AQPs Expressions And Their Controls Of Lung And Kidney Tissues In Lung-Qi Deficiency Model Rats

Objective1. To observe the changes of urine volume, specific gravity andcontents of ET,IL-1β,TNF-αin blood in the model group. The aimwas to supply and consummate the objective index in Lung-Qi deficiencymodel rats.2. To observe the expressions of AQP1, AQP2, AQP4 in lung and kidneytissues. The aim was to provide experimental evidence for thetraditional Chinese medical theory of "the kidney governs watermetabolism", "the lung being in charge of regulating the waterpassage" in physiologic and pathologic body fluid metabolism.3. To observe the contents of ET,IL-1β,TNF-αin blood in Lung-Qideficiency model rats and to study the relationship with the expressionof ET,IL-1β,TNF-αand AQPs in lung and kidney tissues. To examinethe mechanism of AQPs expressions in lung and kidney tissues and detectthe relationship with lung and kidney in water metabolism, the aim wasto prove that the lung and the kidney are related in traditional Chinesemedical theory.Method1. Twenty healthy Wistar rats were randomly divided into control groupand lung-Qi deficiency model group(model group) with 10 rats each group.The rats of lung Qi deficiency were simulated by injectinglipopolysaccharide into the trachea twice together with cigarettesmoking. Behavior was observed and Body weights were measured with electrical scale 2h after rats tsking food on the 1 d, 14 d and 28 d.Measures of urine volume and specific gravity: 24 h urine volumes weremeasured with graduated cylinder since 28 d, and urine specific gravitywas detected with refractometer. After 28 d, the ET,IL-1β,TNF-αconcentrations in blood was measured and the morphological changeswere observed by HE staining.2. Immunohistochemistry and Western blot analysis were used toobserve the expression of AQP1, AQP2, AQP4 in lung and kidney tissues.RT-PCR was applied to analyze the expression of AQP1, AQP2, AQP4 mRNAin lung and kidney tissues.3. blood Sample and lung tissues were taken and the concentrationof ET, IL-1β, TNF-αwere measured by radioimmunoassay. Correlationsbetween ET, IL-1β, TNF-αin the blood and AQPs expressions wereanalyzed in lung and kidney tissues.4. Statistical data analysisThe data were expressed as (?)±s. Independent-samples T Test ofSPSS10.0 was employed in the group comparison, and correlationanalysis was also used.Results1. Evaluation of Lung-Qi deficiency modelThe model rats showed coughing, breath lessness, wheezing, humidand minute red in the nasal part, depression, action retardation,immobilility, hair burnishness and shedding, and meanwhile withgradually appetite decrease, softening and thin stool. The increaseamplification of body weight in model rats was progressly lesser thanthat in the control group, and there were obviously differences of thebody weight between the two groups on 14d, 28d (P＜0.01). Compared withthose of the control group, urine volume decreased markedly and urinedensity increased obviously in the rats of the model group (P＜0.01). Plasma ET concentration in the model group on the 28d was higherthan that in the control group (P＜0.01), and so were the serumIL-1βand TNF-α(P＜0.01). The pathological changes of the lung tissuein the model group seen with naked eyes and light microscope wereconsistent with those of chronic bronchitis and emphysema, and edemawas seen in local alveoli. 2. AQPs expressions in lung and kidney tissues in the Lung-Qideficiency model ratsAQP1 expression in the lung tissue mainly appeared in theendothelium of peripheral alveoli and capillary endothelium of thebronchi, and its expression in the model group was obviously weakerthan that in the control group, and so was AQP1 mRNA (P＜0.01). AQP1expression in the kidney tissue mainly showed on the apical membraneand basal membrane of the proximal tubule and thin segment ofdescending limb of the Henle' loop, and AQP1 expression in the controlgroup was greater than that in the control group, and so was AQP1 mRNA(P＜0.01). AQP2 expression in the lung tissue showed negation, whilein the kidney tissue showed on the apical membrane and cytoplasmicvesicles of the chief cells of the collecting tube in the innermedullary, which shown much more positive than that in the controlgroup, so was AQP2 mRNA (P＜0.01); AQP4 expression of the lung tissueappeared on the basal membranes of endothelium in the trachea, bronchi,which was higher than that in the control group, and so was the AQP4mRNA expression (P＜0.01), while AQP4 expression in the kidney tissueappeared on the basal membranes of the chief cells in the nearly 2/3collecting tube of the inner medullary, which was weaker than that inthe control group, and so was AQP4 mRNA expression (P＜0.01).3. ET,IL-1β,TNF-αconcentrations of lung tissue in the model groupET,IL-1β,TNF-αconcentrations of lung tissue in the model groupwere markedly higher than those in the control group (P＜0.01).4. Correlations between ET, IL-1β, TNF-αin the blood and AQPsexpressions in lung and kidney tissuesThere were positive correlations between blood ET,IL-1β,TNF-αconcentrations and AQP1, AQP2 expressions in the kidney tissue, AQP4expression in the lung tissue, and there were negative correlationsbetween blood ET, IL-1β, TNF-αconcentrations and AQP4 expressionin the kidney tissue and APQ1 expression in the lung tissue, which implyblood concentration increases of the ET, IL-1β, TNF-αmightupregulate AQP1, AQP2 expressions in the kidney tissue and AQP4expression in the lung tissue, and also might downregulate AQP4expression in the kidney tissue and AQP1 expression in the lung tissue.There exited the inter-correlations of blood concentrations of ET,IL-1βand TNF-αin the model group: there was correlations between ET and IL-1β, TNF-α(0. 01＜P＜0.05), while the correlation betweenthe IL-1βand TNF-αwas P＜0.01, which implied that one increase ofET, IL-1βand TNF-αmight promote the other secretions, and thecorrelation between the IL-1βand TNF-αwas more closely.Conclusions1. There are obvious decrease in urine volume and significantincrease in urine specific gravity of rats of Lung-Qi deficiency model.2. In the Lung-Qi deficiency model rats, AQP1 expression downregulated and AQP4 expression up regulated in the lung tissue. AQP1and AQP2 expressions up regulated and AQP4 expression down regulatedin the kidney tissue.3. Concentrations of ET, IL-1βand TNF-αincreased obviously inboth blood and lung tissue in the Lung-Qi deficiency model rats.4. Concentration increases in blood ET, IL-1βand TNF-αcan bethe standards of Lung-Qi deficiency model rats.5. One of the substance basis of relationship between Lung andKidney in body fluid metabolism may be the AQPs retro-expression inthe lung and kidney tissues.6. The substance basis of Lung is charged in dispersing outward anddownward may be relative to the release of ET, IL-1βand TNF-αby lung.