The Experimental Study On The Anti-tumor Mechanism Of Clove Inhibiting The Activity Of Matrix Metalloproteinase 14 And 13

Tumour is seriously to threaten in health and vital safety of human, lots of lifves of tumour people have been deprived. So primary task of health organization and research master is very important. In-depth study the pathology mechanism , and seek to prevent from diffused. This text is external experiment to look for the new drug of guarding against tumour and cure tumour. The course of tumour invasion and metastasis is complicated, it is not a invariable equation of singlenesss, therefore, we don't only research the metastasis of microcosmic course, including every detail, but also exploit new medicine that can inhibit the tumor metastasis. The procedures contain tumour cells going across the protective screen of ECM, going across blood vessel wall, thereby tumour cells enter into the microenvironment of parasitifer. By a mass of medicine experiment, the ability of tumour cells of invadeing and attacking is related to ptotease. For the past few years, person pay more attention to the tumor metastasis. Cancer cells invade and attack process is not a changeless formula, is very complesity. As a result, an extremely urgency is MMP is making certain tumour invade and attack, containing every detail, at the same time, seek effective drug. Go by medicine test attesting, the ability of cancer cells invade and attack is closely related to protease who lead and produce .This protease can decomprose ECM and BM. It is name is Matrix Metalloproteinase(MMPs). In the process of tumour cells invade and attack, professors found that MMPs is very important, and the research of tissue inhibitors of metallo proteinases is at a premium.When the environment of human body is pathology condition, in especial tumour condition, the MMPs regulateing and controling ECM exceed the routine scope. The MMPs excessively express themselves, excessively break down ECM, seriously destroy the structure and constituent. The other one key point is the balance of MMPs and TIMPs, the balance can keep human body in normal. The imbalance of MMPs and TIMPs is important reason that ECM is broken down, is not only the mark of early stage soaking, but also is very important in tumour cells of invadeing and attacking. At present, it does not have enough evidence to proof themselves effective means , that take precautions against and cure tumour. But traditional Chinese medicine still gets along with initial phase. We need forceful proof to testify themselves. Now traditional Chinese medicine has itself distinctive preponderance.Traditional Chinese medicine consider that pathogeny of tumorigenesis : Firstly, the six evils, visualization statement is livelihood environment, contain inate laws of things and chemistry factors that could be carcinogenic, when human body function is feebleness. Qi activity going up and down is chaos and confusion, the move of Qi and blood is hindered, result in occurring tumour at last. Secondly, anger, happiness, think, sad and grief lead to the stagnation of Qi activity, blood and phlegmatic malaria, Qi activity going up and down is chaos and confusion, the move of Qi and blood is hindered. Thirdly, Zang and Fu divide into the day after tomorrow QI and innate QI, innate QI summarize to power of immunity of human body oneself, it is insufficient, is hypoimmunity. So QI and Blood is insufficient, superadd external fators influence human body, the positive activity is confined. Injure human body health.Traditional Chinese medicine have been a most important way of anticancer, and a single drug of traditiongal Chinese medicine have been carried out. Sapor of clove is pungent and lukewarm , go back to spleen, stomach, lungs, kidney. The efficacy of warming middle energizer descend adverse-rising and reinforce the kidney yang is master. In clinical practice, the effect of clove is very broad . It can treat hiccup, vomit, regurgitation, Xie Li, chest and abdomen paint, and so on. The modern research achievements make known that, clove can prevent and cure digestive tract squamous cancer and breast cancer. This article research the mechanism of curing the tumour.Objectives:In the experiment, record and analyse the index of water extract of clove, whether exist restraining from MMPs or not. If exist restraining from MMPs, the bull and bear of restraining effect, it will offer the research proof according to the theory proof and basic experiment of traditional Chinese medicine water extract of clove treating tumourMethods:Add 10g of Prepared drug in pieces of clove to distilled water 50ml, and then, put them in water bath with the temperature being 100℃. Put them out the water bath afer 120 minutes, proceed colling at normal temperature condition. Filter the Chinese medicine, put some filtrate in freeze-drying bottle, finish freeze-drying. Obtain powdery substance, use dissolvent with distilled water, deploy needful consistence. Supply the medicine for the experiment one and experiment two.Experiment 1: Mix 1μl MMP-14 with enzyme buffer solution (deploy standard: every 1μl enzyme use 50μl buffer solution, set final volume 50μl without negligible factor), deploy seval group according to requisite fraction, respectively drip in WLISA plate have selected locations, then, put them in constant temperature foster box , set 37℃fou 30 minutes, let enzyme activity optimal. Continue the reagent deploying, Mix 0.2μl DQ-gelatin with buffer solution (deploy standard: every 0.2μl enzyme use 50μl buffer solution, set final volume 50μl without negligible factor).Deploy the Prepared drug in pieces of clove with 10μg/ml,1.25μg/ml,0.5μg/ml,0.165μg/ml,0.05μg/ml for using. Drip respectively the Prepared drug in pieces of clove in WLISA plate (without matched group), at the same time, put 50μl zymolyte in every plate. Put the WLISA plate in Microplate system , the medicin final volum is 100μl without negligible factor. When the Bio-Red Flx-800 Microplate system detects, can arouse wavelength of 460nm, emission wavelength 520nm. Detect 15 minutes, the rake ratio of fluoredcence intensity variation curve is the vitality of degradation zymolyte in bull and bear. Experiment 2: Mix 1μl MMP-13 with enzyme buffer solution (deploy standard: every 1μl enzyme use 50μl buffer solution, set final volume 50μl without negligible factor), deploy seval group according to requisite fraction, respectively drip in WLISA plate have selected locations, then, put them in constant temperature foster box , set 37℃fou 30 minutes, let enzyme activity optimal. Continue the reagent deploying, Mix 0.2μl DQ-gelatin with buffer solution (deploy standard: every 0.2μl enzyme use 50μl buffer solution, set final volume 50μl without negligible factor).Deploy the Prepared drug in pieces of clove with 5μg/ml,2.5μg/ml,1.25μg/ml,1μg/ml,0.5μg/ml for using. Drip respectively the Prepared drug in pieces of clove in WLISA plate (without matched group), at the same time, put 50μl zymolyte in every plate. Put the WLISA plate in Microplate system , the medicin final volum is 100μl without negligible factor. When the Bio-Red Flx-800 Microplate system detects, can arouse wavelength of 460nm, emission wavelength 520nm. Detect 15 minutes, the rake ratio of fluoredcence intensity variation curve is the vitality of degradation zymolyte in bull and bear.Principles:When enzyme resolve zymolyte, it can issue different intensity fluorescence, use FLX-800 Microplate system to deploy the rake ratio of fluoredcence intensity variation curve. At the experiment, the zymolyte have deal with fluoredcence. The rake ratio of fluoredcence intensity variation curve is the vitality of degradation zymolyte in bull and bear. Results:Prepared drug in pieces of clove have the better effect of inhibiting MMP-14 , and have the effect of inhibiting MMP-13. That is to say , in the experimental result, help people to seek new drug to prevent and cure tumour. The rake ratio of fluoredcence intensity variation curve is the vitality of degradation zymolyte in bull and bear.Conclusion:Prepared drug in pieces of clove can restrain the liveness of the MMP-14 and MMP- 13. The experimental result help clinical experiment to develop, help human to search new drug from natural pharmaceuticals, and the future of MMPs/TIMPs is immeasurable.