生物活性透明质酸对脂多糖诱导的人树突状细胞和巨噬细胞炎症应答作用  

Bioactive hyaluronic acid(B-HA) on lipopolysaccharide-induced inflammatory response in human dendritic cells and macrophages

作　　者：尤娜[1] 褚萨萨 朱进 张馨[3] 杨志国[3] 汪茂荣[1,3] 

机构地区：[1]安徽医科大学附属解放军八一临床学院,合肥市230032 [2]南京军区疾控中心流行病与微生物研究所,南京市210002 [3]解放军第81医院全军肝病中心,南京市210002

出　　处：《中华实验和临床感染病杂志（电子版）》2017年第1期14-19,共6页Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition)

基　　金："十二五"军队特需药品保密专项课题(No.2013ZJ09J13110-05B)

摘　　要：目的探讨经人透明质酸酶PH20特异处理的重组HA,即生物活性透明质酸(B-HA)对脂多糖(LPS)诱导的人树突状细胞(DC)和巨噬细胞炎症应答的影响。方法 DC细胞和人急性单核细胞白血病单核细胞株(THP-1)经不同浓度的B-HA预先处理,加入LPS刺激不同时间,收集细胞和培养上清,采用实时荧光定量PCR和ELISA方法检测细胞因子mRNA和蛋白表达水平的变化。结果 THP-1细胞实验中,与LPS组相比,B-HA 20 mg/ml不仅能够显著降低促炎因子肿瘤坏死因子-α(TNF-α)和白细胞介素-8(IL-8)的表达量,同时能够显著增加抗炎因子白细胞介素-10(IL-10)的表达量。其细胞中TNF-α、IL-8和IL-10 mRNA表达水平分别为24.41±1.94、35.76±4.19和14.99±0.07,而LPS组分别为38.48±1.47、62.7±1.93和2.27±0.16,差异均具有统计学意义(t=5.781、P=0.0290,t=5.695,P=0.0300,t=15.20、P=0.0040)。细胞上清中三者蛋白表达水平分别为(1 609.00±75.66)pg/ml、(213.10±9.14)pg/ml和(496.3±41.92)pg/ml,LPS组分别为(3 018±102.8)pg/ml、(587.4±3.140)pg/ml和(243.3±20.23)pg/ml,差异均具有统计学意义(t=11.04、P=0.0080,t=38.74、P=0.0007,t=5.434、P=0.0320)。在人外周血分化的DC细胞实验中,B-HA对细胞因子TNF-α、IL-8和IL-10的表达均无显著影响(P均>0.05)。结论 B-HA能有效调节人巨噬细胞炎性因子的表达,抑制其炎症反应;对人DC细胞的炎症反应无显著影响,提示B-HA对人炎性免疫细胞的炎症反应作用具有选择性。Objective To investigate the effects of hyaluronidase PH20 processed recombinant human hyaluronic acid fragments（bioactive HA,B-HA） on lipopolysaccharide（LPS）-induced inflammatory response in human dendritic cells（DC） and macrophages.Methods DC cells and human acute monocytic leukemia cells（THP-1） were pretreated with different concentrations of B-HA in absence or unabsence of LPS for different times.The levels of inflammatory cytokines in culture medium were measured by ELISA and inflammatory cytokines mRNA of cultured cells was detected by real-time PCR.Results Compared with LPS-only group,the expression of proinflammatory cytokines TNF-a and IL-8 were significantly inhibited by B-HA 20 mg/ml in THP-1 cells,while the expression of anti-inflammatory cytokine IL-10 was significantly elevated.The mRNA levels of TNF-a,IL-8 and IL-10 were 24.41 ± 1.94,35.76 ± 4.19 and 4.99 ± 0.07 in B-HA treated group,respectively,while the LPS-only group were 38.48 ± 1.47,62.7 ± 1.93 and 2.27 ± 0.16,with significant differences（t = 5.781,P = 0.0290;t = 5.695,P = 0.0300;t = 15.20,P = 0.0040）.The TNF-a,IL-8 and IL-10 protein levels in culture medium were（1 609.00 ± 75.66） pg/ml,（213.10 ± 9.14） pg/ml and（496.30 ± 41.92） pg/ml,while the LPS-only group were（3 018 ± 102.8） pg/ml,（587.4 ± 3.140） pg/ml and（243.3 ± 20.23） pg/ml,with significant differences（t = 11.04,P = 0.0080;t = 38.74,P = 0.0007;t = 5.434,P =0.0320）.Unfortunately,no effect of B-HA was observed in human PBMC differentiated DC cells（all P〉0.05）.Conclusions B-HA was effective on regulating inflammatory cytokines and could inhibit the inflammatory response in human macrophage.There was no significant effect on the inflammatory response in human PBMCdifferentiated DC cells.The effect of B-HA on regulating inflammatory response is selective.

关 键 词：生物活性透明质酸 脂多糖 炎症反应 树突状细胞 人急性单核细胞白血病单核细胞株 

分 类 号：R392[医药卫生—免疫学]