肽聚糖与氨磷汀对10Gy全身照射小鼠的协同救治作用

发布时间：2018-05-01 06:13

本文选题：肽聚糖 + 氨磷汀　；参考：《苏州大学》2012年硕士论文

【摘要】：目的探讨肽聚糖(peptidoglycan,PGN)与氨磷汀(amifostine,WR2721)对接受10Gy X射线(IR)全身照射小鼠的救治作用。方法将C57BL/6J小鼠随机分为IR+肽聚糖给药组、IR+氨磷汀给药组、IR+肽聚糖+氨磷汀给药组、IR组、未处理组，于照射前0.5小时按150mg/kg给予氨磷汀，，照后24小时给予肽聚糖，除未处理组外其余各组给予10Gy(2Gy/min)X射线全身照射。(1)记录30天内各组小鼠的体重，比较不同处理组间30天生存率。(2)于照后1.25天、第4天、第9天、第16天、第25天、第40天分批处死各组小鼠，取血、股骨、肠标本，予苏木素-伊红染色，并利用D-木糖吸收率、粪便形成计数、结肠炎症评分、肠道隐窝干细胞标记计数实验，比较各时间点各组小鼠肠道功能损伤程度。(3)应用CD34标记流式细胞计数、骨髓干细胞集落培养、细胞因子测定，反应不同处理对小鼠造血的影响。(4)提取联合给药组、未处理组及IR组小鼠小肠蛋白，应用Western-blot法分析各组小鼠肠道中TLR2蛋白水平。(5)将TLR2-/-小鼠随机分为联合给药组、未处理组、IR组，重复上述实验。结果(1)联合给药组C57BL/6J小鼠30天生存率达到50%，两组单独给药组都在9天内死亡，而IR组则在5天内全部死亡，联合给药组与其余各组间区别有统计学差异(p0.01)。(2)各组小鼠照后5天内体重均有明显下降，联合给药组小鼠体重逐渐稳定，但30天内并未恢复到照前水平，而其余各组小鼠体重持续下降，直至死亡。联合给药组C57BL/6J小鼠骨髓与肠道经数星期的恢复，在照后40天几乎恢复到正常水平，而其余各组均显示明显且不可恢复的骨髓、肠道损伤。(3)联合给药组小鼠木糖吸收率保持在60%以上，而其余各组均下降极快(p0.05)；联合给药组小鼠肠道保有部分吸收功能，而其他组粪便形成能力不足(p0.05)；联合给药组小鼠结肠炎症程度评分仅为3.71±0.57，而IR组评分达到5.57±1.34(p0.05)；联合给药组小鼠保存较少的肠道隐窝干细胞，而其余各组到照后第4天仅存极少的隐窝干细胞。(4)利用CD34标记流式细胞计数，可见各照射组均有少量CD34标记的造血干细胞，但照后4天内，受照各组小鼠骨髓干细胞集落培养实验均无集落形成，照后一周左右开始，联合给药组小鼠造血逐渐得到恢复，在照后40天其集落形成数与未处理组并无统计学差异；但联合给药未有效改善该组小鼠血中细胞因子水平。(5)TLR2-/-小鼠联合给药组与IR组各指标均无统计学差异。结论(1)10Gy(2Gy/min)X射线全身照射可造成C57BL/6J与TLR2-/-小鼠骨髓与肠道的严重损伤，迅速死亡。(2)肽聚糖与氨磷汀联合用药可保护肠道隐窝干细胞，小鼠可维持一定的肠道吸收功能，度过照后损伤最重阶段。联合用药组小鼠造血干细胞有少许保留，为后期造血恢复奠定了基础。(3)肽聚糖或氨磷汀的单独应用均不能对受照小鼠起到有效的救治作用。
[Abstract]:Objective to investigate the therapeutic effects of peptidoglycann (PGNN) and amphotin amifostine (WR2721) on mice exposed to 10Gy X ray irradiation. Methods C57BL/6J mice were randomly divided into IR peptidoglycan group and IR aminophosphastine group. The untreated group was treated with 150mg/kg 0.5 hours before irradiation and peptidoglycan 24 hours after irradiation. The body weight of the mice in each group was recorded during the 30-day period except the untreated group. The 30-day survival rate was compared between the different treatment groups. The survival rate was 1.25 days, 4 days, 9 days, 16 days, 25 days after exposure. On the 40th day, the mice were killed in batches, blood, femur and intestine were collected and stained with hematoxylin and eosin, and the D-xylose absorption rate, fecal formation count, colitis score and intestinal crypt stem cell labeling test were used. To compare the degree of intestinal dysfunction in each group with CD34 labeled flow cytometry, bone marrow stem cell colony culture, cytokine determination, and the effects of different treatments on hematopoiesis in mice. Small intestine protein of untreated group and IR group were analyzed by Western-blot method. TLR2-r-mices were randomly divided into two groups: untreated group and untreated group, IR group. The above experiment was repeated. Results 1) the 30-day survival rate of the C57BL/6J mice in the combined administration group was 50. The two groups died within 9 days, while the IR group died within 5 days. There was significant difference between the combined administration group and the other groups. (p0.01) the body weight of the mice in the combined administration group decreased significantly within 5 days after irradiation, but the body weight of the mice in the combined administration group gradually stabilized, but did not return to the pre-irradiation level within 30 days. The rest of the mice continued to lose weight until they died. The bone marrow and intestinal tract of C57BL/6J mice in the combined administration group recovered almost to normal level 40 days after irradiation, while the other groups showed obvious and unrecoverable bone marrow. The xylose absorption rate of mice in the combined administration group was above 60%, while the other groups decreased rapidly (p0.05), and the intestinal absorption function of the mice in the combined administration group was partially retained. However, the fecal formation ability of other groups was insufficient (p 0.05), the colitis degree of mice in combined administration group was only 3.71 卤0.57, and the score of IR group was 5.57 卤1.34 p0.05.The mice in combined administration group had fewer intestinal crypt stem cells. On the 4th day after irradiation, there were only a few crypt stem cells in the other groups. (4) CD34 labeled flow cytometry was used to count the hematopoietic stem cells, and a small amount of CD34 labeled hematopoietic stem cells were found in each irradiation group, but within 4 days after irradiation, there were only a few hematopoietic stem cells in each group. There was no colony formation in bone marrow stem cell colony culture of irradiated mice. The hematopoiesis of mice in combined administration group was gradually recovered from about one week after irradiation, and there was no statistical difference between the colony formation number and untreated group at 40 days after irradiation. However, the combined administration did not improve the level of cytokines in the blood of mice. There was no significant difference between the two groups. Conclusion whole body irradiation with 10 Gy / min X ray can cause serious damage in bone marrow and intestine of C57BL/6J and TLR2-r-mice. The combination of peptidoglycan and amphotin can protect intestinal crypt stem cells and maintain certain intestinal absorption function in mice. Pass the most severe damage after irradiation. In combination group, hematopoietic stem cells were reserved, which laid a foundation for hemopoietic recovery in late stage.) Peptidoglycan or amphostatin alone could not effectively cure irradiated mice.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R818

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