组胺对人角质形成细胞增殖、凋亡和分化的影响

南方医科大学学报 ›› 2005, Vol. 25 ›› Issue (10): 1286-1289.

组胺对人角质形成细胞增殖、凋亡和分化的影响

冉立伟¹, 谭卫明², 谭升顺¹, 张茹³, 王万卷¹

1. 西安交通大学第二医院皮肤科, 陕西, 西安, 710004;
2. 西安交通大学第二医院消化科, 陕西, 西安, 710004;
3. 西安交通大学第二医院口腔医学院颌面成形科, 陕西, 西安, 710004

出版日期:2005-10-20 发布日期:2005-10-20
基金资助:
收稿日期:2005-3-1。
作者简介:冉立伟(1974- ),女,医学博士,电话:029-87727402,E-mail:wlbrlw@163.com
通讯作者:谭升顺,教授,博士生导师,电话:029-88063053,E-mail:wlbrlw@163.com

Effects of histamine on proliferation, apoptosis and differentiation of human keratinocytes

RAN Li-wei¹, TAN Wei-ming², TAN Sheng-shun¹, ZHANG Ru³, WANG Wan-juan¹

1. 西安交通大学第二医院皮肤科, 陕西, 西安, 710004;
2. 西安交通大学第二医院消化科, 陕西, 西安, 710004;
3. 西安交通大学第二医院口腔医学院颌面成形科, 陕西, 西安, 710004

Online:2005-10-20 Published:2005-10-20

摘要/Abstract

摘要： 目的探讨组胺对体外培养人角质形成细胞(HKC)增殖、凋亡和分化的影响及其机制。方法采用MTT法和蓝染色法检测组胺对HKC体外生长的影响,流式细胞仪检测细胞周期和细胞早期凋亡,细胞DNA梯度降解法检测凋亡,激光扫描共聚焦显微镜结合钙荧光探针技术检测HKC胞内游离Ca²⁺浓度([Ca²⁺]_i),SABC免疫细胞化学法检测HKC分化标志物角蛋白10(K10)及内披蛋白表达。结果高浓度组胺抑制HKC生长,以10^-4mol/L抑制率最大(细胞存活率65.6%);低浓度组胺则促进HKC生长,以10^-8mol/L作用显著(细胞存活率130.7%)。10^-4mol/L组胺致HKCG0/G1期比例增高30.97%,S期比例减少73.81%,抑制G1/S期转换,并明显促进细胞凋亡,早期凋亡率18.64%明显高于对照组(5.60%,P<0.05)。10^-4mol/L组胺使HKC[Ca²⁺]_i上升58.9%,H2组胺受体拮抗剂西咪替丁则使[Ca²⁺]_i下降24.5%。10^-4mol/L组胺下调HKCK10及内披蛋白表达,但与对照组无显著性差异(P>0.05)。结论高浓度组胺阻抑HKC细胞周期进程、诱导[Ca²⁺]_i增加及其显著的促凋亡作用可能是使HKC体外生长受抑的部分机制。在生理条件下,组胺可能具有调节表皮组织更新的作用;在炎症、损伤等病理条件下,大量的组胺可能抑制表皮的再生和KC的分化。

Abstract: Objective To investigate the effects of histamine on the proliferation, apoptosis and differentiation of human keratinocytes (HKC) and the mechanisms. Methods The effect of histamine on the growth of HKC in vitro was examined by MTT assay and trypan blue exclusion assay. Cell cycle analysis and early apoptosis analysis by double staining with Annexin V-FITC and PI were carried out by flow cytometry. DNA ladder assay was performed for the detection of cell apoptosis. HKC free calcium concentration ([Ca²⁺]_i) was measured by laser scanning confocal microscopy in combination with calcium fluorescence probe Fluo-3/AM. HKC differentiation markers keratin 10 (K10) and involucrin was detected by streptavidin- biotin complex immunocytochemical assay. Results Histamine at high concentration inhibited the proliferation of HKC with cell viability ratio of 65.6% at 1×10^-4 mol/L, while histamine at low concentrations promoted proliferation of HKC with the cell viability ratio of 130.7% at 1×10^-8 mol/L. Histamine at 1×10^-4 mol/L altered cell cycle distribution of HKC with an increase in G0/G1-phase cell population to 30.97%, a decrease in S-phase population to 73.81% and inhibition of G1/S switching. Histamine at 1×10^-4 mol/L induced obvious apoptosis of HKC with early apoptosis ratio of 18.64% as compared with the control (5.60%, P<0.05). Histamine 1×10^-4 mol/L induced an increase of HKC [Ca²⁺]_i up to 58.9% and cimetidine (an H2 receptor antagonist) decreased HKC [Ca²⁺]_i down to 25.4%. Histamine at this concentration down-regulated the expressions of K10 and involucrin of HKC but these changes were not significantly different from those of the control (P>0.05). Conclusions Histamine at high concentrations inhibits cell cycle progress of HKC, mediates cell apoptosis and induces the increase of [Ca²⁺]_i, which might be a partial explanation for growth arrest of HKC elicited by histamine. Histamine may regulate epidermal tissue turnover under physiological conditions, whereas under pathological circumstances of the skin as in trauma and inflammation, histamine at high concentrations may inhibit the regeneration of epidermis and the differentiation of HKC.

中图分类号:

R392.12

冉立伟¹, 谭卫明², 谭升顺¹, 张茹³, 王万卷¹. 组胺对人角质形成细胞增殖、凋亡和分化的影响[J]. 南方医科大学学报, 2005, 25(10): 1286-1289.

RAN Li-wei¹, TAN Wei-ming², TAN Sheng-shun¹, ZHANG Ru³, WANG Wan-juan¹. Effects of histamine on proliferation, apoptosis and differentiation of human keratinocytes[J]. Journal of Southern Medical University, 2005, 25(10): 1286-1289.

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