醛固酮腺瘤与单侧肾上腺增生患者组织KCNJ5基因突变的临床研究

发布时间：2018-05-07 02:33

本文选题：KCNJ5 + 突变　；参考：《新疆医科大学》2015年博士论文

【摘要】：目的:原发性醛固酮增多症是临床上导致继发性高血压最常见的病因之一,究其根本的原因是由于肾上腺皮质出现的病变,导致醛固酮过多的分泌并且抑制了肾素一血管紧张素系统的活性的综合征。醛固酮腺瘤与单侧肾上腺增生是两种原发性醛固酮增多症的亚型,由于二者有着相似的临床表型及较好的手术治疗效果,对二者有无相同的发病机制,较少有研究。2011年choil等首次报道散发性醛固酮腺瘤患者有36.4%KCNJ5基因体系突变,引起该领域学者的广泛关注,该研究在我国目前报道较少。了解本地区醛固酮腺瘤与单侧肾上腺增生患者KCNJ5基因的突变情况以及KCNJ5基因突变后对醛固酮腺瘤与单侧肾上腺增生临床表型的影响,为进一步探索二者的发病机制及更好的管理和控制原发性醛固酮增多症及继发性高血压提供依据。方法:1)收集46例醛固酮腺瘤与14例单侧肾上腺患者组织KCNJ5基因测序:①提取醛固酮腺瘤(n=46)、单侧肾上腺增生(14例)患者组织的总DNA及RNA;提取相应的患者外周血DNA作为对照;②DNA经PCR扩增KCNJ5基因后正反向测序,再将相应的RNA逆转录为cDNA后行PCR扩增基因正向测序,检测上述不同组织中KCNJ5基因的序列;③获取46例醛固酮腺瘤及14例单侧肾上腺患者KCNJ5基因信息。2)收集46例醛固酮腺瘤及14例单侧肾上腺患者年龄、血压等一般临床资料;收集电解质、血脂等生化资料、心脏超声以及多导睡眠监测的相关指标:①分析醛固酮腺瘤患者KCNJ5基因不同突变型与临床表现型之间的关系;②对比KCNJ5基因突变对二者临床表型的影响。结果:1)46例醛固酮腺瘤与14例单侧肾上腺患者组织KCNJ5基因测序结果:①46例醛固酮腺瘤与14例单侧肾上腺患者组织KCNJ5基因均发现G151R、L168R、S209T及L102Q四个位点突变,其中S209T和L102Q为新的基因突变,而与其相匹配外周血的基因测序正常;国内外尚无报道;②19.57%醛固酮腺瘤患者(9/46例)的肿瘤组织中存在p.G151R和/或p.L168R的杂合突变;21.43%单侧肾上腺增生患者(3/14例)增生组织中为P.G151R或P.L168R突变,其中p.G151R突变分别是c.451GA或c.451GC,为杂合突变;③26.07%醛固酮腺瘤患者(9/46例)的肿瘤组织中存p.s209t突变,64.29%单侧肾上腺增生患者(9/14例)组织中存在p.s209t突变;30.43%例醛固酮腺瘤患者(14/46例)的肿瘤组织中存p.l102q突变;71.43%单侧肾上腺增生患者(10/14例)组织中存在p.l102q突变;④肾上腺醛固酮腺瘤组织中存在kcnj5基因g151r的杂合突变、l168r的突变、s209t与l102q突变,突变率为65.21%(30/46);单侧肾上腺增生组织中存在kcnj5基因g151r的杂合突变、l168r的突变、s209t与l102q突变,突变率为71.43%(10/14例)。2)46例醛固酮腺瘤与14例单侧肾上腺患者组织kcnj5基因突变的临床特点:①在apa患者中kcnj5基因突变与未突变患者比较平均收缩压差异有统计学意义(p=0.042)。两组血钾比较差异有统计学意义(p=0.048),两组在年龄、性别、醛固酮水平、肾功能、心脏超声相关指标比较差异无统计学意义(p0.05);在单侧肾上腺增生患者中kcnj5基因突变与未突变患者比较平均收缩压及舒张压差异有统计学意义(p值分别为0.032和0.016)。两组在年龄、性别、醛固酮水平、血钾、肾功能、心脏超声等相关指标比较差异无统计学意义(p0.05);②46例醛固酮腺瘤患者中kcnj5基因g151r、l168r、s209t及l102q突变与未突变比较g151r突变较未突变组在平均收缩压、平均舒张压、血钾水平、高密度脂蛋白及左室质量指数差异有统计学意义(p值分别为0.005、0.003、0.023和0.004);l168r突变较未突变组在肿瘤直径比较差异有统计学意义(p=0.027);s209t突变较未突变组各项指标比较均差异无统计学意义(p0.05);l102q突变较未突变组在平均舒张压、醛固酮水平、血钾水平、高密度脂蛋白及射血分数差异有统计学意义(p值分别为0.045、0.025、0.030、0.024和0.002);③46例醛固酮腺瘤与14例单侧肾上腺增生患者kcnj5基因g151r+l168r、s209t及l102q突变比较:醛固酮腺瘤患者g151r+l168r突变较单侧肾上腺增生患者在平均收缩压、平均舒张压及坐位醛固酮水平高,差异有统计学意义(p值分别为0.007、0.009和0.033);醛固酮腺瘤患者s209t突变较单侧肾上腺增生患者bmi和胆醇水平高,差异有统计学意义(p分别为0.025和0.005);醛固酮腺瘤患者l102q突变与单侧肾上腺增生患者平均舒张压、坐位醛固酮水平、总胆固醇水平及射血分数比较,差异有统计学意义(p值分别为0.022、0.027和0.033);④在46醛固酮腺瘤患者中有1例男性患者合并中度阻塞性睡眠呼吸暂停低通气综合征,14例单侧肾上腺增生患者合并4例重度阻塞性睡眠呼吸暂停低通气综合征,其中3名男性,1名女性。结论:1)肾上腺醛固酮腺瘤组织中存在kcnj5基因g151r的杂合突变、l168r的突变、s209t与l102q突变,突变率为65.21%(30/46),其中s209t与l102q突变为新发现的突变位点,与既往的研究不同,提示kcnj5基因的突变可能与人种及地域有关;2)kcnj5基因突变加重了apa患者的临床症状,其中g151r突变影响最大,之后依次为l102q、l168r及s209t;3)单侧肾上腺增生患者与醛固酮腺瘤存在相同KCNJ5基因G151R的杂合突变、L168R的突变、S209T与L102Q突变,突变率为71.43%(10/14),高于醛固酮腺瘤的突变率,其中S209T与L102Q为新发现的突变位点,既往文献尚未报到;4)KCNJ5基因突变对醛固酮腺瘤患者的临床表型影响较单侧肾上腺增生患更大,其中G151R突变影响最大,之后依次为L102Q、L168R及S209T;5)单侧肾上腺增生患者与醛固酮腺瘤存在相同KCNJ5基因突变,影响程度不同,其发生机制仍需进一步研究;6)KCNJ5基因突变是否与OSAHS的发生有关,仍需进一步的探讨。
[Abstract]:Objective: primary aldosteronism is one of the most common causes of secondary hypertension. The fundamental reason is that the adrenocortical lesions are caused by the hyperaldosterone secretion and the inhibition of the renin's activity of angiotensin system. The aldosterone adenoma and unilateral adrenal hyperplasia are two. The subtype of primary aldosteronism, because the two have similar clinical phenotypes and good surgical treatment effect, there is no same pathogenesis for the two, and the first report of.2011 choil, which is the first report of sporadic aldosteronadenoma, has the mutation of the 36.4%KCNJ5 gene system, which has aroused extensive attention of the scholars in this field. At present, there are few reports in China. To understand the mutation of KCNJ5 gene in aldosterone adenoma and unilateral adrenal hyperplasia in the local area and the effect of KCNJ5 gene mutation on the clinical phenotype of aldosterone adenoma and unilateral adrenal hyperplasia in order to further explore the pathogenesis of the two and to better manage and control the primary aldosteronism. And to provide basis for secondary hypertension. Methods: 1) the KCNJ5 gene of 46 aldosterone adenomas and 14 unilateral adrenal patients was sequenced. (1) the aldosterone adenoma (n=46) was extracted, the total DNA and RNA of the tissues of the unilateral adrenal hyperplasia (14 cases) were obtained, and the corresponding peripheral blood DNA was extracted as the control; and the KCNJ5 gene was amplified by PCR by PCR. Sequence, then reverse transcription of the corresponding RNA to cDNA after PCR amplification gene sequencing to detect the sequence of KCNJ5 gene in the above different tissues; (3) to obtain 46 aldosterone adenoma and 14 cases of unilateral adrenal patients with KCNJ5 gene.2) to collect 46 aldosterone adenomas and 14 cases of unilateral adrenal patients' age, blood pressure and other general clinical data; collect electricity. Biochemical data such as degradation, blood lipid, cardiac ultrasound and polysomnography: (1) analysis of the relationship between different mutations of KCNJ5 gene in aldosterone adenoma and clinical phenotype; (2) the effect of KCNJ5 gene mutation on the clinical phenotype of two cases. Results: 46 cases of aldosterone adenoma and 14 cases of unilateral adrenal gland patients, KCNJ5 Gene sequencing results: (1) 46 aldosterone adenomas and 14 cases of unilateral adrenal patients with KCNJ5 gene were found to have four mutations in G151R, L168R, S209T and L102Q, of which S209T and L102Q were new gene mutations, and the gene sequence matched with the peripheral blood was normal; there was no report at home and abroad; (2) the swelling of the 19.57% aldosterone adenoma patients (9/46 cases) There was a heterozygous mutation of p.G151R and / or p.L168R in the tumor tissue; 21.43% the hyperplastic (3/14) patients with unilateral adrenalic hyperplasia were P.G151R or P.L168R mutations, of which p.G151R mutations were c.451GA or c.451GC, as heterozygous mutation, and (3) the tumor tissues of 26.07% aldosterone adenomas (9/46 cases) had p.s209t mutations and 64.29% unilateral adrenal glands increased. There was a p.s209t mutation in the tissue of 9/14, and p.l102q mutation in 30.43% cases of aldosterone adenoma (14/46), and p.l102q mutation in the tissues of 71.43% unilateral adrenal hyperplasia (10/14); (4) the heterozygous mutation of the kcnj5 gene g151r, the mutation of l168r, s209t and l102q process in the adrenal aldosterone adenoma tissue Mutation rate was 65.21% (30/46), and there was a heterozygous mutation of kcnj5 gene g151r in unilateral adrenal hyperplasia, l168r mutation, s209t and l102q mutation, mutation rate of 71.43% (10/14 case).2) 46 aldosterone adenomas and 14 cases of unilateral adrenal patients with kcnj5 gene mutation: (1) kcnj5 gene mutation and non mutation in APA patients The difference of the average systolic pressure was statistically significant (p=0.042). There was a significant difference between the two groups (p=0.048). There was no significant difference between the two groups in age, sex, aldosterone level, renal function, and echocardiography (P0.05). In the patients with mono adrenal hyperplasia, the kcnj5 gene mutation was compared with that of the non mutated patients. The differences in mean systolic and diastolic pressure were statistically significant (P values were 0.032 and 0.016 respectively). There was no significant difference in age, sex, aldosterone level, potassium, renal function, and cardiac ultrasound (P0.05) in two groups (P0.05); (2) the kcnj5 gene g151r, l168r, s209t and l102q mutations were compared with the non mutation in the patients with aldosterone adenoma. There was significant difference in mean systolic pressure, mean diastolic pressure, blood potassium level, high density lipoprotein and left ventricular mass index (P value 0.005,0.003,0.023 and 0.004), and the difference between l168r mutation and non mutation group was statistically significant (p=0.027), and the s209t mutation was compared with that of the non mutation group. The difference was not statistically significant (P0.05), and there was a significant difference in mean diastolic pressure, aldosterone level, blood potassium level, high density lipoprotein and ejection fraction (P value of 0.045,0.025,0.030,0.024 and 0.002), and 46 cases of aldosterone adenoma and 14 cases of unilateral adrenal hyperplasia, g151r+l168r, s209t, s209t, s209t, s209t, s209t, s209t, and s209t. Compared with the l102q mutation, the g151r+l168r mutation in the aldosterone adenoma patients was higher than that of the unilateral adrenal hyperplasia in the mean systolic pressure, the mean diastolic pressure and the sitting aldosterone level, and the difference was statistically significant (P was 0.007,0.009 and 0.033, respectively), and the s209t mutation of the aldosterone adenoma was higher than that of the unilateral adrenal hyperplasia patients with BMI and bile alcohol. The difference was statistically significant (P was 0.025 and 0.005 respectively); the difference in average diastolic blood pressure, the level of aldosterone, total cholesterol level and ejection fraction in patients with aldosterone adenoma was statistically significant (P was 0.022,0.027 and 0.033), and 1 of the 46 aldosterone adenomas were male patients. Combined with moderate obstructive sleep apnea hypopnea syndrome, 14 cases of unilateral adrenal hyperplasia combined with 4 cases of severe obstructive sleep apnea hypopnea syndrome, including 3 men and 1 women. Conclusion: 1) there is a heterozygous mutation of the kcnj5 gene g151r in the adrenal aldosterone adenoma, the mutation of l168r, the mutation of s209t and l102q, and the mutation of the adrenal aldosterone adenoma. The mutation rate was 65.21% (30/46), in which the mutation of s209t and l102q was a newly discovered mutation site, which was different from previous studies, suggesting that the mutation of the kcnj5 gene may be related to the race and region; 2) the mutation of the kcnj5 gene aggravated the clinical symptoms of the patients with APA, in which the g151r mutation was most affected, followed by l102q, l168r and s209t; 3) unilateral adrenal hyperplasia. The patients and aldosterone adenomas have the same KCNJ5 gene G151R heterozygous mutation, L168R mutation, S209T and L102Q mutation, and the mutation rate is 71.43% (10/14), which is higher than the mutation rate of aldosterone adenoma, of which S209T and L102Q are newly discovered mutation sites, the previous literature has not been reported; 4) the clinical phenotype effect of the KCNJ5 gene mutation on the aldosterone adenoma patients. More than unilateral adrenal hyperplasia, G151R mutation was most affected, followed by L102Q, L168R and S209T; 5) there was a same KCNJ5 gene mutation in the patients with unilateral adrenal hyperplasia and aldosterone adenoma. The mechanism still needs further study. 6) whether the mutation of the KCNJ5 gene is related to the occurrence of OSAHS, it still needs to be further studied. Discuss.

【学位授予单位】：新疆医科大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R544.1;R586.24

【相似文献】