参苓白术散对COPD大鼠下丘脑及胃组织Ghrelin,Obestatin的影响

目的:观察参苓白术散干预对肺脾两虚型慢性阻塞性肺疾病(COPD)模型大鼠生长激素释放肽(Ghrelin),肥胖抑制素(Obestatin)蛋白表达的影响,基于Ghrelin/Obestatin信号调节通路从分子水平探讨培土生金法治疗COPD的作用机制和科学内涵。方法:60只SPF级Wistar大鼠随机分为空白组、肺脾两虚模型组、参苓白术散组(根据药物干预时间不同,分为28,35,42 d组),每组12只。采用病证结合造模法复制肺脾两虚型COPD大鼠,空白组大鼠吸入空气+气管内注射等量生理盐水+灌服等量生理盐水;模型组及参苓白术散组采用烟熏+气管内滴注脂多糖+灌服冰冷番泻叶浸液建立肺脾两虚型COPD大鼠模型;参苓白术散组给予参苓白术散灌胃,分别于药物干预第28,35,42天处死大鼠,利用免疫组化法及蛋白免疫印迹法测定大鼠下丘脑组织及胃组织中Ghrelin,Obestatin蛋白表达变化。结果:与空白组比较,肺脾两虚模型组大鼠下丘脑及胃组织中Ghrelin,Obestatin蛋白表达及积分吸光度均显著降低(P0.01);与肺脾两虚模型组比较,参苓白术散组(28,35,42天)大鼠胃、下丘脑组织Ghrelin,Obestatin蛋白表达及积分吸光度均有不同程度升高(P0.05)。结论:参苓白术散可调节肺脾两虚型COPD模型大鼠能量代谢,改善其营养不良状况,其机制可能通过调节Ghrelin,Obestatin动态平衡从而维持机体能量稳态来实现。

Effect of Shenling Baizhusan on Expression of Ghrelin and Obestatin in Hypothalamus and Gastric Tissue of COPD Rats

Objective:To discuss the effect of Shenling Baizhusan on Ghrelin and Obestatin protein expression of in the rat models with chronic obstructive pulmonary disease (COPD), and investigate its mechanism and scientific connotation in treating COPD based on the Ghrelin/Obestatin signal regulating pathway from the molecular level. Method:The 60 SPF Wistar rats were randomly divided into blank control group, lung and spleen deficiency model group and Shenling Baizhusan groups (28 days group, 35 days group and 42 days group according to different drug intervention time), 12 in each group. Lung and spleen deficiency COPD rat models were established by using the disease and syndrome combination method. In blank control group, the rats inhaled air, intratracheally injected with the same amount of normal saline, and orally took the same amount of normal saline. The rats in model group and Shenling Baizhusan groups were treated with smoking+intratracheal lipopolysaccharide+oral administration of ice and cold senna leaf infusion to build pulmonary and spleen deficiency type COPD rat model. Shenling Baizhusan was given by intragastric administration, and the rats were sacrificed at 28,35,42 days after drug intervention. Immunohistochemistry and Western blot were used to detect the expression changes of Ghrelin and Obestatin in hypothalamus and gastric tissues of rats. Result:As compared with normal blank group, the Ghrelin and Obestatin protein expression and integral absorbance in stomach and hypothalamus were significantly decreased in model group (P<0.01). As compared with model group, the Ghrelin and Obestatin protein expression and integral absorbance in stomach and hypothalamus were increased in different degrees in Shenling Baizhusan groups (P<0.05). Conclusion:The Shenling Baizhusan can regulate the energy metabolism of the lung and spleen deficiency type COPD rats and improve the malnutrition condition. Its mechanism may be associated with the regulation of Ghrelin and Obestatin dynamic balance to maintain body energy homeostasis.