【作者】 高宏伟；

【导师】 牛远杰；

【作者基本信息】 天津医科大学 ， 外科学 泌尿外（专业学位）， 2018， 博士

【摘要】 目的:通过合成具有新型高生物兼容性,能够兼容光、声敏一体化的分子探针,以光动力治疗效果作为对照,明确声动力疗法对细胞的杀伤作用,并在活体水平上探讨声动力疗法用于前列腺癌和前列腺增生的治疗。为良性前列腺增生(BPH)、去势抵抗性前列腺癌(CRPC),或晚期前列腺癌等良、恶性前列腺疾病的治疗探索一种高效、组织穿透性深,并兼具良好临床应用前景的新方法。方法:(1)、以人源白蛋白为模板,合成新型白蛋白-原卟啉IX(HSA-PPIX),验证其作为光、声敏一体化的分子探针,表征该探针的各项物理及化学属性,检测并对比HSA-PPIX在不同辐照条件下(激光和超声)的单线态氧产率。(2)、以HSA-PPIX作为光、声敏探针,对PC3细胞和C42细胞(去势抵抗性人前列腺癌细胞)、LNCaP细胞(雄激素依赖性前列腺癌细胞)、MB49细胞(鼠源膀胱癌细胞)、KYSE140细胞(人食管癌细胞)、BPH1细胞(人前列腺增生细胞)分别进行光动力及声动力治疗,以光动力学疗法(PDT)结果作为对照,探讨不同超声辐照对不同细胞系的杀伤作用。(3)、建立前列腺癌种植瘤小鼠模型,瘤内注射PPIX后,进行声动力学疗法(SDT)治疗,比较SDT治疗前后前列腺肿瘤的变化和特点,探讨SDT疗法用于治疗前列腺癌的可行性。(4)、建立前列腺增生大鼠模型,超声监视下对大鼠增生前列腺进行PPIX注射,以体外直接超声辐照的方式对其进行SDT治疗,探讨SDT用于治疗前列腺增生的可行性和治疗效果。结果:(1)、本研究成功制备了超小、水溶性及生物兼容性良好的光、声敏一体化的分子探针HSA-PPIX,通过检测激光和超声处理后单线态氧产率,两种激发态均可产生单线态氧,其中,光激发单线态氧产率为49%,超声激发的单线态氧产率为35%。(2)、以HSA-PPIX作为光、声敏剂,可对不同肿瘤细胞、前列腺癌细胞及正常前列腺增生细胞具有杀伤作用,其中超声辐照2分钟,超声强度为1W/cm~2,距离为0.5cm,最大可杀伤23%肿瘤细胞,与激光(照射时间2分钟,能量为10J/cm~2,照射距离为0.5cm,可杀伤18.7%肿瘤细胞),激光辐照比超声辐照对肿瘤细胞的杀伤力更大,但SDT能在相对较远的距离和阻隔下实现对细胞的杀伤作用,而同条件下,激光的效率明显降低。(3)、本研究成功构建了PC3前列腺癌小鼠皮下种植瘤模型,通过瘤内直接注射PPIX后,以3只荷瘤鼠为一组,与对照组相比,SDT治疗组的肿瘤抑制率为37.1%。(4)、本研究成功构建前列腺增生大鼠模型12只,经体外直接激发注射于前列腺内的PPIX两次后(间隔2周),每5天进行大鼠前列腺B超检查,连续观察大鼠前列腺体积变化,结果显示,大鼠模型的前列腺体积呈线性下降,治疗30天后,大鼠前列腺标本平均体积为0.84ml,重量为0.82g,对大鼠前列腺标本进行病理学检查,结果显示,经过体外直接SDT治疗BPH后,前列腺组织内存在大片状的固缩坏死。结论:(1)、本研究成功合成了新型光、声敏一体化分子探针HSA-PPIX,该探针有效解决了经典光敏剂PPIX的水溶性和生物相融性的问题,结果显示该探针可分别通过激光或超声激发产生单线态氧,通过进一步提高该探针的超声激发单线态氧产率,有望未来实现临床转化。(2)、本研究证实,声动力疗法对前列腺癌的治疗可行性,声动力治疗具有激发距离远、可穿透深部组织等独特优势,与光动力治疗比较,SDT可较好解决PDT治疗中的组织穿透力差,有创光纤导入及光毒性的局限,有望为晚期前列腺癌的治疗提供一种全新、高效、便捷的定性治疗手段。(3)、本研究首次将SDT疗法用于BPH治疗,研究结果证实SDT可有效缩小大鼠前列腺增生组织,且无明显并发症,该方法具有简便定向、易于重复、操作简单等特点,有望未来用于高龄或者身体条件差,不能耐受手术治疗的BPH患者,为前列腺增生提供了一种较为新颖的治疗方法。更多还原

【Abstract】 Objective:To synthesize new high bio-compatible light and sound-sensitive integrated molecular probes by using photodynamic therapy as control,we clarified the killing effect of sonodynamic therapy in vitro,and explored acoustic-dynamic therapy for both prostate cancer and benign prostatic hyperplasia in vivo.And we found a new method for the treatment of prostate diseases such as CRPC and BPH with high efficiency for deep tissue penetration and good clinical application prospects.Methods:（1）Usinghumanalbuminproteinastemplate,anovel albumin-protoporphyrin IX（HSA-PPIX）was synthesized and verified as a molecular probe for integration of light and sound sensitivity,and both the physical properties and chemical properties of the probe were characterized.The singlet oxygen yield of HSA-PPIX under different irradiation conditions（laser and ultrasound）was measured and compared.（2）We used HSA-PPIX as light,acoustic sensitive probe and PC3 cells,C42 cells,LNCaP cells,MB49 cells,KYSE140 cells,and BPH1 cells were used for photodynamic and acoustic therapy.PDT treatment was used as a control to investigate the killing effect of different ultrasound irradiation on different cell lines.3)We established mouse model of prostate cancer xenografts.After intratumoral injection of PPIX,SDT treatment was performed to compare the changes and characteristics of prostate tumors before and after the treatment and to explore the feasibility of SDT therapy for the treatment of prostate cancer.（4）We established 12 rats model for prostatic hyperplasia.Under the ultrasound monitoring,rats with prostatic hyperplasia were injected with PPIX,and the SDT was treated by direct ultrasonic irradiation in vitro to explore the feasibility and therapeutic effect of SDT for the treatment of benign prostatic hyperplasia.Results:（1）In this study,a light,sonic and sensitive molecular probe HSA-PPIX with ultra-small,water-soluble and biocompatible properties was successfully established.By detecting the singlet oxygen yield after laser and sonication treatment,both excited states can produce singlet oxygen,wherein the photoexcited singlet oxygen yield is 49%,and the ultrasonically excited singlet oxygen yield is 35%.2)HSA-PPIX can be used as a light and sound sensitive agent to kill different tumor cells,prostate cancer cells and normal prostatic hyperplasia cells.Ultrasound irradiation for 2 minutes,ultrasonic intensity of 1W/cm^2,distance of 0.5cm,the maximum tumor cells killing rate was 23%,and laser irradiation with 2 minutes irradiation time,energy 10J/cm²,irradiation distance of 0.5cm,the maximum tumor cells killing rate was 18.7%compared with ultrasound irradiation.However,SDT can achieve cell killing effect at relatively long distances and barriers,and under the same conditions,the efficiency of laser is significantly reduced.（3）This study successfully constructed a subcutaneous implanted tumor model of PC3 cells（prostate cancer cells）.In the group of 3 tumor-bearing mice,the tumor inhibition rate was 37.1%in the SDT-treated group compared with the control group by direct injection of PPIX into the tumor.（4）In this study,12 rat models of benign prostatic hyperplasia were successfully constructed.After directly injecting PPIX into the prostate twice（interval 2 weeks）,ultrasound examination for the rat prostate was performed every 5 days,and the prostate volume change was observed continuously.The results showed that the prostate volume of the rat model decreased linearly.After 30 days of treatment,the rat prostate specimen volume was 0.84 ml and the weight was 0.82 g.Pathological examination of rat prostate specimens showed that there was a large piece of pyknosis and necrosis in prostate tissue after direct SDT treatment.Conclusion:（1）In this study,we successfully synthesized a novel optical and acoustic-sensitive integrated molecular probe HSA-PPIX,which effectively solved the problem of water solubility and bio-blending of its classic light and sensitizer PPIX.2)This study confirms the feasibility of acoustic-dynamic therapy for the treatment of CRPC,which HSA the unique advantages of long-distance excitation and deep tissue penetration.3)This study was the first to use SDT therapy for BPH therapy.更多还原