Effect Of Allyl Benzene Series Compounds From Guizhi Decoction On Enzyme Families About PGE₂ Synthesis

The study of material basis and functional mechanism of Chinese Medical Formulae is the key to the modernization of Chinese materia medica in basic research.It is a good entry to reveals inherent characteristics of Chinese materia medica based on modern science and technology, and to achieve the modernization of the Chinese materia medica theory. Chinese Medical Formulae constitute of active ingredients groups according to certain requirements of compatibility,act through a multi-target,multi-channel integration. It can give traditional Chinese medicine theory contemporary scientific connotation, and improve the academic level of Chinese medicine, to reveal the scientific basis of Chinese Medical Formulae about action characteristics, mechanism and the theory of compatibility of prescriptions. There is important practical significance to explore universal laws of the active ingredients groups of Chinese Medical Formulae,as it has distinctive rules and modes of action different from chemical drugs.In the process of many years of study on the material basis and mechanism of action of GuiZhi Decoction, Our laboratory found that the antipyretic active site,Fraction A (Fr.A) contains 50-100 compounds, which allyl benzene compounds accounted for the Fr.A total weight of 49.2%. In these allyl benzene compounds, six main compounds accounted for 99.4 of the total amount of allyl benzene compounds,they were cinnamic aldehyde(67.11 %), o-methoxy cinnamic aldehyde(4.37%),cinnamic alcohol(27.57%), o-methoxy cinnamic alcohol(0.20%), cinnamic acid(0.39%), o-methoxy cinnamic acid(0.37%).Repeated experiments confirmed that the above-mentioned six kinds allyl benzene compounds combined according to the natural ratio of GuiZhi Decoction retained GuiZhi Decoction's antipyretic role well in "rat fever model induced by subcutaneous injection of dried yeast",and showed good effect in "IL-1?-induced brain microvascular endothelial cells to secrete PGE2 experimental model". Further study showed that the 6 allyl benzene compounds presented Mutual antagonism on some links of inhibiting the synthesis of PGE2 (such as: inhibition of sPLA2 and cPLA2 activity), and mutual synergies on some links (such as: inhibition of pp-38 MAPK protein expression and COX-2 activity), and presented a good synergistic effect in the terminal effect, inhibition of PGE2 synthesis. This phenomenon aroused our strong interest.Phytochemical research shows that they were different oxidation states of the series of compounds ubiquitous in plants. Knowledge of drug metabolism suggests that these compounds often translate along the direction from the reduced state to oxidation state in the organism's metabolic processes. After comprehensive analysis, a working hypothesis of this paper was formed: the series of compounds are universal exist in Chinese Medical Formulae, it is an important basis of Chinese Medical Formulae that the pharmacological activity, ratio, transformation, and mutual synergistic or antagonistic of its members.This study investigate the Pharmacological characteristics of the allyl benzene series compounds from GuiZhi Decoction and relationship between them at the subtype level of the enzymes related to PGE2 synthesis, and explore the rationality of our working hypothesis.From the experimental results, we conclude that:1. The intensity order of of the effects of the allyl benzene series compounds from GuiZhi Decoction is: cinnamic aldehyde> o-methoxy cinnamic aldehyde> cinnamon alcohol> o-methoxy cinnamon alcohol> cinnamic acid> o-anisidine cinnamon acid,combined with the knowledge of drug metabolism, these compounds in the body's metabolic processes, often along the direction from the 'reduced form' to 'oxidation state'. We conclude that: cinnamon alcohol / o-methoxy cinnamon alcohol is oxidized to cinnamic aldehyde / o-methoxy cinnamic aldehyde, and pharmacological activity increased; cinnamic aldehyde / o-methoxy cinnamic aldehyde is oxidized to cinnamic acid / o-anisidine cinnamon acid, reduced pharmacological activity. This shows the metabolism of these series compounds was not always along the direction of lower pharmacological activity. The phenomena is the material basis of why Chinese Medical Formulae have durable efficacy.2. The Allyl benzene series Compounds from GuiZhi Decoction combined according to the natural ratio of GuiZhi Decoction, showed a mutual weakening relationship on inhibition of sPLA2 family subtypes, and mutually reinforcing relationship on COX-2 and PGES family subtypes, in the terminal effect of the inhibition of PGE2 secretion, has a good synergy relationship. It can be seen that the synergy relationship of PGE2 inhibition mainly due to the downstream enzymes of PGE2 synthesis. Chinese Medical Formulae active ingredients were not always show mutual synergism relationship.3. Cinnamic aldehyde has the strongest inhibition effect on sPLA2 (2E) and sPLA2(12A), cinnamon alcohol has the strongest inhibition effect on sPLA2 (5) and sPLA2(12B). They showed a mutual weakening relationship when combined, but they have different strength on subtypes of some target enzyme family, make the inhibition tend to saturation.4. When the ratio of cinnamic acid and o-methoxy cinnamic acid were improved,the Allyl benzene series Compounds showed mutually reinforcing relationship on most of the links. We can speculate that the Allyl benzene series Compounds were metabolized from 'reduced form' to 'oxidation state', the extent of activity lowered was less than active component. This may also be a reason to why Chinese Medical Formulae have durable efficacy.5. The activity of o-methoxy cinnamon alcohol, cinnamic acid and o-anisidine cinnamon acid are very weak or even contrary to the goal efficacy, but they were very low proportion in the 6 compounds, were only 0.20%, 0.39%, 0.37%, hundreds of times lower than cinnamic aldehyde. Their influence is limited, but their proportion of cases is important content of the study of material basis and functional mechanism of Chinese Medical Formulae.There were many series of compounds in a Chinese Medical Formulae,they play their pharmacological effects respectively, and influence each other at the same time,intertwined together to form the overall effect of the prescription. The interaction between different groups of series of compounds is more complex, more in-depth new topic.The article's main innovation is: "Raise the series of compounds as important basis of Chinese Medical Formulae, and investigate its mechanism at PGE2 biosynthetic pathway enzyme family subtype level." It is a new attempt.