Wang Xiaoying, Lu Yuxin, Cheng Xiaochen, Zhu Xuefeng, Li Dujuan, Duan Haiying, Hu Shenhui, Xiao Fengjun, Du Li, Zhang Qinglin
College of Chemistry and Environmental Sciences, Hebei University, Baoding, Hebei, China.
Beijing Institute of Radiation Medicine, Beijing, China.
Curr Drug Deliv. 2024;21(5):763-774. doi: 10.2174/1567201820666230508120720.
Most patients who undergo radiotherapy develop radiation skin injury, for which effective treatment is urgently needed. MnSOD defends against reactive oxygen species (ROS) damage and may be valuable for treating radiation-induced injury. Here, we (i) investigated the therapeutic and preventive effects of local multiple-site injections of a plasmid, encoding human MnSOD, on radiation-induced skin injury in rats and (ii) explored the mechanism underlying the protective effects of pMnSOD.
The recombinant plasmid (pMnSOD) was constructed with human cytomegalovirus (CMV) promoter and pUC-ori. The protective effects of pMnSOD against 20-Gy X-ray irradiation were evaluated in human keratinocytes (HaCaT cells) by determining cell viability, ROS levels, and ferroptosisrelated gene expression. In therapeutic treatment, rats received local multiple-site injections of pMnSOD on days 12, 19, and 21 after 40-Gy γ-ray irradiation. In preventive treatment, rats received pMnSOD injections on day -3 pre-irradiation and on day 4 post-irradiation. The skin injuries were evaluated based on the injury score and pathological examination, and ferroptosis-related gene expression was determined.
In irradiated HaCaT cells, pMnSOD transfection resulted in an increased SOD2 expression, reduced intracellular ROS levels, and increased cell viability. Moreover, and expression was significantly upregulated, and erastin-induced ferroptosis was inhibited in HaCaT cells. In the therapeutic and prevention treatment experiments, pMnSOD administration produced local SOD protein expression and evidently promoted the healing of radiation-induced skin injury. In the therapeutic treatment experiments, the injury score in the high-dose pMnSOD group was significantly lower than in the PBS group on day 33 post-irradiation (1.50. 2.80, < 0.05). In the prevention treatment experiments, the skin injury scores were much lower in the pMnSOD administration groups than in the PBS group from day 21 to day 34. , and were upregulated in irradiated skin tissues after pMnSOD treatment, while was downregulated.
The present study provides evidence that the protective effects of MnSOD in irradiated HaCaT cells may be related to the inhibition of ferroptosis. The multi-site injections of pMnSOD had clear therapeutic and preventive effects on radiation-induced skin injury in rats. pMnSOD may have therapeutic value for the treatment of radiation-induced skin injury.
大多数接受放射治疗的患者会发生放射性皮肤损伤，迫切需要有效的治疗方法。锰超氧化物歧化酶（MnSOD）可抵御活性氧（ROS）损伤，可能对治疗放射性损伤具有重要价值。在此，我们（i）研究了局部多点注射编码人MnSOD的质粒对大鼠放射性皮肤损伤的治疗和预防作用，以及（ii）探讨了pMnSOD保护作用的潜在机制。
用人类巨细胞病毒（CMV）启动子和pUC-ori构建重组质粒（pMnSOD）。通过测定细胞活力、ROS水平和铁死亡相关基因表达，评估pMnSOD对20 Gy X射线照射的人角质形成细胞（HaCaT细胞）的保护作用。在治疗性治疗中，大鼠在40 Gy γ射线照射后的第12、19和21天接受pMnSOD局部多点注射。在预防性治疗中，大鼠在照射前第-3天和照射后第4天接受pMnSOD注射。根据损伤评分和病理检查评估皮肤损伤情况，并测定铁死亡相关基因表达。
在受照射的HaCaT细胞中，pMnSOD转染导致SOD2表达增加、细胞内ROS水平降低和细胞活力增加。此外，HaCaT细胞中 和 的表达显著上调，且抑制了erastin诱导的铁死亡。在治疗和预防治疗实验中，给予pMnSOD可产生局部SOD蛋白表达，并明显促进放射性皮肤损伤的愈合。在治疗性治疗实验中，高剂量pMnSOD组在照射后第33天的损伤评分显著低于PBS组（1.50. 2.80， < 0.05）。在预防性治疗实验中，从第21天到第34天，pMnSOD给药组的皮肤损伤评分远低于PBS组。pMnSOD治疗后，受照射皮肤组织中 、 和 上调，而 下调。
本研究提供了证据表明MnSOD在受照射的HaCaT细胞中的保护作用可能与抑制铁死亡有关。pMnSOD的多点注射对大鼠放射性皮肤损伤具有明显的治疗和预防作用。pMnSOD可能对治疗放射性皮肤损伤具有治疗价值。
