健脾益肺Ⅱ号减少香烟暴露联合脂多糖气道滴注诱导的肺组织凋亡的机制研究

Mechanism of JYⅡ on Protecting Lung Tissue from Apoptosis Induced by Cigarette Smoke Exposure Combined with LPS Instillation

目的:探讨健脾益肺Ⅱ号减少香烟暴露联合脂多糖气道滴注诱导的肺组织凋亡的机制。方法:SD大鼠经香烟烟雾暴露8 w联合脂多糖(LPS)重复滴注(d7,d21),建立COPD大鼠模型。实验分为正常组、模型组、健脾益肺Ⅱ号12. 15 g/kg、24. 3 g/kg组及氨茶碱60 mg/kg组。观察和记录各组大鼠一般状态,HE染色法观察各组大鼠肺部病理改变,Western blot法检测各组大鼠肺组织中Akt、p-Akt、Bax和Bcl-2的蛋白表达,并检测和记录大鼠血液中白细胞总数(WBC)计数和中性粒细胞(NEU),单核细胞(MONO),淋巴细胞(Lymph)的数量。结果:与正常组相比,模型组大鼠一般状态较差,表现为呼吸急促,口鼻分泌物增多,而健脾益肺Ⅱ号组大鼠及氨茶碱组大鼠一般状态相对较好。HE染色结果显示,健脾益肺Ⅱ号(12. 15 g/kg、24. 3 g/kg)可明显改善模型大鼠肺泡结构的破坏,而氨茶碱作用不明显。Western blot结果表明,健脾益肺Ⅱ号12. 15 g/kg明显增加了模型大鼠血液中WBC、NEU和MONO细胞数量,明显降低模型大鼠肺组织p-Akt/Akt比值,并下调模型大鼠肺组织Bax蛋白表达,健脾益肺Ⅱ号24. 3 g/kg明显增加了模型大鼠血液中NEU和MONO细胞数量;氨茶碱对凋亡信号通路无影响。结论:健脾益肺Ⅱ号可通过下调Bax表达,减少肺组织细胞的凋亡和破坏。而下调Bax表达的原因,可能与血液中白细胞数目增多有关。

Objective : To investigate the mechanism of JYⅡ on apoptosis of lung tissues induced by cigarette smoke exposure combined with LPS intratracheal instillation. Methods: SD rats were exposed to cigarette smoke for 8 weeks,and were intratracheal-instilled with LPS on day7 and day21 to establish COPD rat model. Rats were divided into the control group,the model group,JYⅡ groups(12. 15 g/kg、24. 3 g/kg) and 60 mg/kg aminophylline group. General conditions of rats were observed and recorded. Pathological changes of lungs were observed by HE staining. Expressions of Akt,p-Akt,Bax,Bcl-2 were tested by western blot. Cell counting of WBC,neutrophils,macrophages and lymphocytes in blood were recorded. Results: Compared with the control group,rat conditions were worse in the model group,especially polypnea and hyper secretion in noses and mouths,while rat conditions in JYⅡ and aminophylline groups were in good state. Compared with the control group,pulmonary alveoli damages were observed in the model group by HE staining,while JYⅡ significantly improved the damages. Compared with the model group,western blot results showed JYⅡ(12. 15 g/kg) significantly increased the numbers of WBC、NEU and MONO in blood,decreased the level of p-Akt/Akt ratio in lung tissues,and down regulated the BAX protein expression in lung tissues,24. 3 g/kg JYⅡ significantly increased the number of NEU and MONO in blood. No significant change in apoptosis signal way was observed in aminophylline group. Conclusion: JYⅡ can reduce apoptosis and injury of lung tissues by down regulating the expression of Bax,which may be related to the increasing the number of white blood cells in rat circulation.