The Correlation Study In The Different TCM Syndromes Of Prolapsed Internal Hemorrhoids And The Expression Of Fibulin Proteins Family

ObjectiveBy observing and comparing expression characterizations of Fibulin family proteins and matrix metalloproteinase-9(MMP9) in the three TCM syndromes of prolapsed internal hemorrhoids, we can explore the relationship between proteins and prolapsed internal hemorrhoids, and explain clinical characteristics of different TCM syndromes of prolapsed internal hemorrhoids, to provide the basis for clinical Differentiation and treatment of TCM.MethodsAfter the examination and approval from the First Affiliated Hospital of Guangzhou University of Chinese Medicine ethics committee and all patients signed informed consent.54 samples of anal cushions tissue taken from prolapsed internal hemorrhoids (including 21 samples with syndrome of dampness-heat diffusing downward,18 samples with syndrome of qi stagnation and blood stasis, and 15 samples with syndrome of deficiency of both qi and blood) as well as 10 samples of normal anal cushions tissue taken from colorectal polyps patients without prolapsed internal hemorrhoid were collected in experiments. The expression and distribution of Fibulin-3, Fibulin-5 and MMP9 were observed from tissue-gene-protein hierarchy by using the HE staining, PCR method, the Western-blot analysis, and the gelatin zymography method. Then we could draw conclusions from comparing the expression and distribution of different TCM syndromes of prolapsed internal hemorrhoids.Result1. the study showed that the gene expression level of Fibulin-3 decreased gradually in the three TCM syndromes of dampness-heat diffusing downward, qi stagnation and blood stasis, and deficiency of both qi and blood, while Fibulin-3 protein expression increased gradually and reached its highest expression level in the syndrome of deficiency of both qi and blood.2. There were no statistically significant differences in the gene expression level of MMP9 in the three types of syndromes. In the syndrome of qi stagnation and blood stasis, MMP9 had relatively higher gene expression level but relatively lower protein expression.3. Same as Fibulin-3, the gene expression level of Fibulin-5 decreased gradually in the three TCM syndromes of dampness-heat diffusing downward, qi stagnation and blood stasis, and deficiency of both qi and blood, while Fibulin-3 protein expression increased gradually and reached its highest expression level in the syndrome of deficiency of both qi and blood.4. In the syndrome of qi stagnation and blood stasis where the protein expression of Fibulin-3 was the lowest compared with the other two syndromes, the MMP-9 protein expression was also the lowest, while the activity trends of MMP-9 were consistent.5. By contrast, in the syndrome of qi stagnation and blood stasis where the protein expression of Fibulin-5 was the highest compared with the other two syndromes, the MMP-9 protein expression was also the highest, while the activity trends of MMP-9 were consistent. Conelusion1. Based on ethics, using from endoscopic polypectomy (without internal hemorrhoids) process to obtain normal anal cushion tissue as control group, from after PPH specimens were obtained for anal cushion tissue as the experimental group, both of which are comparative.2. The expression levels of Fibulin-3, Fibulin-5 and MMP9 in internal hemorrhoids tissues are higher than in anal cushion with all three TCM syndromes of dampness-heat diffusing downward, qi stagnation and blood stasis, and deficiency of both qi and blood. The proteins expression levels are relatively low with syndrome of qi stagnation and blood stasis, and positively relate to the severity of prolapse with syndromes of dampness-heat diffusing downward, and deficiency of both qi and blood.3. In prolapsed internal hemorrhoids, the increase in the protein expression level of FBNL-3 might cause the increse in MMP9 activity, accelerate the disintegration of elastic fibers which results in the pathologic shift of anal cushion.4. Increasing protein expression level of Fibulin-5 could decrease the support elasticity of anal cushion which can give rise to prolapsed internal hemorrhoids.5. The effects of Fibulin-3 and Fibulin-5 on internal hemorrhoids with syndrome of qi stagnation and blood stasis might be different with syndromes of dampness-heat diffusing downward, and deficiency of both qi and blood.