香菇多糖通过调控miR-221表达对肺结核大鼠巨噬细胞凋亡的影响  

Effect of Lentinan on Macrophage Apoptosis in Pulmonary Tuberculosis Rats by Regulating the Expression of miR-221

作　　者：沈凌筠 李莉 王戈 曾海燕 骆鹏举 朱兴玉 SHEN Ling-yun;LI Li;WANG Ge;ZENG Hai-yan;LUO Peng-ju;ZHU Xing-yu(Yunnan Infectious Disease Clinical Medical Center/Kunming Third People's Hospital,Kunming Yunnan 650000,China;Basic Medicine College of Kunming Medical University,Kunming Yunnan 650000,China)

机构地区：[1]云南省传染性疾病临床医学中心/昆明市第三人民医院,云南昆明650000 [2]昆明医科大学基础医学院,云南昆明650000

出　　处：《中医药导报》2022年第3期18-23,29,共7页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：昆明市卫生健康委员会卫生科研课题项目(2021-03-02-003)。

摘　　要：目的:探讨香菇多糖通过调控miR-221表达对肺结核大鼠巨噬细胞凋亡的影响。方法:采用尾静脉注射结核分枝杆菌悬液的方法建立肺结核大鼠模型,随机分为模型组、香菇多糖组、miR-221 agomir阴性对照组、miR-221 agomir组、香菇多糖+miR-221 agomir组,每组12只,另选取12只SD大鼠作为对照组,尾静脉注射0.9%NaCl溶液,经药物干预后,检测各组大鼠肺组织结核分枝杆菌菌落数;HE染色检测各组大鼠肺组织病理形态变化;流式细胞实验检测各组大鼠肺泡巨噬细胞凋亡情况;使用试剂盒测量各组大鼠肺组织ROS、MDA水平及血清炎症因子IL-6、IL-17水平;qRT-PCR法检测各组大鼠肺组织miR-221表达水平;免疫印记法检测各组大鼠肺组织凋亡蛋白Caspase-9、Bax表达水平。结果:与对照组比较,模型组、miR-221 agomir阴性对照组大鼠肺组织结构形态破坏严重,出现大量结核结节,肺间质水肿且炎症细胞浸润明显,肺泡结构变形,可见坏死细胞脱落,结核分枝杆菌菌落数、巨噬细胞凋亡率、肺组织ROS与MDA水平、血清炎症因子IL-6与IL-17水平、肺组织miR-221及凋亡蛋白Caspase-9、Bax表达水平均明显升高(P<0.05)。与模型组、miR-221 agomir阴性对照组、香菇多糖+miR-221 agomir组比较,香菇多糖组大鼠肺组织上述病理损伤均减轻,结核分枝杆菌菌落数、巨噬细胞凋亡率、肺组织ROS与MDA水平、血清炎症因子IL-6与IL-17水平、肺组织miR-221及凋亡蛋白Caspase-9、Bax表达水平均明显降低(P<0.05);miR-221 agomir组大鼠肺组织上述病理损伤均加重,结核分枝杆菌菌落数、巨噬细胞凋亡率、肺组织ROS与MDA水平、血清炎症因子IL-6与IL-17水平、肺组织miR-221及凋亡蛋白Caspase-9、Bax表达水平均明显升高(P<0.05)。miR-221 agomir阴性对照组大鼠肺组织上述病理损伤及各指标水平与模型组比较,差异无统计学意义(P>0.05)。结论:香菇多糖可通过下调miR-221表达,抑制结�Objective: To investigate the effect of lentinan on the apoptosis of macrophages in pulmonary tuberculosis rats by regulating the expression of miR-221. Methods: The pulmonary tuberculosis rat model was established by injecting mycobacterium tuberculosis suspension into the tail vein. The rats were randomly divided into model group, lentinan group, miR-221 agomir negative control group, miR-221 agomir group, and lentinan +miR-221 agomir group, with 12 rats in each group, another 12 SD rats were injected with 0.9% sodium chloride solution through the tail vein, set as the control group. After intervention with drugs, the number of mycobacterium tuberculosis colonies in the lung tissues of rats in each group was detected. The pathological and morphological changes in the lung tissues of rats in each group were detected by HE staining. The apoptosis of rat alveolar macrophages in each group was detected by flow cytometry. The levels of ROS, MDA and serum inflammatory factors IL-6 and IL-17 in the lung tissues of rats in each group were measured by the kit. The expression level of miR-221 in the lung tissues of rats in each group was determined by qRT-PCR experiment;and the expression levels of apoptotic proteins caspase-9 and Bax in the lung tissues of rats in each group were detected by immunoblotting experiment. Results: Compared with the control group, rats in the model group and the miR-221agomir negative control group had severely damaged lung tissue morphology, with a large number of tuberculosis nodules, pulmonary interstitial edema, obvious inflammatory cell infiltration, alveolar structure deformation, and shedding necrosis cells, the number of Mycobacterium tuberculosis colonies, the rate of macrophage apoptosis,the levels of ROS and MDA in lung tissue, the levels of serum inflammatory factors IL-6 and IL-17, the ex-pression levels of miR-221 and the apoptotic protein caspase-9 and Bax in lung tissue were significant increased(P<0.05). Compared with the model group, the miR-221 agomir negative control grou

关 键 词：肺结核 香菇多糖 MIR-221 巨噬细胞 凋亡 大鼠 

分 类 号：R285.5[医药卫生—中药学]