Glucurondation Profing Of Yinzhihuang Herbal Components And The Cytotoxic Effects On HeLa Cell

Yin-zhi-huang,a herbal formula containing complex natural Chinese medicine ingredients,most of which show anti-inflammatory,anti-oxidation,anti-bacterial,anti-cancer and other pharmacological effects.But the mechanism of actions is not clear.On the other hand,the main reason for low applicability of flavonoids is the low bioavailability in the body,in particular,the first pass effect of its glucuronide metabolism.Therefore,it is necessary to study the metabolic mechanism of different types of polyphenols in Chinese medicine,or to explore the structure-activity relationship of their structural analogues,which will help to search for safe and effective anti-tumor traditional Chinese medicine.The main contents of this paper include:1)Six flavonoids ingredients in Yin-zhi-huang injection with similar chemical structure,including emodin,physcione,chrysophanol,chlorogenic acid,geniposide and ferulic acid were selected as the substrate to study the metabolic activity of glucuronidases in vitro.The characteristics of glucuronidation reactions on different types of hydroxyl groups were investigated.We found that the main glucuronidation sites of emodin and its related derivatives occurred on C3.2)The main flavonoid/glycoside and its aglycone in Yin-zhi-huang injection,including quercetin-hyperoside-rutin,luteolin-luteoloside,baicalein-baicalin,wogonin-wogonoside were selected as the UGTs substrates.A method for the determination of metabolites by HPLC-UV and LC-MS/MS was established.The spatial stereoselectivity of glucuronidation of flavonoids was discussed.It was found that wogonoside and baicalin which have no hydroxyl on the B ring were not prone to glucuronidation,but they were easily hydrolyzed into the aglycones.However flavonoid glycosides that have hydroxyl on the B ring such as hyperoside,rutin and luteolin can be combined with one or two glucuronides and these substances are also susceptible to partial enzymatic hydrolysis to produce their aglycones.3)The toxic effects of the active ingredients of Yin-zhi-huang and its glucuronidation metabolites on cervical cancer cells were detected by CCK-8.Among the 15 kinds of Yin-zhi-huang monomers,quercetin,luteolin,baicalein,wogonin,emodin and physcione could inhibit the proliferation of HeLa cells in vitro.The inhibitory rate of emodin on cell proliferation was dose-dependent and the maximum inhibition rate was 62.9%.The cell inhibition was almost 100%when the concentration of physcione was more than 10 μM and the effect of metabolism on cells almost disappeared after glucuronidation.The inhibition of cells significant after ferulic acid metabolism.It is predicted that emodin and ferulic acid possessed the most potential as a candidate compound for the treatment of cervical cancer cells.