Suzuki Y, Nomura J, Koyama J, Horii I
Shiseido Research Center, Yokohama, Japan.
Arch Dermatol Res. 1994;286(5):249-53. doi: 10.1007/BF00387596.
The effects of protease inhibitors on cell dissociation were studied in vitro in order to examine the involvement of proteases in stratum corneum desquamation. Stratum corneum sheet (peeled from human backs after sunburn) was incubated in a detergent mixture containing 8 mM N,N-dimethyldodecylamine oxide, 2 mM sodium lauryl sulphate and 60 micrograms/ml kanamycin with or without protease inhibitors, and the number of released cells was counted after incubation for 48 h. Cell dissociation was inhibited strongly by antipain or aprotinin, but not at all by N-[N-(L-3-transcarboxyoxiran-2-carbonyl)-L-leucyl]-agmatin, N-ethylmaleimide or pepstatin, which suggests that only serine proteases are associated with desquamation. Furthermore, leupeptin and chymostatin each reduced cell dissociation about half as effectively as aprotinin or antipain, while a mixture of leupeptin and chymostatin prevented stratum corneum dissociation as potently as antipain or aprotinin. In addition, the activity of chymotrypsin-like protease in scaly skin was higher than that in normal skin, as we have previously found for trypsin-like protease. These results suggest that both trypsin-like and chymotrypsin-like serine proteases are involved in stratum corneum desquamation.
为了研究蛋白酶在角质层脱屑中的作用，在体外研究了蛋白酶抑制剂对细胞解离的影响。将角质层薄片（晒伤后从人体背部剥离）在含有8 mM N,N-二甲基十二烷基氧化胺、2 mM十二烷基硫酸钠和60微克/毫升卡那霉素的去污剂混合物中孵育，添加或不添加蛋白酶抑制剂，孵育48小时后计算释放细胞的数量。细胞解离受到抗蛋白酶或抑肽酶的强烈抑制，但N-[N-(L-3-反式羧基环氧乙烷-2-羰基)-L-亮氨酰]-胍丁胺、N-乙基马来酰亚胺或胃蛋白酶抑制剂对其没有抑制作用，这表明只有丝氨酸蛋白酶与脱屑有关。此外，亮抑蛋白酶肽和抑糜蛋白酶肽各自降低细胞解离的效果约为抑肽酶或抗蛋白酶的一半，而亮抑蛋白酶肽和抑糜蛋白酶肽的混合物阻止角质层解离的效力与抗蛋白酶或抑肽酶相同。另外，正如我们之前对胰蛋白酶样蛋白酶的发现，鳞状皮肤中类胰凝乳蛋白酶的活性高于正常皮肤。这些结果表明，胰蛋白酶样和类胰凝乳蛋白酶样丝氨酸蛋白酶都参与角质层脱屑。
