川椒方含药血清对大鼠嗜碱性白血病细胞脱颗粒的影响

Impact of Drug-Containing Serum of Chuanjiao Formula on the Degranulation of RBL-2H3 Cells

目的探讨川椒方治疗变态反应性结膜炎的可能作用机制。方法 SD大鼠20只,随机分为川椒方低、中、高剂量含药血清组和空白血清组,每组5只。川椒方低、中、高含药血清组分别以含川椒方生药0.334、0.668、1.336 g/ml的水煎剂20 ml/kg灌胃,空白血清组给予相同剂量的生理盐水灌胃,每日2次,连续给药3日,制备SD大鼠含药血清。筛选不同浓度含药血清对大鼠嗜碱性白血病细胞(RBL-2H3)的最佳作用浓度,并筛选含药血清最佳作用时间。实验再分为模型组、川椒方治疗高、中、低剂量组、酪氨酸激酶抑制剂(PP2)治疗组、对照组。川椒方高、中、低剂量含药血清组按筛选所得浓度和作用时间每孔分别给予含药血清40μl;PP2治疗组于加抗原前30 min给予终浓度为1μM的PP2 40μl;对照组于加抗原前2 h给予10%空白血清40μl,酶联免疫吸附(ELISA)法检测β-氨基己糖苷酶释放率。结果川椒方浓度为10%时细胞活力较好,故本实验选择了10%浓度血清作为实验浓度。川椒方高、中剂量组含药血清作用120 min时,释放量最少(P<0.01),故选用120 min作为作用时间。与正常组比较,模型组和对照组β-氨基己糖苷酶释放率升高明显(P<0.01);川椒方各剂量组及PP2治疗组β-氨基己糖苷酶释放率下降(P<0.01)。结论川椒方治疗变态反应性结膜炎的作用机制可能与抑制肥大细胞脱颗粒有关。

Objective To study the possible mechanism ot G tmanjtao rormuta ~ t^jr ~ allergic conjunctivitis （AC）. Methods Totally 20 SD rats were randomlly divided into the blank group, the low, medium, and high dose CJF groups, each with 5 rats. Rats in the low, medium, and high dose CJF groups were given CJF 20 ml/kg at the dose of 0. 334, 0. 668, 1. 336 g/ml by gavage, twice a day for 3 days. Rats in the blank group were given same amount of normal saline by garage, twice a day for 3 days. Then drug-containing serum was made. The optimal concentration of serum on rat basophilic leukemia cells （ RBL-2H3 ） was screened and the optimal action time were screen as well. Experiments subdivided into model group, high , medium and low dose CJF group, tyrosine kinase inhibitor （PP2） treatment group and control group. The low, medium, and high dose CJF groups were given drug-containing serum 40 μl each well using the screened serum concentration and time. PP2 treatment group were given 40 p2 of PP2 with the concentration of 1 μM 30min before adding the antigen. The control group were given 10% blank serum 40 μl 2 hours before adding the antigen. Enzyme-linked immunosorbent assay （ELISA） was applied to detect hexosaminidase release rate. Results With the concentration of 10% CJF contained serum, ceils showed better viability, so in this study, the concentration of 10% was chosen as the experimental concentration. Cell incubation with serum 120 min in medium, and high dose CJF groups showed the least degranula- tion （P 〈 0.01 ）, so 120 rain was chosen as the incubation time. Compared with the normal group, [3-hexosaminidase release rates of the model group and the control group increased significantly （ P 〈 0.01 ）. The β-hexosaminidase release rates of each dose CJF group and PP2 treatment group declined （P 〈 0.01 ）. Conclusion The mechanism of CJF in treating allergic conjunctivitis may be related to the inhibition of degranulation in mast cells.