中华预防医学杂志
中華預防醫學雜誌
중화예방의학잡지
CHINESE JOURNAL OF
2010年
7期
612-616
,共5页
蒿芩清胆汤%肺炎%病毒性%湿热%治疗结果%免疫组织化学
蒿芩清膽湯%肺炎%病毒性%濕熱%治療結果%免疫組織化學
호금청담탕%폐염%병독성%습열%치료결과%면역조직화학
HAOQIN QINGDAN DECOCTION%Pneumonia,viral%DAMPNESS-HEAT%Treatment outcome%Immuno-histochemistry
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 .
目的 在動物模型的基礎上,探討蒿芩清膽湯在小鼠機體免疫調節中的信號傳導機製,為該藥物治療濕熱證打下理論基礎.方法 採用隨機數字錶法將40隻由美國國立衛生研究院(National Institute of Health,NIH)培育而成的小鼠(1980年引進我國,簡稱NTH小鼠)分為正常組8隻、流行性感冒(簡稱流感)病毒組(單純病毒感染)8隻、複閤模型組(高脂飲食+濕熱環境+病毒感染)8隻、暘性藥物組(模型組+病毒唑)8隻、蒿芩清膽湯組(模型組+蒿芩清膽湯)8隻.計算各組小鼠肺指數及肺指數抑製率;提取小鼠腹腔巨噬細胞用逆轉錄PCR(RT-PCR)法檢測TLR2(Toll樣受體-2)及NF-κB mRNA(覈因子κB-mRNA)的錶達;採用酶聯免疫吸附測定法(ELISA)測定標本中白介素-4(IL-4)、榦擾素-γ(IFN-γ)的濃度,求齣IFN-γ/IL-4比值用于瞭解機體Th1/Ih2(Th1類與Th2類細胞比值)類細胞的平衡狀態.結果 正常組、病毒組、複閤模型組、暘性藥物組和蒿芩清膽湯組小鼠的肺指數分彆為(0.79±0.11)%、(1.93±0.38)%、(1.41±0.26)%、(1.10±0.26)%、(1.02±0.16)%;暘性藥物組和蒿芩清膽湯組小鼠的肺指數和複閤模型組相比均有所減低,有統計學意義(t=0.322,P＜0.05).RT-PCR的結果顯示,正常組、病毒組、複閤模型組、暘性藥物組和蒿芩清膽湯組TLR2 mRNA的錶達分彆是0.145±0.017、0.991±0.149、0.903±0.124、0.257±0.032和0.413±0.031;蒿芩清膽湯組、暘性藥物組比複閤模型組的小鼠低,均有統計學意義(t=0.013,F=73.467,P＜0.05).正常組、病毒組、複閤模型組、暘性藥物組和蒿芩清膽湯組NF-κB mRNA的錶達分彆是0.075±0.015、379±0.019、0.291±0.012、0.169±0.026和0.175±0.033;暘性藥物組、蒿芩清膽湯組與複閤模型組相比均有所減低(t=0.422,F=112.834,P＜0.05).正常組、病毒組、複閤模型組、暘性藥物組和蒿芩清膽湯組小鼠血清中IFN-γ含量分彆為(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;蒿芩清膽湯組和暘性藥物組血清中IFN-γ含量比複閤模型組減低,有統計學意義(t=0.201,F=5.390,P＜0.05).正常組、病毒組、複閤模型組、暘性藥物組和蒿芩清膽湯組小鼠血清中IL-4含量分彆為(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;各組間差異無統計學意義(t=0.414,F=0.505,P＞0.05).Th1/Th2比值(即:IFN-γIL-4)分彆為:2.02±0.19、2.38±0.10、2.36±0.14、2.22±0.17和2.07±0.15;與複閤模型組相比,蒿芩清膽湯組和暘性藥物組均呈減低趨勢,但無統計學意義(t=0.587,F=3.684,P＞0.05).結論 蒿芩清膽湯對流感病毒性肺炎濕熱證療效顯著,它通過降低膜錶麵受體TLR2的錶達來抑製NF-κB的活化,降低血清IFN-γ水平,使Th1/Th2類細胞趨于平衡;降低肺泡內炎癥反應,從而達到減少肺組織損傷的治療目的 .
목적 재동물모형적기출상,탐토호금청담탕재소서궤체면역조절중적신호전도궤제,위해약물치료습열증타하이론기출.방법 채용수궤수자표법장40지유미국국립위생연구원(National Institute of Health,NIH)배육이성적소서(1980년인진아국,간칭NTH소서)분위정상조8지、류행성감모(간칭류감)병독조(단순병독감염)8지、복합모형조(고지음식+습열배경+병독감염)8지、양성약물조(모형조+병독서)8지、호금청담탕조(모형조+호금청담탕)8지.계산각조소서폐지수급폐지수억제솔;제취소서복강거서세포용역전록PCR(RT-PCR)법검측TLR2(Toll양수체-2)급NF-κB mRNA(핵인자κB-mRNA)적표체;채용매련면역흡부측정법(ELISA)측정표본중백개소-4(IL-4)、간우소-γ(IFN-γ)적농도,구출IFN-γ/IL-4비치용우료해궤체Th1/Ih2(Th1류여Th2류세포비치)류세포적평형상태.결과 정상조、병독조、복합모형조、양성약물조화호금청담탕조소서적폐지수분별위(0.79±0.11)%、(1.93±0.38)%、(1.41±0.26)%、(1.10±0.26)%、(1.02±0.16)%;양성약물조화호금청담탕조소서적폐지수화복합모형조상비균유소감저,유통계학의의(t=0.322,P＜0.05).RT-PCR적결과현시,정상조、병독조、복합모형조、양성약물조화호금청담탕조TLR2 mRNA적표체분별시0.145±0.017、0.991±0.149、0.903±0.124、0.257±0.032화0.413±0.031;호금청담탕조、양성약물조비복합모형조적소서저,균유통계학의의(t=0.013,F=73.467,P＜0.05).정상조、병독조、복합모형조、양성약물조화호금청담탕조NF-κB mRNA적표체분별시0.075±0.015、379±0.019、0.291±0.012、0.169±0.026화0.175±0.033;양성약물조、호금청담탕조여복합모형조상비균유소감저(t=0.422,F=112.834,P＜0.05).정상조、병독조、복합모형조、양성약물조화호금청담탕조소서혈청중IFN-γ함량분별위(7434.06±323.27)pg/ml、(8679.77±198.70)pg/ml、(8068.78±113.8)pg/ml、(7454.66±301.30)pg/ml화(7484.56±229.85)pg/ml;호금청담탕조화양성약물조혈청중IFN-γ함량비복합모형조감저,유통계학의의(t=0.201,F=5.390,P＜0.05).정상조、병독조、복합모형조、양성약물조화호금청담탕조소서혈청중IL-4함량분별위(3701.74±256.00)pg/ml、(3569.64±161.35)pg/ml、(3530.88±334.63)pg/ml、(3481.84±282.25)pg/ml화(3618.00±262.16)pg/ml;각조간차이무통계학의의(t=0.414,F=0.505,P＞0.05).Th1/Th2비치(즉:IFN-γIL-4)분별위:2.02±0.19、2.38±0.10、2.36±0.14、2.22±0.17화2.07±0.15;여복합모형조상비,호금청담탕조화양성약물조균정감저추세,단무통계학의의(t=0.587,F=3.684,P＞0.05).결론 호금청담탕대류감병독성폐염습열증료효현저,타통과강저막표면수체TLR2적표체래억제NF-κB적활화,강저혈청IFN-γ수평,사Th1/Th2류세포추우평형;강저폐포내염증반응,종이체도감소폐조직손상적치료목적 .
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.
