摘要
目的 在动物模型的基础上,探讨蒿芩清胆汤在小鼠机体免疫调节中的信号传导机制,为该药物治疗湿热证打下理论基础.方法 采用随机数字表法将40只由美国国立卫生研究院（National Institute of Health,NIH）培育而成的小鼠（1980年引进我国,简称NTH小鼠）分为正常组8只、流行性感冒（简称流感）病毒组（单纯病毒感染）8只、复合模型组（高脂饮食＋湿热环境＋病毒感染）8只、阳性药物组（模型组＋病毒唑）8只、蒿芩清胆汤组（模型组＋蒿芩清胆汤）8只.计算各组小鼠肺指数及肺指数抑制率;提取小鼠腹腔巨噬细胞用逆转录PCR（RT-PCR）法检测TLR2（Toll样受体-2）及NF-κB mRNA（核因子κB-mRNA）的表达;采用酶联免疫吸附测定法（ELISA）测定标本中白介素-4（IL-4）、干扰素-γ（IFN-γ）的浓度,求出IFN-γ/IL-4比值用于了解机体Th1/Ih2（Th1类与Th2类细胞比值）类细胞的平衡状态.结果 正常组、病毒组、复合模型组、阳性药物组和蒿芩清胆汤组小鼠的肺指数分别为（0.79±0.11）%、（1.93±0.38）%、（1.41±0.26）%、（1.10±0.26）%、（1.02±0.16）%;阳性药物组和蒿芩清胆汤组小鼠的肺指数和复合模型组相比均有所减低,有统计学意义（t=0.322,P＜0.05）.RT-PCR的结果显示,正常组、病毒组、复合模型组、阳性药物组和蒿芩清胆汤组TLR2 mRNA的表达分别是0.145±0.017、0.991±0.149、0.903±0.124、0.257±0.032和0.413±0.031;蒿芩清胆汤组、阳性药物组比复合模型组的小鼠低,均有统计学意义（t=0.013,F=73.467,P＜0.05）.正常组、病毒组、复合模型组、阳性药物组和蒿芩清胆汤组NF-κB mRNA的表达分别是0.075±0.015、379±0.019、0.291±0.012、0.169±0.026和0.175±0.033;阳性药物组、蒿芩清胆汤组与复合模型组相比均有所减低（t=0.422,F=112.834,P＜0.05）.正常组、病毒组、复合模型组、阳性药物组和蒿芩清胆汤组小鼠血清中IFN-γ含量分别为（7434.06±323.27）pg/ml、（8679.77±198.70）pg/ml、（8068.78±113.8）pg/ml、（7454.66±301.30）pg/ml和（7484.56±229.85）pg/ml;蒿芩清胆汤组和阳性药物组血清中IFN-γ含量比复合模型组减低,有统计学意义（t=0.201,F=5.390,P＜0.05）.正常组、病毒组、复合模型组、阳性药物组和蒿芩清胆汤组小鼠血清中IL-4含量分别为（3701.74±256.00）pg/ml、（3569.64±161.35）pg/ml、（3530.88±334.63）pg/ml、（3481.84±282.25）pg/ml和（3618.00±262.16）pg/ml;各组间差异无统计学意义（t=0.414,F=0.505,P＞0.05）.Th1/Th2比值（即：IFN-γIL-4）分别为：2.02±0.19、2.38±0.10、2.36±0.14、2.22±0.17和2.07±0.15;与复合模型组相比,蒿芩清胆汤组和阳性药物组均呈减低趋势,但无统计学意义（t=0.587,F=3.684,P＞0.05）.结论 蒿芩清胆汤对流感病毒性肺炎湿热证疗效显著,它通过降低膜表面受体TLR2的表达来抑制NF-κB的活化,降低血清IFN-γ水平,使Th1/Th2类细胞趋于平衡;降低肺泡内炎症反应,从而达到减少肺组织损伤的治疗目的 .
Objective To explore the immunoregulation existing signal transduction mechanism, to evaluate the role of lay its experimental basis By using Haoqin Qingdan decoction for treaments on the mouse models Methods A total of 40 NIH Mice were randomly divided into five groups： conol group, virus group（infecting by influenza virus） ,complex model group（richly fatty and sweet diet ＋ Humid heat environment ＋infecting by influenza virus）,virazole group（mouse of model group was treated by virazole）,and Haoqin Qingdan decoction group（mouse of complex model group was treated by decoction of Haoqin Qingdan）.When the complex model was estabhshed,determination of the mice lung indexes in each group and calculate the inhibition of lung indexes.The level of TLR2 mRNA and NF-κB mRNA expressions of peritoneal macrophages in each group of mice were quantitated by reverse transcription-polymerase chain reaction （RT-PCR）.The level of IL-4 and IFN-γin mouse serum was detected by ELISA to calculate the Th1/Th2（IFN-γ/IL-4）.Results The lung index of control group,virus group, complex model group, virazole group and Haoqin Qingdan decoction group were separately：（0.79±0.11）%,（1.93 ±0.38）%,（1.41 ±0.26）%,（1.10 ±0.26）% and （1.02 ±0.16）%; The mice of virazole group and Haoqin Qingdan decoction group lung index were decreased （ t = 0.322, P 〈 0.05 ）.TLR2 mRNA expression The results showed that the control group,virus group,complex model group,virazole group and Haoqin Qingdan decoction group were： 0.145 ±0.017,0.991±0.149,0.903 ±0.124,0.257 ±0.03 and 0.413 ±0.031; Compared to the complex model group,Haoqin Qingdan decoction group and virazole group were decreased （t =0.422,F = 112.834,P 〈 0.05 ）.Control group, virus group, complex model group, virazole group and Haoqin Qingdan decoction group NF-κB mRNA expression were separately： 0.075 ±0.148,0.379 ±0.019,0.291 ±0.012,0.169±0.026 and 0.175 ±0.033; the expression in virazole group and Haoqin Qingdan decoction group were decreased （t = 0.422, F = 112.834, P 〈 0.05 ）.The level of IFN-γin mice serum of control group, virus group,complex model group,virazole group and Haoqin Qingdan decoction group were： （7434.06 ±323.27） pg/ml,（8679.77 ±198.70）pg/ml,（8068.78 ±113.8） pg/ml, （7454.66 ±301.30） pg/ml and （7484.56 ±229.85） pg/ml respectively; the IFN-γ level in serum of Haoqin Qingdan decoction group and virazole group were decreased （t =0.201 ,F =5.390,P 〈0.05）.Each group of mice IL-4 contents were （3701.74 ±256.00） pg/ml, （3569.64 ±161.35） pg/ml, （3530.88 ±334.63）pg/ml,（3481.84 ±282.25） pg/ml and （3618.00 ±262.16 ） pg/ml; there were no significant difference between each group（t =0.414,F =0.505 ,P 〉0.05）.Th1/Th2 type cells in state of equilibrium （means IFN-γ/IL-4） were： 2.02 ±0.19,2.38 ±0.10,2.36 ±0.14,2.22 ±0.17 and 2.07 ±0.15; and complex model group Haoqin Qingdan decoction group and virazole group were decreased,and there was no significant difference observed （ t = 0.587, F = 3.684, P 〉 0.05 ）.Conclusion The effect of Haoqin Qingdan decoction on treatment of damp-heat syndrome of pneumonia infected by influenza virus was observed.Through reducing the expressions of TLR2,it decreases the levels of NF-κB mRNA and the proportionality of Th1/Th2 are obviously descend（P 〈0.05）.Haoqin Qingdan decoction can reduce the lung index and relieve the pathogenic changes.
出处
《中华预防医学杂志》
CAS
CSCD
北大核心
2010年第7期612-616,共5页
Chinese Journal of Preventive Medicine
基金
国家自然科学基金-广东省联合基金（U0632009）
关键词
蒿芩清胆汤
肺炎
病毒性
湿热
治疗结果
免疫组织化学
HAOQ1N QINGDAN DECOCTION
Pneumonia,viral
DAMPNKSS-HEAT
Treatmentoutcome
Immuno-histochemistry
作者简介
潘沅现工作单位：510282广州，南方医科大学第二附属医院检验科
通信作者：罗炳德，Email：gw@fimmu．com
