OSAHS患者及CIH大鼠血清PTX3、HIF-1α水平变化研究

发布时间：2018-05-18 15:22

  本文选题：阻塞性睡眠呼吸暂停低通气综合征 + 慢性间断性低氧　； 参考：《中南大学》2012年硕士论文

【摘要】：目的： 通过观察慢性间断性低氧大鼠模型血清中PTX3、HIF-1α的水平,并观测在抗氧化剂N-乙酰半胱氨酸(N-acetylcysteine, NAC)干预后上述因子的含量变化和中重度OSAHS患者血清中PTX3、HIF-1α的水平及其在CPAP治疗后上述因子含量变化,探讨慢性间断缺氧对OSAHS患者血浆PTX3、HIF-1α的水平影响及可能机制。 方法：实验分为两部分。 第一部分：通过临床多导睡眠监测收集病例,设立正常对照组(A组)和中重度OSAHS(B组)、中重度OSAHS治疗组(C组)3个实验组。记录OSAS组患者的呼吸暂停低通气指数(Apnea Hypopnea Index, AHI)、最低血氧饱和度(Lowest Oxygen Saturation, LS02)以及血氧饱和度低于90%时间占总睡眠时间百分比(T-SaO290%),并于次日晨起睡眠监测结束后采集静脉血,自然静置1小时后用离心机以3000r／min离心10分钟,然后收集标本上层液置于Eppendorf管中。标本置于-70℃低温冰箱保存,待批量以ELISA法分别测定血清中PTX3、HIF-1α浓度。中重度OSAHS治疗组(C组)患者给予CPAP治疗1月后,再次行多导睡眠监测并次日晨起睡眠监测结束后采集静脉血,待批量以ELISA法分别测定血清中PTX3、HIF-1α浓度。 第二部分：24只SD雄性大鼠采用随机排列表法分为慢性间断低氧组(CIH1)、慢性间断低氧+NAC干预组(CIH2)、慢性间断低氧+生理盐水组(CIH3)、和常氧对照组(NC组),每组6只。CIH1、CIH2、CIH3三组大鼠均置同—自制有机玻璃舱内,给予低氧循环即压缩空气和氮气(180s为—循环,舱内氧浓度最低达6%-8%,维持20-25s,然后氧浓度恢复至21%,如此低氧循环7h/d)；NC组大鼠常规饲养即常氧(Fi0221%)。CIH2组给予NAC(800mg.kg-1.d-1)腹腔注射,CIH3组予生理盐水(4m1.kg-1.d-1)腹腔注射。均于实验后第六周处死各组大鼠。取心脏血,静置2小时,予离心机以1000转/分*10分钟分离血清,分装后置—70℃冰箱储存待检。以ELISA法分别测定血清中PTX3、HIF-1α浓度。 结果： 第一部分：中重度OSAHS患者血清PTX3与HIF-1α含量较对照组升高(P0.05),PTX3、HIF-1α水平呈正相关(r=0.281,P0.05),中重度OSAHS患者血清PTX3、HIF-1α水平与呼吸暂停低通气指数(AHI)正相关(r分别为0.311,0.485,P0.05)、与血氧饱和度低于90%时间占总睡眠时间百分比正相关(r分别为0.349,0.444,P0.05),与最低血氧水平负相关(r分别为-0.327,-0.535,P0.05)。经CPAP治疗,中重度OSAHS患者血清PTX3、HIF-1α水平与治疗前比较明显降低(P0.05)。 第二部分：第六周时,CIH1、CIH3组大鼠与NC组比较血清PTX3、HIF-1α显著升高(P0.05)；CIH2组与CIH1、CIH3组比较,血清PTX3、HIF-1α水平降低(P0.05)。 结论： 1.血清PTX3、HIF-1α水平在中重度OSAHS患者中明显升高,与中重度OSAHS患者的AHI及低氧程度正相关,且通过CPAP治疗后其水平明显降低。提示中重度OSAHS患者血清PTX3、HIF-1α水平升高可能与CIH有关,纠正CIH能降低其水平。 2.CIH大鼠血清PTX3、HIF-1α水平升高,进一步证实,慢性间断低氧是导致血清PTX3、HIF-1α水平升高的重要原因。 3.抗氧化剂NAC干预后可降低CIH大鼠血清PTX3、HIF-1α水平,提示CIH致血清PTX3、HIF-1α水平升高的机制可能与氧化应激有关。
[Abstract]:Objective:
By observing the level of PTX3, HIF-1 alpha in the serum of chronic intermittent hypoxia rat model, and observing the changes of the above factors after the intervention of the antioxidant N- acetylcysteine (N-acetylcysteine, NAC) and the level of PTX3, HIF-1 a in the serum of the moderate and severe OSAHS patients and the changes of the above factors after the treatment of CPAP, the chronic discontinuity was discussed. Effects of hypoxia on plasma levels of PTX3 and HIF-1 alpha in patients with OSAHS and possible mechanisms.
Methods: the experiment was divided into two parts.
The first part was to collect cases by clinical polysomnography and set up a normal control group (group A) and moderate to severe OSAHS (group B), and 3 experimental groups (group C) with moderate and severe OSAHS. The apnea hypopnea index (Apnea Hypopnea Index, AHI), the lowest oxygen saturation (Lowest Oxygen), and oxygen saturation were recorded in the OSAS group. The percentage of the total sleep time (T-SaO290%) was lower than 90%, and the venous blood was collected after the end of the morning sleep monitoring. After 1 hours, the centrifuge was centrifuged with 3000r / min for 10 minutes, and then the superfluid was collected in the Eppendorf tube. The specimens were stored at -70 centigrade cryogenic refrigerator, and the blood was determined by ELISA method respectively. In the middle and severe OSAHS treatment group (group C), the patients in the middle and severe OSAHS treatment group (group C) were treated with polysomnography again after January, and the venous blood was collected after the end of the morning sleep monitoring, and the concentration of PTX3 and HIF-1 a in the serum was measured by ELISA in the group of patients with moderate and severe OSAHS treatment (group C) after January.
The second part: 24 SD male rats were divided into chronic intermittent hypoxia group (CIH1), chronic intermittent hypoxia +NAC intervention group (CIH2), chronic intermittent hypoxia + physiological saline group (CIH3), and normal oxygen control group (NC group), each group of 6.CIH1, CIH2, CIH3 three rats were placed in the same homemade plexiglass capsule and given hypoxic circulation or compression. Air and nitrogen (180s - cycle, the lowest oxygen concentration in the cabin reached 6%-8%, maintained 20-25s, then the oxygen concentration was restored to 21%, so oxygen cycle 7h/d); the rats in group NC were given normal oxygen (Fi0221%).CIH2 group given NAC (800mg.kg-1.d-1) intraperitoneal injection, and the CIH3 group was injected with saline (4m1.kg-1.d-1) intraperitoneally. All of them were killed at sixth weeks after the experiment. Group rats. Take the heart blood for 2 hours, give the centrifuge to separate the serum with 1000 / *10 minutes and separate the post - 70 - C refrigerator to be checked. The concentration of PTX3 and HIF-1 a in serum is measured by ELISA method.
Result锛，

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