本文選題：原發(fā)性高血壓 + 內(nèi)皮素-1��； 參考：《南昌大學(xué)》2016年博士論文

【摘要】：目的：原發(fā)性高血壓(essential hypertension, EH)是一種多基因遺傳與環(huán)境因素相互作用的慢性疾患,且是冠心病、腦卒中等心腦血管疾病及終末期腎病的獨(dú)立危險(xiǎn)因素。許多研究證實(shí),在人類的基因中存在著高血壓病的易感基因。但目前為止,人們還沒有發(fā)現(xiàn)某個(gè)人類基因的多態(tài)性與高血壓的發(fā)生直接相關(guān)。既往的研究表明,內(nèi)皮素-1(ET-1)表達(dá)增多能夠增強(qiáng)動(dòng)脈血管的阻力,高血壓病患者血漿中ET-1水平明顯高于血壓正常者的人群,而且靶器官的損害更嚴(yán)重,提示ET-1可能會(huì)增加高血壓病發(fā)生的風(fēng)險(xiǎn)；另外,人們還發(fā)現(xiàn)在高血壓患者的動(dòng)脈壁內(nèi)有ET-1受體基因的過表達(dá),這可能表明ET-1導(dǎo)致高血壓發(fā)生的機(jī)制是通過影響內(nèi)皮功的能障礙或促進(jìn)血管平滑肌細(xì)胞增殖的方法來實(shí)現(xiàn)的;使用ET-1受體拮抗劑能夠降低慢性高血壓患者的血壓,從而進(jìn)一步證實(shí)ET-1與高血壓存在明顯的相關(guān)性。既往的研究也表明,一氧化氮(NO)在心血管疾病中發(fā)揮了重要作用,一氧化氮是在一氧化氮合酶(NOS)催化下生成的。三種不同的一氧化氮合酶在心血管疾病中的作用各不相同,通過對一氧化氮合酶的干預(yù)將成為心血管疾病治療的新策略。鑒于上述的研究結(jié)果提示ET-1、eNOS與高血壓的發(fā)生有關(guān),因此,ET-1、eNOS基因已被認(rèn)為是高血壓病的高危候選基因。篩查原發(fā)性高血壓相關(guān)基因的單核苷酸多態(tài)性,進(jìn)行基因分型與疾病相關(guān)性研究及與降壓療效關(guān)系研究是對EH進(jìn)行早期診斷、預(yù)防和治療的基礎(chǔ)。本論文分兩章探討：.初步探討內(nèi)皮素-1基因多態(tài)性與江西漢族人群原發(fā)性高血壓及硝苯地平降壓療效的相關(guān)性。二.探討內(nèi)皮型一氧化氮合酶基因多態(tài)性與江西漢族人群原發(fā)性高血壓及硝苯地平降壓療效的相關(guān)性方法：第一部分：對確診為高血壓病的423例患者和114例健康志愿者空腹靜脈抽血分別用于血脂、血糖及ET-1濃度的檢測及全基因組的提取,并對所有受試對象的ET-1基因中rs5370 (Lys198Asn)和rs2071942(G8002A)兩個(gè)位點(diǎn)位進(jìn)行基因分型及測序;同時(shí),收集受試對象身高、體重、吸煙等資料。其中281例入選者皆為高血壓初次確診或已停用降壓藥2周以上,抽取靜脈血后,給予口服硝苯地平控釋片30 mg/d,共2周。分別按照收縮壓及舒張壓的降壓效果,選取降壓效果最好及最差的各50例病人,這樣,共156例原發(fā)性高血壓病人成為硝苯地平控釋片降壓療效的研究對象。分別按收縮壓和舒張壓,對兩極端組病人基因多態(tài)性與降壓效果進(jìn)行分析。第二部分：對確診為高血壓病的423例患者和114例健康志愿者空腹靜脈抽血分別用于血脂、血糖及eNOS濃度等檢測及全基因組的提取,并對所有受試對象的eNOS基因中T786C (rs2070744)和G894T (rs1799983)兩個(gè)位點(diǎn)位進(jìn)行基因分型及測序;同,收集受試對象身高、體重、吸煙等資料。其中281例入選者皆為高血壓初次確診或已停用降壓藥2周以上,抽取靜脈血后,給予口服硝苯地平控釋片30 mg/d,共2周。分別按照收縮壓及舒張壓的降壓效果,選取降壓效果最好及最差的各50例病人,這樣,共156例原發(fā)性高血壓病人成為硝苯地平控釋片降壓療效的研究對象。分別按收縮壓和舒張壓,對兩極端組病人基因多態(tài)性與降壓效果進(jìn)行分析。結(jié)果：第一部分：(1)兩組患者中,除性別及年齡無差別外,高血壓組患者的血脂、空腹血糖、血壓及體重指數(shù)均明顯高于健康對照組,兩組有統(tǒng)計(jì)學(xué)差異(P0.01),此外高血壓組患者吸煙人數(shù)明顯高于健康對照組(P0.001)。(2)ET-1基因rs2071942和rs5370的多態(tài)性檢測符合Hardy-Weinberg平衡(p0.05),表明所選取的受試對象來自檢個(gè)較大的、處于隨機(jī)分配平衡狀態(tài)的群體,具有一定代表性。(3)高血壓患者中,rs5370位點(diǎn)的純合了G/G、T/T及雜合子G/T的基因型頻率分別為73.0%、2.3%及24.7%,T等位基因頻率為14.6%,但與健康對照組相比,無論在基因型頻率還是等位基因頻率均有統(tǒng)計(jì)學(xué)差異。 (4)高血壓組rs2071942位點(diǎn)的G等位基因頻率為66.6%,明顯低于健康對照組的76%,兩組差異明顯,P值為0.007,表明A等位基因的頻率增高可能與高血壓的發(fā)病相關(guān)；以G/G與G/A+A/A進(jìn)行比較,發(fā)現(xiàn)高血壓組與對照組之間亦有顯著的統(tǒng)計(jì)學(xué)差異(P=0.001),表明G/G型純合子的個(gè)體可能能夠抵抗高血壓的發(fā)生(5)ET-1基因rs2071942和rs5370的基因型分布分布均與性別、血壓無相關(guān)性。(6)無論以收縮壓還是以舒張壓來分,ET-1基因rs5370位點(diǎn)多態(tài)性與硝苯地平控釋片降壓效果的關(guān)系,均無顯著相關(guān)性。(7)在高血壓組及健康對照組內(nèi),超重者rs2071942基因型G/G頻率有升高趨勢,而攜帶A等位基因的基因型有下降趨勢,但均無統(tǒng)計(jì)學(xué)意義(P=0.067和P=0.057)。(8)在高血壓患者中rs5370位點(diǎn)攜帶T等位基因的基因型的吸煙者比例明顯高于G/G純合子患者,提示rs5370位點(diǎn)攜帶T等位基因的吸煙者可能與高血壓有關(guān)。(9)高血壓患者血清ET-1濃度與健康對照組相比有顯著的統(tǒng)計(jì)學(xué)意義(P0.05)第二部分：(1)除性別及年齡無差別外,高血壓組患者的血脂、空腹血糖、血壓及體重指數(shù)均明顯高于健康對照組,兩組有統(tǒng)計(jì)學(xué)差異(P0.01),此外高血壓組患者吸煙人數(shù)明顯高于健康對照組(P0.001)。(2) eNOS基因T786C和G894T的多態(tài)性檢測符合Hardy-Weinberg平衡(p0.05),表明所選取的受試對象來自一個(gè)較大的、處于隨機(jī)分配平衡狀態(tài)的群體,具有一定代表性。(3)eNOS基因啟動(dòng)子T786C位點(diǎn)T/T, T/C和C/C基因型在高血壓組分別為22.22%,51.53%,26.47%,在健康對照組分別為37.72%,53.51%,8.77%,此位點(diǎn)高血壓病組與健康對照組的基因分布頻率差異有顯著性。(4)eNOS基因第七外顯子第894位點(diǎn)G/G, G/T和T/T基因型在高血壓組分別為69.28%,22.46%,8.26%,在健康對照組分別為82.46%,14.91%,2.63%,此位點(diǎn)基因分布頻率在高血壓病組與對照組的差異有顯著性。(5)當(dāng)T786C、 G894T兩位點(diǎn)基因型為CC+TT或TC+TT時(shí),經(jīng)計(jì)算患原發(fā)性高血壓的OR值明顯大于基因型為TT+GG者。(6)高血壓患者血清e(cuò)NOS濃度與健康對照組相比降低,有顯著的統(tǒng)計(jì)學(xué)意義(P0.05),但無論高血壓組還是健康對照組,組內(nèi)不同eNOS G894T基因型間血漿eNOS濃度比較無顯著差異(P0.05)。 (7)eNOS的G894T多態(tài)性與硝苯地平控釋片降壓效果,無論以收縮壓還是以舒張壓來分,均無顯著相關(guān)性。結(jié)論：第一部分(1)高血壓患者ET-1基因rs2071942的A等位基因頻率明顯增高,提示ET-1基因A等位基因也許可以作為江西漢族人群高血壓易感基因的遺傳標(biāo)志;(2) ET-1rs5370多態(tài)性與硝苯地平控釋片降壓療效無相關(guān)性；(3)高血壓病的高危因素可能會(huì)影響ET-1基因多態(tài)性的改變;(4)rs5370位點(diǎn)攜帶有T等位基因的吸煙者可能更易患高血壓。第二部分(1) eNOS基因啟動(dòng)子T786C及第七外顯子第894位點(diǎn)基因多態(tài)性與原發(fā)性高血壓的發(fā)病有一定程度的相關(guān)性；(2) eNOS基因啟動(dòng)子T786C位點(diǎn)T/T及第七外顯子894位點(diǎn)G/G基因型是原發(fā)性高血壓的保護(hù)性基因;(3) eNOS基因T786C和G894T變異在原發(fā)性高血壓的發(fā)病中可能有一定程度的協(xié)同作用；(4)在江西漢族高血壓人群中,,G894T多態(tài)性與血漿eNOS濃度無顯著相關(guān)性；(5) eNOS G894T多態(tài)性與硝苯地平控釋片降壓療效無相關(guān)性。
[Abstract]:Objective: essential hypertension (EH) is a chronic disease with multiple genetic and environmental interaction, and is an independent risk factor for coronary heart disease, cerebral cerebral vascular disease and end-stage renal disease. Many studies have confirmed that there is a susceptible gene for hypertension in the human basis. It has not been found that the polymorphism of a human gene is directly related to the occurrence of hypertension. Previous studies have shown that the increase in the expression of endothelin -1 (ET-1) can enhance the resistance of arterial blood vessels. The level of ET-1 in plasma of hypertensive patients is significantly higher than that of people with normal blood pressure, and the damage of target organs is more serious, suggesting that ET-1 may be more serious. The risk of hypertension is increased; in addition, the overexpression of the ET-1 receptor gene in the arterial wall of the hypertensive patients may be found, which may indicate that the mechanism of ET-1 induced hypertension is achieved through a method of affecting the energy barrier of endothelial function or promoting the proliferation of vascular smooth muscle cells; the use of the ET-1 receptor antagonist can be used. There is a significant correlation between ET-1 and hypertension. Previous studies have also shown that nitric oxide (NO) plays an important role in cardiovascular disease, and nitric oxide is produced under the catalysis of nitric oxide synthase (NOS). Three different nitric oxide synthases are in cardiovascular disease. The effects of the nitric oxide synthase will become a new strategy for the treatment of cardiovascular diseases. In view of the results suggested that ET-1, eNOS is associated with the occurrence of hypertension, the ET-1, eNOS gene has been considered as a high risk candidate for hypertension. The study of the relationship between genotyping and disease and the relationship with the antihypertensive effect is the basis for the early diagnosis, prevention and treatment of EH. This paper is divided into two chapters: the correlation of endothelin -1 gene polymorphism and the effect of primary hypertension and nifedipine in Jiangxi Han population. Two. The correlation method between the gene polymorphism of nitric oxide synthase and the effect of primary hypertension and nifedipine in Jiangxi Han population: Part 1: the detection of blood lipid, blood glucose and ET-1 concentration and the extraction of whole genome were used in 423 patients with essential hypertension and 114 healthy volunteers, respectively. The two loci of rs5370 (Lys198Asn) and rs2071942 (G8002A) in the ET-1 gene of the subjects were genotyping and sequencing. At the same time, the height, weight, smoking and other data of the subjects were collected. 281 of the subjects were all first diagnosed with hypertension or more than 2 weeks of antihypertensive drugs had been discontinued. After the extraction of venous blood, oral Nifedipine Controlled Release Tablets 30 was given. Mg/d for 2 weeks. According to the effect of systolic and diastolic blood pressure, 50 patients with the best and worst antihypertensive effect were selected, so that 156 patients with essential hypertension became the research object of the antihypertensive effect of Nifedipine Controlled Release Tablets. According to the systolic and diastolic pressure, the gene polymorphism and the effect of blood pressure in the two extremes were divided. The second part: 423 patients with essential hypertension and 114 healthy volunteers with fasting venous blood were used to detect the blood lipid, blood glucose and eNOS concentration and to extract the whole genome, and the gene typing and sequencing of the two loci of T786C (rs2070744) and G894T (rs1799983) in all the subjects of the subjects were sequenced and sequenced. 281 of the 281 patients were given the first diagnosis of hypertension or more than 2 weeks of antihypertensive drugs. After the extraction of venous blood, the oral Nifedipine Controlled Release Tablets was given 30 mg/d for 2 weeks. In accordance with the effect of systolic and diastolic blood pressure, the best and worst 50 patients were selected, respectively. A total of 156 patients with essential hypertension became the research object of the antihypertensive effect of Nifedipine Controlled Release Tablets. According to the systolic and diastolic pressure, the gene polymorphism and antihypertensive effect of the two extremist groups were analyzed. The first part: (1) among the two groups, the blood lipid and fasting blood sugar in the hypertensive group were not different except for the sex and age. The blood pressure and body mass index were significantly higher than those in the healthy control group (P0.01). In addition, the number of smokers in the two groups was significantly higher than that in the healthy control group (P0.001). (2) the polymorphism of ET-1 gene rs2071942 and rs5370 conformed to Hardy-Weinberg balance (P0.05), indicating that the selected subjects were from a larger test. (3) in hypertensive patients, the rs5370 locus was G/G, the genotype frequencies of T/T and heterozygote G/T were 73%, 2.3% and 24.7%, and the T allele frequencies were 14.6%, but there were statistically significant differences in genotype frequencies and allele frequencies compared with those of the healthy control group (4). The G allele frequency of rs2071942 loci in the hypertension group was 66.6%, which was significantly lower than that of the healthy control group, which was significantly lower than that in the healthy control group. The two groups were significantly different, and the P value was 0.007. The higher frequency of the A allele might be associated with the pathogenesis of hypertension. Compared with G/G and G/A+A/A, there were significant statistical differences between the hypertension group and the control group (P=0.001). The results showed that the individuals of G/G homozygote may be able to resist hypertension (5) ET-1 gene rs2071942 and rs5370 genotype distribution distribution were not related to sex and blood pressure. (6) no matter in systolic or diastolic pressure, there was no significant correlation between the rs5370 locus polymorphism of the ET-1 gene and the effect of Nifedipine Controlled Release Tablets's antihypertensive effect. (7) in the hypertension group and the healthy control group, the frequency of rs2071942 genotype G/G increased in the overweight people, but the genotype of A allele was decreased, but there was no statistical significance (P=0.067 and P=0.057). (8) the genotype of T allele carrying T allele in the hypertensive patients was significantly higher than that of G/G homozygote The smokers who suggest that the rs5370 locus carrying T allele may be related to hypertension. (9) the serum ET-1 concentration in hypertensive patients has significant statistical significance (P0.05) compared with the healthy control group (P0.05): (1) the blood lipid, fasting blood glucose, blood pressure and body mass index of the hypertensive patients are obviously Gao Yujian except for sex and age. In the control group, there was a statistical difference between the two groups (P0.01). In addition, the number of smokers in the hypertension group was significantly higher than that in the healthy control group (P0.001). (2) the polymorphism of eNOS gene T786C and G894T conformed to Hardy-Weinberg balance (P0.05), indicating that the selected subjects were from a larger group in a random distribution equilibrium state. (3) the T786C locus T/T of the eNOS gene promoter, T/C and C/C genotypes were 22.22%, 51.53%, 26.47% in the hypertension group, respectively 37.72%, 53.51%, 8.77% in the healthy control group. The gene distribution frequency difference between the hypertensive group and the healthy control group was significant. (4) the 894th loci of eNOS gene seventh exon G/G, G/T and T/T. The genotype in the hypertension group was 69.28%, 22.46%, 8.26%, respectively, 82.46%, 14.91%, 2.63% in the healthy control group. The difference in the frequency of gene distribution between the hypertensive group and the control group was significant. (5) when T786C, G894T two loci genotype was CC+TT or TC+TT, the OR value of the patients with essential hypertension was significantly greater than that of the genotype TT. (6) (6) the serum concentration of eNOS in patients with hypertension was lower than that of the healthy control group. There was significant statistical significance (P0.05), but there was no significant difference in the concentration of eNOS between the different eNOS G894T genotypes in the hypertensive group and the healthy control group (P0.05). (7) the G894T polymorphism of eNOS and the effect of Nifedipine Controlled Release Tablets on the antihypertensive effect, There is no significant correlation between the systolic pressure or diastolic pressure. Conclusion: the first part (1) the A allele frequency of ET-1 gene rs2071942 in hypertensive patients is significantly higher, suggesting that the A allele of the ET-1 gene may be a genetic marker for the susceptibility gene of hypertension in Jiangxi Han population; (2) ET-1rs5370 polymorphism and nifedipine controlled release (3) high risk factors of hypertension may affect the change of ET-1 gene polymorphism; (4) smokers with T allele in rs5370 loci may be more susceptible to hypertension. Second (1) eNOS gene promoter T786C and seven exon 894th loci gene polymorphisms are associated with the onset of essential hypertension The degree of correlation; (2) eNOS gene promoter T786C site T/T and seven exon 894 locus G/G genotype is a protective gene for essential hypertension; (3) the eNOS gene T786C and G894T variation may have a certain degree of synergy in the pathogenesis of essential hypertension; (4) the polymorphism of G894T in Jiangxi Han hypertension population There was no significant correlation with plasma eNOS concentration; (5) eNOS G894T polymorphism was not associated with Nifedipine Controlled Release Tablets hypotensive effect.

【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R544.1

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