Chronic Obstructive Pulmonary Disease In The Rat Diaphragm Cell Apoptosis And Fiber Types Change And Intervention Study

ObjectiveBy rat diaphragm fibers in COPD classification changes and diaphragmaticapoptosis to investigate the pathogenesis of diaphragmatic fatigue.Materials and methods40male Wistar rats (initially weighing200±10g)were randomly divided intohealthy control group and COPD group, with20rats in each group. The rats in themodel group were exposed to5%smoke (every day two times, each time30minutes,interval8hours, for28days) in a sealed box(47.0cm×31.5cm×49.0cm), and200uglipopolysaccharide (Sigma, USA) was injected intratracheally on the1st and14th day,injection day without smoke. On the29th day the rats were sacrificed.Toconfirming the occurrence of chronic obstructive pulmonary disease by HE stainingof lung tissue of rats in two groups, To onfirmed the presence of diaphragmaticmuscle cell apoptosis of model group by electron microscope, Fas protein andCaspase-3gene expression of diaphragmatic muscle cell were detected by SABC andRT-PCR, and diaphragm myofibers classification dyeing by ATPase histochemicaldrops of staining and Nicotinamide adenine dinucleotide-Tetrazolium reductase.ResultsOn the28th day, COPD rats modeling success by confirmed of HE staining of ratlung tissue, destruction of alveolar walls and enlargement of alveolar space wereobserved on COPD group, the diaphragmatic muscle cell apoptosis rate, Fas proteinand Caspase-3gene expression level are higher than the health in control group (P <0.01), compared to healthy controls, model group of type I fiber proportion ofincrease and Ⅱ type fiber proportion of reduce (P <0.01). ConclusionsAbnormal apoptosis of diaphragmatic muscle cell and fiber type of change werepresent in rats with COPD. Caspase-3and Fas/FasL way were involved in thediaphragmatic muscle cell apoptosis. Diaphragm cell apoptosis and the regulation offiber type of change in the late patients with COPD diaphragmatic muscle fatiguecure difficult. ObjectiveThrough the establishment of COPD rat model,to discuss the excessive apoptosisof diaphragmatic muscle in the fatigue of diaphragmatic muscle,to explore theadjustment way of apoptosis,and through the intervention of Tiotropium Bromide toimprove the excessive apoptosis of diaphragmatic muscle in COPD rats.Methods60male Wistar rats are randomly divided into control con, COPD model group andTiotropium Bromide intervention group, each group has20rats.Model rats are put inseal smoked box（47.0cm×31.5cm×49.0cm）, and the smoke concentration is5%,2times a day,30minutes every time,8hours an interval, and28days in all.In1daysand a day14,it will inject lipopolysaccharide200ug into endotracheal.The rats ofcontrol con are put in airtight box but not to smoke.In1days and a day14,it willinject physiological saline200ug into endotracheal.30minutes Before smoked,theTiotropium Bromide intervention group need inhale Tiotropium Bromide throughAtomization （0.12mmol/L）.29days later，the rats will be made to death to take thediaphragmatic muscle and the lungs,and to detect the pathology of the lung tissue,tomeasure the apoptosis rate of diaphragmatic muscle cell, the expression of Fas protein、Caspase-3and Fasl gene. Diaphragm myofibers classification dyeing byATPase histochemical drops of staining and Nicotinamide adeninedinucleotide-Tetrazolium reductase.Results28days later,in the COPD model rats, the apoptosis rate of diaphragmatic musclecell, the expression of Fas protein、Caspase-3and Fasl gene,are higher than thehealthy controls（P<0.01）),In Tiotropium Bromide group, the pathologicalchanges,the expression of Fas protein、Caspase-3and Fasl gene, are less than COPDmade module（P<0.01）.ConclusionsIn the occurrence of COPD process,the diaphragmatic muscle cell has excessiveapoptosis,C aspase-3and Fas/FasL approaches are involved in the control ofapoptosis in diaphragmatic muscle cell,Tiotropium Bromide can intervene theapoptosis of diaphragmatic muscle cell,and reduce the level of apoptosis indiaphragmatic muscle cell.