吸烟对高密度脂蛋白抗氧化功能的影响

发布时间：2018-05-27 13:33

本文选题：吸烟急性期 + 长期吸烟　；参考：《北京协和医学院》2011年硕士论文

【摘要】：背景和目的高密度脂蛋白(high density lipoprotein, HDL)通过促进胆固醇逆转运、抗氧化和抗炎症等功能,在抗动脉粥样硬化中发挥了重要的作用.在临床上,通过检测血清高密度脂蛋白胆固醇(HDL-C),间接反映人体循环血液中HDL的水平.人们很早就发现,HDL-C的水平和冠心病风险负相关,并且将提高HDL-C水平作为抗动脉粥样硬化治疗的靶标之一.然而最近的研究表明,HDL的功能在急性应激或慢性炎症状态等条件下将被削弱甚至发生逆转.于是人们认识到,HDL的功能比水平更重要. 脂质氧化是动脉粥样硬化起始的核心事件,通过诱导泡沫细胞生成和凋亡而形成粥样斑块.吸烟是动脉粥样硬化的独立危险因素.大量研究表明,吸烟能够使低密度脂蛋白胆固醇(LDL-C)的水平升高而使HDL-C水平降低,并且戒烟能够使血脂成分改善.然而,人们对吸烟过程中HDL的功能变化尚不清楚.本研究的目的,即针对既往无吸烟史和有长期吸烟史的健康人群,测定其在单次吸烟前后HDL抗氧化功能的改变,并且尝试探寻其机制. 方法 1.收集20名无吸烟史和22名长期吸烟的健康受试者吸烟前后的血清样本,将血样分成4组：无吸烟史组单次吸烟前、无吸烟史组单次吸烟后、长期吸烟组单次吸烟前、长期吸烟组单次吸烟后,冻存于-80℃待用 2.通过密度梯度超速离心方法分离高密度脂蛋白,BCA法测定其浓度,琼脂糖凝胶电泳鉴定其纯度 3.酶联免疫吸附法(ELISA)检测HDL上淀粉样蛋白A(SAA)的含量 4.将各个受试者的LDL分别通过检测紫外光(波长234nm)下OD值,以评价LDL基础氧化程度；之后加入铜离子氧化,记录氧化曲线及各个参数,以氧化速度达峰时间(t at Vmax)和最大OD值(ODmax)分别评估LDL的氧化易感性和最终氧化程度 5.将无吸烟史组单次吸烟前低密度脂蛋白等比例混合,作为正常态LDL(LDLpool).先将各个受试者的HDL分别通过检测紫外光(波长234nm)下OD值,以评价HDL基础氧化程度;再将其与LDLpoo等比例混合,加入铜离子氧化,记录氧化曲线及各个参数,以迟滞时间(Tlag)和氧化速度达峰时间评估HDL抗氧化性能 6.荧光法测定血清对氧磷酶-1(PON-1)活性结果 1.无吸烟史组和长期吸烟组单次吸烟后心率升高(67.2±13.4对71.5±9.9,p0.05；74.8±10.5对78.4±9.2,p0.05),收缩压明显升高(116.2±13.4对119.8±13.3,p0.01；123.4±2.8对126.2±3.9,p0.01),舒张压明显升高(71.0±8.9对76.8±10.0,p0.01；70.8±8.0对74.2±7.5,0.01). 2.长期吸烟组胆红素,无论是总胆红素或直接胆红素均较无吸烟史组明显降低(15.15±5.42对10.80±4.05,p0.01；5.07±2.20对3.53±1.21,p0.01),亦低于无吸烟史组单次吸烟后(14.37±7.05对10.80±4.05,p0.05；4.84±2.59对3.53±1.21,p0.05). 3.长期吸烟组单次吸烟后尿酸升高(273.7±56.9对323.8±49.4,p0.05). 4.无吸烟史组单次吸烟后,SAA水平明显升高(10.07±4.37对15.83±8.50,p0.01). 5.长期吸烟组LDL基础OD值,明显高于无吸烟史组及其单次吸烟后(0.209±0.025对0.276±0.054,p0.01；0.217±0.052对0.276±0.054,p0.01). 6.长期吸烟组LDL氧化速度达峰时间低于无吸烟史组及其单次吸烟后(52.5±15.2对45.0±8.7,p0.05；53.9±14.9对45.0±8.7,p0.01)；长期吸烟组吸烟后最大OD值升高(0.580±0.162对0.707±0.118,p0.05). 7.无吸烟史组或长期吸烟组单次吸烟后迟滞时间均有所下降(96.0±14.4对88.5±10.8,p0.05；106.0±15.6对89.4±20.3,p0.05),长期吸烟者迟滞时间高于无吸烟史组及其单次吸烟后(96.0±14.4对106.0±15.6,p0.05；89.4±20.3对106.0±15.6,p0.01)；长期吸烟组吸烟前后(下降)差值,相比无吸烟史组增大. 8.长期吸烟组氧化速度达峰时间高于无吸烟史组及其单次吸烟后(135.5±16.7对145.4±14.0,p0.05；128.1±17.7对145.4±14.0,p0.01). 结论 1.单次吸烟后,无吸烟史者和长期吸烟者,其机体均处于急性应激状态. 2.长期吸烟者其机体处于慢性氧化状态,胆红素等内源性抗氧化物质被过度消耗. 3.单次吸烟使无吸烟史者和长期吸烟者HDL抗氧化能力均出现降低. 4.长期吸烟者HDL抗LDL氧化能力,较无吸烟史者增高,但是仍然不能纠正长期吸烟所导致的慢性氧化状态. 5.长期吸烟者在急性应激时,HDL抗氧化性能降低更明显.
[Abstract]:Background and Purpose

High density lipoprotein ( HDL ) plays an important role in anti - atherosclerosis by promoting cholesterol reversal , anti - oxidation and anti - inflammation . It has been found that HDL - C is inversely correlated with coronary heart disease risk by detecting serum high density lipoprotein cholesterol ( HDL - C ) . However , recent studies have shown that the function of HDL is impaired or even reversed in acute stress or chronic inflammatory conditions .

Lipid oxidation is a core event of atherosclerosis . Smoking is an independent risk factor for atherosclerosis . Smoking is an independent risk factor for atherosclerosis . A large number of studies have shown that smoking enables low density lipoprotein cholesterol ( LDL - C ) levels to lower HDL - C levels , and smoking cessation can improve the blood lipid component .

method

1 . Serum samples before and after smoking in 20 healthy subjects without smoking history and 22 long - term smoking were collected , and the blood samples were divided into 4 groups : before the single smoking in the smoking history group , after a single smoking in the smoking history group , after a single smoking in the long - term smoking group , after a single smoking in the long - term smoking group , the long - term smoking group was frozen at - 80.degree . C . for use .

2 . High density lipoprotein was isolated by density gradient hypervelocity centrifugation . The purity of high density lipoprotein was determined by BCA method , and its purity was determined by agarose gel electrophoresis .

3.Enzyme - linked immunosorbent assay ( ELISA ) to detect the content of amyloid A ( SAA ) in HDL

4 . The LDL of each subject was measured by detecting the OD value of ultraviolet light ( wavelength of 254 nm ) , respectively , to evaluate the degree of oxidation of LDL base ;
After the oxidation of copper ions , the oxidation curve and the parameters were recorded , the oxidation susceptibility and final oxidation degree of LDL were assessed by oxidation rate up to peak time ( t at Vmax ) and maximum OD value ( ODmax ) .

5 . The low density lipoprotein was mixed with low density lipoprotein in non - smoking history group as normal low density lipoprotein ( LDLpool ) . HDL of each subject was first measured by detecting the OD value of ultraviolet light ( wavelength of 254 nm ) to evaluate the degree of oxidation of HDL base , and then mixed with LDLpoo and other ratios , then added with copper ion oxidation , recorded oxidation curve and various parameters , and the anti - oxidation performance of HDL was assessed with hysteresis time ( Tlag ) and oxidation speed up to peak time .

6 . Determination of serum paraoxalidase - 1 ( PON - 1 ) activity by fluorimetry

Results

1 . After single smoking , the heart rate was increased ( 67.2 卤 13.4 vs 71.5 卤 9.9 , p < 0.05 ) .
74.8 卤 10.5 ( 78.4 卤 9.2 , p < 0.05 ) , systolic blood pressure was significantly increased ( 116.2 卤 13.4 vs 119.8 卤 13.3 , p0.01 ) ;
123.4 卤 2.8 ( 126.2 卤 3.9 , p0.01 ) and diastolic blood pressure increased significantly ( 71.0 卤 8.9 vs 76.8 卤 10.0 , p0.01 ) ;
70.8 卤 8.0 vs 74.2 卤 7.5 , 0.01 ) .

2 . The bilirubin of long - term smoking group , whether total bilirubin or direct bilirubin , was significantly lower than that in non - smoking history group ( 15.15 卤 5.42 vs 10.80 卤 4.05 , p0.01 ) .
5.07 卤 2.20 vs 3.53 卤 1.21 , p0.01 and 14.37 卤 7.05 to 10.80 卤 4.05 , p0.01 respectively after single smoking in non - smoking history group ( 14.37 卤 7.05 vs 10.80 卤 4.05 , p0.01 ) ;
4.84 卤 2.59 vs 3.53 卤 1.21 , p0.05 ) .

3 . After single smoking in the long - term smoking group , uric acid increased ( 273.7 卤 56.9 vs 323.8 卤 49.4 , p0.01 ) .

4 . After single smoking , the SAA level increased significantly ( 10.07 卤 4.37 vs 15.83 卤 8.50 , p0.01 ) .

5 . The OD value of LDL in long - term smoking group was significantly higher than that of non - smoking history group and its single smoking ( 0.209 卤 0.025 vs 0.276 卤 0.054 , p0.01 ) .
0.217 卤 0.052 vs 0.276 卤 0.054 , p0.01 ) .

6 . LDL oxidation rate of long - term smoking group was lower than that of non - smoking history group and single smoking group ( 52.5 卤 15.2 vs 45.0 卤 8.7 , p < 0.05 ) .
53.9 卤 14.9 ( 45.0 卤 8.7 , p0.01 ) ;
The maximum OD value after smoking in the long - term smoking group increased ( 0.580 卤 0.162 vs 0.707 卤 0.118 , p < 0.05 ) .

7 . After a single smoking group or long - term smoking group , the lag time was decreased ( 96.0 卤 14.4 vs 85.5 卤 10.8 , p < 0.05 ) .
106.0 卤 15.6 ( 89.4 卤 20.3 , p < 0.05 ) , the lag time of long - term smokers was higher than that in the non - smoking history group and its single smoking ( 96.0 卤 14.4 vs 106 . 0 卤 15.6 , p . 05 ) .
89.4 卤 20.3 ( 106.0 卤 15.6 , p0.01 ) ;
The difference of smoking before and after smoking in the long - term smoking group increased compared with the non - smoking history group .

8 . The oxidation rate of long - term smoking group was higher than that of non - smoking history group and its single smoking ( 135.5 卤 16.7 vs 145.4 卤 14.0 , p < 0.05 ) .
128.1 卤 17.7 ( 145.4 卤 14.0 , p0.01 ) .

Conclusion

1 . After a single smoking , there were no smokers and long - term smokers , and all of them were in an acute stress state .

2 . Long - term smokers whose organism is in chronic oxidation state , bilirubin and other endogenous antioxidant substances are overconsumed .

3 . There was a decrease in the anti - oxidation ability of non - smokers and long - term smokers .

4 . Long - term smoker HDL anti - LDL oxidation resistance , higher than non - smoking history , but still cannot correct chronic oxidation state caused by long - term smoking .

5 . In the case of acute stress , the anti - oxidation performance of HDL was lower .
【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

【共引文献】