CAJ | 학술논문

目的：探讨大黄素对病毒性心肌炎（VMC）小鼠心肌的保护作用及其分子机制。方法55只雄性BALB/c小鼠随机分为对照组（n=15）、模型组（n=20）及大黄素组（n=20），模型组、大黄素组小鼠腹腔接种0.1 ml内含柯萨奇病毒B3（CVB3）的Eagle's液建立VMC模型，对照组仅注射Eagle’s液，于接种当天，大黄素组以3 mg/ml大黄素溶液0.1 ml灌胃，其余2组以0.1 ml蒸馏水灌胃，1次/d，共21 d，记录实验期间小鼠死亡数目，比较各组死亡率。第7天每组处死5只小鼠，取心脏，测定病毒滴度。第22天称体质量（BW）后处死全部小鼠，收集外周血，剥离心脏，称心脏质量（HW），计算HW/BW，行HE染色计算心肌病理积分，采用荧光实时定量PCR、Western blotting分别检测心肌白介素-23（IL-23）、白介素-17（IL-17）mRNA和蛋白表达，通过酶联免疫吸附法测定血清IL-23、IL-17浓度，流式细胞术分析Th17细胞频率，利用Western blotting测定心肌细胞核内核因子-κB（NF-κB）p65表达，ELISA分析心肌白介素-1β（IL-1β）、白介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）含量。结果大黄素组死亡率、心肌病理积分及病毒滴度较模型组减少（P<0.05）。模型组HW/BW、心肌IL-23及IL-17 mRNA与蛋白表达水平、血清IL-23和IL-17浓度、Th17细胞频率、胞核NF-κB p65表达水平及心肌IL-1β、IL-6、TNF-α含量显著高于对照组（P<0.01），与模型组比较，大黄素组上述指标明显降低（P<0.05）。结论大黄素可能通过抑制IL-23/IL-17炎症轴、Th17细胞增殖及病毒复制发挥抗VMC作用。
목적：탐토대황소대병독성심기염（VMC）소서심기적보호작용급기분자궤제。방법55지웅성BALB/c소서수궤분위대조조（n=15）、모형조（n=20）급대황소조（n=20），모형조、대황소조소서복강접충0.1 ml내함가살기병독B3（CVB3）적Eagle's액건립VMC모형，대조조부주사Eagle’s액，우접충당천，대황소조이3 mg/ml대황소용액0.1 ml관위，기여2조이0.1 ml증류수관위，1차/d，공21 d，기록실험기간소서사망수목，비교각조사망솔。제7천매조처사5지소서，취심장，측정병독적도。제22천칭체질량（BW）후처사전부소서，수집외주혈，박리심장，칭심장질량（HW），계산HW/BW，행HE염색계산심기병리적분，채용형광실시정량PCR、Western blotting분별검측심기백개소-23（IL-23）、백개소-17（IL-17）mRNA화단백표체，통과매련면역흡부법측정혈청IL-23、IL-17농도，류식세포술분석Th17세포빈솔，이용Western blotting측정심기세포핵내핵인자-κB（NF-κB）p65표체，ELISA분석심기백개소-1β（IL-1β）、백개소-6（IL-6）、종류배사인자-α（TNF-α）함량。결과대황소조사망솔、심기병리적분급병독적도교모형조감소（P<0.05）。모형조HW/BW、심기IL-23급IL-17 mRNA여단백표체수평、혈청IL-23화IL-17농도、Th17세포빈솔、포핵NF-κB p65표체수평급심기IL-1β、IL-6、TNF-α함량현저고우대조조（P<0.01），여모형조비교，대황소조상술지표명현강저（P<0.05）。결론대황소가능통과억제IL-23/IL-17염증축、Th17세포증식급병독복제발휘항VMC작용。
Objective To explore the effects of emodin in myocardial protection in mice with viral myocarditis (VMC) and explore molecular mechanisms. Methods Fifty-five male 4-week-old BALB/c mice were randomly divided into control group (n=15), model group (n=20), and emodin group (n=20). The mice in model and emodin groups were inoculated with 0.1 ml Eagle's solution containing coxsackievirus B3 intraperitoneally, and those in the control group were given only 0.1 ml Eagle's solution. From the day of inoculation, the mice in emodin group received intragastric administration with 0.1 ml of 3 mg/ml emodin solution once daily for 21 consecutive days, and those in the control and model groups received 0.1 ml distilled water only. On day 7 after inoculation, 5 mice from each group were sacrificed to determine the viral titers in the cardiac tissues. All the mice were sacrificed on day 22 for measurement of the heart weight and histopathological inspection of the heart with HE staining. The mRNA and protein expression levels of myocardial interleukin-23 (IL-23) and interleukin-17 (IL-17) were detected by real-time quantitative PCR and Western blotting, respectively, and serum IL-23 and IL-17 levels were examined using enzyme linked immunosorbent assay (ELISA). Th17 cell frequencies were analyzed by flow cytometry. The expression levels of myocardial nuclear factor-κB (NF-κB) p65 in the cardiomyocyte nuclei were examined using Western blotting, and myocardial interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α) contents were detected by ELISA. Results The mortality, myocardial histopathologic scores and virus titers in emodin group were all significantly lower than those in the model group (P<0.05). The heart-to-body weight ratio, myocardial IL-23 and IL-17 expressions, serum IL-23 and IL-17 levels, Th17 cell frequencies, cardiomyocyte nuclear NF-κB p65 expression, and myocardial contents of IL-1β, IL-6 and TNF-αwere all significantly increased in the model group as compared to the control group (P<0.01) but reduced significantly in emodin group as compared to model group (P<0.05). Conclusion Emodin can protect against VMC by inhibiting IL-23/IL-17 inflammatory axis, Th17 cell proliferation and viral replication in mice.