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河南中医药大学 第二附属医院,郑州 450000
Received:05 June 2019,
Published Online:21 October 2019,
Published:20 February 2020
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Hong-en BIAN, Tuan-ying CHEN. Therapeutic Effect and Mechanism of Chaige Qinlian Tang on Young Mouse MRSA Pneumonia Model[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(4): 23-28.
Hong-en BIAN, Tuan-ying CHEN. Therapeutic Effect and Mechanism of Chaige Qinlian Tang on Young Mouse MRSA Pneumonia Model[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(4): 23-28. DOI: 10.13422/j.cnki.syfjx.20200303.
目的:
2
观察柴葛芩连汤对肺炎幼年小鼠(幼鼠)模型的治疗作用,并探讨其相关的作用机制。
方法:
2
以金黄色葡萄球菌经幼鼠鼻孔缓慢滴入小鼠鼻腔内的方法复制肺炎模型,造模成功后将幼鼠随机分为模型组、克林霉素组、柴葛芩连汤高、低剂量组,并以假手术组作为阴性组。造模后当天开始给药,柴葛芩连汤高、低剂量200,100 mg·kg
-1
,克林霉素120 mg·kg
-1
。每天观察幼鼠状况,5 d后对每组幼鼠的肺脏进行菌落计数,采用酶联免疫吸附测定(ELISA)检测每组小鼠的肺脏灌洗液中白细胞介素(IL)-16,肿瘤坏死因子(TNF)-
α
的表达水平,采用实时荧光定量聚合酶链式反应(Real-time PCR)及蛋白免疫印迹法(Western blot)测量幼鼠肺中肿瘤坏死因子受体(TNFR)1,半胱氨酸的天冬氨酸蛋白水解酶(Caspase)-3,Caspase-7 mRNA及蛋白的表达,并观察肺脏的病理学改变。
结果:
2
与假手术组比较,造模幼鼠呼吸状态及活动状态较差,给药后均有不同程度改善,其中以柴葛芩连汤高剂量组存活率较高。与假手术组比较,模型组幼鼠肺内菌落计数均有所增加;与模型组比较,克林霉素组、柴葛芩连汤高剂量组肺内菌落计数改善较明显(
P
<
0.05,
P
<
0.01)。与假手术组比较,模型组IL-16,TNF-
α
的表达水平及肺中TNFR1,Caspase-3,Caspase-7 mRNA和蛋白的表达水平均显著升高(
P
<
0.01),而与模型组比较,克林霉素组、柴葛芩连汤高、低剂量组IL-16,TNF-
α
的表达水平及肺中TNFR1,Caspase-3,Caspase-7 mRNA和蛋白的表达水平均明显下降(
P
<
0.05,
P
<
0.01),且肺脏病理学改变有所改善,克林霉素组、柴葛芩连汤高剂量组改善较明显。
结论:
2
柴葛芩连汤对金黄色葡萄球菌肺炎幼鼠模型具有一定的治疗作用,该作用可能与调控TNFR1,Caspase-3,Caspase-7通路,减少IL-16,TNF-
α
的分泌,增强对金黄色葡萄球菌的清除有关。
Objective:
2
To explore the therapeutic effect and mechanism of Chaige Qinlian Tang on pneumonia in young mice.
Method:
2
The pneumonia model was duplicated by slowly dripping Staphylococcus aureus into the nasal cavity of mice.After successful modeling
the mice were randomly divided into model group
clindamycin group
and high and low-dose Chaige Qinlian Tang groups
with sham operation group as negative control group.The rats were given 200 mg·kg
-1
high-dose Chaige Qinlian Tang
100 mg·kg
-1
low-dose Chaige Qinlian Tang and 120 mg·kg
-1
clindamycin.The mice were observed every day.Colonies were counted in the lungs of each group five days later.The expression levels of interleukin(IL)-16
tumor necrosis factor (TNF)-
α
in lung lavage fluid of each group were determined by enzyme linked immunosorbent assay (ELISA). Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to measure the expression levels of IL-16
TNF-
α
in lung lavage fluid of each group.The expressions of tumor necrosis factor receptor (TNFR) 1
Caspase-3 and Caspase-7 in lung and the pathological changes of lung were observed.
Result:
2
Compared with the sham operation group
the respiratory state and the activity state of the model mice were worse
and the survival rate was higher in the high-dose Chaige Qinlian Tang group.Compared with the sham operation group
the pulmonary colony counts in the model group and treatment groups were increased
compared with the model group
the lung colony counts in clindamycin group and high-dose Chaige Qinlian Tang group were improved significantly (
P
<
0.05
P
<
0.01). Compared with the control group
the expression levels of IL-16
TNF-
α
TNFR1
Caspase-3
Caspase-7 mRNA and protein in the lung of model group and treatment groups were significantly increased (
P
<
0.01). Compared with model group
the expression levels of IL-16
TNF-
α
and TNFR1
Caspase-3
Caspase-7 in the lung of clindamycin group and high and low-dose Chaige Qinlian Tang groups were significantly increased (
P
<
0.01). The expression levels of protein and mRNA were significantly decreased (
P
<
0.05
P
<
0.01)
and the pathological changes of lung were improved
especially in clindamycin group and high-dose Chaige Qinlian Tang group.
Conclusion:
2
Chaige Qinlian Tang has a certain therapeutic effect on Staphylococcus aureus pneumonia in young mice.This effect may be related to regulating TNFR1
Caspase-3 and Caspase-7 pathways
reducing the secretion of IL-16 and TNF-alpha
and enhancing the clearance of staphylococcus aureus.
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