双黄连颗粒对大鼠肺炎支原体肺炎模型中TNF-α、IL-6和NF-κB的影响

Effects of Shuanghuanglian Granules on TNF-α,IL-6 and NF-κB in Mycoplasma Pneumoniae Pneumonia Model in Rats

目的:研究双黄连颗粒对肺炎支原体感染大鼠血清或肺组织中肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6和核因子(NF)-κB等炎性相关因子的影响,进一步研究双黄连颗粒的药理作用机制。方法:制备大鼠肺炎支原体肺炎模型,60只SD大鼠随机分为空白对照组、模型组和双黄连低、中、高剂量组及阳性药对照组六组,在连续感染3 d后开始给药。连续给药7 d后采集大鼠血清及肺组织样本,肺组织经HE染色后于光学显微镜下观察病变情况;酶联免疫吸附实验(ELISA)法检测血清中TNF-α、IL-6水平;Westernblot检测肺组织中磷酸化NF-κB p65的表达。结果:空白对照组大鼠血清中TNF-α、IL-6水平分别为(0.96±0.23)ng/mL、(451.43±33.42)ng/mL,磷酸化NF-κB p65相对含量为0.16±0.05;模型组大鼠肺组织病理评分为(4.50±1.32)分,血清中TNF-α、IL-6的水平分别为(1.84±0.42)ng/mL、(603.29±63.57)ng/mL,磷酸化NF-κB p65相对含量为1.07±0.06,与空白对照组相比均升高。双黄连低、中、高剂量组血清TNF-α水平分别为(1.62±0.32)ng/mL、(1.47±0.31)ng/mL、(1.24±0.25)ng/mL,血清IL-6水平分别为(551.56±50.93)ng/mL、(509.57±49.92)ng/mL、(487.14±44.15)ng/mL,磷酸化NF-κB p65相对含量分别为0.43±0.15、0.12±0.07、0.10±0.05,与模型组相比均呈剂量依赖性下降。结论:双黄连颗粒能够通过降低TNF-α、IL-6水平,抑制NF-κB信号通路异常活化,从而减轻肺组织中的炎性症状,从而使肺组织恢复正常的形态和生理功能。

Objective:To study the effects of Shuanghuanglian granules on inflammatory factors such as tumor necrosis factor(TNF)-α,interleukin(IL)-6 and nuclear factor(NF)-κB in serum or lung tissues of rats infected with Mycoplasma pneumoniae,so as to further study the pharmacological mechanism of Shuanghuanglian granules.Methods:Mycoplasma pneumoniae pneumonia model in rats was established.Sixty SD rats were randomly divided into the blank control group,the model group,the Shuanghuanglian low,medium,and high dose group and the positive control group.The administration was started after 3 d of continuous infection.After 7 d of continuous administration,serum and lung tissues were collected,and the lung tissues were stained with HE to observe the lesions under an optical microscope.The serum levels of TNF-αand IL-6 were detected by enzyme-linked immunosorbent assay(ELISA).Westernblot was adopted to detect the expression of phosphorylated NF-κB p65 in lung tissues.Results:The serum levels of TNF-αand IL-6 in the blank control group were(0.96±0.23)ng/mL,(451.43±33.42)ng/mL,and the relative content of phosphorylated NF-κB p65 was 0.16±0.05.The lung tissue pathological score of rats in the model group was(4.50±1.32)points,the serum levels of TNF-αand IL-6 were respectively(1.84±0.42)ng/mL and(603.29±63.57)ng/mL,and the relative content of phosphorylated NF-κB p65 was 1.07±0.06,which were higher than those of the blank control group.Serum levels of TNF-αin the Shuanghuanglian low,medium,and high dose group were respectively(1.62±0.32)ng/mL,(1.47±0.31)ng/mL and(1.24±0.25)ng/mL,and serum levels of IL-6 were respectively(551.56±50.93)ng/mL,(509.57±49.92)ng/mL and(487.14±44.15)ng/mL,the relative content of phosphorylated NF-κB p65 were respectively 0.43±0.15,0.12±0.07 and 0.10±0.05,compared with the model group,the dose-dependent decrease was observed.Conclusion:Shuanghuanglian granules can reduce the inflammatory symptoms in the lung tissues by reducing the levels of TNF-αand IL-6,inhibit the abnormal activation of the NF-κB signaling pathway,thereby restoring the normal morphology and physiological functions in lung tissues.