A Study On The Association Of Tumor Necrosis Factors And Their Gene Polymorphism With Endometriosis Of Women In Guangdong

BackgroundEndometriosis is a frequently and commonly encountered disease in women of child-bearing age, which may affect generation and easily relapse, and is very difficult to treat clinically. Many researchers had conducted various studies in order to explore causes, development and related factors of endometriosis, finally without definite conclusions. The existing studies indicated that endometriosis may be a genetic autoimmune disease caused by multiple sites genes interaction, which is involved in cytokine adjustment to immune response. The cytokines such as tumor necrosis factor-alpha (TNF-α) and tumor necrosis factor-beta( TNF-β)may induce a great variety of cytokines including interleukin-1(IL-1), interleukin-6(IL-6) and themselves, etc. as some inflammatory mediators that interact and compose network to regulate cooperatively, resulting in enhancing generation and differentiation of palingenetic vessel. At the same time, excessive Tumor necrosis factor(TNF)express may influence immune state of the body. At present, the studies of immune response-related cytokines genes have been taken attention frequently. TNF gene is one of the genes involved in autoimmune disease, which has been arisen interest of the researchers now. Current studies showed that TNF gene polymorphism might be highly involved in many autoimmune diseases, especially TNF level in HLA-linked diseases. TNF gene consists of multiple sites of gene polymorphism, and domestic and international studies indicated that -308 site in promotor region of TNF-aand +252 site in No. 1 intron of TNF-βcontaimng G/A mononucleotide replacement polymorphism was related to expressing amount of TNF-α and TNF-β in serum and abdominal cavity liquid, but correlation between other sites of polymorphism and TNF level was not significant. Therefore, the indices including mononucleotidepolymorphism in -308 site in promoter region of TNF-a and +252 site in No. 1 intron of TNF-p were researched in this study. With further studies on correlation between cytokines and endometriosis, TNF have been playing an important role in causing of endometriosis. But, relation between TNF gene polymorphism and endometriosis remains unclear. The object of the study is to preliminarily explore relation between TNF gene polymorphism and causes of endometriosis in women of Guangdong province.Objective1. To research change condition of expressing amount of TNF-a and TNF-P in serum before operation, in serum and abdominal cavity liquid after operation in the patients with endometriosis and without endometriosis, and to explore effects of TNF-a and TNF-p on causes of endometriosis and possibility of serving as indices of relapsing and evaluation of curative effect after operation of endometriosis.2. To study polymorphism in -308 site in promoter region of TNF-a gene and +252 site in No. 1 intron of TNF-p gene in patients with endometriosis and without endometriosis, and to explore relation between polymorphism of TNF-a gene and genetic susceptibility of endometriosis, and try to explore pathogenic mechanism of endometriosis in gene level, and find out essential foothold of treatment of endometriosis.Methods1. By enzyme-linked immunosorbent assay (ELISA), serum before operation in 82 patients with endometriosis, serum after operation in 73 patients with endometriosis, and express of TNF-a and TNF-P in abdominal cavity liquid in 42 patients with endometriosis were measured, and serum before and after operation in 68 patients without endometriosis and abdominal cavity liquid in 40 patients without endometriosis were assigned as control group.2. By polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and Allele-Specific Amplification (ASA) methods, polymorphism in -308 site in promoter region of TNF-a gene and +252 site in No. 1 intron of TNF-p gene in 82 patients with endometriosis (endometriosis group)and 80 patients without endometriosis(control group), was measured, and relation between TNF gene polymorphism and endometriosis risk was analyzed. Bothgroups of women belong of Han nationality of Guangdong province.Results1. The results show that, level of TNF-a and TNF-P in serum in endometriosis group is significantly increased compared with control group ( P<0.01). expressing amount of two TNF appearing increased following worsening of patients' condition. The level of TNF-a and TNF-P in abdominal cavity liquid is significantly higher than that in control group ( P<0.01) .The level of TNF-a and TNF-P in abdominal cavity liquid is significantly higher than that in serum ( P <0.01). The level of TNF-a and TNF-P in serum after operation is decreased following clearing of endometriosis focus, especially that in III and IV phase is decreased more significantly than that in I and II phase(PO.01) . The difference of level of TNF-a and TNF-P in abdominal cavity liquid between proliferative phase and secretion phase, is no statistical significance.2. The results show TNF-P+252 site allele and genotype distribution between endometriosis group and control group indicate significant difference(x2=6.562, P <0.05; x2=6.562, P <0.05), and relative risk of suffering from endometriosis caused by allele A is increased by 1.793 times. The risk of suffering from endometriosis in person carrying AA genotype is higher than that in person carrying GG genotype by 3.33 times(x2=6.562, P<0.05). TNF-a-308 site allele and the distribution of genotype frequency show no significant difference between two groups (x2=0.5429, x2=0.640 P i^>0.05) . But TNF-a-308 site and variant genotype of TNF-p+252 site has synergetic effect, and OR value of TNF-a gene-308 site GA and TNF-p gene+252 site GG combined genotype is 2.445, and OR value of TNF-a gene-308 site GA and TNF-P gene+252 site AA combined genotype is increased to 10.99(P<0.05). The TNF-P level in serum in endometriosis group with TNF-P gene+252 site AA genotype is significantly increased compared with GG genotype (t=2.029 P<0.05), while TNF-a and TNF-p level in serum shows no statistical significance in patients with other genotypes in TNF-p gene+252 site in endometriosis group and control group. Similarly, TNF-a and TNF-P level in serum shows no statistical significance in patients with all sorts of genotypes in TNF-a gene-308 site in endometriosis group and control group.Conclusions1. TNF play an important role in causing of endometriosis, level of which in serum and abdominal cavity liquid is increased following severity of disease condition, as well as have important effects of evaluation of follow-up after operation and operation effects detection to some extent.2. AUele A in TNF-p gene+252 site can significantly increase relative risk of suffering from endometriosis in Guangdong women. TNF-p AA genotype might be one of genetic susceptible factors of suffering from endometriosis in Guangdong women, and single existing polymorphism of TNF-a gene+308 site might be not significantly involved in endometriosis, but has synergetic effect to TNF-p gene+252 site, so coexisting of two sites of point mutation might be one of causes that can enhance occurence risk of endometriosis. TNF-P gene+252 site AA genotype might be enhance TNF-p level in serum in patients with endometriosis.