| Objective: Early in vitro cell culture experiments found that lipoprotein (a)[LP (a)]impair endothelial progenitor cells (EPCs) biological function. The survival,migration, adhesion, into vessel-like structures, colony formation of EPCs is damagedafter a certain concentration of LP (a) treatment, especially in the300mg/mLconcentration. LP (a) impairs EPCs biological functions are the most significantbelow this concentration, and has obvious concentration effect. High LP (a)hyperlipidemia is considered to be an independent risk factor for atherosclerosis, butaccording to the in vitro cell experimental results there is no reported about the highLP (a) hyperlipidemia patient [HLPCAD] monitoring the EPCs function. The high LP(a) hyperlipidemia might lead to the patient EPCs dysfunction; hence develop high LP(a) hyperlipidemia patient EPCs functional analysis is necessary. This study intends tocollect HLP (a) CAD and low-LP (a) coronary heart disease [LLPCAD] Cardiologycoronary heart disease blood from clinical, separating its circulation EPCs, in vitroanalysis of the EPCs function to provide an experimental basis for the treatment ofHLPCAD. Important regulations as the miRs play on EPCs function, this studyintends to analyze HLPCAD and LLPCAD EPCs expression spectrum of EPCsdifferences, hence identify some miRs that play promote/inhibit function of EPCs, tofind some endogenous small molecules regulation EPCs function.Methods: We get HLPCAD23cases with people there was no significant differencein age, gender, body mass index, smoking, blood pressure, blood sugar, blood lipids[LP (a) except], creatinine, uric acid, glomerular filtration rate, drug use etc after thescreening, and LLPCAD27cases, venous blood5mL/times, differential adherenceisolated EPCs, get CD34+/AC133+/VEGFR-2EPCs and counted byimmunofluorescence, ac-LDL-DiI swallowed and lectin binding Identification. MTTassay EPCs survival and proliferation, Modified Boyden chamber migration test,Gelatin slide method to measure the adhesion, Observed and counted on a single cell hybridoma clones dish, measuring tubular structures formed on Matrigel matrix length.Extracting total RNA, refer to the instruction to use miRNeasy mini kit (QIAGEN)miRs expression spectrum analysis.Results: The number of circulating EPCs in HLPCAD patients is significantly than inLLPCAD patients (109.4±13.89vs384.0±37.07, P=0.0023); MTT analysis resultsshow that the OD value of LLPCAD group is0.77±0.057, and HLPCAD OD valueis0.23±0.045(P=0.0018), this showed HLP (a) CAD patients EPCs was of poorliving conditions, it is further reflected in the apoptosis detection. HLPCAD EPCsapoptosis were significantly higher than LLPCAD group (4.1±0.81vs14.9±3.31, P=0.035). HLPCAD patients and LLPCAD patients EPCs in the adhesion (25.3±4.63vs78.6±6.88, P=0.0030), migration (22.0±2.65vs56.0±4.93, P=0.0037),(2.4±0.41vs11.0±1.37, P=0.0003), tubular structure formation (7.4±1.24vs33.3±2.62, P=0.0001) were significantly impaired. Chip shows miRs expression changesrelated to EPCs (greater than1.5times), miR-126expression of HLPCAD groupEPCs decreased nearly10times, miR-221/222expression increased2.3times, what’sinteresting is miR-34a rise but not declined about4times. However on the expressionof the EPCs miR-34a is concerned, still have large gap with miR-126(1265.5vs430.5). Fluorescence quantitative PCR detection of EPC miR-126expression showedHLPCAD group EPCs miR-126expression levels were significantly lower than theLLPCAD (125.0±22.07vs25.8±6.49, P=0.0125, n=3).Conclusions:①HLPCAD EPCs function is significantly impaired compared withLLPCAD EPCs. The survival ability, adhesion, migration, colony forming ability andblood vessel-like structures forming ability of EPCs is reduced, but a significantincrease in the number of apoptotic cells.②HLPCAD miR-126expression level was significantly lower than the LLPCAD... |
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