發(fā)布時(shí)間：2018-05-01 06:13

  本文選題：肽聚糖 + 氨磷汀�。� 參考：《蘇州大學(xué)》2012年碩士論文

【摘要】：目的探討肽聚糖(peptidoglycan,PGN)與氨磷汀(amifostine,WR2721)對(duì)接受10Gy X射線(IR)全身照射小鼠的救治作用。 方法將C57BL/6J小鼠隨機(jī)分為IR+肽聚糖給藥組、IR+氨磷汀給藥組、IR+肽聚糖+氨磷汀給藥組、IR組、未處理組，于照射前0.5小時(shí)按150mg/kg給予氨磷汀，，照后24小時(shí)給予肽聚糖，除未處理組外其余各組給予10Gy(2Gy/min)X射線全身照射。(1)記錄30天內(nèi)各組小鼠的體重，比較不同處理組間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天分批處死各組小鼠，取血、股骨、腸標(biāo)本，予蘇木素-伊紅染色，并利用D-木糖吸收率、糞便形成計(jì)數(shù)、結(jié)腸炎癥評(píng)分、腸道隱窩干細(xì)胞標(biāo)記計(jì)數(shù)實(shí)驗(yàn)，比較各時(shí)間點(diǎn)各組小鼠腸道功能損傷程度。(3)應(yīng)用CD34標(biāo)記流式細(xì)胞計(jì)數(shù)、骨髓干細(xì)胞集落培養(yǎng)、細(xì)胞因子測(cè)定，反應(yīng)不同處理對(duì)小鼠造血的影響。(4)提取聯(lián)合給藥組、未處理組及IR組小鼠小腸蛋白，應(yīng)用Western-blot法分析各組小鼠腸道中TLR2蛋白水平。(5)將TLR2-/-小鼠隨機(jī)分為聯(lián)合給藥組、未處理組、IR組，重復(fù)上述實(shí)驗(yàn)。 結(jié)果(1)聯(lián)合給藥組C57BL/6J小鼠30天生存率達(dá)到50%，兩組單獨(dú)給藥組都在9天內(nèi)死亡，而IR組則在5天內(nèi)全部死亡，聯(lián)合給藥組與其余各組間區(qū)別有統(tǒng)計(jì)學(xué)差異(p0.01)。(2)各組小鼠照后5天內(nèi)體重均有明顯下降，聯(lián)合給藥組小鼠體重逐漸穩(wěn)定，但30天內(nèi)并未恢復(fù)到照前水平，而其余各組小鼠體重持續(xù)下降，直至死亡。聯(lián)合給藥組C57BL/6J小鼠骨髓與腸道經(jīng)數(shù)星期的恢復(fù)，在照后40天幾乎恢復(fù)到正常水平，而其余各組均顯示明顯且不可恢復(fù)的骨髓、腸道損傷。(3)聯(lián)合給藥組小鼠木糖吸收率保持在60%以上，而其余各組均下降極快(p0.05)；聯(lián)合給藥組小鼠腸道保有部分吸收功能，而其他組糞便形成能力不足(p0.05)；聯(lián)合給藥組小鼠結(jié)腸炎癥程度評(píng)分僅為3.71±0.57，而IR組評(píng)分達(dá)到5.57±1.34(p0.05)；聯(lián)合給藥組小鼠保存較少的腸道隱窩干細(xì)胞，而其余各組到照后第4天僅存極少的隱窩干細(xì)胞。(4)利用CD34標(biāo)記流式細(xì)胞計(jì)數(shù)，可見(jiàn)各照射組均有少量CD34標(biāo)記的造血干細(xì)胞，但照后4天內(nèi)，受照各組小鼠骨髓干細(xì)胞集落培養(yǎng)實(shí)驗(yàn)均無(wú)集落形成，照后一周左右開始，聯(lián)合給藥組小鼠造血逐漸得到恢復(fù)，在照后40天其集落形成數(shù)與未處理組并無(wú)統(tǒng)計(jì)學(xué)差異；但聯(lián)合給藥未有效改善該組小鼠血中細(xì)胞因子水平。(5)TLR2-/-小鼠聯(lián)合給藥組與IR組各指標(biāo)均無(wú)統(tǒng)計(jì)學(xué)差異。 結(jié)論(1)10Gy(2Gy/min)X射線全身照射可造成C57BL/6J與TLR2-/-小鼠骨髓與腸道的嚴(yán)重?fù)p傷，迅速死亡。(2)肽聚糖與氨磷汀聯(lián)合用藥可保護(hù)腸道隱窩干細(xì)胞，小鼠可維持一定的腸道吸收功能，度過(guò)照后損傷最重階段。聯(lián)合用藥組小鼠造血干細(xì)胞有少許保留，為后期造血恢復(fù)奠定了基礎(chǔ)。(3)肽聚糖或氨磷汀的單獨(dú)應(yīng)用均不能對(duì)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 鹵0.57, and the score of IR group was 5.57 鹵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.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R818

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本文編號(hào)：1828073