The Study Of The Protective Effect And Mechanism Of Preconditioning With Cinnabar Phenic Acid B On The Cardiac Cell

In order to study the protective effect and mechanism of preconditioning with cinnabar phenic acid B on the cardiac cell, based on the mechanism of anoxic-preconditioning, we establish the damaging model of anoxic-reoxygenation on the cell by the technique of cultivating cardiac microvascular endothelial cell (CMEC) and cardiac cell in vitro. To observe the protective effect on the cell with the cinnabar phenic acid B - preconditioning, we use the biochemistry examination such as the rate of live cell(MTT), lactate dehydrogenase(LDH^ superoxide dismutase(SOD^ nitric oxide(NO^ endothelin (ET) ect and ultrastructure as the evaluating criterion. By the means of medicine such as chelerythrine, a kind of an inhibitor of protein kinase C (PKC), glibenclamide, a kind of an inhibitor of the ATP-sensitive potassium channel (KATp), and from the point of view of the intrinsic protective substance, PKC, and the express of mRNA of the heat shock protein 70 (HSP70) we probe into the protective mechanism on the cell with the cinnabar phenic acid B -preconditioning.In addition, in order to search the best ratio of compatibility of Chinese medicine, enhance the clinical curative effect, enrich the scientific connotation of the description compatibility theory and advance the progress of the Chinese medical modernization, we studied the relationship between the varieties of the effective substance composition in a description and its bio-effect by the injury cell model of CMEC by anoxic-reoxygenation using all composition of the prepared herbs (the paired description elan-sen root and pseudo-ginseng ) and its effective composition.Result:(1) Under the normal culturing condition, cinnabar phenic acid B canenhance the livability of both of cells and the activity of SOD significantly, reduce the release of LDH, keep the ultrastructure, have the tendency of decreasing the secret of ET and the release of NO of CMEC.(2) Hypoxic preconditioning can strengthen the cell endurance of the later damage of longer time anoxic-reoxygenation, which can be observed through the increase of the cell livability and the SOD activity, and the decrease of the LDH activity. At the same time, it can obviously decrease the level of ET and NO of CMEC by anoxic-reoxygenation and improve the change of the ultrastructure.( 3) The protective effect on the cell by cinnabar phenic acid B is similar with the effect by anoxic-reoxygenation. For CMEC, this function will be expressed as the early and delayed protection phases.(4) Glibenclamide, a kind of an inhibitor of KATP, cannot eliminate the HPC protective effect of CMEC. Chelerythrine, a kind of selective inhibitor of PKC, can eliminate the HPC protective effect of CMEC that can be recurred by the preconditioning of cinnabar phenic acid B.(5) During the early preconditioning, cinnabar phenic acid B can improve the mRNA espression of HSP70 and PKC on the anoxic-reoxygenation injured CMEC. The effect is better than HPC obviously. During the delayed preconditioning, cinnabar phenic acid B also can improve the mRNA expression of HSP70, and it has the plus effect with HPC.( 6 ) Under the normal or the anoxic cultivating conditions, elan-sen root can improve the reproduce proliferation of CMEC while pseudo-ginseng inhibits it. Among all the compatibility groups, the proliferation effect of the group of only elan-sen root is the most obvious (compare with other groups p<0.01 p<0.05), especially under the anoxic condition. All the compatibility groups can obviously decrease the release of NO and the secret of ET of CMEC. The best groups decreasing the secret of ET is the compatibility of 10:1 and 10:6. No matter the normal conditions or the anoxic conditions, all groups greatly promote the activity of CMEC, decrease the release of LDH. The effect the group of 2:8 decreasing thesecret of ET and the level of NO is the best. Especially under the anoxic condition, the effect of only cinnabar phenic acid B and only tanshinone II -A is not better than the compatibilit...