应用小鼠腘窝淋巴结试验评价5种清热类中药注射液致敏潜能

Experimental Research of Sensitization Potential of Five Kinds of Traditional Chinese Medicine Injection with Clearing-heat Function by Mice Popliteal Fossa Lymph Node Assay

摘要:

目的：评价5种清热类中药注射液（TCMI）引起的BALB/c小鼠腘窝淋巴结反应，探讨清热类注射液潜在的致敏性。方法：70只雌性BALB/c小鼠随机分为阳性致敏组（氯化汞），阴性对照组（生理盐水）及各中药注射液组，每组10只。实验第1天，小鼠一侧单次足跖部sc 50 μL/只生理盐水或50 μg/只氯化汞，各中药注射液组按临床高剂量的4倍给药，sc于小鼠一侧足跖部分别为清开灵注射液0.96 mL·kg^-1、穿心莲注射液0.96 mL·kg^-1、肿节风注射液1.92 mL·kg^-1、柴胡注射液0.96 mL·kg^-1、板蓝根注射液0.48 mL·kg^-1；6 d后处死，摘取两侧腘窝淋巴结（PLN），称重，计算质量指数（weights index，WI）；每组将4只小鼠的PLN用于病理学检查，其余6只制备单细胞悬液，计算细胞指数（cells index，CI），比较PLN的变化。结果：与小鼠未处理侧淋巴结相比，氯化汞组（WI=3.82±0.93，CI=7.40±0.63），肿节风注射液（WI=2.21±0.15，CI=5.23±0.18），清开灵注射液（WI=2.13±0.10，CI=5.17±0.18）及穿心莲注射液组（WI=2.06±0.14，CI=5.04±0.27），与小鼠未处理侧淋巴结相比，均可引起显著的腘窝淋巴结反应，且WI>2和CI>5（P<0.05或P<0.01）；与生理盐水组（WI=1.10±0.14，CI=1.23±0.11）比较，PLN质量及细胞计数显著升高，且足跖肿胀明显（P<0.05或P<0.01）；病理学检查，生发中心明显，副皮质区内高内皮质静脉（HEV）横断面增多。而柴胡组（WI=1.39±0.16，CI=3.15±0.21）和板蓝根组（WI=1.38±0.13，CI=3.25±0.36）相应的变化不明显，反应未达到阳性标准。结论：肿节风、清开灵及穿心莲注射液具有潜在致敏性；柴胡及板蓝根注射液无潜在的致敏性。

Abstract:

Objective: To study sensitization potential of five kinds of traditional Chinese medicine injections (TCMI) with clearing-heat function by using the BALB/c popliteal lymph node assay. Method: Seventy female BALB/c mice were randomly divided into positive group (mercuric chloride), negative control group (normal saline) and the traditional Chinese medicine injection (TCMI) groups, mice were injected subcutaneously (50 μL) with drugs into the right pad of hind foot. Dosages of each animal from normal saline group and mercuric chloride group were 50 μL 0.9%NaCl and 50 μg HgCl₂, respectively. The traditional Chinese medicine (TCM) injection groups, were given 1.92 mL·kg^-1 Zhongjiefeng injection(ZJF), 0.96 mL·kg^-1 Qingkailing injection(QKL), 0.96 mL·kg^-1 Chuanxinlian injection (CXL), 0.96 mL·kg^-1 Chaihu(CH) and 0.48 mL·kg^-1 Banlangen(BLG). The animals were killed on the 6th day, the popliteal lymph nodes(PLN) were isolated and weighed respectively to calculate the PLN weight index(WI).Then the PLN from four of ten mice in each group were fixed with paraformaldehyde solution for histopathologic examination;the other PLN were calculated cell index(CI). Result: Compared to untreated sides, WI and CI of the PLN from mercuric chloride group (WI=3.82±0.93, CI=7.40±0.63), Zhongjiefeng injection(ZJF, WI=2.21±0.15, CI=5.23±0.18), Qingkailing injection(QKL, WI=2.13±0.10, CI=5.17±0.18) and Chuanxinlian injection (CXL, WI=2.06±0.14, CI=5.04±0.27) reached to WI>2 and CI>5(P<0.05 or P<0.01)and Compared to normal saline groups(WI=1.10±0.14, CI=1.23±0.11), their PLN were significantly enlarged, cells were increased.And their pathological examination showed that treated side PLN were enlarged with remarkable germinal center and increased high endothelial venules proliferation.And the responses in Chaihu injection (CH, WI=1.39±0.16, CI=3.15±0.21) or Banlangen injection (BLG, WI=1.38±0.13, CI=3.25±0.36)were unreached to positive response standard, corresponding change was not obvious. Conclusion: ZJF, QKL and CXL have the allergenic potential the sensitization of CH or BLG is not obvious.