基质金属蛋白酶8-799C/T基因多态性与颈动脉斑块易损性的关联分析

目的 探讨我国浙江地区汉族人群基质金属蛋白酶-8 (matrix metalloproteinase-8,MMP-8)启动子基因-799C/T多态与颈动脉斑块易损性的关系.方法 451例脑梗死患者根据颈动脉超声分为易损斑块组(共135例)和稳定斑块组(共316例).采用酶联免疫吸附法测定两组血清MMP-8水平,并采用聚合酶链反应-限制性片段长度多态性方法分析两组MMP-8启动子基因-799C/T多态的基因型.结果 易损斑块组发病48 h内血清MMP-8水平为(0.86±0.16)ng/μL,稳定斑块组为(0.80±0.13) ng/μL,两者差异有统计学意义(t=2.894,P=0.004).易损斑块组CT+ TT基因型频率为73.3％,稳定斑块组为54.7％,两者差异有统计学意义(x2=13.65,P=0.000);T等位基因频率在易损斑块组为48.1％,在稳定斑块组为33.5％,两者差异有统计学意义(x2=17.14,P=0.000).CC基因型组血清MMP-8水平为(0.79±0.13) ng/μL,TT基因型组血清MMP-8水平为(0.92±0.11) ng/μL,两者比较差异有统计学意义(t=3.141,P=0.001).结论 MMP-8启动子基因-799C/T多态可能与浙江地区汉族人颈动脉易损斑块发生的倾向有关,T等位基因可能是颈动脉斑块易损的遗传易患性标志之一.     

支气管哮喘患儿血清中IL-17水平表达及IL-17 A基因152G/A位点单核苷酸多态性分析

目的了解支气管哮喘患儿血清中白细胞介素17(Interleukin-17,IL-17)水平表达情况,并探讨IL-17A基因152G/A位点单核苷酸多态性(single-nucleotide polymorphism,SNP)分布特征与深圳地区支气管哮喘患儿易感的相关性。方法收集2016年7月～2018年5月来院就诊并确诊为支气管哮喘患儿80例为哮喘组,同时选择同期健康体检儿童120名为对照组,采用免疫组化ESLIA法检测血清中IL-17水平,采用聚合酶链式反应-限制性片段长度多态性(PCRRFLP)法检测IL-17A基因152G/A位点SNP多态性,并对两组检测结果进行统计分析。结果哮喘组儿童血清中IL-17水平为3198.54±241.92ng/L,明显高于对照组健康儿童的982.74±69.38ng/L,差异有统计学意义(t=6.1854,P0.05)。男性哮喘患儿血清中IL-17水平为3067.29±219.63ng/L,略低于女性哮喘患儿的3385.76±256.08ng/L,但差异无统计学意义(t=1.3175,P0.05)。哮喘组患儿血清中IL-17A基因152G/A位点AA基因型检出率为26.25%,明显高于对照组健康儿童的11.67%,差异有统计学意义(χ~2=3.9105,P0.05);哮喘组患儿A等位基因频率为50.63%,明显高于对照组健康儿童的37.08%,差异有统计学意义(χ~2=3.0524,P0.05)。结论支气管哮喘患儿血清中IL-17水平明显高于健康儿童,且IL-17A基因152G/A位点分布呈多态性,其中AA基因型检出率和A等位基因频率明显高于健康儿童,可能是深圳地区支气管哮喘患儿发病的易感基因。     

去甲肾上腺转运蛋白基因多态性与高血压合并心力衰竭的关系

目的探索汉族人群去甲肾上腺转运蛋白(norepinephrine transporter,NET)基因(SLC6A2)启动子3、2多态性与高血压病(essential hypertension,EH)合并心力衰竭(heart failure,HF)的关系。方法按年龄、性别和居住地配对原则收集3组受试者:对照组(n=176,健康体检者),EH组(n=176,心功能正常的EH患者)和EH-HF组(n=176,EH合并NYHAⅢ～Ⅳ级心功能患者),应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)检测SLC6A2启动子3和启动子2多态性。结果EH-HF组SLC6A2启动子3AG/GG基因型分布频率(41.48%)高于EH组(26.70%)和对照组(22.16%),3组SLC6A2启动子2GG、GC和CC基因型分布差异无统计学意义。以EH组为参照系,调整混杂因素后,SLC6A2启动子3AG/GG基因型发生心衰的OR值为1.905(95%CI:1.138～3.188,P=0.014);以SLC6A2启动子3A-G/启动子2单倍体为参比基线,单因素分析SLC6A2启动子3G-C/启动子2单倍体发生心衰的OR值为2.744(95%CI:1.390～5.417,P=0.004),而且携带SLC6A2启动子3AG/GG基因型的EH-HF患者血浆脑钠肽水平明显高于AA基因型患者(P<0.001)。结论携带SLC6A2启动子3G等位基因及启动子3G-C/启动子2单倍体型患者与EH-HF有关,可能是EH-HF分子遗传学基础之一。     

基质金属蛋白酶-3基因多态性与脑卒中亚型的相关性

目的 探讨中国北方汉族人群基质金属蛋白酶-3(matrix metalloproteinase-3,MMP-3)基因多态性与缺血性脑卒中(ischemic stroke,IS)亚型的相关性.方法 应用病例对照研究,选取289例急性缺血性脑卒中患者(发病≤3d)和175名同期健康体检者.卒中组按急性卒中治疗低分子肝素试验病因分型法分为大动脉粥样硬化性(large artery atherosclerotic,LAA)脑卒中185例,小动脉闭塞性(small artery occlusion,SAO)脑卒中104例.选取MMP-3基因rs3025058(-11715A/6A),rs522616(-709A/G)及rs679620(133A/G)3个常见单核苷酸多态性(single nucleotide polymorphisms,SNPs)位点,应用聚合酶链反应限制性片段长度多态性或直接测序法对MMP-3基因3个SNP位点进行基因分型.结果 3个位点存在不完全连锁不平衡,且基因多态性均与LAA型脑卒中相关(P＜0.05).在显性模型中,rs3025058位点5A5A+5A6A基因型携带者患LAA型脑卒中的风险是6A6A基因型携带者的1.72倍(P=0.017,OR=1.72,95％CI:1.10～2.69);rs522616位点GG+ AG基因型携带者患LAA型脑卒中的风险是AA基因型携带者的0.52倍(P=0.005,OR=0.52,95％CI:0.33～0.82);rs679620位点AA+ GA基因型携带者患LAA型脑卒中的风险是AA基因型携带者的1.55倍(P=0.042,OR=1.55,95％CI:1.01～2.37).但是,3个SNPs基因型和等位基因频率在对照组与SAO型脑卒中之间差异无统计学意义(P＞0.05).另外,LAA组5A-A-A及6A-A-A单倍型高于对照组,差异有统计学意义(P＜0.05),而6A-G-G单倍型显著低于对照组(P＜0.01).结论 MMP-3血清水平在LAA型脑卒中急性期增高明显,SAO型脑卒中次之;中国北方汉族人群MMP-3基因rs3025058,rs522616及rs679620多态性可能与LAA型脑卒中易感性相关.     

TGF-β1基因启动子-800G/A、-509C/T多态性与食管癌的研究

目的研究转化生长因子β1(TGF-β1)基因启动子多态性各等位基因及基因型在食管癌患者中的分布频率,初步分析其基因型及血清水平与食管癌的相关性.方法采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术,检测118例食管癌患者和130例正常对照组TGF-β1的基因多态性,包括TGF-β1基因启动子-800G/A、-509C/T位点,同时采用ELISA检测血清TGF-β1水平.结果食管癌患者血清TGF-β1水平显著高于对照组(P＜0.01),TGF-β1基因-800G/A位点多态性在食管癌组和正常人群中的分布差异无显著性(P＞0.05),而TGF-β1基因-509C/T多态性各等位基因及基因型频率在两组人群中的分布差异存在显著性(P＜0.05);等位基因频率的相对风险分析发现,T等位基因携带者患食管癌的风险是C等位基因的1.624倍(OR=1.624,95%CI1.134～2.324),携带T等位基因的食管癌患者血清TGF-β1水平显著高于不携带者(50.97±8.91μg/LVS44.23±8.54μg/L,P＜0.01).结论TGF-β1基因-509C/T多态性与食管癌的发病具有相关性,其中T等位基因可能是食管癌发病的遗传易感基因;携带T等位基因的个体可能通过促进TGF-β1的高度表达进而增加了食管癌的发病风险.     

多囊卵巢综合征患者25-(OH)VD、AMH水平表达及AMH和其Ⅱ型受体基因多态性分析

目的了解深圳地区多囊卵巢综合征(PCOS)患者血清中25-羟基维生素D[25-(OH)VD]、抗苗勒管激素(AMH)水平及AMH和其Ⅱ型受体(AMHR II)基因多态性,并探讨其与PCOS发病之间的相关性。方法选取2019年1月至2020年7月在深圳市龙华区人民医院及各社康服务中心妇产科就诊并确诊为PCOS患者107例为PCOS组,并选择同期来医院检查的健康育龄妇女95例为对照组,分别检测血清中25-(OH)VD和AMH水平,同时采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)法对AMH G/T和AMHR II A/T基因多态性进行检测,并对检测结果进行统计分析。结果 PCOS组患者血清中25-(OH)VD水平为32.97±6.41nmol/L,明显低于对照组的63.84±12.60nmol/L,而AMH水平为17.68±6.84ng/ml,明显高于对照组的5.28±1.97ng/ml,差异有统计学意义(P0.05);PCOS组患者AMH G/T基因TT基因型和T等位基因检出率分别为39.26%和52.80%,明显高于对照组的28.42%和42.73%,差异均有统计学意义(P0.05);PCOS组患者AMHRⅡA/T基因TT基因型及T等位基因检出率分别为22.43%和38.32%,明显高于对照组的11.58%和26.32%,差异有统计学意义(P0.05);携带AMH基因TT基因型的PCOS患者血清中AMH水平为21.85±7.63ng/ml,明显高于AT基因型的14.96±5.42ng/ml,且AT基因型明显高于AA基因型的9.25±2.78ng/ml,而TT基因型的25-(OH)VD低于AT基因型,且AT基因型低于TT基因型,不同基因型PCOS患者血清中AMH、25-(OH)VD水平之间差异均有统计学意义(P0.05);PCOS患者25-(OH)VD水平明显降低,而AMH水平明显升高,经Spearman相关性分析,结果显示两者呈负相关(r=-0.6275,P0.05)。结论深圳地区PCOS患者血清中25-(OH)VD水平明显降低,而AMH水平明显升高,且两者呈负相关。同时PCOS患者AMH G/T基因TT基因型和AMHRⅡA/T基因TT基因型检出率明显升高,可能是该地区PCOS发病的遗传易感基因。     

MMP-12、-13基因多态性与上皮性卵巢癌发病风险的关联研究

目的 探讨MMP-12、-13基因启动子区功能多态性与上皮性卵巢癌发病风险的关系.方法 应用聚合酶链反应-限制性片段长度多态性方法 检测300例上皮性卵巢癌患者和300名对照妇女的MMP-12-82A/G及MMP-13-77A/G单核甘酸多态性(single nucleotide polymorphism,SNP)的基因型和等位基因频率分布情况.结果 上皮性卵巢癌组中MMP-12-82A/G SNP的A、G等位基因频率和AA、AG基因型频率与对照组相比差异有统计学意义(P=0.004;P=0.003);与AA基因型比较,AG基因型可显著增加上皮性卵巢癌的发病风险(OR=2.81,95%CI:1.38～5.74).MMP-13-77 A/G SNP的等位基因及基因型频率在上皮性卵巢癌组和对照组中分布差异无统计学意义(P=0.06和P=0.15),但根据病理类型分层分析发现,与GG基因型相比,AA基因型可显著增加浆液性及粘液性上皮性卵巢癌的发病风险(OR=1.93,95%CI:1.05～3.53;OR=5.16,95%CI:1.62～16.44).结论 MMP-12-82 A/G和MMP-13-77A/G多态性位点可能为上皮性卵巢癌或特定病理类型上皮性卵巢癌发病的独立风险因素.     

MMP-12、-13基因多态性与上皮性卵巢癌发病风险的关联研究

目的 探讨MMP-12、-13基因启动子区功能多态性与上皮性卵巢癌发病风险的关系.方法 应用聚合酶链反应-限制性片段长度多态性方法 检测300例上皮性卵巢癌患者和300名对照妇女的MMP-12-82A/G及MMP-13-77A/G单核甘酸多态性(single nucleotide polymorphism,SNP)的基因型和等位基因频率分布情况.结果 上皮性卵巢癌组中MMP-12-82A/G SNP的A、G等位基因频率和AA、AG基因型频率与对照组相比差异有统计学意义(P=0.004;P=0.003);与AA基因型比较,AG基因型可显著增加上皮性卵巢癌的发病风险(OR=2.81,95%CI:1.38～5.74).MMP-13-77 A/G SNP的等位基因及基因型频率在上皮性卵巢癌组和对照组中分布差异无统计学意义(P=0.06和P=0.15),但根据病理类型分层分析发现,与GG基因型相比,AA基因型可显著增加浆液性及粘液性上皮性卵巢癌的发病风险(OR=1.93,95%CI:1.05～3.53;OR=5.16,95%CI:1.62～16.44).结论 MMP-12-82 A/G和MMP-13-77A/G多态性位点可能为上皮性卵巢癌或特定病理类型上皮性卵巢癌发病的独立风险因素.     

纤维蛋白原Bβ多态基因FGB-455G/A多态性与动脉粥样硬化性脑梗塞的关系

目的　探讨纤维蛋白原 (fibrinogen ,FGB )Bβ多态基因FGB 45 5G/A多态性在中国汉族人群中的分布及其与动脉粥样硬化性脑梗塞 (atheroscleroticcerebralinfarction ,ACI)的关系。方法　应用聚合酶链反应和限制性内切酶片段长度多态性技术检测了 13 2例ACI患者和 14 8名年龄、性别相匹配的正常对照者的Bβ基因FGB -4 5 5G/A多态性 ,比浊法测定血浆纤维蛋白原水平。 结果　血浆纤维蛋白原水平在ACI组显著高于正常对照组 (P <0 .0 0 1)。各组内A等位基因携带者血浆纤维蛋白原水平显著高于GG基因型者 ,且更易受吸烟等环境因素影响。Bβ基因FGB -4 5 5G/A多态性分布在患病组和对照组均符合Hardy Weinberg平衡定律。A等位基因频率在ACI组 ( 0 .2 5 8)显著高于正常对照组 ( 0 .15 2 ) (P <0 0 5 )。Logistic回归分析发现 ,存在此突变位点者发生ACI的危险提高了 65 .3 %。结论　Bβ基因FGB -4 5 5G/A多态性中的A等位基因可能是中国人缺血性脑血管疾病的遗传易感标志之一。     

MMP-12、-13基因多态性与上皮性卵巢癌发病风险的关联研究

目的 探讨MMP-12、-13基因启动子区功能多态性与上皮性卵巢癌发病风险的关系.方法 应用聚合酶链反应-限制性片段长度多态性方法 检测300例上皮性卵巢癌患者和300名对照妇女的MMP-12-82A/G及MMP-13-77A/G单核甘酸多态性(single nucleotide polymorphism,SNP)的基因型和等位基因频率分布情况.结果 上皮性卵巢癌组中MMP-12-82A/G SNP的A、G等位基因频率和AA、AG基因型频率与对照组相比差异有统计学意义(P=0.004;P=0.003);与AA基因型比较,AG基因型可显著增加上皮性卵巢癌的发病风险(OR=2.81,95%CI:1.38～5.74).MMP-13-77 A/G SNP的等位基因及基因型频率在上皮性卵巢癌组和对照组中分布差异无统计学意义(P=0.06和P=0.15),但根据病理类型分层分析发现,与GG基因型相比,AA基因型可显著增加浆液性及粘液性上皮性卵巢癌的发病风险(OR=1.93,95%CI:1.05～3.53;OR=5.16,95%CI:1.62～16.44).结论 MMP-12-82 A/G和MMP-13-77A/G多态性位点可能为上皮性卵巢癌或特定病理类型上皮性卵巢癌发病的独立风险因素.     

Association of matrix metalloproteinase 3 promoter genotype with disease outcome in rheumatoid arthritis

Matrix metalloproteinases (MMPs) are implicated in joint destruction in rheumatoid arthritis (RA). We investigated whether the 5A/6A polymorphism within the MMP-3 (stromelysin-1) gene promoter region is associated with disease outcome in 254 patients with established RA. Patients homozygous for the MMP-3 6A allele had more radiographic damage (measured by Larsen score) than those with other genotypes (109.8 vs 91.1, P=0.04). Patients with the 6A/6A genotype also had more functional impairment and higher serum proMMP-3 levels, although only the latter was significant (P=0.002). A possible association was found between homozygosity for the 6A allele and carriage of the RA-associated HLA-DRB1 shared epitope (SE). Combination of these factors was associated with more severe disease than the SE alone. The data suggest that the MMP-3 6A/6A genotype is associated with worse RA outcome, and that this genotype may have an additive effect with the SE on disease severity.     

Influence of variations across the MMP-1 and -3 genes on the serum levels of MMP-1 and -3 and disease activity in rheumatoid arthritis

Matrix metalloproteinases (MMPs) are involved in joint destruction in rheumatoid arthritis (RA), and are strongly associated with levels of inflammation. To understand the relationship between MMP-1 and -3 variants and MMP levels in RA, we investigated the genotypic and haplotypic relationships of the MMP-1 and -3 genes with circulating levels of these MMPs. The genotypes of single-nucleotide polymorphisms (SNPs) rs1799750 (1G/2G, MMP-1 promoter), rs495366 (G/A, intergene), rs679620 (A/G, MMP-3 coding region) and rs3025058 (5A/6A, MMP-3 promoter) were determined in 430 RA patients. Each polymorphism was associated with serum levels of MMP-1 (P trend <0.0001 for each SNP), with haplotype 1G-G-A-5A associated with the highest level. The intergenic and MMP-3 SNPs were associated with MMP-1 levels independent of the MMP-1 promoter SNP. The MMP-3 SNPs were associated with serum MMP-3 level (P trend <0.0001 for each SNP), and were each associated with mean time-averaged disease activity (DAS28) in patients followed up for 2 years (P=0.003). Our findings indicate that several closely linked polymorphisms in the MMP-1-MMP-3 loci have an important role in determining the circulating levels of these MMPs in RA, and that MMP-3 polymorphism is associated with the level of disease activity over time.     

高血压病患者基质金属蛋白酶3基因多态性与左室重构的关系

 目的：探讨高血压病患者基质金属蛋白酶3（MMP-3）基因多态性与左室重构的关系。方法：入选190名高血压病患者，彩色多普勒检测心脏相关指标，酶联免疫吸附法检测血清MMP-3水平，采用多聚酶链反应（PCR）和测序方法检测MMP-3基因启动子-1512位点5A和6A等位基因。结果：高血压伴左室肥厚108人，不伴左室肥厚82人；MMP-3基因型测序结果6A纯合子128人，5A纯合子9人，5A/6A杂合子53人；携带5A基因者血清MMP-3水平比携带6A基因者高，但差异无显著性；血清MMP-3水平与左室肥厚指标正相关；携带5A基因高血压患者发生左室肥厚的机率是携带6A基因者的2倍。结论：MMP-3基因多态性影响血清MMP-3水平的表达，从而对高血压病患者左室重构的发生、发展产生一定的作用。     

Association of the matrix metalloproteinase-3 (MMP-3) promoter polymorphism with celiac disease in male subjects

Matrix metalloproteinases (MMPs), such as stromelysin-1 (MMP-3), are a family of enzymes important in resorption and remodeling of the extracellular matrix whose degradation may play a role in the villous atrophy characteristic of celiac disease (CD). We investigated the association between the polymorphism at position -1171 of the MMP-3 gene and susceptibility to CD in 225 Italian patients and 170 controls previously typed for human leukocyte antigen (HLA) class II genes. We also evaluated sex differences in the effect of this polymorphism on disease risk. A male-specific association of the 5A/6A polymorphism with CD was observed. The frequencies of 6A allele and 6A/6A genotype in affected male subjects were significantly above those observed both in male controls (p = 4.1 x 10(-3) and p = 3.4 x 10(-3); odds ratio = 2.4, 95% confidence interval 1.3-4.3) and in female patients (p = 2.7 x 10(-4) and p = 6.2 x 10(-4)). This is the first demonstration of a sex-specific association between the MMP-3 promoter polymorphism and CD. Homozygosity for the 6A allele appears as a risk factor for CD only in men, which is different from the HLA susceptibility alleles that confer a higher risk in women.     

Heterozygote AG variant of -596 A/G IL-6 gene polymorphism is a marker for cutaneous T-cell lymphoma (CTCL)

The aim of the study was to investigate possible associations of IL-6 gene polymorphisms (−596 A/G and −174 C/G) with cutaneous T-cell lymphoma (CTCL). In the case-control study, genotype distributions and allelic frequencies in two promoter IL-6 gene polymorphisms in the group of 63 Czech patients with CTCL were compared to those of 105 control non-CTCL subjects matched for age and sex. The IL-6 gene polymorphisms were determined by PCR with following restriction analysis. A significant difference of −596 A/G IL-6 genotype distribution was found between the CTCL patients and the controls (P = 0.002) with almost threefold odds ratio for the heterozygote (AG) genotype in CTCL patients (OR = 2.64, P = 0.002). No significant differences in genotype distribution and/or allelic frequency of functional −174 C/G IL-6 gene polymorphism were observed. The double heterozygote AGCG of both IL-6 promoter polymorphisms was associated with CTCL (OR = 2.24, 95% confidence interval 1.17–4.28, P = 0.01). Thus, the heterozygote variant of −596 A/G promoter IL-6 polymorphism could be considered as a genotype marker for CTCL.     

Matrix metalloproteinases in premature coronary atherosclerosis: influence of inhibitors, inflammation, and genetic polymorphisms

Matrix metalloproteinases (MMPs) are thought to participate in the pathogenesis of coronary artery disease (CAD), particularly in the occurrence of acute coronary syndrome (ACS). Little is known about human in vivo MMP regulation in CAD. The expression and regulation of MMPs and their tissue inhibitors (TIMPs) were evaluated in premature CAD. The distribution of MMP-3 5A/6A and MMP-9 C/T promoter polymorphisms and MMP-9 A/G exon-6 polymorphism were investigated in 200 consecutive male premature CAD patients (aged < or = 55 years) and 201 age-matched male blood donors. Plasma concentrations/activities of MMP-2 and MMP-9 were also measured, as were plasma concentrations of MMP-3, TIMP-1, and TIMP-2 in 80 patients (49 with ACSs and 31 with stable CAD) and 40 controls. Inflammation markers were also obtained. MMP genetic polymorphism distributions did not vary between patients and controls and did not seem to influence their respective MMP plasma levels. Patients showed increased MMP-9 and TIMP-1 concentrations and decreased TIMP-2 concentration and MMP-2 total activity (all P < or = 0.002). Overall, TIMP-1 correlated with C-reactive protein (CPR) (r = 0.594, P < 0.001) and haptoglobin (r = 0.276, P = 0.005), whereas MMP-2 activity correlated inversely with haptoglobin (r = -0.195, P = 0.032). Blood glucose correlated positively with TIMP-1 concentration (r = 0.711, P < 0.001) and negatively with MMP-2 activity (r = -0.250, P = 0.006). In conclusion, MMP and TIMP plasma levels in premature CAD are linked to clinical presentation and markers of inflammation and metabolic disorders rather than to genetic polymorphisms.