| Primary liver cancer is malignant tumor which origin from hepatocyte or epithelial cells of bile duct .It is highly malignant,rarely diagnosed during the early stage,prognosis is bad .It is reported that PLC's mortality rate is 44/100,000 by the Ministry of Public Hea1th in1995, taking the second place among all of the malignant tumors .As in early stage,operation is still considered the best treatment .However,when was found,the majority of the patients of PLC are already in mid stage or even advanced disease,or complicated by Cirrhosis of liver,so miss their opportunities of resection.interventiona1 therapy and other local therapy,such as TACE,PEI,RFA,always can not be done due to the great mass or poor liver function .TCM treatment with Syndrome Differentiation and Disease Differentiation,has both effections of strengthening body resistance and anticancer,is becoming one important alternative to treat PLC,it is widespread in China,Inversely,the treatment of Traditional Chinese Medicine for this disease has the long history and the effect is definite. Primary liver cancer belongs to"meteorism","accumulation","icterus","abdominal mass", and"flank pain"in the Traditional Chinese Medicine。According to TCM the pathogenesis of PLC related to reception of moist and toxic heat pathogen,long-term eating and drinking without temperance,being addicted to drink excess and internal injury caused by excess of seven emotions and so on which caused the organism yin–yang disequilibrium. Healthy energy deficiency and decline, evil factor affecting health to take the chance of assault the liver, depression of liver-QI, hinder the activities of qi ,blood runs with no smooth, sputum and stagnant blood tie with eath other, to form the abdominal mass dand at last the liver cancer happened .The principal etiopathogenisis is stagnation of vital energy and blood stasis,humid heat tie together,hepatosplenic disorder, healthy energy deficiency and decline etc.The main reason of primary liver cancer of endoretention of damp heat type is transverse invasion of hyperactiver liver-QI to overcome oil, weakness of the spleen and stomach , dyssplenism,liquid and frain cannot transportation and transformation as normal,all these can make liquid and moist to stop and gather,then to form phlegm or stagnant blood intersect with phlegm.Long time they will heat-syndrome resulting from exogenous and endogenous evils, humid heat fumigate ,to stay stagnant the liver meridian, hepatochlic hygropyrexia ,so in the advanced stage of PLC we may see the following symptoms: jaundice,flank expand,bitter taste of mouth,foul breath,oliguria with reddish urine,red tongue,musci yellow and greasy,present humid heat image.Scutellariae barbatae as a commonly used Chinese crude drug ,it educe an important effect in therapy primary liver cancer of endoreten- tion of damp heat type .Now doctors utilize Scutellariae barbatae cure primary liver cancer also gain the better curative effect Scutellariae Barbatae D.Don,named as Indian skullcap herb with root,violetlotus,bingtou herb,toothbrush herb and banxuehua as well,is used extensively as a medicinal material spread almost all over China inclinics and therefore it is worth researching since its chemical compositions include flavonoids, terpene,polysaccharide,alkaloid and sterides. As a traditional Chinese medicine,Scutellaria barbata D .Don plays an important role in detumescence, allay fever, detoxification , antitumor and so on. Epecially,antitumor function is emphasized .In clinic, usually uses Scutellaria barbata D. Don ,mixed some other traditional Chinese medicine, to treat liver cancer, stomach cancer ,rectum cancer ,lung cancer ,woman tumour and so on.The invasion and metastasis of tumor cell is a complex process .But the matrix metalloproteinases(MMPs) produced by tumor cell play a critical role in metastasizing to remote organs and invading surrounding tissues,for MMPs can degrade extracellular matrix(ECM) and basement membrance which the tumor cells must traverse . MMPs are a family of more than 20 enzymes that are intimately involved in tissue remodelling.These zinc-containing endopeptidases consist of several subsets of enzymes.In normal steady state tissues ,the expression of MMPs is microcrystalline.But MMPs are thought to be essential not only to the embryogenesis and the generation of cell,but also to the invasive processes of ulceration﹑tissue repair﹑angiogenesis﹑inflammatory reaction and tumor metastasis. At present , the discovered MMPs have more than 20 kinds.According to their different hydrolytic substrate,it may divided into six kinds: collagenase﹑gelatinase﹑elastase,stroma-lysin,basilaris substantia hydratase﹑MT- MMPs and others .Numerous studies have shown that there was a close association between expressions of various members of the MMPs and inflammation of membrana mucosa of Primary liver cancer.It containes MMP-1﹑MMP-2﹑MMP-3,MMP-7,MMP-9and mmp-10et al. Meanwhile,the research indicate that when the expression of MMPs improved, the expression of TIMPs also increased. So it is consider that disequilibrium of MMPs/TIMPs was the major factor of invasion andmetastasis of primary liver cancer.Thus, the inhibition of MMPs may represent a novel therapeutic approach for the treatment of Primary liver cancer.。The aim of our experiment is to look for the MMPs'inhibitor from the Traditional Chinese Medicine Scutellariae Barbatae and to provid necessary foundment of theory and experiment for the pharmaco- mechanism of the Scutellariae Barbatae treat primary liver cancer.The experiment is on the basis of enzyme that can degradate the substrate (DQ-gelatin) which is marked with fluorophore and quench group. Fluorophore and quench group may dissociate from the substrate then fluoresce when enzyme cut the substrate.Then we may definite the activity of enzyme from detecting the change of fluorescence intensity. Take 3g Scutellariae Barbatae, boil﹑filter and freeze drying it,then prepare to be solution.The density of the different solutions are 0.5mg/ml﹑ 1.0mg/ml﹑ 1.5mg/ml﹑ 2.0mg/ml﹑3.0mg/ml.Then put the quantitative mmp-16 into the solutions of different density and put them into 96-hole borad.After that put the buffer solution into the mixed liquor(50mM HEPES,0.2M NaCl,10mM CaCl2,20μM ZnSO4, 0.05 % Brij-35).The blank group dose not add the aqueous extract of Scutellariae Barbatae.Put it into incubator for 30 minutes.After that,add 0.2μg DQ-gelatin into these solutions.The final reaction system of each group is 100μl. Detect for 10 minutes.The change of fluorescence intensity represent the capability of enzyme degradate the substrate. Inhibit activity calculate:Vi represent the change of fluorescence intensity of catastaltic group. V0 represent the change of fluorescence intensity of non-catastaltic group.%I =1-Vi / V0.when %I=50%,it is the IC50 of inhibitory enzymatic activity.We receive the result that the variations of fluorescence intensity of Scutellariae Barbatae extract groups in the different concentrations are all more diminished than the control groups'. And with the density increaseing, the intensity of fluorescence can be week.On the besis of formula calculation,IC50=35.5μg/ml.On the whole, aqueous extract of Scutellariae Barbatae have the inhibitory activity to MMP-16,and accompany with the increasing of the density, inhibitory action also reinforce.It is proved that the Scutellariae Barbatae has the activity of inhibiting MMPs .From considerable clinical observations about Scutellariae Barbatae treat Primary liver cancer and experiments about MMPs,We can presume that it may be the Molecule mechanism of therapeutic action of Scutellariae Barbatae to cure PLC.It will provide a new theoretical and experimental basis for studying Primary liver cancer. . |
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