the whole receipt of BYHWD. 40(2) Naojian Tablet can significantly reduce the expression of hippocampus Cyclins B1and Cyclins B2in cerebral ischemia model rats, and its effect is equivalent to the whole receipt of BYHWD.(3) The chemical compositions of tR=20.649,6.474and26.404min in the fingerprinting spectrum of Naojian Tablet may play a key role in regulation of cerebral ischemic injury and intervention of cyclin, which were speculated as its primary efficacy material basis.(4) There is no significant difference between Naojian Tablet and BYHWD on the absorption, distribution and elimination of ferulic acid and ligustilide, but the absorption, distribution and elimination of senkyunolide A has significant difference, on which a follow-up in-depth investigation can be carried on.(5) The spectrum-effect relationship of traditional Chinese medicine compound can be objectively evaluated based on PCA-GRA method, which is a good method worth learning.(6) The pharmacokinetic mathematical model established based on curve fitting method combined with matlab programming can effectively and objectively evaluate the absorption, distribution and elimination of drug in body, providing a new thought for the follow-up study of pharmacokintrics.(7) Naojian Tablet have a good advantage in the treatment of cerebral ischemia, according with the needs of stroke patients, which provides a theoretical basis for indicating theclinical rational medication. |
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