在本研究中我們發現活體外細胞毒性試驗方面，MTT reduction assay以不同濃度(100、50、10、1ug/mL)之六十六種中草藥與小鼠纖維母細胞株(Mus musculus fibroblast cell line, L929)混合培養24時後，以IC50=100μg/mL為臨界值，予以判別中草藥誘發細胞毒性之輕重程度。其結果顯示五加皮、青蒿、艾葉、北黃耆、白朮、柴胡、肉桂、桂枝、薑黃、莪朮、白薇、香附、麻黃、甘草、女貞子、紫草、厚朴、紫蘇、丹參、五味子、苦參、蒲公英、乾薑等二十三種中草藥之IC50均小於100μg/mL具有細胞毒性。Neutral red uptake (NRU) assay以不同濃度(100、50、10、1ug/mL)之六種中草藥與L929細胞混合培養24時後，以IC50=100μg/mL為臨界值，予以判別中草藥誘發細胞毒性之輕重程度。其結果顯示魚腥草、乾薑等二種中草藥之IC50均小於100μg/mL具有細胞毒性。活體內皮膚原發刺激性試驗方面，patch test結果顯示魚腥草、乾薑等二種中草藥之PII(primary irritation index)分別為0.8±1.1、0.6±0.9，均屬於slight irritation之刺激等級。活體外及活體內試驗相關性方面，Neutral red uptake assay(R=-0.915)判別中草藥毒性及刺激性之相關性是遠高於MTT reduction assay(R=-0.461)，故往後判別中草藥之毒性方式則可先以細胞毒性之Neutral red uptake assay來篩選，以有效及快速地判別中草藥毒性及符合人道之觀點。
The emergence of cosmetics provides us an access to prevention of aging and maintenance of youthfulness. Based on consumers functional demanding in cosmetics as well as the expectation of multiple effects from one single product, such as moisturizing, whitening, anti-aging, and sunscreen, nowadays the ingredients tend to be more complicated and multiple. The adoption of natural materials has inevitably become one of the key factors in cosmetics development. In the previous documents and literature concerning herbs, a lot of Chinese herbal medicines and natural products made of animals possess the functions to prevent or cure skin diseases and sunscreen, as well as an increase of the nutrition of dermal. Moreover, they contribute to mend the deficiencies of skin, such as drying, mottling, acnes, and wrinkles. Also, they can strengthen the elasticity of skin, reduce the keratosis pilaris in dermal and pigmentation, and prevent the sebum function from degenerating. Therefore, the application of Chinese herbal medicines to cosmetics has been viewed as a current in a nature-oriented world.
As a matter of fact, composition or addition of Chinese herbal medicines to cosmetics has taken the lead in modern society, and its wide usage in cosmetology has also been prevailing worldwide. Though there is a lot of written information about Chinese herbal medicines cosmetology in ancient Chinese medicine records, its toxicity has not been closely examined yet. Traditionally, cosmetics safety evaluations are primarily conducted on animals and human bodies. However, animal welfare groups criticize these deeds as inhuman so that researchers decide to substitute cell levels for animals and human bodies. Therefore, in this study, I intend to conduct a cytotoxicity test to evaluate the possible toxicity existing in Chinese herbal medicines with cosmetology effects. By so doing, I wish to establish a complete and fast safety evaluation model system on Chinese herbal medicines.
In this study, we see that in vitro cytotoxicity test, MTT reduction assay mixes sixty-six kinds of Chinese herbal medicines of different concentrations (100, 50, 10, 1μg/mL) with Mus musculus fibroblast cell line (L929) for twenty-four hours, then it marks IC50=100 μg/mL as critical value to determine the degree of cytotoxicity. The result indicates that the values of IC50 in Acanthopanax gracilistylus, Artemisia apiacea, Artemisia argyi, Astragalus mongholic, Atractylodes macroce, Bupleurum chinense, Cinnamomum cassia, Cinnamomum cassia, Curcuma longa, Curcuma zedoaria, Cynanchum atratum, Cyperus rotundus, Ephedra sinica, Glycyrrhiza uralensis, Ligustrum lucidum, Lithospermum erythrorhizon, Magnolia officinals, Perilla frutescens, Salvia miltiorrhiza, Schizandra chinensis, Sophora flavescens, Taraxacum mongolicum and Zingiber officinale are all less than 100 μg/mL, which suggests that they all contain cytotoxicity. On the other hand, Neutral red uptake (NRU) assay mixes six kinds of Chinese herbal medicines of different concentrations (100, 50, 10, 1μg/mL) with L929 cells for twenty-four hours, then it marks IC50=100μg/mL as critical value to determine the degree of cytotoxicity. The outcome shows that the values of IC50 in Houttuynia cordata and Zingiber officinale are both less than 100 μg/mL, which suggests that they both contain cytotoxicity. In vivo dermal primary irritation test, patch test reveals that the PII (primary irritation index) in Houttuynia cordata and Zingiber officinale are 0.8±1.1, 0.6±0.9 respectively, which fall in the slight irritation category. For the correlation of in vitro and in vivo test, Neutral red uptake assay (R=-0.915) presents far more accurately than MTT reduction assay (R=-0.461) in judging the correlation of toxicity and irritation of Chinese herbal medicines. Therefore, we can take Neutral red uptake assay of cytotoxicity as a first priority to judge whether certain Chinese herbal medicines contain toxicity or not, which may well ensure effectiveness and quickness and meet the request of humanity.