护肤生物制品辅料中重组人表皮细胞生长因子抑制剂的筛选

Screening of recombinant human epidermal growth factor inhibitor in subsidiary materials of biological products for skin care

目的筛选护肤生物制品辅料中重组人表皮细胞生长因子(recombinant human epidermal growth factor,rhEGF)抑制剂。方法以BALB/3T3细胞构建模型,采用MTT法对配制好的rhEGF及单组分辅料样品进行生物活性检测,筛选出对rhEGF活性有影响的辅料。通过建立的动物模型,观察rhEGF对皮肤切割伤的促愈合作用,进一步验证筛选出的辅料对rhEGF生物活性的影响。结果经细胞模型筛选的25种辅料中,大分子透明质酸钠和黄原胶显著抑制rh EGF活性。动物模型进一步验证rhEGF具有促兔表皮切割伤愈合的作用,平均愈合时间较生理盐水组提前1～3 d,并呈现良好的剂量效应,随剂量升高,促愈合作用进一步增强,剂量为100μg/m L时,促创面愈合效果最佳。0. 4%黄原胶组和10μg/mL rhEGF+0. 4%黄原胶联合给药组兔创面愈合时间均较生理盐水组晚1 d,黄原胶抑制rhEGF促皮肤切割伤愈合的作用,与细胞模型结果一致。0. 4%透明质酸钠组和10μg/m L rhEGF+0. 4%透明质酸钠联合给药组的兔创面愈合时间均较生理盐水组提前2 d,并且与10μg/mL rhEGF组愈合时间相同,与细胞模型结果不一致。结论黄原胶在细胞和动物模型中均可抑制rhEGF活性,为rhEGF在护肤生物制品中的应用配方提供了理论依据。

Objective To screen recombinant human epidermal growth factor(rh EGF)inhibitor in subsidiary materials of biological products for skin care. Methods BALB/3 T3 cell model was established and used for determination of biological activities of rh EGF and each of its subsidiary materials by MTT assay,based on which the subsidiary materials influencing the activity of rhEGF were screened. The promoting effect of rhEGF on healing of skin cutting wound was observed in established animal model to further verify the effect of screened subsidiary materials on biological activity of rhEGF. Results Of the 25 kinds of subsidiary materials screened with cell model,sodium hyaluronate with a high relative molecular mass and xanthan gum(XAN) significantly inhibited the activity of rhEGF. However,in the further verification in animal model,rhEGF promoted the healing of skin cutting wound. The mean healing time of skin cutting wound of the rabbits treated with rhEGF was 1 ~ 3 d shorter than that with physiological saline as a control. The promoting effect was dose-dependent,which was satisfactory at a dosage of 100 μg/mL. The healing times of rabbits treated with 0. 4% XAN and with 10 μg/mL rhEGF + 0. 4% XAN were 1 d longer than those with physiological saline,indicating that XAN inhibited the promoting effect of rhEGF on healing of skin cutting wound,which was consistent with the test result in cell model. However,the healing times of rabbits treated with 0. 4% sodium hyaluronate and with10 μg/mL rh EGF + 0. 4% sodium hyaluronate were 2 d shorter than that with physiological saline,and consistent with that treated with 10 μg/mL rh EGF,which was inconsistent with the test result in cell model. Conclusion XAN inhibited the activity of rhEGF in both cell and animal models,which provided a theoretical basis for application of rh EGF in biological products for skin care.