| The number of patients with chronic heart failure (CHF) caused by various reasons is increasing year by year. Those with acute decompe-nsated heart failure (ADHF) caused by acute infection, hemodynamic changes, myocardial toxic substances, etc, have become the main group of hospitalizd patients with heart failure. The main inducement of ADHF is acute infection, the infection of Gram-negative bacteria. The lipopolysa-ccharide (Lps) plays a key role in it. The patients with chronic heart failure can preserve their life depending on drug administration. When acute exac-erbation of heart failure occurs, it is necessary to deliver them to hospital for active treatment and even rescue. The treatment method has changed from the traditional drug treatment with cardiotonic, diuresis and vasodilator to intervention therapy with biological method such as marrow mesenchymal stem cells (MSCs), erythropoietin (EPO) and hepatocyte growth factorn (HGF). The HGF is a powerful myocardium trophic factor, which has the functions of anti-apoptosis, anti-inflammatory factors, anti-fibrosis, and promoting angiogenesis and stem cell homing. The bone marrow derived mesenchymal stem cells (BMSCs), which has a powerful paracrine effect, can secrete a variety of bioactive factors. Under the hypoxia and low-nutrition condition, the growth factors secreted from BMSCs significantly increased, and then the HGF secreted from BMSCs/Ad-HGF conditioned medium (CM) significantly increased by using Ad-HGF transfection techniques. A preliminarily attempt was made to determine protective effect of the early intervention of BMSCs/Ad-HGF-CM on heart function of the patients with ADHF. The experiment steps:(1) Mouse bone marrow mononuclear cells were isolated and put into the5%CO2incubator at37℃for72hours. Subsequently, half volume of fluid was replaced until cell converge rate reached80%. Cellular morphology was observed by microscopy and cell surface-markers was assayed by flow cytometry. The third-generation BMSCs was transfected with Ad-HGF plasmid. The culture medium was removed after48hours and these cells were cultured in the hypoxia and serum-free medium for24hours, and HGF expression were assayed with ELISA. Results showed that, primary BMSCs presented a vortex state before being non-transfected, BMSCs-positive expression of CD29, CD73and sca-1and negative expression of CD45were determined through flow cytometric detection, and the results were consistent with the MSCs surface markers. As while as determined by ELISA, the HGF expression of BMSCs/Ad-HGF conditioned medium (BMSCs/Ad-HGF-CM) was sign-ificantly higher than that of BMSCs-CM (P<0.001).(2)CHF models were constructed by injecting doxorubicin (Dox) into the tail vein and ADHF models were induced by Lps injections. Then BMSCs/Ad-HGF-CM were injected in the same way, to observe whether they can improve heart fun-ction of ADHF mice. Fifty male Kuming mice of with the same age weighing (20±1.0) g were divided into five groups randomly:the Nor, ADHF, HGF, BMSCs-CM and BMSCs/Ad-HGF-CM groups. Mice of ADHF, HGF, BMSCs-CM and BMSCs/Ad-HGF-CM groups were injected with Dox,5mg/kg/week, once per week for consecutive six weeks, with the total dose of30mg-kg-1to construct CHF models. In the7th week, Lps (5mg·kg-1) were injected to construct ADHF models. Then the three treatment groups were given HGF, BMSCs-CM, and BMSCs/Ad-HGF-CM while the Nor group were administered the same amount of physiological saline. The mice of each group were examined with echocardiogram, while plasma BNP/IL-6level was measured, myocardium H.E.stain was performed, Bcl-2and Bax expressions were assayed with the immu- nohistochemical method. Results indicated that compared with the ADHF group, FS%increased (P<0.05), the plasma IL-6level significantly decreased (P<0.01), Bcl-2proteins increased (P<0.05), Bax proteins decreased (P<0.05) and the plasma BNP level significantly decreased (P<0.01) in the BMSCs/Ad-HGF-CM group. Compared with the ADHF group, FS%increased (P<0.05), the IL-6level decreased (P<0.05), Bax expressions decreased (P<0.05), Bcl-2expressions increased (P<0.05) and BNP decreased (P<0.05) in HGF and BMSCs-CM groups. Hema to-xylin and eosin stain showed that BMSCs/Ad-HGF-CM, HGF and BMSCs-CM groups had less inflammatory cell infiltration and less edema in the myocardial interstitium, and the number of myocardial cells more than ADHF group. but there were no significant differences among the three groups. Echocardiographies showed no statistical dif-ferences among the three treatment groups.The experiment results implied that:(1) Through observation of cell morphology, the cell marker assay and green immunofluorescence examination, it was found that BMSCs maintained the initial cell mo-rphology and the special expressions. ELISA showed that HGF expressions were significantly higher in the BMSCs/Ad-HGF culture medium than in the single BMSCs culture medium. Moreover, the paracrine function of BMSCs was enhanced through the conditional culture technique. After the mice of ADHF group were injected HGF, BMSCs-CM and BMSCs/Ad-HGF-CM, inflammatory factors, pro-apoptotic genes and BNP decrease, anti-apoptotic genes increased, and the heart function was improved according to ultrasound examination results. But there were no statistical differences between the BMSCs-CM group and the HGF group. Compared to HGF and BMSCs-CM groups, the inflammatory reaction reduced, the myocardial apoptosis and BNP decreased and the heart function was improved in the BMSCs/Ad-HGF-CM group. However, echocardiographies showed no significant differences among the three groups. H.E.stain indicated that the myocardical tissues had no significant pathological changes. Therefore, it is supposed that HGF, BMSCs-C and BMSCs/Ad-HGF-CM may have positive effects on ADHF in early period by antiinflammatory factors and anti-cardiomyocyte apoptosis, and the effect of BMSCs/Ad-HGF-CM was superior to HGF and BMSCs-CM. |
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