Correlation Of Aromatic Resuscitation Drugs And P-gp

Recent literatures reported that resuscitation-inducing aromatic herbs could penetrate and assist other drugs in passing through the BBB, and play an important role in the treatment of brain disorders. It provides a theoretical basis for the meridian distribution of resuscitation-inducing aromatic herbs in TCM theory. P-glycoprotein, a drug efflux pump, distributes widely in BBB and many other parts of the body. Generally, resuscitation-inducing aromatic herbs take an important role in inhibiting drug’s uptake, drug distribution, accelerating drug elimination. It is speculated that there was a certain correlation between resuscitation-inducing aromatic herbs and P-glycoprotein was investigated in this thesis. The results not only provide evidence for elaborating mechanism of resuscitation-inducing aromatic herbs, but also screen drugs as P-glycoprotein inhibitors, and then provide basis for new drug development. The main contents are listed as follows:1. Caco-2cell was selected in this experiement. The content of rhodamine-123which is affected by resuscitation-inducing aromatic herbs in cells, was determinated by flow cytometry, thereby studying the effects of drugs on P-glycoprotein function at the cellular level. The result have shown that fluorescence intensity of rhodamine-123in benzaldehyde, vanillinv and the β-asarone groups increased significantly compared with the control group, and it is suggested that benzaldehyde, vanillin and β-asarone could obviously inhibit P-glycoprotein efflux function at the cellular level, which reveals inhibiting the rhodamine-123efflux in Caco-2cells.2. Everted intestinal sacs were made, and content of rhodamine-123content which is affected by resuscitation-inducing aromatic herbs in cells was determinated by HPLC. The results showed that: resuscitation-inducing aromatic herbs could inhibit rhodamine-123efflux from in the jejunum and ileum. The Suppression intensity of resuscitation-inducing aromatic herbs in the jejunum:shichangpu Volatile oil﹥borneol﹥α-asarone﹥vanillin﹥Styrax volatile oil﹥β-asarone﹥musk ketone﹥benzaldehyde﹥benzoin volatile oil; that of resuscitation-inducing aromatic herbs in the ileum: β-asarone﹥benzoin volatile oil﹥benzaldehyde﹥vanillin﹥α-asarone﹥Styrax volatile oil﹥musk ketone﹥Shichangpu volatile Oil﹥borneol. The results indicated that the drug could inhibit P-glycoprotein drug efflux function in vitro tissue levels. The action time of the drugs, the binding rate and distribution of P-gp in the rat gut probablely contributed to different inhibition rate of the jejunum and ileum.3. Effects of α-asarone, β-asarone, benzoin, vanillin, musk ketone, borneol, shichangpu volatile oil, benzoin essential oil and volatile oil Styrax, benzoin essential oil on the MDRl gene expression of Caco-2cells were investigated by Real-Time PCR. In genetic lever, the mechanism of these drugs effecting on P-glycoprotein was studied. The reducing rates of MDRl expression were:musk ketone﹥shichangpu volatile oil﹥β-asarone﹥benzoin volatile oil﹥borneol﹥α-asarone﹥benzoin﹥vanillin﹥Styrax volatile oil. The results showed that these drugs could reduce the expression level of MDR1gene in P-glycoprotein, which may be one of mechanism of resuscitation-inducing aromatic herbs inhibiting P-glycoprotein.4. Effects of a-asarone, β-asarone, benzoin, vanillin, musk ketone, borneol, shichangpu volatile oil, benzoin essential oil and volatile oil Styrax, benzoin essential oil on the P-gp expression levels were detected by Flow cytometric, thus researching mechanism of action of drugs to P-gp in protein lever. The reducing rates of MDR1expression were:styrax Essential Oil﹥benzaldehyde﹥borneol﹥benzoin volatile oil﹥β-asarone﹥shichangpu Volatile Oil﹥vanillin﹥musk ketone﹥α-asarone. The results showed that these drugs reduced the expression lever of P-glycoprotein in different degrees, which suggested that it may be one of mechanism of resuscitation-inducing aromatic herbs inhibiting P-glycoprotein.