Chinese Journal of Tissue Engineering Research ›› 2024, Vol. 28 ›› Issue (14): 2173-2179.doi: 10.12307/2024.303
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Wu Xiaodan1, 2, 3, 4, Wang Zhiguo1, 2, 3, 4, Zhan Ying1, 2, 3, 4, Zhang Guoxu1, 2, 3, 4
Received:
2023-02-16
Accepted:
2023-04-12
Online:
2024-05-18
Published:
2023-07-28
Contact:
Zhang Guoxu, Master, Chief physician, General Hospital of Northern Theater Command, Shenyang 110016, Liaoning Province, China
About author:
Wu Xiaodan, Master, Pharmacist in charge, General Hospital of Northern Theater Command, Shenyang 110016, Liaoning Province, China
CLC Number:
Wu Xiaodan, Wang Zhiguo, Zhan Ying, Zhang Guoxu. Establishment of an acute radioactive skin injury model induced by 32P-beta ray radiation and the mechanism of injury[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(14): 2173-2179.
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2.1 实验动物数量分析 选取69只SD大鼠,随机分为30 Gy、45 Gy、60 Gy β射线组(n=21)及对照组(n=6),造模后,30,45和60 Gy β射线组分别于照射后第7,15,30,45,60天各取3只大鼠切取皮肤组织观察损伤情况,每组中6只大鼠继续留观直到实验结束,实验过程中无意外死亡,所有大鼠均进入结果分析。 2.2 不同剂量β射线对SD大鼠体质量的影响 30 Gy、45 Gy以及60 Gy β射线单次局部照射后,各组大鼠饮食饮水正常,体质量呈逐渐增加趋势,各剂量组无统计学差异。与对照组相比,60 Gy剂量组在照射后第30天体质量有所下降(P < 0.05),之后逐渐恢复,见图 1。"
照射后第7天,各组大鼠照射部位均出现干性脱皮,45,60 Gy β射线组开始出现少量充血性红斑。照射后第15天,30 Gy β射线组出现少量片状红斑,45,60 Gy β射线照射部位皮肤红肿,出现显著性片状红斑,结痂,创面扩大。照射后第30天,30 Gy β射线组出现显著性片状红斑,创面结痂;45 Gy β射线组照射野皮肤结痂融合,创面扩大不明显;60 Gy β射线组照射野皮肤结痂加深、融合,有稀薄渗出液。照射后第45天,45 Gy β射线组出现痂皮脱落,60 Gy β射线组出现少量痂皮脱落,创面有收缩趋势,同时可观察到30 Gy β射线组痂皮脱落创面基本恢复。照射后第50天,45 Gy β射线组可见大量新生上皮部分覆盖创面,60 Gy β射线组可见少量痂皮脱落。照射后第60天,45 Gy β射线组大鼠创面基本恢复,未见毛发生长;60 Gy β射线组大鼠创面仍有大部分结痂未脱落,且创面溃疡反复迁延不愈。各组大鼠创面愈合时间及愈合率见表1,2。"
2.4 苏木精-伊红染色和Masson染色观察β射线对大鼠皮肤组织病理学损伤 苏木精-伊红染色和Masson染色结果显示:对照组大鼠皮肤表皮层细胞排列清晰,皮肤附属器结构完整。照射后15 d,30 Gy β射线组表皮细胞、毛囊上皮细胞肿胀,可见到坏死细胞,附属器减少,周围少量炎症细胞浸润;45 Gy以及60 Gy β射线组表皮可见小灶状缺失灶,浅表糜烂加深,真皮内胶原纤维出现水肿、融解、断裂、排列紊乱,其内有散在炎症细胞浸润。照射后30 d,30 Gy β射线组真皮内胶原纤维肿胀,胶原纤维断裂,溶解,排列紊乱;45 Gy β射线组可见上皮细胞变性坏死,真皮层胶原纤维断裂,炎性细胞浸润;60 Gy β射线组真皮层大量断裂,成纤维细胞很少,大量炎性细胞浸润。照射后45 d,30 Gy β射线组有新生上皮覆盖创面,新生肉芽组织明显,胶原纤维增生明显,排列较为整齐;45 Gy β射线组可见大量胶原纤维增生及上皮组织向创面中心爬行,胶原纤维丰富、真皮深层及皮下组织水肿较轻;60 Gy β射线组可见少量胶原纤维增生,真皮层可见大量炎性细胞浸润。照射后60 d,45 Gy β射线组有大量新生上皮覆盖创面,胶原纤维增生明显,排列整齐;60 Gy β射线组真皮层结缔组织有少量增生,皮肤表层结构有明显缺损。见图3,4。"
2.8 免疫组化学检测P53、Bax及Bcl-2蛋白表达水平 照射后第15天,3 组大鼠皮肤组织中P53及Bax蛋白的表达均呈阳性,60 Gy及45 Gy β射线组P53、Bax 蛋白的表达水平高于30 Gy β射线组;照射后第30 天,各组表达均呈增长趋势,60 Gy及45 Gy β射线组大鼠皮肤组织 P53、Bax 蛋白的表达水平达峰值,且60 Gy β射线组高于45 Gy β射线组(P < 0.05),45 Gy β射线组高于30 Gy β射线组(P < 0.05);照射后第45 天,60 Gy β射线组P53、Bax 蛋白的表达仍呈较高水平,30 Gy及45 Gy β射线组P53、Bax 蛋白表达明显减弱(P < 0.05)。照射后第15 天,3 组大鼠皮肤组织中Bcl-2 蛋白的表达均呈阳性;照射第30 天,各组Bcl-2表达均呈下降趋势,60 Gy及45 Gy β射线组大鼠皮肤组织Bcl-2蛋白的表达水平达低峰值,且60 Gy β射线组显著低于30 Gy及45 Gy β射线组(P < 0.05),见图7;照射后第45 天,60 Gy β射线组Bcl-2蛋白的表达仍呈较低水平,30 Gy及45 Gy β射线组P53、Bax 蛋白表达显著增加(P < 0.05)。"
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