月华胶囊对耐多药结核鼠Th1/Th2型细胞因子等影响的研究
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摘要
目的：①观察月华胶囊对耐多药结核鼠的治疗效果；②从分子免疫学角度探讨月华胶囊对细胞因子调控的作用；③了解月华胶囊对抗痨化疗药物肝肾损害的影响。
方法：昆明种小鼠尾部静脉注射耐多药结核杆菌建立感染模型，随机分为四组，(设立空白对照组)连续灌服相应的药物28天(空白对照组、模型组灌生理盐水)。观测：①体重、体温一般指标的变化；②肺病变指数、肺脏器指数、结核结节、吞噬细胞吞噬结核杆菌指数肺组织形态方面的变化；③胸腺脏器指数、免疫组化法观测血液中CD4~+、CD8~+细胞以及ELISA法观察各组血清IFN-γ、TNF-α、IL-2、IL-10细胞免疫指标的变化；④病理切片法观察月华胶囊对抗痨化学药物肝肾损害的作用。
结果：①与模型组比较：抗痨二线药物组、月华胶囊组、月华胶囊加抗痨二线药物组均能明显改善耐多药结核小鼠体温上升、体重降低(P＜0.01～0.05)；均能明显降低肺病变指数、肺脏器指数、坏死结核结节数、吞噬结核杆菌指数(P＜0.01)；均能显著提高胸腺指数，明显增加T细胞亚群中CD4T细胞数量，使Th1型细胞因子IFN-γ、TNF-α、IL-2含量上升，使Th2型细胞因子IL-10含量显著降低(P＜0.01)。②各治疗组间比较：月华胶囊组与抗痨二线药物组之间除体重外，其它各指标比较差异无显著性(P＞0.05)；月华胶囊加抗痨二线药物组优于月
华胶囊组和杭疥二线药物组(P<0.时一0.05)。③与杭疥二线药物组比
较，经病理组织学观察，月华胶囊加杭疥二线药物组肝脏多发性坏死、
肾小球坏死病变明显减轻。
结论:①月华胶囊在动物实验中具有治疗耐多药结核病的效应;②
月华胶囊和抗疹二线药合用对耐多药结核鼠模型的治疗有协同增效的
作用;③月华胶囊主要通过调节机体的细胞免疫功能来杀灭体内结核杆
菌，增加T细胞亚群数量，促进Thl型细胞因子IFN一Y、IL一2的分泌，
把TNF一a调控在一定的范围，降低工L一10含量促使ThZ细胞向Thl细胞
漂移，形成结核性肉芽肿，防止结核杆菌扩散;④月华胶囊对抗痔化疗
所致肝肾病理损害有一定的保护作用。
Objective: (1)To observe the therapeutic efficacy of Yue-hua Capsule to rats with MDR-TB. (2)To assess the mechanism of action of the regulation of Yue-hua Capsule to cytokine TH1/TH2 by molecular immunology.(3)To comprehend the effect of yue-hua Capsule to the injury of liver and kidney by antituberculosis chemotherapeutics.
Method: Build the infection model by injection Multi-drug resistant Bacillus tuberculosis (normal saline)into the vein of tail of Kunming genera rats and then group them into 4 groups random(Build the blank group). Drench them corresponding drugs 28 days continuously(Drench NS to the blank group , the model group).(1)To observe the change of weight and temperature, (2) To observe the change of lung organ exponent, thymus organ exponent, lung affection exponent, color, excreta, tubercler, the composition that macrophage phagocytize tubercler, etc.(3)To observe the change of CD4+
CD8+ in blood by immunohistochemistry staining method and To observe the change of IFN- y , TNF- a , IL-2, IL-10 in serum by ELISA assay. By pathological section, observing the effect of yue-hua Capsule to the injury to liver and kidney by antituberculosis chemotherapeutics drug.
Result: To campare with model group: the latter line antituberculisis grug group, Yue-hua Capsule group and the Yue-hua Capsule&the latter line antituberculosis drug group all could make multi-drug resistant-tuberculosis rats' body temperature descent, and make those body weigh increase (P<0.01 ~ 0.05 ) . And they could evidently descent the lung pathologic change exponent, lung organ exponent, necrotic tubercler and macroohage pathagocytize tubercler(p<0.01). They could improve the thymus organ exponent, increase the number of CD4I\ CDS in T cell subpopulation, make the content of IFN- Y TNF- a > IL-2 belonging to Thl cell factor increase, and make the content of IL-10 belonging to Th2 cell factor discrease(p<0.01). To compare with respective therapying group: There was no significant different in the therapeutic efficacy between Yue-hua Capsule group and the latter line antituberculosis drug group(p>0.05); Yue-hua Capsule&the latter line antituberculosis drug group had better efficacy than Yue-hua Capsule group and the latter line antituberculosis drug group ( P<0.01 ~ 0.05 ) . (3)To compare with the latter line antituberculosis
group ( P<0.01 ~ 0.05 ) . To compare with the latter line antituberculosis drug group: to observe by the result of pathological histology, liver multiple thanatosis and glomerulus lesion had lessened in Yue-hua Capsule group and the latter line antituberculosis drug group.
Conclusion: (Din the animal experiment, we can get good therapeutic efficacy from using Yue-hua Capsule to treat MDR-TB. Yue-hua Capsule&the latter line antituberculosis drug made the effect enhance on therapying the Multi-drug resistam iberculosis rats model. (3)Yue-hua Capsule could kill Bacillus tuberculosis in body by elevate the immune function of body, increase the number of T cell subpopulation, promote the secretion of IFN- Y and IL-2 belonging to Thl cell factor, regulate TNF-a to the certain range, discrease the content of IL-10, promote Th2 cell's transformation to Thl cell, form the scrofulous gumma to prevent the defusing of Bacillus tuberculosis. Yue-hua Capsule had the certain protecting effect on liver and kidney pathologic lesion induced by antiphtisy and chemotherapy.
方法：昆明种小鼠尾部静脉注射耐多药结核杆菌建立感染模型，随机分为四组，(设立空白对照组)连续灌服相应的药物28天(空白对照组、模型组灌生理盐水)。观测：①体重、体温一般指标的变化；②肺病变指数、肺脏器指数、结核结节、吞噬细胞吞噬结核杆菌指数肺组织形态方面的变化；③胸腺脏器指数、免疫组化法观测血液中CD4~+、CD8~+细胞以及ELISA法观察各组血清IFN-γ、TNF-α、IL-2、IL-10细胞免疫指标的变化；④病理切片法观察月华胶囊对抗痨化学药物肝肾损害的作用。
结果：①与模型组比较：抗痨二线药物组、月华胶囊组、月华胶囊加抗痨二线药物组均能明显改善耐多药结核小鼠体温上升、体重降低(P＜0.01～0.05)；均能明显降低肺病变指数、肺脏器指数、坏死结核结节数、吞噬结核杆菌指数(P＜0.01)；均能显著提高胸腺指数，明显增加T细胞亚群中CD4T细胞数量，使Th1型细胞因子IFN-γ、TNF-α、IL-2含量上升，使Th2型细胞因子IL-10含量显著降低(P＜0.01)。②各治疗组间比较：月华胶囊组与抗痨二线药物组之间除体重外，其它各指标比较差异无显著性(P＞0.05)；月华胶囊加抗痨二线药物组优于月
华胶囊组和杭疥二线药物组(P<0.时一0.05)。③与杭疥二线药物组比
较，经病理组织学观察，月华胶囊加杭疥二线药物组肝脏多发性坏死、
肾小球坏死病变明显减轻。
结论:①月华胶囊在动物实验中具有治疗耐多药结核病的效应;②
月华胶囊和抗疹二线药合用对耐多药结核鼠模型的治疗有协同增效的
作用;③月华胶囊主要通过调节机体的细胞免疫功能来杀灭体内结核杆
菌，增加T细胞亚群数量，促进Thl型细胞因子IFN一Y、IL一2的分泌，
把TNF一a调控在一定的范围，降低工L一10含量促使ThZ细胞向Thl细胞
漂移，形成结核性肉芽肿，防止结核杆菌扩散;④月华胶囊对抗痔化疗
所致肝肾病理损害有一定的保护作用。
Objective: (1)To observe the therapeutic efficacy of Yue-hua Capsule to rats with MDR-TB. (2)To assess the mechanism of action of the regulation of Yue-hua Capsule to cytokine TH1/TH2 by molecular immunology.(3)To comprehend the effect of yue-hua Capsule to the injury of liver and kidney by antituberculosis chemotherapeutics.
Method: Build the infection model by injection Multi-drug resistant Bacillus tuberculosis (normal saline)into the vein of tail of Kunming genera rats and then group them into 4 groups random(Build the blank group). Drench them corresponding drugs 28 days continuously(Drench NS to the blank group , the model group).(1)To observe the change of weight and temperature, (2) To observe the change of lung organ exponent, thymus organ exponent, lung affection exponent, color, excreta, tubercler, the composition that macrophage phagocytize tubercler, etc.(3)To observe the change of CD4+
CD8+ in blood by immunohistochemistry staining method and To observe the change of IFN- y , TNF- a , IL-2, IL-10 in serum by ELISA assay. By pathological section, observing the effect of yue-hua Capsule to the injury to liver and kidney by antituberculosis chemotherapeutics drug.
Result: To campare with model group: the latter line antituberculisis grug group, Yue-hua Capsule group and the Yue-hua Capsule&the latter line antituberculosis drug group all could make multi-drug resistant-tuberculosis rats' body temperature descent, and make those body weigh increase (P<0.01 ~ 0.05 ) . And they could evidently descent the lung pathologic change exponent, lung organ exponent, necrotic tubercler and macroohage pathagocytize tubercler(p<0.01). They could improve the thymus organ exponent, increase the number of CD4I\ CDS in T cell subpopulation, make the content of IFN- Y TNF- a > IL-2 belonging to Thl cell factor increase, and make the content of IL-10 belonging to Th2 cell factor discrease(p<0.01). To compare with respective therapying group: There was no significant different in the therapeutic efficacy between Yue-hua Capsule group and the latter line antituberculosis drug group(p>0.05); Yue-hua Capsule&the latter line antituberculosis drug group had better efficacy than Yue-hua Capsule group and the latter line antituberculosis drug group ( P<0.01 ~ 0.05 ) . (3)To compare with the latter line antituberculosis
group ( P<0.01 ~ 0.05 ) . To compare with the latter line antituberculosis drug group: to observe by the result of pathological histology, liver multiple thanatosis and glomerulus lesion had lessened in Yue-hua Capsule group and the latter line antituberculosis drug group.
Conclusion: (Din the animal experiment, we can get good therapeutic efficacy from using Yue-hua Capsule to treat MDR-TB. Yue-hua Capsule&the latter line antituberculosis drug made the effect enhance on therapying the Multi-drug resistam iberculosis rats model. (3)Yue-hua Capsule could kill Bacillus tuberculosis in body by elevate the immune function of body, increase the number of T cell subpopulation, promote the secretion of IFN- Y and IL-2 belonging to Thl cell factor, regulate TNF-a to the certain range, discrease the content of IL-10, promote Th2 cell's transformation to Thl cell, form the scrofulous gumma to prevent the defusing of Bacillus tuberculosis. Yue-hua Capsule had the certain protecting effect on liver and kidney pathologic lesion induced by antiphtisy and chemotherapy.
引文
[1] 中国防痨杂志专刊．2002，(24)2，65
[2] 汤红明，刘君炎，高立芬．ErN—Y与TNF联合治疗结核茵感染小鼠疗效及机理的研究[J]．免疫学杂志，2001，17(6)：457—459．
[3]. Mohammed KA, Nasreen N.[J].ward MJ Am J Respir Crit CareMed, 1999,1 9(5Pt,1): 1653-1659.
[4]. AmeixaC, FriedllandJS. [J]. Infect Immun,2001,69(4): 2470-2476.
[5] Driksen U,Hattcnhorst U,SchneiderP ctal. [J]. Blood,1998,(92)4: 1097-11033
[6] 唐神结，综述，肖和平，审校．结核病免疫研究进展[J]．国外医学内科学分册。2002，29(9)369—374
[7] 阎邦首．中草药抗结核作用的实验方法．中华结核病和呼吸系疾病杂志．1980．3 (2)123
[8] 涂叔云．卞如耀，陈修主编．药理实验方法学．第二饭，北京人民卫生出版社．1994．1．1392
[9] 陈虹，罗永艾，周波．实验性糖尿病结核动物模型的建立[J]．中国防痨杂志。2002。24(8)192-194
[10] 徐善荪月华丸治疗肺结核咯血64例疗效观察工企医刊1997，10(1)．-59-59
[11] 王花端．月华丸为主治疗复制性肺结核100例中医研究．1999，10(6)37
[12] 梁秋，李师，中西医结合治疗耐多药肺结核病的探讨 辽宁中医杂志 2003，30(3)221-222
[13] 李鲁铭．中药月华丸对预防抗痨药引起的药物性肝炎的影响 中国中西医结合杂志 2003，23(3)233-234
[14] 欧阳建军，伍参荣，白宇乾，月华胶囊对耐多药结核分支杆菌的抑菌实验”湖南中医学院学报．2003，23(5)．-14-15，22
[15] 中国防痨杂志．月华胶囊对耐多药结核病的临床研究。第八期，已付版费。
[16] 向阳，易晋华等．肺结核的中医治疗现状及展望 浙江中西医结合杂志．2001 (11)4、261-264
[17] Tchou-Wong KM，Harkin TJ,Chi C,et al．[J]Am J ResPir Crit Care Med，1997，156(6)：1996-2002．
[18] 张立群，马伟路．抗结核保护性免疫机制研究的若干进展[J]．中华呼吸与结核杂志，2000，23(3)：183—185
[19] Kitamura T, Ucsia K, Tanaka N, et al. [J]. Am J Respir Crit Care Med, 2000, 162(2Pt1): 6588—662.
[2] 汤红明，刘君炎，高立芬．ErN—Y与TNF联合治疗结核茵感染小鼠疗效及机理的研究[J]．免疫学杂志，2001，17(6)：457—459．
[3]. Mohammed KA, Nasreen N.[J].ward MJ Am J Respir Crit CareMed, 1999,1 9(5Pt,1): 1653-1659.
[4]. AmeixaC, FriedllandJS. [J]. Infect Immun,2001,69(4): 2470-2476.
[5] Driksen U,Hattcnhorst U,SchneiderP ctal. [J]. Blood,1998,(92)4: 1097-11033
[6] 唐神结，综述，肖和平，审校．结核病免疫研究进展[J]．国外医学内科学分册。2002，29(9)369—374
[7] 阎邦首．中草药抗结核作用的实验方法．中华结核病和呼吸系疾病杂志．1980．3 (2)123
[8] 涂叔云．卞如耀，陈修主编．药理实验方法学．第二饭，北京人民卫生出版社．1994．1．1392
[9] 陈虹，罗永艾，周波．实验性糖尿病结核动物模型的建立[J]．中国防痨杂志。2002。24(8)192-194
[10] 徐善荪月华丸治疗肺结核咯血64例疗效观察工企医刊1997，10(1)．-59-59
[11] 王花端．月华丸为主治疗复制性肺结核100例中医研究．1999，10(6)37
[12] 梁秋，李师，中西医结合治疗耐多药肺结核病的探讨 辽宁中医杂志 2003，30(3)221-222
[13] 李鲁铭．中药月华丸对预防抗痨药引起的药物性肝炎的影响 中国中西医结合杂志 2003，23(3)233-234
[14] 欧阳建军，伍参荣，白宇乾，月华胶囊对耐多药结核分支杆菌的抑菌实验”湖南中医学院学报．2003，23(5)．-14-15，22
[15] 中国防痨杂志．月华胶囊对耐多药结核病的临床研究。第八期，已付版费。
[16] 向阳，易晋华等．肺结核的中医治疗现状及展望 浙江中西医结合杂志．2001 (11)4、261-264
[17] Tchou-Wong KM，Harkin TJ,Chi C,et al．[J]Am J ResPir Crit Care Med，1997，156(6)：1996-2002．
[18] 张立群，马伟路．抗结核保护性免疫机制研究的若干进展[J]．中华呼吸与结核杂志，2000，23(3)：183—185
[19] Kitamura T, Ucsia K, Tanaka N, et al. [J]. Am J Respir Crit Care Med, 2000, 162(2Pt1): 6588—662.