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Chinese Journal of Burns and Wounds logoLink to Chinese Journal of Burns and Wounds
. 2026 May 20;42(5):477–485. [Article in Chinese] doi: 10.3760/cma.j.cn501225-20250628-00284

谷胱甘肽对小鼠皮肤急性光损伤的作用及其机制

Effects of glutathione on acute cutaneous photodamage in mice and its mechanism

Qi Wang 1,2, Ling Zhou 3, Yin Deng 1, Zijin Qiu 1, Xia Wu 1, Chuan Zhao 1,2,*
PMCID: PMC13234706  PMID: 42209186

Highlights

(1) It was demonstrated that glutathione significantly alleviated acute cutaneous photodamage induced by combined irradiation of ultraviolet B and ultraviolet A in mice, with a clear dose-dependent protective effect.

(2) It was also demonstrated that glutathione reduced the inflammatory response in the skin of mice with acute photodamage, inhibited excessive epidermal hyperkeratosis, promoted a more regular arrangement of collagen fibers, and gradually restored the dermal structure to normal.

Keywords: Ultraviolet rays, Glutathione, Skin, Inflammation, Oxidative stress, Photodamage, Wound repair

Abstract

Objective

To investigate the effects of glutathione on acute cutaneous photodamage in mice and its mechanism.

Methods

This study was a group-designed experimental study. Fifteen 8-week-old male C57BL/6 mice were shaved on the dorsal skin and divided into blank control group, model group, low-dose intervention group, medium-dose intervention group, and high-dose intervention group using a random number table, with 3 mice in each group. Mice in model group received daily combined irradiation of ultraviolet B and ultraviolet A on the dorsal skin to cause acute photodamage, followed by intraperitoneal injection of phosphate-buffered saline (PBS). Mice in blank control group were subjected to sham injury without ultraviolet irradiation and received daily intraperitoneal injection of PBS only. Mice in low-dose intervention group, medium-dose intervention group, and high-dose intervention group received daily ultraviolet irradiation as the model group, followed by intraperitoneal injection of 50, 100, and 200 mg/kg of glutathione, respectively. Two hours after the last injection on day 7 post injury (hereinafter referred to as day 7 post injury), the color and morphology of the dorsal skin of mice in each group were observed grossly. Then, dorsal skin tissue was excised for the following assays. Hematoxylin-eosin staining was used to examine the structure of the stratum corneum, epidermis, and dermis, the morphology of appendages (hair follicles, sweat glands, and sebaceous glands), the presence of hemorrhage, inflammatory cell infiltration, and the thickness of epidermis were also measured. Masson staining was used to detect collagen fiber deposition in the skin tissue. Western blotting was used to detect the protein expressions of inflammation-related proteins (interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α (TNF-α), and matrix metalloproteinase 1 (MMP-1)) in the skin tissue.

Results

On day 7 post injury, compared with that in blank control group, the skin of mice in model group showed extensive scaling, redness, swelling, and crusting. Compared with that in model group, the severity of photodamage of the skin of mice in low-dose intervention group, medium-dose intervention group, and high-dose intervention group was alleviated successively. On day 7 post injury, compared with that in blank control group, the skin tissue structure of mice in model group was disorganized, characterized by thickened and detached stratum corneum, increased number and disordered arrangement of epidermal cell layers, dermal edema, abnormal morphology of appendages (hair follicles, sweat glands, sebaceous glands), scattered hemorrhagic foci, and extensive inflammatory cell infiltration. Compared with that in model group, the degree of tissue disorganization in the skin of mice in low-dose intervention group, medium-dose intervention group, and high-dose intervention group was alleviated successively. On day 7 post injury, the epidermal thickness of the skin of mice in model group was (116.4±6.4) μm, which was significantly greater than (20.9±1.6) μm in blank control group (P < 0.05). Compared with that in model group, the epidermal thickness of the skin of mice in medium-dose intervention group and high-dose intervention group ((77.7±5.6) μm and (56.9±0.8) μm, respectively) was significantly decreased (with P values both < 0.05). On day 7 post injury, compared with that in blank control group, the skin tissue of mice in model group showed a significant increase in collagen fiber content and disorganized fiber arrangement, indicating a certain degree of collagen fiber proliferation. Compared with that in model group, the arrangement of collagen fibers in the skin tissue of mice in low-dose intervention group, medium-dose intervention group, and high-dose intervention group became progressively more regular, and the overall tissue structure gradually recovered to normal. On day 7 post injury, compared with those in blank control group, the protein expression levels of IL-1β, IL-6, TNF-α, and MMP-1 in the skin tissue of mice in model group were significantly increased (with P values all < 0.05). Compared with those in model group, the protein expression levels of IL-1β and IL-6 in the skin tissue of mice in low-dose intervention group were significantly decreased (with P values both < 0.05), while the protein expression levels of IL-1β, IL-6, TNF-α, and MMP-1 in the skin tissue of mice in medium-dose intervention group and high-dose intervention group were significantly decreased (with P values all < 0.05).

Conclusions

Glutathione can significantly alleviate acute cutaneous photodamage induced by combined irradiation of ultraviolet B and ultraviolet A in mice, with a clear dose-dependent protective effect, and its mechanism may be related to inhibiting the expression of inflammatory factors, reducing MMP-1-mediated collagen degradation, and improving dermal collagen structure.

Keywords: Ultraviolet rays, Glutathione, Skin, Inflammation, Oxidative stress, Photodamage, Wound repair

紫外线辐射是诱发急性光损伤、光老化和皮肤癌的最常见外部因素，研究者们了解到其潜在分子机制为DNA损伤和活性氧生成增加[1-2]。光老化是一个复杂的持续过程，临床表现为皮肤衰老、萎缩、松弛、色素沉着、毛细血管扩张、粗糙及色斑形成。长波紫外线（ultraviolet A）波长为320~400 nm，可渗透到真皮层，是引起光老化的主要诱因，其特点是导致皮肤形成皱纹、色素沉着和弹性丧失。中波紫外线（ultraviolet B）波长为290~320 nm，作用于皮肤时通过形成6-4环丁烷嘧啶二聚体和嘧啶6-4啶基酮光产物对DNA造成直接损害，从而导致晒伤。长波紫外线和中波紫外线暴露均会增加基底细胞癌、鳞状细胞癌和黑色素瘤的形成风险[3]。近期研究显示，线粒体功能障碍可导致电子传递链异常和产生更多活性氧，导致氧化应激反应。氧化应激反应会进一步激活NOD样受体热蛋白结构域相关蛋白3（NOD-like receptor pyrin domain-containing 3 protein，NLRP3）炎症小体，从而加剧细胞内的炎症反应，并可能导致线粒体DNA损伤[4]，最终使皮肤表皮层增厚、角化异常，真皮层水肿渗出、毛细血管扩张、胶原组织减少、胶原纤维排布混乱等[5]。

谷胱甘肽是一种重要的内源性抗氧化剂，主要分布于细胞质中，具有抗氧化、解毒、调节代谢及保护细胞等多种生物学功能。在紫外线诱导的细胞DNA损伤过程中，线粒体自身无法合成谷胱甘肽，因此必须依赖从细胞质中高效转运谷胱甘肽以维持其氧化还原稳态。还有研究显示，谷胱甘肽的直接前体——γ-谷氨酰半胱氨酸（γ-glutamylcysteine，γGC）在细胞氧化还原调控中可能比谷胱甘肽本身发挥更为关键的作用，尤其在保护线粒体免受活性氧过度损伤方面具有重要功能[6]。

此外，谷胱甘肽可显著减轻中波紫外线辐照引起的细胞毒性，恢复细胞抗氧化防御能力，并抑制MAPK通路的激活。同时，谷胱甘肽处理可降低细胞凋亡率，降低裂解的酪蛋白酶3及聚腺苷二磷酸核糖聚合酶的表达水平[6]。谷胱甘肽还能减少中波紫外线暴露后磷酸化组蛋白H2A组蛋白家族成员X的积累，从而减轻DNA损伤，维持基因组稳定性[7]。本研究团队进一步探讨谷胱甘肽对中波紫外线与长波紫外线联合诱导小鼠皮肤急性光损伤的作用及其机制。

1. 材料与方法

本成组设计实验研究所涉及的动物实验通过重庆市沙坪坝区陈家桥医院（重庆医药高等专科学校附属医院）伦理委员会审批（批号：CJQYY-ECKY-2024016），并遵循该单位和国家有关实验动物管理和使用的规定。

1.1. 动物及主要仪器和试剂来源

15只8周龄健康无特殊病原体级、体重22~26 g雄性C57BL/6小鼠，购自北京斯贝福生物技术有限公司（动物合格证号：110324251101866671），实验动物生产许可证号：SCXK（渝）2022-0005、使用许可证号：SYXK（渝）2022-0009。所有小鼠饲养于无特殊病原体级屏障环境，环境温度约22 ℃，相对湿度（50±10）%，并采用12 h明暗交替光照周期，保证充足食物和饮水。

谷胱甘肽粉剂购自上海皓元生物医药科技有限公司，HE染色试剂盒、Masson染色试剂盒购自北京索莱宝科技有限公司，兔抗小鼠IL-1β单克隆抗体、兔抗小鼠IL-6单克隆抗体、兔抗小鼠TNF-α单克隆抗体、兔抗小鼠基质金属蛋白酶1（matrix metalloproteinase 1，MMP-1）单克隆抗体购自美国CST公司，鼠抗小鼠β肌动蛋白单克隆抗体购自武汉赛维尔生物科技有限公司，辣根过氧化物酶（horseradish peroxidase，HRP）标记的山羊抗兔IgG多克隆抗体、HRP标记的山羊抗鼠IgG多克隆抗体购自武汉爱博泰克生物科技有限公司。

Pannoramic SCANⅡ型病理切片扫描仪购自匈牙利3DHISTECH Kft厂家，Tanon 5200 Multi型多色荧光凝胶成像仪购自上海天能生命科学有限公司，T12 40W+长波紫外线-340高功率增强型紫外灯（波长范围300~420 nm）购于北京医献堂医疗科技有限公司。

1.2. 小鼠分组及大体观察与标本获取

取15只小鼠，剃去背部毛发后按随机数字表法分为空白对照组、模型组、低剂量干预组、中剂量干预组、高剂量干预组，每组3只。除空白对照组外，其余各组小鼠背部皮肤每日接受中波紫外线联合长波紫外线照射2 h，连续7 d，累计剂量为65 J/cm3，以构建急性光损伤模型；空白对照组小鼠不予紫外线照射做假伤处理，即除不打开紫外灯外，其余操作步骤及时间与其他组别相同。每日紫外线照射结束后，空白对照组与模型组小鼠均经腹腔注射150 μL的PBS；低剂量干预组、中剂量干预组、高剂量干预组小鼠则分别经腹腔注射剂量为50、100、200 mg/kg的谷胱甘肽（质量浓度分别为5、10、20 g/L，溶剂为PBS）。伤后第7天末次注射后2 h时（下称伤后7 d），大体观察各组小鼠背部皮肤色泽、形态，然后常规麻醉小鼠并脱颈处死。切取小鼠背部皮肤组织，一部分储存在-80 ℃冰箱，一部分用40 g/L的多聚甲醛固定后进行石蜡包埋，备用。

1.3. 小鼠皮肤组织的病理情况检测

取空白对照组、模型组、低剂量干预组、中剂量干预组、高剂量干预组小鼠皮肤的石蜡组织，制作厚5 μm的切片，然后将切片脱蜡复水。常规行HE染色，经梯度乙醇脱水、二甲苯透明后用中性树胶封片。使用病理切片扫描仪于50倍放大倍数下检测皮肤组织的角质层、表皮层和真皮层结构，毛囊、汗腺、皮脂腺等附属器官的形态，有无出血现象及炎症细胞浸润等情况，并采用ImageJ软件（美国国立卫生研究院）统计表皮厚度。样本数为3。

1.4. 小鼠皮肤组织中的胶原纤维沉积情况检测

取空白对照组、模型组、低剂量干预组、中剂量干预组、高剂量干预组小鼠皮肤的石蜡组织，制作厚度为5 μm的切片，然后将切片进行脱蜡复水。按照Masson染色试剂盒说明书进行操作，使用病理切片扫描仪于50倍放大倍数下检测皮肤组织中胶原纤维沉积情况。

1.5. 小鼠皮肤组织中的炎症相关蛋白的蛋白表达情况检测

取空白对照组、模型组、低剂量干预组、中剂量干预组、高剂量干预组小鼠冰冻皮肤组织，经研磨裂解后提取总蛋白。常规采用蛋白质印迹法检测各炎症相关蛋白的表达。其中一抗为兔抗小鼠IL-1β单克隆抗体（稀释比为1∶1 000）、兔抗小鼠IL-6单克隆抗体（稀释比为1∶1 000）、兔抗小鼠TNF-α单克隆抗体（稀释比为1∶1 000）、兔抗小鼠MMP-1单克隆抗体（稀释比为1∶1 000）、鼠抗小鼠β肌动蛋白单克隆抗体（稀释比为1∶5 000），二抗为HRP标记的山羊抗兔IgG多克隆抗体、HRP标记的山羊抗鼠IgG多克隆抗体（稀释比均为1∶10 000）。采用化学发光成像系统获取图像，采用ImageJ软件分析目的条带灰度值，以β肌动蛋白为内参照计算各炎症相关蛋白的相对表达水平。将空白对照组各蛋白表达水平设为1，计算其他各组相应蛋白的蛋白表达水平。样本数为5。

1.6. 统计学处理

采用SPSS 24.0统计软件对数据进行分析。计量资料数据均符合正态分布，以x ± s表示，组间总体比较采用单因素方差分析，多重比较采用Bonferroni法。所有检验为双侧检验，以P < 0.05为差异有统计学意义。

2. 结果

2.1. 大体观察小鼠皮肤外观

伤后7 d，与空白对照组相比，模型组小鼠皮肤出现大范围皮屑、红肿、痂壳；与模型组相比，低剂量干预组、中剂量干预组、高剂量干预组小鼠皮肤的光损伤程度依次减轻，尤其是高剂量干预组小鼠皮肤症状得到明显改善，仅表现为轻微红肿。见图 1。

图 1.

5组小鼠伤后7 d皮肤外观。1A、1B、1C、1D、1E.分别为空白对照组、模型组、低剂量干预组、中剂量干预组、高剂量干预组，图 1B中小鼠皮肤组织的光损伤程度最严重，图 1C、1D、1E光损伤程度依次减轻，其中图 1E的外观最接近图 1A

Skin apearance of 5 groups of mice on day 7 post injury

注：模型组小鼠背部皮肤每日接受中波紫外线联合长波紫外线照射，然后经腹腔注射磷酸盐缓冲液（PBS）；空白对照组小鼠不照射紫外线致假伤，仅每日经腹腔注射PBS；低剂量干预组、中剂量干预组、高剂量干预组小鼠每日同模型组接受紫外线照射后分别经腹腔注射50、100、200 mg/kg的谷胱甘肽

图 1

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2.2. 小鼠皮肤组织的病理情况

伤后7 d，与空白对照组相比，模型组小鼠皮肤组织结构紊乱，角质层增厚、剥脱，表皮层细胞层数增多、排列紊乱，真皮层水肿，毛囊、汗腺、皮脂腺等附属器官形态异常，可见散在出血灶及大量炎症细胞浸润。与模型组相比，低剂量干预组、中剂量干预组、高剂量干预组小鼠皮肤组织结构紊乱程度依次减轻，表皮层依次趋于规整，真皮层水肿及炎症细胞浸润情况依次减轻。伤后7 d，空白对照组、模型组、低剂量干预组、中剂量干预组、高剂量干预组小鼠皮肤的表皮厚度分别为（20.9±1.6）、（116.4±6.4）、（104.4±8.6）、（77.7±5.6）、（56.9±0.8）μm，总体比较，差异有统计学意义（F=146.626，P < 0.001）。与空白对照组相比，模型组小鼠皮肤的表皮厚度明显增厚（P < 0.001）；与模型组相比，低剂量干预组小鼠皮肤的表皮厚度无明显变化（P=0.230），中剂量干预组、高剂量干预组小鼠皮肤的表皮厚度明显变薄（P值均 < 0.001）。见图 2。

图 2.

5组小鼠伤后7 d皮肤组织病理情况苏木精-伊红×50。2A、2B、2C、2D、2E.分别为空白对照组、模型组、低剂量干预组、中剂量干预组、高剂量干预组，图 2B中的小鼠皮肤组织结构紊乱，角质层增厚、剥脱，表皮层细胞层数增多、排列紊乱，真皮层（箭头所示）水肿，毛囊、汗腺、皮脂腺等附属器官形态异常，可见散在出血灶及大量炎症细胞浸润，图 2C、2D、2E中的小鼠皮肤组织结构紊乱程度依次减轻，表皮层依次趋于规整，真皮层水肿及炎症细胞浸润情况依次减轻

Pathological condition of skin tissue of 5 groups of mice on day 7 post injury

注：模型组小鼠背部皮肤每日接受中波紫外线联合长波紫外线照射，然后经腹腔注射磷酸盐缓冲液（PBS）；空白对照组小鼠不照射紫外线致假伤，仅每日经腹腔注射PBS；低剂量干预组、中剂量干预组、高剂量干预组小鼠每日同模型组接受紫外线照射后分别经腹腔注射50、100、200 mg/kg的谷胱甘肽

图 2

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2.3. 小鼠皮肤组织中的胶原纤维沉积情况

伤后7 d，与空白对照组相比，模型组小鼠皮肤组织中胶原纤维含量显著增加，且纤维排列紊乱，呈现出一定程度的胶原纤维增生现象。与模型组相比，低剂量干预组、中剂量干预组、高剂量干预组小鼠皮肤组织中胶原纤维排列依次趋于规则，整体组织结构逐渐恢复至接近正常皮肤。见图 3。

图 3.

5组小鼠伤后7 d皮肤组织中胶原纤维沉积情况Masson ×50。3A、3B、3C、3D、3E.分别为空白对照组、模型组、低剂量干预组、中剂量干预组、高剂量干预组，图 3B中的小鼠皮肤组织中胶原纤维大量增生、排列疏松紊乱，胶原沉积明显增多，图 3C、3D、3E中小鼠皮肤组织中胶原纤维沉积与排列紊乱程度依次减轻

Collagen fiber deposition in skin tissue of 5 groups of mice on day 7 post injury

注：模型组小鼠背部皮肤每日接受中波紫外线联合长波紫外线照射，然后经腹腔注射磷酸盐缓冲液（PBS）；空白对照组小鼠不照射紫外线致假伤，仅每日经腹腔注射PBS；低剂量干预组、中剂量干预组、高剂量干预组小鼠每日同模型组接受紫外线照射后分别经腹腔注射50、100、200 mg/kg的谷胱甘肽；箭头指示胶原纤维

图 3

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2.4. 小鼠皮肤组织中炎症相关蛋白的蛋白表达

伤后7 d，与空白对照组相比，模型组小鼠皮肤组织中IL-1β、IL-6、TNF-α和MMP-1的蛋白表达水平显著升高（P值均 < 0.001）；与模型组相比，低剂量干预组小鼠皮肤组织中IL-1β、IL-6的蛋白表达水平均明显降低（P值分别为0.020、0.005）而TNF-α、MMP-1的蛋白表达水平变化差异无统计学意义（P值均 > 0.999），中剂量干预组小鼠皮肤组织中IL-1β、IL-6、TNF-α和MMP-1的蛋白表达水平均显著降低（P值分别为 < 0.001、0.002、0.024、0.012），高剂量干预组小鼠皮肤组织中IL-1β、IL-6、TNF-α和MMP-1的蛋白表达水平均显著降低（P值分别为 < 0.001、 < 0.001、 < 0.001、0.002）。见图 4、表 1。

图 4.

蛋白质印迹法检测的5组小鼠伤后7 d皮肤组织中炎症相关蛋白的蛋白表达水平

Protein expression levels of inflammation-related proteins in skin tissue of 5 groups of mice on day 7 post injury detected by Western blotting

注：条带图上方的1、2、3、4、5分别指空白对照组、模型组、低剂量干预组、中剂量干预组、高剂量干预组；模型组小鼠接受中波紫外线联合长波紫外线照射后经腹腔注射磷酸盐缓冲液（PBS）；空白对照组不进行紫外线照射致假伤，仅经腹腔注射PBS；低剂量干预组、中剂量干预组、高剂量干预组小鼠每日同模型组接受紫外线照射后分别经腹腔注射50、100、200 mg/kg的谷胱甘肽

图 4

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表 1.

5组小鼠伤后7 d皮肤组织中炎症相关蛋白的蛋白表达水平比较

Comparison of protein expression levels of inflammation-related proteins in skin tissue of 5 groups of mice on day 7 post injury

组别 样本数 白细胞介素-1β 白细胞介素-6 肿瘤坏死因子-α 基质金属蛋白酶1
注：模型组小鼠背部皮肤每日接受中波紫外线联合长波紫外线照射，然后经腹腔注射磷酸盐缓冲液（PBS）；空白对照组小鼠不照射紫外线致假伤，仅每日经腹腔注射PBS；低剂量干预组、中剂量干预组、高剂量干预组小鼠每日同模型组接受紫外线照射后分别经腹腔注射50、100、200 mg/kg的谷胱甘肽；与空白对照组相比，aP < 0.05；与模型组相比，bP < 0.05
空白对照组 5 1.00±0.00 1.00±0.00 1.00±0.00 1.00±0.00
模型组 5 5.97±0.17a 11.17±1.56a 3.62±0.18a 5.74±1.20a
低剂量干预组 5 4.17±0.44b 5.86±0.64b 3.69±0.40 5.56±1.35
中剂量干预组 5 2.88±1.02b 5.03±2.37b 2.60±0.27b 2.61±0.08b
高剂量干预组 5 2.31±0.38b 1.80±0.24b 1.80±0.47b 1.76±0.63b
F值 37.983 28.727 41.566 19.669
P值 < 0.001 < 0.001 < 0.001 < 0.001
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3. 讨论

研究显示，谷胱甘肽氨基酸前体可显著提高人表皮KC中谷胱甘肽水平，使离体皮肤经紫外线照射后DNA损伤减轻、屏障蛋白表达升高[8]。相关报道显示，谷胱甘肽处理可降低DNA损伤标志物磷酸化组蛋白H2A组蛋白家族成员X表达，相较于抗坏血酸等无解毒作用的抗氧化剂，具有更优的基因修复作用[9]。值得注意的是，紫外线诱导色素沉着的相关靶点可与抗氧化通路协同作用，减轻色素沉着并改善KC光老化，其核心机制涉及氧化应激、DNA损伤、端粒缩短、褪黑素减少及自噬异常。DNA损伤及端粒缩短相关遗传性疾病常伴随皮肤老化与色素异常，提示二者存在密切关联，而谷胱甘肽可显著抑制黑色素合成[10-12]。由此可见，谷胱甘肽可从内部改善皮肤防御系统，减少外界环境对皮肤屏障的累积性损伤[13-14]。

3.1. 谷胱甘肽对小鼠皮肤表皮层的保护

KC为表皮主要细胞成分，约占表皮细胞总数的90%，该类细胞起源于基底层，经增殖、分化后向表层迁移并最终形成角质层，同时合成角蛋白以维持皮肤机械强度。角质层细胞与脂质共同构成皮肤物理屏障，减少水分流失并抵御外界刺激[15]。

中波紫外线可直接被DNA吸收，导致DNA双螺旋结构扭曲；若损伤未通过核苷酸切除修复系统及时修复，可诱发基因突变，促进癌变发生。紫外线还可激活KC中Toll样受体（Toll-like receptor，TLR）及NLRP3炎症小体，释放IL-1β、IL-18等促炎性细胞因子，上调MMP-1、MMP-3表达。此外，紫外线可刺激KC分泌α-黑色素细胞刺激素，激活黑色素细胞酪氨酸酶，促进黑色素合成，长期暴露可导致色素分布不均，形成老年斑等[16-17]。

本研究中，除空白对照组外，其余各组小鼠背部皮肤均出现局部红肿、皮屑及结痂等急性光损伤表现。随谷胱甘肽给药剂量升高，皮肤损伤程度依次减轻、范围缩小，光泽度改善。HE染色显示，模型组小鼠表皮层明显增厚，角质层角化过度伴角化不全，皮下毛细血管扩张充血，皮肤附属器官减少，基底层细胞受损且排列紊乱；随谷胱甘肽给药剂量增加，上述病理改变逐渐减轻。表皮厚度统计结果也验证了上述现象。这表明，谷胱甘肽可抑制紫外线所致表皮层增厚，稳定角化过程，对急性光损伤状态下的皮肤表皮层具有保护作用。

3.2. 谷胱甘肽对小鼠皮肤真皮层的保护

中波紫外线与长波紫外线均可激活MMP，抑制DNA修复途径，加速皮肤老化[18-20]。过量长波紫外线可促进MMP合成与分泌，诱导巨噬细胞、中性粒细胞等炎症细胞浸润，导致胶原纤维及弹力纤维减少、断裂。胶原纤维在维持皮肤强度及弹性中具有重要作用，其降解可致皮肤弹性下降、皱纹加深、松弛及色素沉着，形成典型光老化表现[21-22]。MMP家族中，MMP-1是降解Ⅰ型、Ⅲ型胶原的关键蛋白酶[23]。研究显示，光损伤状态下炎症因子、氧自由基等均可诱导MMP表达升高，导致真皮层胶原降解[24]。

本研究中的Masson染色结果显示，与空白对照组相比，模型组小鼠背部皮肤胶原纤维含量明显升高，排列紊乱，呈现胶原纤维增生表现。随谷胱甘肽给药剂量增加，真皮层胶原纤维排列逐渐接近正常，高剂量干预组小鼠皮肤组织形态与空白对照组最为接近。蛋白质印迹法检测结果也显示，模型组小鼠背部皮肤组织中MMP-1蛋白表达升高，而谷胱甘肽可呈剂量依赖性地降低MMP-1蛋白表达。这提示，谷胱甘肽可通过抑制MMP-1蛋白表达，减轻中波紫外线与长波紫外线所致的真皮层损伤，对急性光损伤状态下的皮肤真皮层发挥保护作用。

3.3. 谷胱甘肽通过减轻炎症反应缓解小鼠皮肤急性光损伤

急性光损伤可诱发明显炎症反应，炎症介质的动态变化是皮肤损伤及修复进程的关键。活性氧所致氧化应激可诱导IL-1β、IL-6、TNF-α等促炎性细胞因子高表达，促进中性粒细胞及巨噬细胞浸润，加剧炎症反应[25-26]。紫外线还可通过诱导活性氧生成及钾离子外流激活NLRP3炎症小体，促进IL-1β成熟与释放[27-29]；同时，高迁移率族蛋白B1等损伤相关分子模式可激活模式识别受体，启动TLR4/核因子κB通路，驱动炎症基因转录[30-32]。

炎症反应可诱发DNA突变，导致皮肤损伤及老化。NLRP3可介导胱天蛋白酶-1裂解IL-1β、IL-18的前体，从而生成成熟的IL-1β及IL-18[33-34]。本实验观察到，中高剂量谷胱甘肽干预后皮肤组织中IL-1β、IL-6、TNF-α及MMP-1的蛋白表达水平均显著降低，其中高剂量干预组下降最为明显。IL-1β为NLRP3炎症小体活化的标志性产物，其表达降低提示谷胱甘肽可通过抑制NLRP3炎症小体激活，减少成熟IL-1β生成。TLR4/核因子κB通路可受促炎性细胞因子、自由基、紫外线等多种因素激活，同时为TNF-α生成的关键启动信号[35-36]。本实验中谷胱甘肽干预后的皮肤组织中TNF-α的蛋白表达显著降低，其中高剂量干预组下降最为显著，提示谷胱甘肽可抑制TLR4/核因子κB通路。炎症反应可诱导MMP-1合成增加，谷胱甘肽可通过恢复氧化还原平衡，降低IL-1β、IL-6、TNF-α及MMP-1等的蛋白表达，抑制TLR4/核因子κB通路激活及NLRP3炎症小体活化，减轻炎症反应。Masson染色结果显示，紫外线照射可导致真皮胶原纤维含量升高且排列紊乱，谷胱甘肽干预可使胶原排列逐渐规整，真皮结构接近正常。上述结果显示，谷胱甘肽可通过抑制炎症反应，发挥对小鼠皮肤急性光损伤的保护作用。

3.4. 局限及小结

本研究存在一定局限性：（1）未进一步探究γGC的抗氧化能力，以及谷胱甘肽与γGC联合应用是否具有更强的保护效果[37]；（2）仅通过大体观察、组织学染色及蛋白质印迹法评估皮肤炎症状态，后续可检测皮肤组织活性氧水平、细胞凋亡情况及ATP含量[38]；（3）可进一步检测炎症相关基因的mRNA水平，明确谷胱甘肽对NLRP3炎症小体的抑制作用[39-40]；（4）铁死亡为铁依赖性、脂质过氧化介导的程序性细胞死亡，与紫外线诱导的皮肤炎症相关，后续可探讨其与皮肤炎症的作用机制，以阐明谷胱甘肽防治皮肤急性光损伤的潜在通路[40]。

综上所述，谷胱甘肽可显著减轻中波紫外线与长波紫外线联合诱导的小鼠皮肤急性光损伤，其作用机制可能与抑制炎症因子表达、减少MMP-1介导的胶原降解、改善真皮胶原结构相关，且保护作用呈明显剂量依赖性。本研究为后续皮肤急性光损伤相关药物研发提供了理论及实验基础。

Funding Statement

重庆市自然科学基金面上项目（2024NSCQ-MSX3514）；重庆医药高等专科学校自然科学基金（ygz2024107）

General Program of Chongqing Natural Science Foundation (2024NSCQ-MSX 3514); Natural Science Foundation of Chongqing Medical and Pharmaceutical College (ygz2024107)

Footnotes

利益冲突  所有作者声明不存在利益冲突

作者贡献声明  王祺：实验设计及操作、论文撰写；周玲、邓银：实验操作、数据整理；邱子津、吴夏：文献查阅、统计学分析；赵川：实验指导、论文修改及经费支持

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