维生素D在豚鼠结核感染中的辅助治疗作用评价

Evaluation of the Effect of Vitamin D on Adjuvant Therapy in the Guinea Pig Model of Tuberculosis

目的建立豚鼠慢性结核病模型,评价维生素D在结核病辅助治疗中的作用c方法采用结核分枝杆菌标准株H37Rv以气溶胶方式成功感染豚鼠48只,按照随机数字表法分为空白对照组8只,每日常规剂量维生素D给药组(VD组)8只,单次高剂量维生素D隔周给药组(VDh组)8只,异烟脐-利福平给药组(H-R组)8只,异烟阱-利福平化疗联合每日常规剂量维生素D给药组(H-R+VD组)8只;异烟脐-利福平化疗联合单次高剂量维生素D隔周给药组(H-R+VDH组)8只。结核感染豚鼠4周后开始给药。分别于给药4周和8周后每组取4只豚鼠称体质量,处死,解剖,观察肺脏和脾脏病变,并进行病理学检查和活菌计数二结果豚鼠感染及给药过程中各组耐受性良好,无死亡。单独维生素D治疗的两组均未见豚鼠肺脏和脾脏活菌计数下降,也未观察到肺脏和脾脏病变减轻。治疗4周H-R组肺脏和脾脏活菌计数分别为(4.98 ± 0.26 ) lgCFU 和(4.02 ± 0.03 ) lgCFU, H-R+VDh 组肺脏和脾脏活菌计数分别为(4.39 ±0.11 ) lgCFU 和(2.30 ± 0.43 )lgCFU, H-R+VDH组较H-R组肺脏和脾脏活菌计数分别下降了 0.591gCFU和1.721gCFU,并观察到脾脏病理改变较之减轻:治疗8周后H-R组肺脏和脾脏活菌计数分别为(3.73 ± 0.23 ) lgCFU和(2.26 ± 0.24 ) lgCFU, H-R+VDH组肺脏活菌计数为(3.21 ± 0.23 ) lgCFU,脾脏达到无菌化,H-R+VDh组较H-R组肺脏和脾脏活菌计数分别下降了0.521gCFU和2.261gCFU,并观察到肺脏病理改变较之减轻。治疗4周后H-R+VD组脾脏活菌计数为(2.36 ±0.10) lgCFU, H-R+VD组较H-R组脾脏活菌计数下降了1.661gCFU。结论在豚鼠慢性结核病模型中,维生素D显示出辅助异烟耕-利福平治疗结核病的作用,可以进一步研究以应用于临床。

Objective To establish the chronic Guinea Pig model of tuberculosis and to evaluate the role of adjuvant therapy with Vitamin D. Methods Forty-eight Guinea Pigs infected with Mycobacterium tuberculosis standard strain (H37Rv) were randomly divided into blank control group, daily regular dosage of vitamin D group, single high dosage of vitamin D group taken every other week, isoniazid and rifampicin group, isoniazid and rifampicin plus daily regular dosage of vitamin D group, and isoniazid and rifampicin plus high dosage of vitamin D taken every other week group according to different treatment plans. There were 8 Guinea Pigs in each group. Four weeks after infection, we initiated treatment. 4 Guinea Pigs in each group were weighed, sacrificed, and dissected 4 weeks and 8 weeks after treatment. Later we counted the colony forming unit (CFU). We also performed the pathological examination in lungs and spleens. Results Our model animals had good tolerance and there was no death in each group throughout the infection and treatment. When compared to the control groups, the CFU counts of lungs and spleens of the Guinea Pigs did not decline, and the pathological changes of lungs and spleens did not improve in the two groups treated with vitamin D alone. Besides, after 4 weeks of treatment the CFU of lungs and spleens were (4.98 土 0.26) lgCFU and (4.02 ± 0.03) lgCFU in the group of isoniazid and rifampicin and (4.39 ±0.11) lgCFU and (2.30 ± 0.43) lgCFU in the group of isoniazid and rifampicin plus a single high dosage of vitamin D. When compared the group of isoniazid and rifampicin plus a single high dosage of vitamin D every other week with the group of isoniazid and rifampicin alone, the CFU counts decreased by 0.59 lgCFU in lungs and 1.72 lgCFU in spleens. After 8 weeks of treatment the CFU of lungs and spleens were (3.73 ±0.23) and (2.26 ± 0.24) lgCFU in the group of isoniazid and rifampicin, and the CFU of lungs were (3.21 ± 0.23) lgCFU and spleens were amicrobic in the group of isoniazid and rifampicin plus a single high dosage of vitamin D. When compared the group of isoniazid and rifampicin plus a single high dosage of vitamin D every other week with the group of isoniazid and rifampicin alone, the CFU counts decreased by 0.52 lgCFU in lungs and 2.26 lgCFU in spleens. Also the pathological changes of spleens in the group of isoniazid and rifampicin plus a single high dosage of vitamin D after 4 weeks of treatment improved and so as the lungs in the same group but of 8 weeks of treatment. The CFU of spleens were (2.36 ±0.10) lgCFU in the group of isoniazid and rifampicin plus daily regular dosage of vitamin D after 4 weeks of treatment. When compared the group of isoniazid and rifampicin plus daily regular dosage of vitamin D with isoniazid and rifampicin alone, the CFU of spleens decreased by 1.66 lgCFU after 4 weeks of treatment. Conclusion In the chronic tuberculosis model of Guinea Pig, vitamin D shows the effect as an adjuvant therapy when combined with isoniazid and rifampicin. It can be further studied to be used for clinical practice.