| Prostatitis is one of the most common urinary system diseases in men,especially in men over the age of 40,which has a serious impact on men's physical and mental health,Prostatitis includes acute prostatitis and chronic prostatitis,and most acute prostatitis eventually transforms into chronic prostatitis,Compared with acute prostatitis,chronic prostatitis has a longer duration,more difficult to cure,therefore,chronic prostatitis has a greater impact on male,it's urgent for us to develop safe and effective method of treatment of chronic prostatitis,At present,antibiotics is the main clinical treatment method in the treatment of chronic prostatitis,but there are many problems in the treatment of antibiotics,including systemic side effects,drug resistance,untargeted treatment lead to a poor effect and so on,Therefore,people are always looking for other ways to treat chronic prostatitis.Studies have shown that mesenchymal stem cells(MSCs)can be specifically targeted to inflammatory tissues and tumor tissues,which is related to the expression of their surface-specific antigens,In addition,mesenchymal stem cells can secrete some anti-inflammatory factors and have certain anti-inflammatory effect,Curcumin(Cur)is a kind of natural polyphenols extracted from curcumin,It has been proved that curcumin has many effects,including anti-oxidation,anti-inflammatory,antibacterial and anti-cancer,Curcumin itself is poorly water-soluble,it's seriously affecting its clinical application,in recent years,the use of some synthetic materials wrapped curcumin greatly improved its water solubility and improve bioavailability,Studies have shown that mesenchymal stem cells have a certain anti-inflammatory effect on chronic bacterial prostatitis,but it require multiple treatments and a large number of cells,Therefore,we envisaged if combining mesenchymal stem cells with curcumin could improve the therapeutic effect on chronic prostatitis.The biggest obstacle to the treatment of prostatitis is the blood-prostate barrier(BPB),a natural biological barrier between blood vessels and prostate tissues made up of vascular endothelial cells and tight junction proteins,BPB prevents harmful substances from entering,at the same time,it also prevented the blood from entering the tissues with drugs for treatment.Some experiments have proved that ultrasound mediated by micro bubbles has certain effect on the opening of blood-prostate barrier.Based on a large number of literatures and achievements,the chronic bacterial prostatitis was used as the inflammatory model,curcumin,bone marrow mesenchymal stem cells,ultrasound are combined to explore the anti-inflammatory effect of chronic prostatitis,Not only the inflammatory tendency and anti-inflammatory effect of mesenchymal stem cells,curcumin anti-inflammatory effect,but also the role of ultrasound opening BPB,In the first part,the mesenchymal stem cells of rat bone marrow were isolated by density centrifugation and identified,Secondly,the mesenchymal stem cells were detected by MTT,Flow cytometry and fluorescence microscopy were used to screen the best parameters for constructing curcumin carrying mesenchymal stem cells,In the second part,the rat model of chronic bacterial prostatitis was established by direct injection of Escherichia coli,In the third part,the tendency of mesenchymal stem cells to prostatic inflammatory tissue was verified by in vivo imaging of small animals,Secondly,we examined whether ultrasound can promote the enrichment of mesenchymal stem cells in prostate tissue,and whether this enrichment is related to the opening of BPB,The levels of inflammatory cytokines were observed to evaluate the anti-inflammatory effects of different treatments on chronic prostatitis,followed by changes in body weight,The security of different treatments was evaluated by observing the appearance and tissue sections of the main organs,The main results of the experiments were as follows:1.Density gradient centrifugation could successfully isolate high purity of bone marrow mesenchymal stem cells,which have a high expression of CD 90 and low expression of CD 45.2.MTT assay showed that curcumin-loaded micelles were less toxic to mesenchymal stem cells than curcumin alone,so curcumin-containing micelles were chosen to construct curcumin-loaded mesenchymal stem cells.3.Flow cytometry and fluorescence microscopy screened out the best parameters and dosing mode of constructing curcumin-loaded mesenchymal stem cells:the concentration of is 100 ?g/mL,incubation time 4 h,Change the supernatant that containing curcumin 100 ?g/mL at 4 hours and reincubate 4 h.4.200 g-220 g of SD male rat prostate tissue directly is injected 100 ?L concentration of 1 × 109cfu/mL of E.coli can successfully establish rat chronic bacterial prostatitis model,prostate tissue inflammatory factor was significantly increased,have a large number of inflammatory cell infiltration,and tissue obvious swelling.5.Live animal imaging showed that bone marrow mesenchymal stem cells showed a significant tendency toward prostatitis,In addition,microbubble-mediated ultrasound can significantly promote the enrichment of mesenchymal stem cells in rat prostate tissue,and this promotion may be related to the opening of BPB.6.The results of anti-inflammatory effect evaluation showed that compared with microbubble mediated ultrasound treatment and Curcumin loaded bone marrow mesenchymal stem cells,microbubble mediated ultrasound combined with Mesenchymal Stem cells or microbubble mediated ultrasound combined with mesenchymal stem cells loaded with curcumin have certain anti-inflammatory effect on CBP,and MUSMMC treatment has obvious anti-inflammatory effect,the treatment effect is similar to normal rats.7.The results of safety evaluation showed that compared with normal rats,the body weight of other treatment groups changed normally,the appearance of the main organs was smooth,the color is normal,and the histological sections show the normal morphology and no lesions,all the treatments are relatively safe for SD rats.In conclusion,the rat bone marrow-derived mesenchymal stem cells isolated from SD rats in this experiment can autonomously tend to the inflammatory prostate site of the CBP model,and microbubble-mediated ultrasound can promote the accumulation of mesenchymal stem cells in prostate tissue to exert anti-inflammatory effects,In addition,mesenchymal stem cells loaded with anti-inflammatory drug curcumin,its anti-inflammatory effect has been greatly improved,Clinically,These experimental results could provide new ideas and methods for the treatment of chronic bacterial prostatitis. |
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