盐酸小檗碱对人牙周膜细胞分泌单核细胞趋化蛋白1影响的体外实验

Effect of berberine hydrochloride on the secretion of monocyte chemoattractant protein-1 from human periodontal ligament cells in vitro

目的研究盐酸小檗碱对体外培养的人牙周膜细胞（periodontal ligament cells，PDLC）分泌单核细胞趋化蛋白1（monocyte chemoattractant protein-1，MCP-1）的影响，旨在为利用天然药物治疗牙周病提供依据。方法从离体牙分离牙周膜组织，体外培养人PDLC。经免疫组化鉴定后的人PDLC首先根据细胞培养液中是否含有10mg／L脂多糖（1ipopolysaccharide，LPS）分为LPS组和无LPS组（NLPS组），然后两组再依据培养液中盐酸小檗碱的终质量浓度（0、0．01、0．02、0．03g／L）分为LPS空白对照组、LPS1、2、3组及NLPS空白对照组和NLPS1、2、3组。应用酶联免疫吸附测定法分别检测各组细胞培养上清液中MCP-1的含量，对多组间结果比较采用方差分析，组间两两比较采用Bonferroni t检验，以双侧P〈0．05为差异有统计学意义。结果NLPS1、2、3组MCP-1含量[分别为（11．33±0．16）、（11．45±0．53）、（11．25±0．14）ng／L]与NLPS空白对照组[（11．32±0．35）ng／L]相比差异均无统计学意义（P值均大于0．05）；LPS1、2、3组MCP-1含量[分别为（38．14±5．34）、（34．15±3．36）、（26．13±2．12）ng／L]与LPS空白对照组[（58．42±1．52）ng／L]相比均显著降低，差异均有统计学意义（P值均小于0．001）；LPS1、2、3组间两两比较差异均有统计学意义（P值均小于0．001）。结论当以LPS作为刺激因子时，盐酸小檗碱抑制体外培养的PDLC分泌MCP-1作用明显，并在一定范围内呈浓度依赖性。

Objective To observe the effects of berberine hydrochloride on the secretion of monocyte ehemoattractant protein-1 （ MCP-1 ） from human periodontal ligament cells （ PDLC ） in vitro culture. Methods Periodontal ligament was isolated from extracted human premolars, and PDLC were cultured in vitro. PDLC were divided into two groups, lipopolysaccharide （LPS） group and non-lipopolysaccharide （NLPS） group. Then in accordance with the final concentrations of berberine hydrochloride in cells culture medium（0,0. 01,0.02,0.03 g/L） ,the groups were subdivided into LPS and NLPS control group, LPS1 and NLPS1 group, LPS2 and NLPS2 group, LPS3 and NLPS3 group. Cellular concentration of MCP-1 of each group was determined by enzyme-linked immunosorbent assay（ELISA）. The data were statistically analyzed. Results The MCP-1 contents were not significantly different between the groups of NLPS1, NLPS2 and NLPS3 [ （ 11.33 ± 0.16 ） , （ 11.45 ± 0.53 ）, （ 11.25 ± 0. 14 ） ng/L, respectively ] and the NLPS control group[ （11.32 ±0.35） ng/L] （P =0. 692,0. 568,0. 524）. MCP-1 contents in the groups of LPS1 ,LPS2 and LPS3 [ respectively （38.14 ± 5.34）, （34.15 ± 3.36）, （26.13 ± 2.12） ng/L] were significantly lower than in LPS control group [（58.42 ± 1. 52） ng/L], P =0. 000,0. 000,P =0. 000. Conclusions The inhibitory effect of berberine hydrochloride on the activities of MCP-1 from PDLC was more significant when PDLC were stimulated with LPS and in a concentration-dependent manner.