腘窝淋巴结试验模型的构建及在清开灵注射液致敏性研究中的应用

目的构建直接和间接腘窝淋巴结试验(d-PLNA和s-PLNA)模型,并用其检测清开灵注射液(QKLI)的致敏性。方法雌性BALB/c小鼠右侧后肢足趾一次性分别sc给予50μl盐酸D-青霉胺(D-Pen)12.5,25.0,37.5,50.0和62.5 mg.kg-1,分别在给药后的第5,7和9天处死小鼠,摘取两侧腘窝淋巴结(PLN),计算PLN质量指数(MI)和细胞指数(CI),确定D-Pen的最低有效剂量和最佳解剖时间。小鼠右侧后肢足趾一次性sc给予D-Pen 37.5 mg.kg-1、QKLI原液、2倍和4倍QKLI原液,7 d后活杀,进行d-PLNA实验。小鼠右侧后肢足趾一次性sc给予D-Pen和QKLI,2个月后,给予亚剂量激发,7 d后活杀进行s-PLNA实验,检测MI和CI。结果根据MI≥2和CI≥5阳性标准判定D-Pen最低有效剂量为37.5 mg.kg-1,最佳解剖时间为给药后第7天。d-PLNA实验结果显示,QKLI原液组处理侧PLN的质量和细胞计数较未处理侧有轻度的升高,但未达到阳性反应判定标准;2倍原液浓度组MI为2.0±1.0,CI为5.4±0.9;4倍原液浓度组MI为3.4±0.4,CI为5.4±0.9,均达到阳性反应判定标准。s-PLNA实验结果显示,2倍原液浓度组MI为2.4±0.6,CI为6.2±0.8;4倍原液浓度组MI为3.2±0.9,CI为8.4±1.8均达到阳性反应判定标准。结论制备了能够用于致敏实验的腘窝淋巴结模型,利用此模型发现QKLI具有诱发过敏反应的可能。

Establishment of popliteal lymph node assay model and application to evaluation of Qingkailing Injection sensitivity

OBJECTIVE To construct BALB/c mice models for direct popliteal lymph node assay(d-PLNA) and secondary popliteal lymph node assay(s-PLNA),and to investigate the sensitizing potential of Qingkailing Injection(QKLI).METHODS Female BALB/c mice were sc given D-penicillamine hydrochloride(D-Pen) 12.5,25.0,37.5,50.0 and 62.5 mg·kg-1 into the right pad of hind foot,50 μl per mouse.Mice were sacrificed on the fifth,seventh and nineth days after injection,respectively,and their bilateral PLN were isolated and weighted to calculate the PLN mass index(MI) and cell index(CI).Dose-response relationship and kinetics of PLN response were studied in order to construct the BALB/c mice models for d-PLNA and s-PLNA.The test compounds used in d-PLNA were injected directly into skin of right hind footpad of mice.Mice in s-PLNA were pre-sensitized by immunization with the test compounds,and 2 months later,mice were challenged with suboptimal dose of the same test compound.And then the sensitizing potential of the different concentrations of QKLI was studied.RESULTS In accordance with the positive standard,MI≥2 and CI≥5,the lowest effective dose of D-Pen was 37.5 mg·kg-1,the optimum examining time was the seventh day after D-Pen given.And then BALB/c mice models for d-PLNA and s-PLNA were constructed under the conditions obtained above.MI and CI of PLN response were ≥2 and ≥5,respectively,in d-PLNA and s-PLNA,in 2 and 4 times of the original QKLI concentration(2×QKLI and 4×QKLI) groups,while the original QKLI group not.In d-PLNA,MI and CI of original QKLI group were 1.8±0.7 and 3.4±1.4,those of 2×QKLI group were 2.0±1.0 and 5.4±0.9,and of 4×QKLI group were 3.4±0.4 and 5.4±0.9.In s-PLNA,MI and CI were 2.4±0.6 and 6.2±0.8 in 2×QKLI group,and 3.2±0.9 and 8.4±1.8 in QKLI 4 group.CONCLUSION QKLI can induce significant PLN response in mice,suggesting it have certain sensitizing potential.