复方双黄连制剂对单核巨噬细胞(RAW264.7) Fc/C3b受体活性和分泌功能的影响

doi:10.16431/j.cnki.1671-7236.2024.01.006

摘要/Abstract

摘要： 【目的】探究复方双黄连制剂对巨噬细胞吞噬能力和分泌功能的影响，为复方双黄连的深度开发及畜禽用药的合理选择提供科学依据。【方法】将金银花、黄芩、连翘、穿心莲干燥粉碎后分别制备浸膏，按照比例制备双黄连口服液（金银花：黄芩：连翘=1：1：2）、复方双黄连口服液（金银花：黄芩：连翘：穿心莲=1：1：2：2）及穿心莲口服液，调节pH为7.0，生药浓度为1 g/mL。3种药物作用于小鼠单核巨噬细胞RAW264.7，采用CCK-8法检测细胞活力，确定3种药物安全浓度，将药物最大安全浓度以二倍稀释法稀释为3个浓度；采用脂多糖（LPS）和氢化考的松琥珀酸钠（HCSS）作用于RAW264.7细胞模拟机体炎症和免疫抑制状态，3种药物作用于这2种状态的细胞和正常细胞，采用YC、EA玫瑰花环法及中性红吞噬法检测巨噬细胞Fc/C3b受体活性和吞噬能力，采用ELISA法检测巨噬细胞分泌能力。【结果】复方双黄连、双黄连、穿心莲的安全浓度分别为0.625~2.5、3.125~12.5和0.625~2.5 mg/mL。不同浓度复方双黄连、双黄连、穿心莲作用于巨噬细胞，细胞吞噬能力均显著高于空白对照组（P<0.05）。复方双黄连可降低LPS巨噬细胞RAW264.7活跃的吞噬能力，增强HCSS巨噬细胞RAW264.7吞噬能力；不同浓度复方双黄连可使活化的巨噬细胞Fc/C3b受体活性下降，低下的Fc/C3b受体活性增强；不同浓度复方双黄连具有促进巨噬细胞分泌一氧化氮（NO）、肿瘤坏死因子-α（TNF-α）、γ-干扰素（IFN-γ）及溶菌酶（LZM）的作用，抑制LPS巨噬细胞NO、TNF-α、IFN-γ及白细胞介素-6（IL-6）的分泌。【结论】复方双黄连可通过活化巨噬细胞Fc/C3b受体活性以增强机体免疫力和抗炎能力，同时对已活化的巨噬细胞分泌功能起到抑制作用，以减少由于免疫功能亢进造成的机体损伤。研究结果为中兽药复方双黄连的深度开发提供了参考依据。

关键词: 复方双黄连; 巨噬细胞Fc/C3b受体; 吞噬能力; 分泌功能

Abstract: 【Objective】 This study was aimed to investigate the effect of compound Shuanghuanglian preparation on the phagocytosis and secretion function of macrophages, and provide scientific basis for the in-depth development of compound Shuanghuanglian and the rational selection of drugs for poultry and livestock.【Method】 Lonicera japonica, Scutellaria baicalensis, Forsythia suspense and Andrographis paniculata were dried and crushed to prepare extracts, and Shuanghuanglian oral solution (Lonicera japonica:Scutellaria baicalensis:Forsythia suspense=1:1:2), compound Shuanghuanglian oral solution (Lonicera japonica:Scutellaria baicalensis:Forsythia suspense:Andrographis paniculata=1:1:2:2), and Andrographis paniculata oral solution were prepared in the proportions of the three drugs, and adjusted to a pH of 7.0, with a concentration of 1 g/mL.Three drugs were applied to the monocyte-macrophage cells in the mice.RAW264.7 cell viability was determined using CCK-8 method to establish the safe concentration of three drugs.The maximum permissible concentration of the drugs was diluted into three concentrations using a two-fold dilution method.Lipopolysaccharide (LPS) and hydrocortisone succinate sodium (HCSS) were employed to simulate the states of inflammation and immunosuppression in the body.The three drugs were used to treat cells in the two aforementioned states, as well as normal cells.The macrophage Fc/C3b receptor activity and phagocytosis capacity were measured using YC and EA rosette method as well as neutral red phagocytosis.Additionally, the macrophage secretion capacity was evaluated via ELISA.【Result】 The safe concentration range of compound Shuanghuanglian, Shuanghuanglian and Andrographis paniculata were 0.625-2.5, 3.125-12.5 and 0.625-2.5 mg/mL, respectively.Different concentrations of compound Shuanghuanglian, Shuanghuanglian and Andrographis paniculata acted on macrophages, and the cell phagocytic ability was significantly higher than that of blank control group (P<0.05).Compound Shuanghuanglian had the ability to reduce the active phagocytosis of LPS macrophage RAW264.7 and enhance the low phagocytosis of HCSS macrophages RAW264.7.Different concentrations of compound Shuanghuanglian could decrease the Fc/C3b receptor activity of activated macrophages and enhance the Fc/C3b receptor activity of low.Different concentrations of compound Shuanghuanglian had the ability to promote macrophages to secrete nitric oxide (NO), tumor necrosis factor-α (TNF-α), γ-interferon (IFN-γ) and lysozyme (LZM), and inhibit the secretion of NO, TNF-α, IFN-γ and interleukin 6 (IL-6) by LPS macrophages.【Conclusion】 Compound Shuanghuanglian could enhance the immunity and anti-inflammatory ability of the body by activating the Fc/C3b receptor activity of macrophages, while inhibiting the secretion function of activated macrophages to reduce the damage caused by hyperimmune function.The results provided a reference basis for the in-depth development of the traditional Chinese veterinary medicine compound Shuanghuanglian.

Key words: compound Shuanghuanglian; macrophage Fc/C3b receptor; phagocytosis; secretory function

中图分类号:

S853.73

史晗, 戴浩然, 郭彬, 刘群. 复方双黄连制剂对单核巨噬细胞(RAW264.7) Fc/C3b受体活性和分泌功能的影响[J]. 中国畜牧兽医, 2024, 51(1): 52-63.

SHI Han, DAI Haoran, GUO Bin, LIU Qun. Effect of Compound Shuanghuanglian Preparation on Fc/C3b Receptor Activity and Secretory Function of Monocyte Macrophages (RAW264.7)[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(1): 52-63.

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