| Background：Currently, Chinese economy and society developed rapidly which changedpeople’s living standards and eating hapits, people eat more high-fat orhigh-cholesterol food than before. Changed eating hapits and faster life pace madepressure increased and physical activities reduced. And the process of aging societywhich increased atherosclerosis incidence year by year. Atherosclerosis is importantpotential factors of cardiovascular diseases. Arteriosclerosis ofen affects men over40ages and post-menopausal women. High blood pressure or high cholesterol or diabetsofen accompany arteriosclerosis which affects more brainworkers. Artheriosclerosis’scomplications become a serious menace to the elderly population.Arteriosclerosisobliterans (ASO) is one of atherosclerosis (AS)’s degenerative pathological changesin lower extremity arterial, whose incidence was gradually increased in recent years.ASO has many clinical manifestations such as the skin temperature of affected limbwas reduced and affected limb has the sensation of chill, the affected limb becomeseasy fatigability or intermittent claudication, the affected limb also has rest pain orulcers or gangrene, so that it become life-threatening part. ASO seriously affected thepatients’ life quality. Doctors tried to help ASO patients by drug, surgical therapy,gene therapy and stem cell transplantation therapy, but the treatment effect are verylimited.So far, there are many explanations of ASO’s pathogenesis such as lipidinfiltration doctrine, thrombosis source doctrine, hemodynamic doctrine, the doctrine of medial smooth muscle cell proliferation, intimal injury and receptortheory and so on. Foreign scholars point that ASO is a kind of chronic disease causedby infection, dyslipidemia and inflammatory diseases work together. Currently themost important thing for the vascular surgery is that study on the nosogenesis andASO’s development in order to find the diagnosis and treatment. Scholars try todetecte and diagnose as early as possible so as to have earlier treatment. In this wayASO’s incidence can be reduced and the patients’ life quality can be improved thathas become an urgent problem.Lipoprotein-associated phospholipase A2(LP-PLA2) is one of thephospholipase A2superfamily which is closely related to atherosclerosis, manyscholars pay attention on LP-PLA2in recent years. LP-PLA2is a new inflammationenzyme which has close relationship with cardiovascular diseases, which cancombine with low-density lipoprotein (LDL) in plasma, it also can hydrolyze LDL toproduce oxidized free fatty acids (ox-FA) and lysophosphatidylcholine (Lyso-PC) andthen accelerate the occurrence and development of atherosclerosis. The study showsthat LP-PLA2participate in all aspects of AS’s occurrence and development and itplays an important role in all the processes of atherosclerotic development includeplaque formation developing and rupture. Recently scholars find pentraxin-3(PTX3)which was a kind of protein pentamer composed by381amino acids. PTX3is the keymember of long pentraxin and is a kind of acute phase reactive protein. Recentlyscholars around the world pay more attention on this subject and studies show thatinflammatory reaction play an important role in the occurrence and development ofAS. PTX3closely associated with vulnerability of coronary atherosclerotic plaqueand high expression of PTX3was found in coronary plaque rupture. However, infemoral artery atherosclerotic plaque of patients with ASO, there is no report onwhether the levels of LP-PLA2and PTX3are changed or whether there is relevancybetween them.Objective:To discuss the changes and their significance of LP-PLA2and PTX3’s expressionlevels in human femoral artery atherosclerotic plaques, study on the relationshipbetween them. What’s more, the paper will try to further study on the underlying mechanism of arteriosclerosis obliterans.Materials and Methods:1. Collect lower limb femoral artery endarterectomy femoral artery atheroscleroticplaque specimens31cases from the Second Affiliated Hospital of ZhengzhouUniversity and the First Affiliated Hospital of Henan Traditional Chinese MedicineCollege of the Vascular Surgery Department. All the cases come from the patientsduring September2011to March2013,(18males and13females, the average age:55.45±9.40years). All cases had histories of lower limb ischemia; all of them hadtests such as lower extremity arterial Doppler angiography (CTA) to confirmed lowerextremity arterial sclerosis obliterans and had surgeries. Select specimens of8caseswhich had femoral artery autologous saphenous vein bypass grafting femoral arteryresection after traumatic injuries hip fracture femoral artery,(5males and3females,average age:50.50±8.85years) as control group. Histories of diseases such as portalhypertension, atherosclerosis, and systemic inflammatory diseases cancer and so onare excluded in all the cases. All the patients concluded and signed “Informed consentfor femoral artery atherosclerotic plaque science”. Each cases were separated to twoparts after they were got, one was fixed with10%formaldehyde, the other one wasrapid put in liquid nitrogen and then transferred to a-80℃refrigerator for standbyapplication.2.Stain specimens for HE and divide femoral artery atherosclerotic plaques intostable group (14cases) and unstable group (17cases) according morphologicaldifferences. Take the other8normal femoral artery cases as control group.3.Assay the LP-PLA2protein and PTX3protein expressions of three groupsspecimens by using SP immunohistochemical and analyze the significance ofLP-PLA2and both PTX3protein changes and the relationship between LP-PLA2andboth PTX3.4. Detect the expression of the three groups LP-PLA2mRNA and PTX3mRNA byusing reverse transcription-polymerase chain reaction (RT-PCR) and analyze thesignificance of PTX3mRNA and LP-PLA2mRNA changes and the relationshipbetween PTX3mRNA and LP-PLA2mRNA.5. Statistical methods: Experimental data was analyzed by SPSS17.0software for statistical analysis and correlation results showed asx±s, and rank sum test by usingthe Kruskal-Wallis nonparametric. The data was handled by pairwise comparisonsamong the three groups using Bonferroni method. In order to analyze LP-PLA2protein and PTX3protein expressions of atherosclerotic plaque group as well asLP-PLA2mRNA and PTX3mRNA expressions correlation by using Spearman. Testlevel α=0.05, with p<0.05was considered statistically significant.Results:1. Immunohistochemical results:1.1LP-PLA2protein plaques are highly expressed in tissues, weak expression inthe control group. LP-PLA2protein expression showed brown particles, mainly in thecytoplasm of inflammatory cells. Differences of three groups LP-PLA2proteinexpressions were statistically significant (χ2=32.554, P=0.000<0.001). Stableplaque group (2.399±0.853) has higher LP-PLA2protein expression than controlgroup (1.043±0.061)(Z=2.191> Z0.017=2.1, P <0.05/3=0.017), Unstable plaquegroup (6.142±0.472) also has higher LP-PLA2protein expression (1.043±0.061)(Z=5.418> Z0.017=2.1, P <0.017) than control group, And unstable plaque group (6.142±0.472) has higher than the stable plaque group (2.399±0.853)(Z=3.736> Z0.017=2.1, P <0.017).1.2. PTX3proteins are highly expressed in plaque tissue; its expression is weakin the control group. PTX3protein brown particles were mainly distributed in thecytoplasm of inflammatory cells. The differences of PTX3protein expression in thethree groups was statistically significant (χ2=32.792P=0.000<0.001). Stableplaque group (3.081±1.191) higher in PTX3protein expression (1.057±0.434)(Z=2.177> Z0.017=2.1, P <0.017), Unstable plaque group (9.107±1.131) PTX3proteinexpression was also higher (1.057±0.434)(Z=5.421> Z0.017=2.1, P <0.017),And unstable plaque group (9.107±1.131) was higher than the stable plaque group(3.081±1.191)(Z=3.768> Z0.017=2.1, P <0.017).1.3. Spearman correlation analysis of plaque tissue expression of LP-PLA2protein and PTX3protein showed a positive correlation between them（rSstablegroup=0.959, r0.05,14=0.538，p<0.05；rSunstable group=0.998, r0.05,17=0.485, p<0.05），Inthe control group, no correlation between the two proteins （|rScontrol group|=|-0.236|, r0.05,8=0.738, p>0.05）.2. RT-PCR results2.1Differences in the expressions of LP-PLA2mRNA three groups werestatistically significant（χ2=31.973，P=0.000<0.001）, LP-PLA2mRNA highestexpression of unstable plaques（0.698±0.053）is higher than the stable plaque group（0.304±0.103）（Z=3.656>Z0.017=2.1， P<0.017）， Stable expression inLP-PLA2mRNA amount (0.304±0.103) is higher than plaque group (0.102±0.037)（Z=2.262>Z0.017=2.1，P <0.017）.2.2Differences in the expression of PTX3mRNA three groups were statisticallysignificant（χ2=32.098，P=0.000<0.001），the highest expression of PTX3mRNAin the unstable plaque group（0.773±0.129），Higher than the stable plaque group(0.358±0.108)（Z=3.673>Z0.017=2.1，P <0.017），Stable plaque group (0.358±0.108) expression levels in PTX3mRNA is higher than the control group (0.126±0.006)（Z=2.218>Z0.017=2.1，P <0.017）.2.3Correlation analysis showed that the expression of plaque tissueLP-PLA2mRNA positively correlated with PTX3mRNA（rS stable group=0.995, r0.05,14=0.538，p<0.05； rS unstable group=0.963, r0.05,17=0.485，p<0.05）, no correlationbetween the expressions in the control group （|rS control group|=|-0.561|, r0.05,8=0.738，p>0.05）.Conclusion:1.The expressions of LP-PLA2and PTX3in human femoral arteryatherosclerotic plaque tissues were up-regulation.2. The high expressions of LP-PLA2and PTX3may be associated with humanfemoral artery atherosclerotic plaque instability.3. LP-PLA2and PTX3may have a synergistic effect in instability of thepathogenesis in human femoral artery atherosclerotic plaque organization. |
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