Interaction Of TLR4 Promoter Polymorphisms And Environmental Risk Factors With Recurrent Spontaneous Abortions From A Population Of Sounthern China

Part I:Toll like receptor 4(TLR4) expression in decidua from missed abortionObjective To study the expression and distribution of TLR4 in decidua of sporadic miscarriage patients.Methods 32 cases of ceased embryo decidua and 32 healthy decidua taken from surgical induced abortions were chosen for cases and controls. Distinguished by HE biopsies, we furtherly departed ceased embryo cases into 16 inflammatory cases and 16 degenerative cases. They were analysed by immune-histochemistry and Western-blotting for detecting the expression and distribution of TLR4.Results We found TLR4 of decidua tissues was significantly higher in the inflammatory ceased embryoes and significantly lower in the degenerative decidua.Conclusions The maintaining sufficient level of TLR4 may be important during early pregnancy. Either higher or lower TLR4 level of utrine decidua might be reason that underlie spontaneous or missed abortions.Part II:Environmental risk factors for unexplained recurrent spontaneous abortion in Southern Chinese womenObjective:To find risk factors for recurrent spontaneous abortion (RSA) in southern Chinese women.Methods:The association between BMI, family abortion history, smoking, ETS(environmental tobacco smoking), alcohol consumption, coffee intake, and RSA were analyzed with a logistic regression model involving 326 patients with recurrent miscarriages and 400 control cases.Results:There was no positive association between smoking, alcohol consumption, coffee intake, and RSA. Short ETS (<1 hour/day) was associated with an increased risk of RSA (adjusted OR,2.30,95% CI=1.50-3.52) and long ETS (≥1 hour/day) had an increased risk (adjusted OR,4.75,95% CI=3.23-6.99).The increased risk of RSA was significant in those with a positive family history (adjusted OR,2.12,95% CI=1.28-3.49) and those with a BMI≥24.0 kg/m2 (adjusted OR, 1.55,95% CI=1.12,2.14).Conclusion:ETS, increased BMI (≥24.0 kg/m2), and a family history are independent risk factors for RSA in southern Chinese women.PartⅢ:Study on two TLR4 promoter SNPs of a population from Southern ChinaObjective:To find if TLR4 promotor SNPs of rs10983755 and ss77136219 answer for Hardy-Weinberg equilibrium and linkage disequilibrium.Methods:240 successful samples without sibship were distilled DNA for analyse Hardy-Weinberg equilibrium and linkage disequilibrium of TLR4 promoter genotypes of rs10983755 and ss77136219.Results:SNPs of 240 controls in our study answered for Hardy-Weinberg equilibrium, and they showed incomplete linkage disequilibrium.Conclusion:Population from our study can represent a real population from Southeren Chinese women, and two genotype points show independent impact on recurrent spontaneous abortions.PartⅣ:The assosiation between TLR4 promoter SNPs and susceptibility of RSA from Southern Chinese womenObjective:To find if TLR4 promotor A/G SNPs of rs10983755 and ss77136219 were related to undendified recurrent spontaneous abortions of Southern Chinese women.Methods:240 healthy controls and 183 cases of RSA without sibship in our study were distilled DNA for TLR4 promoter genotypes of rs10983755 and ss77136219. Genotypes frequencies were compared between cases and controls.Results:SNPs of TLR4-rs 10983755 were not different between cases and controls, but TLR4-ss77136219-GG genotypes were siginificantly higher in cases group.Conclusion:Maybe TLR4-ss77136219 A→G SNP relate to URSA in Southern Chinese Women.Part V:Study of interactions between TLR4 promoter SNPs and environmental factors and RSA from Southern Chinese womenObjective:To find if TLR4-rs10983755 and ss77136219 SNPs and environmental factors interacted on RSA susceptibility.Methods:240 healthy controls and 183 cases of RSA without sibship were distilled DNA for TLR4 promoter genotypes of rs10983755 and ss77136219. Interactions of genotypes and environmental factors were analysed between cases and controls.Results:GG and GA genotypes of TLR4-ss77136219 and older than 30 years old interacted for URSA in our population.Conclusion:URSA is a complex disease maybe due to interactions of genes and environmental risk factors.Part VI:Study of TLR4 Promoter SNPs on genetic transcription and interpretationObjective:To find if TLR4-ss77136219 A→G SNPs effected a change on TLR4-mRNA and protein expressions.Methods:24 miscarriage embryo decidua cases taken from RSA patients were divided into three groups (AA/GA/GG) by TLR4-ss77136219 genotypes. They were analysed by real-time PCR for detecting TLR4-mRNA and Western-blotting for detecting the protein expressions of TLR4. Results:TLR4-mRNA and protein level of RSA patients with TLR4-ss77136219-GG and GA genotypes were siginificantly higher than AA genotypes.Conclusion:A→G SNPs of TLR4-ss77136219 increased the expressions of TLR4-mRNA and protein.