| Ulcerative colitis is an universal disease in the world .The attack rate of Ulcerative colitis is 5-12/100,000.The prevalence rate is 50-150/100,000.The number of female patients is larger than male patients.The age of onset distributes in double hump.The first hump is in 15 to 30 years old,and the second hump is in 50 to 70 years old. The morbidity of the first hump is more than the second. The patients of 15% to 40% have the family clan of chronic ulcerative clotis or Crohn's disease. The clinical case of Ulcerative colitis is general and in up-tendency. Therefore, there are pay close attention to Ulcerative colitis'invasion and therapeutics at present. Ulcerative colitis is a frequently encountered disease in clinic.The cardinal symptom is abdominal pain,bowel complaint,myxo-pus and bloody stools. The medical treatment of modern medicine for Ulcerative colitis mainly is symptomatic treatment. Meanwhile,the serious Ulcerative colitis is treated by surgical intervention.But there are much malpractice in managements of modern medicine just like expensive charge﹑major injury and sequelae .Inversely,the treatment of Traditional Chinese Medicine for this disease has the long history and the effect is definite.Ulcerative colitis belongs to"decanta"in the Traditional Chinese Medicine.It is recognized that the principal etiopathogenisis is being invaded by exogenous pathogen,unhygienic diet,weakness of the spleen and stomach or decline of the fire from the gate of life.The disease has intimate relationship with pathogenic damp,which is trended to hinder the spleen and stomach to cause the abnormal function of hoisting the spleen and stomach. In the case of internal damage, the weakness of the spleen and stomach take a key point. Because spleen controlling digestion, if deficiency-weakness of spleen-QI,it may cause clear QI failing to ascend and metaplasia endogenous wetness-evil , clear QI in the lower part of the body which at last cause diarrhea. Because of chronicity ulcerative colitis recurrent attacks,we may see the following symptoms : mucous or jel appearance of stool,unclearly abdominal pain,chilly and philo- calefaction,feel cold of waist and knees,deep and thin of the pulses.These are dialectical the Yang asthenia syndrome.The fundamental pathogenesis is asdthenic splenonephro-yang.Therapeutically,ulcerative colitis of Yang asthenia type apply warm and tonics medicine to make up for the deficiency of spleen and kidney. Classical prescription or recipe just like YOUGUI pills, Strobal Bolus for Regulating Middle Energizer et al, which were used to cure Ulcerative colitis of Yang asthenia type obtain better effects.Now doctors utilize Cassia bark cure ulcerative colitis of Yang asthenia type also gain the better curative effect.Cassia bark, which belongs to the spleen meridian﹑the kidney meridian﹑the heart meridian and the liver meridian,is acrid-sweet flavor﹑hot-natured.It has the effects of tonicing the fire of the Gate of Life﹑eliminating cold to stop pain﹑warming the meridian to promot the blood circulation. It is the key medicine to cure the decline of the fire from the Gate of Life. Cassia bark has complicated ingredient,which include amino acids,polypeptide,carbohydrate,organic acid,Phenols,1,2-benzopyrone lactone,anthoxanthin,anthraquinone,saponin,cardiotonic glycoside,aetherolea.The researches indicated that aqueous extract of Cassia bark and ether extract of Cassia bark all had the effects to refrain ulceration, which include water immersion stress ulcer of mouse﹑cold stimulate stress ulcer﹑hydrochloric acid or 5-HTA stimulate stress ulcer of rat .The cause of Ulcerative colitis is very complicated.The existing investigation report that matrix metalloproteinase(MMP) take a significance role in the origin of this disease. 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 five kinds: collagenase﹑ gelatinase﹑ 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 Ulcerative colitis.It containes MMP-1﹑MMP-2﹑MMP-3and mmp-9 et al.There was appear the same result at the mRNA and protein level.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 inflammation of membrana mucosa.Thus, the inhibition of MMPs may represent a novel therapeutic approach for the treatment of ulcerative colitis.On account of above-mentioned, the aim of our experiment is to look for the MMPs'inhibitor from the Traditional Chinese Medicine Cassia bark.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 Cassia bark, boil﹑filter and freeze drying it,then prepare to be solution.The density of the different solutions are 0.4mg/ml﹑ 1.2mg/ml﹑ 2.0mg/ml﹑ 2.8mg/ml﹑3.6mg/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 Cassia bark.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 Cassia bark 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 Cassia bark have the inhibitory activity to MMP-16,and accompany with the increasing of the density, inhibitory action also reinforce.It is proved that the Cassia bark has the activity of inhibiting MMPs .From considerable clinical observations about Cassia bark treat Ulcerative colitis and experiments about MMPs,We can presume that it may be the Molecule mechanism of therapeutic action of Cassia bark to cure UC.It will provide a new theoretical and experimental basis for studying Ulcerative colitis .
|
PDF Full Text Request