Through the method of clinical case control study, to explore the expression and genetic polymorphism of KLF14 gene (rs4731702 and rs972283) and SR-B1 gene (rs5888 and rs838880) in patients with cholesterol gallstones, to analyze the correlation between KLF14 gene and SR-B1 gene, and to study whether the gene locus of KLF14 gene and SR-B1 gene are different and correlated with blood lipid indexes, gender, nationality and environmental factors in patients with cholesterol gallstones.

Objective through the method of clinical case control study, to explore the expression and genetic polymorphism of KLF14 gene (rs4731702 and rs972283) and SR-B1 gene (rs5888 and rs838880) in patients with cholesterol gallstones, to analyze the correlation between KLF14 gen e and SR-B1 gene, and to study whether the gene locus of KLF14 gene and SR-B1 gene are different and correlated with blood lipid index es, gender, nationality and environmental factors in patients with chole sterol gallstones.

Methods A total of 200 patients undergoing elective laparoscopic cholecystectomy in our department were randomly selected, and the cholesterol gallstones group (100 cases) was taken as the case group according to the inclusion criteria. A total of 100 cases in the non-gallstone group were used as the control group. The DNA from peripheral venous blood of all subjects was extracted, and SNP typing was performed on the KLF14 gene (rs4731702, rs972283) and SR-B1 gene (rs5888, rs838880) in the case group and the control group by direct sequencing. To compare the genotypic and allelic frequencies of the KLF14 gene (rs4731702, rs972283) and SR-B1 gene (rs5888, rs838880) between the two groups, to explore whether there is an association between them, and to analyze the association between the locus of the two genes and blood lipid indicators as well as gender, ethnicity, and environmental factors in patients with gallbladder cholesterol gallstones.

Results 1. Serum levels of triglyceride (TG), total cholesterol (TCH), low density lipoprotein (LDL), and apolipoprotein B(Apo-B) in the case group were significantly higher than those in the control group; The levels of high density lipoprotein (HDL) and apolipoprotein AI(Apo-AI) were significantly lower than those in the control group (P < 0.05). 2. The distribution differences of allele frequency and genotype frequency at rs4731702 of 2.KLF14 gene in case group and control group were statistically significant (P < 0.05); The difference in frequency distribution of alleles C and T was statistically significant (P < 0.05), and the frequency of allele C was higher than that of allele T, suggesting that allele C might increase the risk of cholesterol gallstones (OR = 1.547). In the case group, TG levels in CC genotype at rs4731702 of KLF14 gene were higher than those in CT and TT genotypes (P < 0.05). HDL, Apo-AI and Apo-AI/Apo-B levels in the CC genotype were lower than those in the CT and TT genotypes (P < 0.05). There were no significant differences in TCH, LDL or Apo-B (P > 0.05). 3. The distribution differences of allele frequency and genotype frequency at rs5888 locus of SR-B1 gene in case group and control group were statistically significant (P < 0.05); There were significant differences in frequency distributions of alleles C and T between the two groups (P < 0.05). The CC genotype at rs5888 of SR-B1 gene in the case group was significantly higher than that of CT+TT genotype (P < 0.0 5), suggesting that the mutant CC genotype might be a risk factor for cholesterol gallstones (OR = 2.279). The frequency of the C allele was higher than that of the T allele, indicating that the C allele might increase the risk of cholesterol gallstones (OR = 1.898). The gene frequency of the TT type gene in the case group was significantly lower than that of the control group, and HDL-C of the TT type gene was significantly higher than those of CC and CT type, while TG and TC H contents were significantly lower than those of CC and CT type (P < 0.05). LDL, Apo-AI, Apo-B and Apo-AI/ Apo-B had no statistically significant difference (P > 0.05). 4. The distribution differences of allele frequency and genotype frequency of rs838880 of SR-B1 gene in case group and control group were statistically significant (P < 0.05). The number of CC genotype at rs838880 of SR-B1 gene in case group was significantly lower than that in control group, which indicated that genotype CC at rs838880 of SR-B1 gene was the protection genotype. The TCH of patients with TT genotype in the case group was significantly higher than those of patients with TC and CC genotype, and the difference was statistically significant (P < 0.05). HDL of patients with CC type gene was significantly higher than those of patients with TC and TT type in this group (P < 0.05). This indicates that the C allele may be associated with an increase in HDL-C in blood. There were no significant differences in TG, LDL, Apo-AI and Apo-B (P > 0.05), and no significant differences in the distribution of allele and genotype frequencies of rs972283 in 5.KLF14 gene between the case group and the control group (P > 0.05). The TG and TCH levels in the GG genotype at rs972283 of the KLF14 gene in the case group were higher than those in the GA and AA genotypes (P < 0.05). HD L, Apo-AI and Apo-AI/ Apo-B levels in the GG genotype were lower than those in the GA and AA genotypes, and the differences were statistically significant (P < 0.05). There was no significant difference between LDL and Apo-B (P > 0.05). 6. Comparison of the genotypes CC, CT and TT of the KLF14 gene rs4731702 in the case group revealed that there was no significant difference among ethnic groups (P>0. 05), while the gender difference was statistically significant (P<0.05). 7.BMI, family history of cholesterol gallstones and exercise habit were the influencing factors of cholesterol gallstones. 8. Patients with BMI > 26 and genotype (KLF14 rs4731702) CC were more likely to develop cholesterol gallstones (OR=16.379) than BMI < 26. Patients with a family history of gallstones with genotype (KLF14 rs4731702)CC had a 6.689-fold increased risk (OR=6.689) of developing CC compared with patients without a family history of gallstones with genotype (KL F14 RS 473702) (P < 0.05). There was no statistical significance (P > 0.05) in the exercise habits of patients with genotype (KLF14 rs4731 702)CC.

Conclusion 1. The single nucleotide polymorphism (SNP) at rs4731702 of 1.KLF14 gene may have gender distribution differences and be related to the susceptibility to cholesterol gallstones, and the carrying of allele C may increase the risk of cholesterol gallstones. 2. 2. The genotype CC of rs5888 in SR-B1 gene may be the risk genotype for the development of cholesterol gallstones. 3. genotype CC of sr-b1 gene rs 838880 may be the protective genotype for cholesterol gallstones, and allele c may be related to the increase of HDL-C in blood. 4. snps of 4.KLF14 and SR-B1 have significant correlation with dyslipidemia and lipoprotein metabolism. 5. Patients with cholesterol gallstones have different degrees of lipid and lipoprotein metabolism abnormalities. 6. The increased 6.BMI and family history of cholesterol gallstones are the risk factors for cholesterol gallstones. Exercise habit is the protective factor of cholesterol gallstones. 7. The interaction between genetic factors and environmental factors promotes the occurrence of cholesterol gallstones.

The authors have declared no competing interest.

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