当归多糖通过抑制角蛋白17表达抑制角质形成细胞的增殖活性

Angelica sinensis polysaccharides impeding the viability of malpighian cells via inhibiting the expression of keratin 17

目的:探讨当归多糖(Angelica sinensis polysaccharides,ASP)对角蛋白17(K17)介导的人角质形成细胞生长和增殖影响。方法:用不同浓度(1,0. 5,0. 1,0. 05 mg/m L) ASP处理人乳头瘤病毒(HPV)11型基因组的HaCaT细胞(HPV11. HaCaT) 24 h,qRT-PCR与Western blot检测K17水平变化,1 mg/m L ASP处理HPV11. HaCaT细胞Western blot检测ki-67和PNCA的表达; MMT试剂盒检测1 mg/m L ASP处理下HPV11. HaCaT细胞增殖活性变化。构建K17过表达(pcDNA3. 1 (+)/K17)载体并转染KC细胞48 h继续用ASP (1 mg/m L)处理24 h;检测K17的水平变化及细胞增殖活性变化。结果:使用1 mg/m L ASP抑制HPV11. HaCaT细胞中K17表达,差异具有统计学意义(P <0. 05),呈浓度依赖趋势。qRT-PCR及Western blot检测发现1 mg/m L ASP使K17、Ki67及PNCA的表达显著下调,差异具有统计学意义(P <0. 05)。ASP作用下,HPV11. HaCaT细胞增殖率均下调(P <0. 01)。pc DNA3. 1(+)/K17对HPV11. HaCaT细胞增殖率的促进作用被ASP阻断,差异具有统计学意义(P <0. 01)。结论:ASP通过抑制K17抑制角质形成细胞增殖,为银屑病和尖锐湿疣基础研究与临床治疗提供前期研究基础。

Objective: To investigate the effects of angelica sinensis polysaccharides(ASP) on the viability of keratin 17(K17)-mediated human keratinocyte.Methods:Different concentrations of ASP were used to treat HPV11.HaCaT for 24 h.The expression of K17 was tested by qRT-PCR and Western blot.HPV11.HaCaT was treated by 1 mg/mL of ASP,and the expression of ki-67 and PNCA was tested by Western blot.The proliferative activity changes of HPV11.HaCaT cells treated with 1 mg/mL of ASP were tested by MMT kit.K17 overexpression vectors(pcDNA3.1(+)/K17)were constructed,and the vectors were transfected in HaCaT for 48 h,and further treated with ASP(1 mg/mL)for 12 h.The level of K17 and proliferative activity were detected.Results:The expression of K17 was significantly decreased in HPV11.HaCaT by ASP(1 mg/mL)treatment(P<0.05)with a concentration dependent trend.The expression of K17,Ki67 and PNCA were also downregulated by 1 mg/mL of ASP(P<0.05).After treated with ASP,the viability rate of HPV11.HaCaT was downregulated(P<0.05).ASP also attenuated the role of pcDNA3.1(+)/K17 on cell viability of KC,with statistically significant difference(P<0.01).Conclusions:ASP impedes keratinocyte viability by inhibiting K17,which provides a basis for the basic research and clinical treatment of psoriasis and condyloma acuminatum.