Anti-aging Effects and Mechanisms of Recombinant Human-derived Collagen on Aging Mouse Induced by D-Galactose
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摘要: 为探究重组人源胶原蛋白（recombinant human-derived collagen，RHC）的抗衰老作用及机制，通过300 mg/kg D-半乳糖腹腔注射构建衰老小鼠模型，并采用不同剂量RHC进行干预，研究衰老模型小鼠皮肤透明质酸、胶原蛋白、脂褐素、羟脯氨酸含量、抗氧化酶活性、与表皮细胞增殖分化和胶原蛋白合成相关的关键基因、蛋白的表达量等变化。结果表明，与衰老模型组（SLM）相比，小鼠经过RHC干预后，皮肤的褶皱程度明显减轻，胶原蛋白含量和含水量显著增加（P<0.05），超氧化物歧化酶（Superoxide dismutase，SOD）含量增加，丙二醛（Malondialdehyde，MDA）含量显著减少（P<0.0001）。与SLM组相比，RHC处理组的抗氧化指标，如谷胱甘肽过氧化物酶（GSH-Px）含量显著增加，而褐质素含量、病理评分均有所降低，且不同剂量处理组之间也有一定的差异性。此外，RHC处理组小鼠皮肤中的关键基因（EGFR、TIMP-1、COL1A2、COL3A1、TGF-βⅠ、TGF-βⅡ）的表达水平呈上升趋势， MMP-1、MMP-3 基因和蛋白的表达降低。综上所述，RHC具有良好的抗皮肤衰老作用，其作用机制可能通过抗氧化、以及改善 EGFR、TGF 信号通路，调节 MMPs和TIMPs 动态平衡，促进表皮细胞的增殖分化和胶原蛋白合成从而起到延缓皮肤衰老的作用。Abstract: In order to explore the anti-aging effect and mechanism of recombinant recombinant human-derived collagen (RHC), the aging mouse model was constructed through 300 mg/kg D-galactose intraperitoneal injection, and different doses of RHC were used to study the changes of hyaluronic acid, collagen, lipofuscin, hydroxyproline content, antioxidant enzyme activity, key genes and protein expression related to epidermal cell proliferation and differentiation and collagen synthesis. The results showed that compared with the aging model group (SLM), the intervention groups that was gavaged RHC had significantly less skin folds, higher collagen content (P<0.05) and superoxide dismutase (SOD), while lower malondialdehyde (MDA) content in the skins (P<0.0001). Compared with the SLM group, antioxidant indicators such as glutathione peroxidase (GSH-Px) content were increased in the RHC intervention groups, while lipofuscin content and pathological scores were reduced, and significant effect were also observed. In addition, the expression levels of key genes and protein (EGFR, TIMP-1, COL1A2, COL3A1, TGF-βⅠ, TGF-βⅡ) were increased, while those of MMP-1, MMP-3 in the skin of mouse were decreased in the RHC intervention groups. In conclusion, RHC has a good anti-aging effect on the skin, and its mechanism may be related to increase antioxidant effect, promote the EGFR, TGF pathway, and modulate the expression balance of MMPs and TIMPs, promote the proliferation and differentiation of epidermal cells, and the synthesis of collagen.
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Key words:
- recombinant human-derived collagen /
- D-galactose /
- aging model /
- anti-aging /
- lipofuscin
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图 3 RHC对小鼠皮肤组织抗氧化指标的测定
Figure 3. Determination of antioxidant indexes of mouse skin tissue by RHC
图 6 组织切片病理及评分
注：A. NC组；B. SLM组；C. LRHC组；D. MRHC组；E. HRHC组；F. 对照组和各剂量处理组的病理评分。
Figure 6. Pathology and integral of tissue sections
表 1 实验分组与设计
Table 1. Experimental group and design
编号 组别 受试物 剂量
（mg/kg）给药
方式给药
周期小鼠
数量NC 正常对照组 − − ig 6 周 12只 SLM 衰老模型组 − − ig 6 周 12只 LRHC RHC低剂量 RHC低剂量 50 ig 6 周 12只 MRHC RHC中剂量 RHC中剂量 500 ig 6 周 12只 HRHC RHC高剂量 RHC高剂量 1000 ig 6 周 12只 DC 胶原蛋白干预组 胶原蛋白 1000 ig 6 周 12只 注：ig表示经口灌胃。 
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表 2 皮肤损伤评分标准
Table 2. Score standard of skin injury
皮肤病理学改变 积分 最高分 A.棘层肥厚（a） 轻度（表皮为正常厚度1.5～3倍） 1 3 中度（表皮为正常厚度3～4倍） 2 重度（表皮为正常厚度4倍以上） 3 B.过度角化 （b）颗粒层增厚 1 1 （c）角层增厚 1 1 C.其他表皮改变 （d）颗粒层缺乏 1 1 （e）角化不全 1 1 （f）表皮细胞空泡化或细胞内水肿或基底细胞液化变性 1 1 （g）海绵形成 棘细胞间水肿 1 2 水疱形成 2 D.表皮坏死 （h）表皮坏死 轻度（占表皮切面的1/3以下） 8 15 中度（占表皮切面的1/3～2/3） 10 重度（占表皮切面的2/3以上） 15 E.真皮变化 （i）真皮结缔组织血管扩张充血或水肿 1 1 （j）胶原纤维变性或解离 1 1 （k）真皮炎细胞浸润 轻度 1 3 中度 2 重度 3
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