| Diabetes mellitus (Diabetes mellitus, DM) worldwide is a worldwide disease, erectile dysfunction (Erectile dysfunction, ED) is one of the common complications of diabetes mellitus. At present in the treatment of ED first choice of drug treatment, but it is a pity that drug for each ED has a good treatment. Sildenafil on diabetes, after radical prostatectomy and serious illness in patients with ED almost invalid. Because the DM induced by ED (DIED), because of its multiple pathogenic mechanisms, the current treatment of the overall effect is not ideal, the urgent need to study new treatments, in order to obtain satisfactory clinical effect.Endothelial progenitor cells (Endothelial progenitor cells, EPCs) is a vascular endothelial cell (Endothelial cells, ECs) precursor cells. Although EPCs has not expressed the mature ECs phenotype, has not formed the vessel, but EPCs can cycle, proliferate and differentiate into mature ECs. EPCs in vascular repair and production process plays an important role. Scholars home and abroad with the rat as a model, treatment with EPCs ED, that can play a certain effect. But the problem is now in stem cell transplantation therapy has many restrictions, mainly stem cell transplantation efficiency is low, the lack of effective targeting.In recent years, ultrasound microbubble Technology (UTMD) is developping very fast, in the clinical application is becoming more and more widely. Ultrasound microbubble technology with less trauma, reproduced the characteristics of high efficiency, in a clinically more and more. Microbubbles and specific reproduced without the combination, using ultrasound irradiation produces effective biological effect, can be reproduced to the target tissue, this should be the effective clinical validation. From this theory, we designed such a topic, use of ultrasound microbubble destruction mediated targeting of transplanted stem cells EPCs, observation of EPCs in rat penile tissue of expression as well as in type1diabetic rats with erectile dysfunction, so as to DIED therapy provides a new train of thought.Our experimental design is divided into the following four parts: Objective To study the isolation, identification and culture of endothelial progenitor cell method. Method First rat bone marrow MNCs, the cells in culture plates, in cell growth factor induced and cultured for2weeks. Observation of cell morphology change and the appraisal. Results Endothelial progenitor cells grew well, with its typical biological characteristics, through immunohistochemistry and fluorescence method to get the effective identification. Conclusion Cells induced by cultured successfully, we need EPCs, as the next step in the animal test ready. Objective By intraperitoneal injection of streptozotocin (STZ) method to establish the rat model of type1diabetes mellitus DIED. Methods90male SD rats were divided into2groups, model of intraperitoneal injection of65mg/kg STZ; control rats, peritoneal injection with the same amount of physiological saline.4D, docking method for the determination of random blood glucose, a week after the detection of the1. According to10W,100u g/kg measurement, by subcutaneous injection method, apomorphine (APO), observing rat erectile state, thus judging model is successful. Results Model rats with blood glucose increased significantly, compared with control rats compared with statistical significance (P<0.01), penile erection, erection frequency was significantly decreased (P<0.05, P<0.01) by histological observation, model and control group had significant differences in capillary count (P<0.01). Conclusion By intraperitoneal injection of STZ induced diabetic rats, induced by APO test screening, can be successfully obtained in type1diabetic erectile dysfunction (DIED) rat animal model. Objective To prepare an efficient transfer capacity ultrasonic microbubbles, and detection of its physical properties and functions. Methods Through the layer adsorption (layer-by-layer, LbL) made by the method of ultrasound microbubble, detection of microbubbles physical properties and functions. Results Contained in group PLL, load (PLLDNA) and load (PLL+DNA+PLL) group of microbubbles, the morphology, particle size and concentration with blank microbubbles have no significant difference (P>0.05); but the load (PLL+DNA+PLLDNA) microbubbles due to large volume, with the blank comparison P<0.05. Conclusion The method of LbL is simple, can be prepared, reproduced better lipid microbubbles. Objective To explore the feasibility of improving the delivery of endothelial progenitor ceils (EPCs) to cavernosum tissue to treating rat diabetes-induced erectile dysfunction by using ultrasonic microbubble destruction. Methods EPCs were derived from mouse bone marrow in vitro. Fifty-four Sprague-Dawley rats were successfully induced into models of diabetes-induced erectile dysfunction (DIED) and were randomly divided into3groups:Control group, EPCs group, and ultrasound+microbubbles+EPCs (US+MB+EPCs) group. All the rats were sacrificed7days after EPCs implantation. The number of penile erections and erection rate were assayed induced by apomorphine (APO) and the vascular density of3groups was recorded after Hematoxylin-Eosin staining. The EPCs expression was detected by immunohistochemistry (IHC) and the VEGF protein was respectively detected by Western blot. Results Penile erections, erection rate, vascular density and EPCs expression were the best in US+MB+EPCs group (P<0.05). And the expression of VEGF protein in US+MB+EPCs group was higher than that in other groups (P<0.05). Conclusion Combination of ultrasound microbubbles and EPCs can enhance the transfection efficiency and targeting of EPCs effectively, and improve the erectile function of DIED rats.
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