姜黄素对结核小鼠复发及p38丝裂原活化蛋白激酶信号通路的影响

Effect of curcumin on recurrence and p38 MAPK signal pathway in tuberculosis mice

目的研究姜黄素对结核小鼠复发及p38丝裂原活化蛋白激酶(p38 MAPK)信号通路的影响。方法将45只C57BL/6小鼠随机分为模型组(n=15)、实验组(n=15)和对照组(n=15),所有小鼠均尾静脉注射结核分枝杆菌H37RV(1×10^(7) CFU·mL^(-1))0.1 mL。4周后,实验组腹腔注射异烟肼10 mg·kg^(-1)+姜黄素60 mg·kg^(-1),对照组腹腔注射异烟肼10 mg·kg^(-1),模型组腹腔注射等量生理盐水,qd,连续干预4周。4周后各组小鼠均每周腹腔注射肿瘤坏死因子-α(TNF-α)抗体50μg,每天1次,连续4周。分别于药物干预结束后及诱导结束后用分枝杆菌中性罗氏培养管检测小鼠肺、脾组织荷菌量;以苏木精-伊红(HE)染色法观察各组小鼠肺组织病理学形态;以蛋白质印迹(WB)法检测小鼠肺组织中p-p38 MAPK蛋白的表达水平。结果药物干预结束后,模型组、对照组和实验组小鼠肺组织中荷菌量分别为(3.68±1.15),(1.13±0.12),(1.02±0.29)LgCFU·mL^(-1),脾组织中荷菌量分别为(4.26±0.39),(2.12±0.13),(2.05±0.11)LgCFU·mL^(-1),与模型组比较,实验组和对照组小鼠肺、脾组织中荷菌量均明显降低(均P<0.05),但实验组和对照组比较差异无统计学意义(P>0.05)。诱导结束后,模型组、对照组和实验组小鼠肺组织中荷菌量分别为(4.43±0.29),(4.03±0.10),(3.72±0.09)LgCFU·mL^(-1),脾组织中荷菌量分别为(4.53±0.44),(3.97±0.22),(4.05±0.21)LgCFU·mL^(-1),小鼠肺组织中小肉芽肿面积比分别为(8.65±0.32)%,(4.21±0.36)%,(3.98±0.52)%,小鼠肺组织中p-p38 MAPK蛋白相对表达量分别为1.47±0.33,1.13±0.35,0.76±0.25,差异均有统计学意义(均P<0.05)。结论姜黄素可通过降低p38 MAPK信号通路蛋白表达而抑制小鼠结核复发。

Objective To investigate the effect of curcumin on the recurrence and p38 mitogen-activated protein kinases(p38 MAPK)signaling pathway in tuberculosis mice.Methods A total of 45 C57 BL/6 mice were randomly divided into model group(n=15),test group(n=15)and control group(n=15).All mice were injected with mycobacterium tuberculosis H37 RV(1×10^(7) CFU·mL^(-1))0.1 mL via tail vein.After 4 weeks,test group was given isoniazid 10 mg·kg^(-1)+curcumin 60 mg·kg^(-1),control group was given isoniazid 10 mg·kg^(-1),model group was given the same amount of normal saline,all mice were intraperitoneally injected,once a day,continuous intervention for 4 weeks;4 weeks later,each mice in 3 groups were given tumor necrosis factor-α(TNF-α)antibody 50μg·week-1 intraperitoneally for 4 weeks,qd.Mycobacterium neutral Roche tube was used to detect the amount of bacteria in lung and spleen tissues of mice after drug intervention and induction;hematoxylin-eosin(HE)staining method was used to observe the pathological morphology of the lung tissue in each group of mice;the expression of p-p38 MAPK protein in lung tissues of mice was detected by Western blot(WB).Results After drug intervention,the amount of bacteria in lung tissues of model group,control group and test group were(3.68±1.15),(1.13±0.12),(1.02±0.29)LgCFU·m L^(-1),in spleen tissues were(4.26±0.39),(2.12±0.13),(2.05±0.11)LgCFU·m L^(-1).Compared with model group,the amount of bacteria in lung and spleen of mice in test group and control group were decreased significantly(all P<0.05),but there was no significant difference between test group and control group(P>0.05).After induction,the amount of bacteria in lung tissue of model group,control group and test group were(4.43±0.29),(4.03±0.10),(3.72±0.09)LgCFU·m L^(-1),which in spleen tissues were(4.53±0.44),(3.97±0.22),(4.05±0.21)LgCFU·m L^(-1),the area ratio of small granuloma in lung tissue of mice were(8.65±0.32)%,(4.21±0.36)%,(3.98±0.52)%,the relative expression levels of p-p38 MAPK protein in lung tissue of mice were 1.47±0.33,1.13±0.35 and 0.76±0.25,all with significant difference(all P<0.05).Conclusion Curcumin can inhibit the recurrence of tuberculosis in mice by reducing the expression of p38 MAPK signaling pathway protein.