| 王璐,周林水,董泉明,等.益气健脾方对香烟烟雾暴露小鼠的肺保护作用及潜在机制[J].浙江中医药大学学报,2023,47(11):1235-1242. |
| 益气健脾方对香烟烟雾暴露小鼠的肺保护作用及潜在机制 |
| Protective Effect of Yiqi Jianpi Recipe on Lung Injury in Mice Exposed to Cigarette Smoke and Its Potential Mechanism |
| DOI：10.16466/j.issn1005-5509.2023.11.001 |
| 中文关键词: 益气健脾 慢性阻塞性肺疾病 香烟烟雾 细胞凋亡 caspase-3 PARP1 |
| 英文关键词: tonifying Qi and invigorating the spleen chronic obstructive pulmonary disease cigarette smoke cell apoptosis caspase-3 PARP1 |
| 基金项目:浙江省中医药科技计划项目(2023ZR020)；浙江省自然科学基金项目(LQ20H270014)；国家自然科学基金项目(82104587) |
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| 中文摘要: |
| [目的] 初步探讨益气健脾方对香烟烟雾(cigarette smoke，CS)暴露小鼠肺损伤的作用及潜在机制。[方法] 60只C57BL/6小鼠随机分为正常组、模型组、预防组、治疗组和防治组。除正常组外，所有小鼠均接受为期4周的CS暴露，其中预防组、防治组分别在暴露初期接受4周及8周的益气健脾方干预，治疗组则于暴露结束后接受4周干预。记录各组小鼠一般情况，苏木精-伊红(hematoxylin-eosin，HE)染色观察肺组织病理学改变；吉姆萨染色观察肺泡灌洗液细胞分类并计数；原位末端转移酶标记(terminal-deoxynucleoitidyl transferase mediated nick end labeling，TUNEL)检测肺组织凋亡细胞DNA片段，免疫印迹检测B淋巴细胞瘤-2(B-cell lymphoma-2，Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X，Bax)、胱天蛋白酶-3(cysteine aspartate proteinase-3，caspase-3)、激活型caspase-3(cleaved-caspase-3)、多聚ADP核糖聚合酶1[poly(ADP-ribose) polymerase 1，PARP1]、激活型PARP1(cleaved-PARP1)蛋白表达。[结果] 至第4周，模型组小鼠生存率仅为72.72%；至第8周，模型组小鼠体质量最低(P<0.05)，肺组织内可见明显炎性细胞浸润及结构破坏。与模型组比较，益气健脾方干预的三组小鼠体质量增加，死亡率降低(P<0.05)，肺部炎症浸润及组织损伤均有所减轻(P<0.05)，其中以防治组改善最为显著(P<0.01)。与正常组比较，模型组小鼠细胞凋亡指数明显增加(P<0.01)，中药干预后则显著减少(P<0.01)。与正常组比较，模型组小鼠肺组织Bcl-2表达减少，Bax表达增多，Bcl-2/Bax比值下调，并且caspase-3、PARP1蛋白及其活化水平均上调(P<0.01，P<0.05)。与模型组比较，经中药干预8周后，小鼠肺组织中Bcl-2表达增加，Bax表达减少，Bcl-2/Bax比值上调，而Bax、caspase-3、cleaved-caspase-3、cleaved-PARP1表达均降低(P<0.05，P<0.01)。[结论] 益气健脾方可有效减轻CS暴露造成的小鼠肺组织损伤及炎症反应，其机制可能与抑制Bcl-2-Bax/caspase-3/PARP1细胞凋亡通路相关。 |
| 英文摘要: |
| [Objective] To investigate the effect of Yiqi Jianpi Recipe (YQJPR) on lung injury in mice exposed to cigarette smoke(CS)， and its potential mechanism. [Methods] Sixty C57BL/6 mice were randomly divided into normal group， model group(CS)， prevention group(CS+P)， treatment group(CS+T) and prevention and treatment group(CS+P+T). Except for normal group， all mice were exposed to CS for 4 weeks， CS+P group and CS+P+T group were subjected to YQJPR for 4 weeks and 8 weeks from CS exposure beginning， while CS+T group received YQJPR for 4 weeks after CS exposure. The general condition of mice in each group was recorded. Hematoxylin-eosin(HE) staining and Giemsa staining were respectively used to observe the lung histopathology and the classification and counts of inflammatory cells in alveolar lavage fluid. Terminal-deoxynucleoitidyl transferase mediated nick end labeling(TUNEL) apoptosis assay was used to detect the DNA fragments of apoptotic cells in lung tissues， and Western blot was used to assess the protein expression of B-cell lymphoma-2(Bcl-2)， Bcl-2 associated X(Bax)， cysteine aspartate proteinase-3(caspase-3)， cleaved-caspase-3， poly(ADP-ribose) polymerase 1(PARP1) and cleaved-PARP1. [Results] At 4th week， the survival rate of mice in model group was only 72.72%; at 8th week， compared with other groups， the mice in model group had decreased body weights with increased mortalities(P<0.05). HE staining showed obvious inflammatory cells infiltrations and structural damages in lung tissue. Compared with model group， the mice in three groups dministrated with YQJPR had gained body weights and reduced mortalities(P<0.05). Meanwhile， their lung inflammatory infiltrations and tissue damages were alleviated(P<0.05). As expected， the pathological changes had most significant improvement in mice with 8-week YQJPR administration(P<0.01). TUNEL apoptosis assay showed that compared with normal group， the apoptotic index in model group was significantly increased(P<0.01)， while in mice with YQJPR， all three groups had a lower apoptosis rate in lung(P<0.01). Western blot showed that compared with normal group， the expression level of Bax was increased and the expression level of Bcl-2 was decreased in model group. The ratio of Bcl-2 to Bax was also decreased， caspase-3， PARP1 and their activation proteins were totally upregulated(P<0.01， P<0.05). Intriguingly， compared with model group，8-week treatment of YQJPR could inhibit the expression of Bax， caspase-3， cleaved caspase-3， cleaved PARP1， whereas promote the expression of Bcl-2 and raise the ratio of Bcl-2 to Bax(P<0.05， P<0.01). [Conclusion] YQJPR could effectively protect the lung injury and inflammation from CS exposure in mice， the mechanism may relate to the inhibition of apoptosis pathway of Bcl-2-Bax/caspase-3/PARP1. |
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