银屑病外周血中B淋巴细胞对HaCaT细胞增殖能力及细胞周期的影响  

Effect of peripheral blood B lymphocytes from psoriasis patients on HaCaT cells proliferation and cell cycle

作　　者：李思彤 白彦萍[1] 王英[1] LI Sitong;BAI Yanping;WANG Ying(Department of Dermatology,China-Japan Friendship Hospital,Beijing100029,China)

机构地区：[1]中日友好医院皮肤科,北京100029

出　　处：《中国医药导报》2024年第35期39-43,共5页China Medical Herald

基　　金：国家自然科学基金青年科学基金资助项目(81803135)。

摘　　要：目的 探讨体外培养条件下,寻常型银屑病患者外周血中CD3~-CD19~+B淋巴细胞对HaCaT细胞增殖能力和细胞周期的影响。方法 纳入2021年3月至4月就诊于中日友好医院皮肤科门诊或住院的15例寻常型银屑病患者作为银屑病组,及同期接受色素痣切除的10名健康志愿者为对照组,采集肝素钠抗凝外周血20 ml。运用流式分选术分选出CD3~-CD19~+的B淋巴细胞,与HaCaT细胞共培养72 h后收集HaCaT细胞。细胞培养时设定空白对照组(仅HaCaT细胞),银屑病组(银屑病外周血B淋巴细胞+HaCaT细胞共培养),健康对照组(健康者外周血B淋巴细胞+HaCaT细胞共培养)。利用CCK-8实验评估细胞增殖活性,并通过流式细胞学技术检测HaCaT的细胞周期分布及Ki-67表达水平。结果 银屑病组及健康对照组HaCaT细胞在450 nm处的光密度值均较空白对照组升高(P<0.01),且银屑病组高于健康对照组(P<0.05)。银屑病组与健康对照组比较,HaCaT细胞Ki-67表达升高(P<0.05)。健康对照组与空白对照组Ki-67表达比较,差异无统计学意义(P>0.05)。与空白对照组及健康对照组比较,银屑病组HaCaT细胞的S期细胞比例增加,G_0/G_1期细胞比例减少,增殖指数升高(P<0.05);G_2/M期细胞比例与空白对照组比较,差异无统计学意义(P>0.05)。健康对照组与空白对照组各期细胞比例比较,差异无统计学意义(P>0.05)。结论 银屑病患者外周血中CD3~-CD19~+B淋巴细胞在体外条件下能调节人角质形成细胞周期、促进角质形成细胞增殖。Objective To investigate the effect of CD3-CD19+B lymphocytes in peripheral blood of patients with psoriasis vulgaris on the proliferation and cell cycle of HaCaT cells in vitro.Methods Fifteen patients with psoriasis vulgaris who were admitted to the Department of Dermatology,China-Japan Friendship Hospital from March to April 2021 were included as the psoriasis group,and 10 healthy volunteers who underwent pigmented nevi resection during the same period were included as the control group,20 ml of anticoagulant peripheral blood of heparin sodium was collected.CD3-CD19+B lymphocytes were separated by flow sorting,and HaCaT cells were collected after 72 h co-culture with HaCaT cells.During cell culture,blank control group(only HaCaT cells),psoriasis group(psoriasis peripheral blood B cells+HaCaT cells co-culture),and healthy control group(healthy peripheral blood B cells+HaCaT cells co-culture)were set.The cell proliferation activity was evaluated by CCK-8 assay,and the cell cycle distribution and Ki-67 expression level of HaCaT were detected by flow cytometry.Results Optical density value of HaCaT cells at 450 nm in psoriasis group and healthy control group were higher than those in blank control group(P<0.01),and that in psoriasis group was higher the in healthy control group(P<0.05).The expression of Ki-67 in HaCaT cells in psoriasis group was higher than that in healthy control group(P<0.05).There was no significant difference in Ki-67 expression between healthy control group and blank control group(P>0.05).Compared with blank control group and healthy control group,the proportion of S-phase HaCaT cells in psoriasis group increased,the proportion of G0/G1 phase HACAT cells decreased,and the proliferation index increased(P<0.05).There was no significant difference in the proportion of G2/M cells compared with the blank control group(P>0.05).There was no significant difference in cell proportion between healthy control group and blank control group(P>0.05).Conclusion In vitro,peripheral blood CD3-CD19+B lymphocy

关 键 词：银屑病 B淋巴细胞 HACAT细胞 CD19 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]