钠钾合金消融治疗小鼠胰腺癌

发布时间：2018-01-13 02:29

本文关键词：钠钾合金消融治疗小鼠胰腺癌　出处：《河北医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:癌症是威胁人类健康的一个重大问题,全世界每年大约有1400万新发癌症患者,并有将近800万人死于癌症。找到一个与癌症分期相关的肿瘤治疗方法至关重要。局部消融技术作为一种微创治疗方法被广泛应用于恶性肿瘤治疗。钠钾合金消融的方法打破传统消融的理念,通过钠钾合金与组织中的水分发生化学反应,释放能量。反应物钠和钾作为人体的基本元素,可以完全被吸收。通过注射器将很少量液态的钠钾合金注射到目标组织,局部产生高温,反应同时生成氢氧根和氢气。氢氧根产生类似于电化学反应中正负极的作用,形成碱性环境,对肿瘤组织起到杀伤作用。另一方面,反应产生的气体在肿瘤组织内部发生气穴效应,依靠气体冲击造成细胞的机械性坏死破裂。目前并没有关于钠钾合金对早期及晚期胰腺癌小鼠疗效的报道,此次采用经皮穿刺钠钾合金消融技术,术后精确记录小鼠肿瘤体积和重量的方法,评估钠钾合金消融对小鼠早期肿瘤和晚期肿瘤的治疗效果,为肿瘤临床治疗提供一个新方向。方法:将Panc02胰腺癌细胞制成浓度为1×107/ml的悬浮液,注射到21只雌性C57BL/6小鼠中,每只注射0.2ml,约2×106个肿瘤细胞,建立肿瘤小鼠模型。每两天测量一次肿瘤体积。小鼠随机分为早期肿瘤组(ET组,肿瘤平均体积约为150mm3),晚期肿瘤组(DT组,肿瘤平均体积约为550mm3)和对照组(CO组,肿瘤平均体积约为550mm3)共3组。1早期肿瘤组7只小鼠在接种肿瘤细胞后第18天,向小鼠肿瘤中心注射15uL钠钾合金进行消融治疗,注射过程用热电偶、红外热像仪监测温度变化。肿瘤消融术后,早期肿瘤组随机选取两只小鼠处死,切取消融区的肿瘤组织制成病理切片。2晚期肿瘤组7只小鼠在接种肿瘤细胞后第25天,向小鼠肿瘤中心注射15uL钠钾合金,同时用热电偶、红外热像仪监测。肿瘤消融术后,晚期肿瘤组随机选取两只小鼠处死,切取消融区的肿瘤组织制成病理切片。3对照组7只小鼠不进行钠钾合金消融,在接种肿瘤细胞后第25天,向小鼠肿瘤中心注射15ul生理盐水,同时用热电偶、红外热像仪监测。注射完成后,随机选取两只小鼠处死,切取肿瘤组织制成病理切片。继续观察17天,剩余小鼠全部处死。切取肿瘤组织并称重,计算抑瘤率。结果:1温度变化温度时间曲线揭示了消融区中心温度变化,小鼠机体正常温度为33℃(t0),当碱金属合金注入到肿瘤中心(t=0),温度在两秒内迅速由33℃升高至85℃。最高温度维持不到1秒钟,逐渐减低,直到恢复至起始温度。反应释放大量热能,温度可以保持在42℃以上长达50秒。红外热像仪放置在小鼠上方10cm处,记录小鼠表面皮肤温度。红色区域为升温区域,注射钠钾合金时,升温范围最大,形状类似肿瘤,越靠近注射点温度越高。此时小鼠其他部位体温基本维持不变。随着消融结束,深红颜色区域即温度为40℃的范围,向注射中心逐渐缩小。肿瘤组织温度升高可以维持约50秒。2病理组织切片早期肿瘤组可以看到明显的凝固性坏死,细胞核碎裂溶解,细胞结构完全消失,呈均质红染。晚期肿瘤组细胞发生不完全坏死,界限分明。部分为完整的肿瘤细胞,细胞核大染色深,细胞核异形性显著。部分为不完全坏死的肿瘤细胞。细胞核碎裂,细胞肿胀坏死。对照组的肿瘤细胞结构完整,异形性明显,细胞核数量多颜色深,染色质丰富。3肿瘤体积测量肿瘤细胞在小鼠腹膜内接种后生长良好,肿瘤体积迅速增大。早期肿瘤组在接受碱金属消融时,肿瘤体积约为180mm3,术后肿瘤体积迅速下降,逐渐形成硬痂,肿瘤几乎完全消失,到消融术后第24天,测量硬痂处肿瘤体积约为33mm3。晚期组肿瘤在消融术后迅速缩小,至260mm3。由于晚期肿瘤体积大、形状不规则,消融不完全,肿瘤体积在消融术后10天开始增长,观察结束时,体积约为560mm3。对照组小鼠体积一直不断增大。在接种后0-20天,对照组肿瘤几乎呈对数生长。20天后肿瘤生长逐渐减缓,实验结束时未消融组肿瘤平均体积为3300mm3。4肿瘤抑瘤率在观察结束时,小鼠被全部处死,取出肿瘤测量重量,早期肿瘤组平均瘤重0.21g,消融治疗的抑瘤率为88.5%(P0.05),有统计学意义。晚期肿瘤组平均瘤重0.58g,消融治疗的抑瘤率为67.6%(P0.05),有统计学意义。结论:1微量的钠钾合金可以释放高能热量,集中作用于肿瘤区域,使肿瘤细胞发生明显的凝固性坏死,迅速消融肿瘤。2钠钾合金对早期肿瘤可以达到完全消融;对晚期肿瘤可以减轻小鼠肿瘤负荷,在一定时间内抑制肿瘤生长。
[Abstract]:Objective: cancer is a major threat to human health, there are about 1400 millionnew cancer patients worldwide each year, and nearly 8 million people died of cancer. To find a method for cancer therapy and cancer staging is crucial. Local ablation is a minimally invasive treatment technology is widely used in the treatment of malignant tumors. Methods the sodium potassium alloy ablation to break the traditional concept of ablation, chemical reaction, the sodium potassium alloy and the water release energy. Reaction of sodium and potassium as the basic element of the human body, can be completely absorbed by the injector. Will is a small amount of liquid sodium potassium alloy injection to the target tissue, local high temperature reaction, at the same time generation of hydroxyl and hydroxyl hydrogen. Similar to those produced positive and negative effect of electrochemical reaction, the formation of alkaline environment to the killing effect on tumor tissue. On the other hand, reaction yield The gas generating cavitation effect in tumor tissue, depending on the gas shock caused by mechanical necrotic cell rupture. There is no report about the effect of sodium potassium alloy in early and advanced pancreatic cancer in mice, the sodium potassium alloy using percutaneous ablation techniques, postoperative accurate recording method of mouse tumor volume and weight. On early mouse tumors and advanced tumor ablation treatment effect evaluation of sodium potassium alloy, providing a new direction for the clinical treatment of tumors. Methods: Panc02 pancreatic cancer cells into the concentration of 1 * 107/ml suspension was injected into the 21 female C57BL/6 mice, each injection of 0.2ml, about 2 x 106 tumor cells and establish tumor models in mice were measured every two days. Once the volume of the tumor. The mice were randomly divided into early tumor group (ET group, the average tumor volume is about 150mm3), advanced tumor group (DT group, the average tumor volume is about 550mm3) and The control group (CO group, the average tumor volume of about 550mm3) a total of 3 group.1 group of 7 mice in the early stage of tumor inoculation of tumor cells after eighteenth days of treatment to the tumor ablation center of mice injected with 15uL sodium potassium alloy, thermocouple for injection process, to monitor the temperature change of infrared thermography. After tumor ablation, early tumor group randomly selected two mice were sacrificed, cut the ablation area of tumor tissue made pathologic slices of.2 cancer group of 7 mice inoculated with tumor cells after twenty-fifth days, mice were injected with 15uL alloy to the tumor center of sodium potassium, and thermocouple, thermal infrared imager monitoring. After tumor ablation, advanced cancer were randomly selected in two mice were sacrificed, cut the ablation zone of the tumor tissue sections were prepared for.3 sodium potassium alloy ablation control group of 7 mice in twenty-fifth days after inoculation of tumor cells, to the center of tumor mice were injected with 15ul saline at the same time, with power Occasionally, infrared monitoring. After injection, two randomly selected mice were sacrificed, cut the tumor tissue sections were prepared. To observe 17 days, the remaining mice were harvested and weighed. The tumor tissue, the inhibition rate was calculated. Results: 1 temperature time curve reveals the ablation zone center temperature changes, normal the temperature is 33 DEG C mice (T0), when the alkali metal alloy is injected into the center of tumor (t=0), the temperature rapidly within two seconds from 33 degrees to 85 degrees. The highest temperature rise remain less than 1 seconds, decreased gradually, until the return to the initial temperature. The reaction release large amounts of heat, the temperature can be maintained at 42 DEG C for more than 50 seconds. The infrared thermal imager is placed in the mice at 10cm above the surface skin temperature, were recorded. The red area for heating area, injection of sodium potassium alloy, the maximum temperature range, similar in shape to tumor, the more close to the injection point. The higher the temperature at this time of mice The temperature of other parts unchanged. With the end of ablation, deep red color region that is temperature range of 40 DEG C, gradually reduced to the injection center. Tumor tissue temperature can last about 50 seconds.2 pathological sections of early tumor group can see obvious coagulation necrosis, cell nuclear fragmentation dissolved, cell structure disappeared completely, is homogeneous red staining. Late tumor group occurred incomplete cell necrosis, clear boundaries. As part of complete tumor cells, nuclear staining deep, nuclear atypia significantly. Part of incomplete necrosis of tumor cells. Nuclear fragmentation, cell swelling and necrosis of tumor cells. The control group structure integrity, shaped significantly more than the number of nuclei the color is deep, rich chromatin were inoculated with.3 tumor volume measurement of tumor cells in mice after intraperitoneal growth, tumor volume increases rapidly. The early tumor group in an alkali metal ablation When the tumor volume is about 180mm3, postoperative tumor volume decreased rapidly, gradually formed a crust, the tumor had almost completely disappeared, twenty-fourth days after ablation, measuring hard callus tumor volume is about 33mm3. in the late group tumor shrinks quickly after ablation, due to 260mm3. advanced tumor volume, irregular shape, no ablation totally, the tumor volume growth in the first 10 days after ablation, the end of the observation volume is about 560mm3., the mice of control group has been increasing in volume. 0-20 days after inoculation, the tumor control group was almost logarithmic.20 days after tumor growth gradually slowed down, at the end of the experiment without ablation group the average tumor volume for tumor inhibition rate in 3300mm3.4 at the end of experiment, the mice were killed, remove the measurement of tumor weight, tumor early group average tumor weight 0.21g, ablation rate of tumor was 88.5% (P0.05), was statistically significant. The average tumor weight of 0. advanced cancer group 58G, ablation rate of tumor was 67.6% (P0.05), there was statistical significance. Conclusion: 1 trace amounts of sodium potassium alloy can release heat energy, concentration in the tumor region, the coagulative necrosis of tumor cells significantly, rapid tumor ablation.2 sodium potassium alloy for early tumor can achieve complete ablation of advanced; the tumor can reduce the tumor load, inhibit tumor growth in a certain period of time.

【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.9

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