| Objective: This paper aims to research on peroxidation of IPF by detecting the blood of IPF induced by bleomycin rats and changes of MDA, SOD, ET-1 and NO in lung tissue, and the effect on pathomorphism of lung tissue as well as the effect of YF to the expression of iNOS under the condition of IPF is observed by adopting advanced technology including HE staining, MASSON staining and immunohisto-chemistry. Intervening with Chinese medicine for invigorating qi, nourishing yin and promoting blood circulation, and to research on the correlation of IPF and deficiency of both qi and yin as well as blood stasis, to discuss the mechanism of invigorating qi, nourishing yin and promoting blood circulation on IPF rats which will provide experimental basis for clinical treatment.Methods: 120 healthy SD rats (paroecious about half) raised in the experimental environment about one week were randomly divided into five groups (24 per-group):①Control group: pyrogen-free normal saline (NS, 200μl per rat) was instilled intratracheally;②model group;③low-dose group;④high-dose group;⑤Prednisone group: bleomycin A5 was instilled intratracheally (5mg·kg-1). From the second day after instillation, control group and model group were filled with NS (10ml/kg),drenched by isotonic Na chloride; low-dose group and high-dose group were drenched with Chinese crude drug elixation differently(12.3g·kg-1 and 24.6 g·kg-1,7 and 14 times the dosage of the adult's clinical drug consumption); prednisone group were drenched with prednisone(6mg·kg-1).Rats in each group were respectively sacrificed on the 7th, 14th and 28th day after instillation. Serum was remained and plasma was separated immediately, serum frozen for test to -20℃. Right lung tissue was made into tissue homogenate which was centrifugated at 4℃for 5 minutes, and then supernatant fluid was extracted and frozen for test to -20℃. Tissue of middle part of left lung (0.5cm×0.5cm)was taken and fixed in 10% homogenate. Polyoxymethylene of 4% was to fix the lung, routine paraffin imbedded and bladed, histomorphology changes of lung tissues were observed by HE and Masson dyeing, and the expression of iNOS on the lung was detected by immunohistochemistry. The changes of MDA, SOD, ET-1 and NO in blood and lung tissue was tested.Results:1 Observation of the common conditionThe control group food and drink normally, react sensitively, the hairs are luster, the urine and stool are normal, have no death; the model group exist diff-degree cough and dyspnea, scrunch, have dispirited mind and withered fur. The high-dose group and low-dose group have the better mind and fur, have smooth anapnea gradually, the appetite and activity increased; the prednisone group have better condition also, but inferior to the high-dose group and low-dose group.2 the contents of MDA and activity of SOD's changes in blood serum and lung tissue of each group2.1 the contents of MDA in serum and lung tissue of each groupThe results were as follows: compared with the same time points of control group, the contents of MDA in serum and lung tissue were increased (all P<0.01).After intervention, compared with model group, the contents of MDA in serum and lung tissue were decreased in each time points of high-dose group(P<0.05或P<0.01); the contents of MDA in lung tissue were decreased on 7th and 28th day of low-dose group; the contents of MDA in serum were decreased in each time points of prednisone group(P<0.05或P<0.01), and in lung tissue were decreased on 7th and 28th day.There is no statistical significance among the three treatment groups (P>0.05).2.2 the activity of SOD in serum and lung tissueThe results were as follows: compared with control group, the activity of SOD in serum was decreased at each time point, in lung tissue was decreased on 7th and 14th day (P<0.01).After intervention, compared with model group, the activity of SOD in serum and lung tissue was increased on 7th and 14th day(P<0.05 or P<0.01), the activity of SOD in serum was increased on 28th day of high-dose group(P<0.01); the activity of SOD in serum was increased on 14th and 28th (P<0.01), the activity of SOD in lung tissue was increased on 7th and 14th day of low-dose group(P<0.05 or P<0.01); the activity of SOD in serum and lung tissue was increased on 7th day of prednisone group(P<0.05 or P<0.01).Compared with prednisone group, the activity of SOD in serum was increased on 14th and 28th (P<0.05), the activity of SOD in lung tissue was increased on 14th day of high-dose group (P<0.05); the activity of SOD in serum was increased on 28th day of low-dose group(P<0.05).3 changes of the contents of ET-1 and NO in blood and lung tissue3.1 the contents of ET-1 in plasma and lung tissueThe results were as follows: compared with control group, the contents of ET-1 in plasma and lung tissue were increased at each time point (P<0.01).After intervention, compared with mode group at same time point, the contents of ET-1 in plasma and lung tissue were increased of the three treatment group (P<0.05或P<0.01).Compared with prednisone group, the contents of ET-1 in lung tissue were decreased on 7th day of the high-dose group (P<0.01); the contents of ET-1 in plasma were decreased on 14th day of the low-dose and high-dose group (P<0.01).3.2 the contents of NO in serum and lung tissueThe results were as follows: compared with model group, the contents of NO in serum and lung tissue of model group were increased at each time point (P<0.01).After intervention, compared with model group at same point, the contents of NO were decreased of each treatment group(P<0.05或P<0.01).Compared with prednisone group, the contents of NO were decreased on 7th day in serum, 14th day in lung tissue and 28th day in serum of the high-dose group(P<0.05或P<0.01); the contents of NO in serum on 7th day were decreased of the low-dose group(P<0.01).4 Observation the change of the lung's histomorphology and pathology4.1 Macroscopic observationThe lung's appearance of control group was normal with color pink, surface smooth, elasticity good and lobes of lung edge sharpness. Of model group, lungs on 7th day were bigger and spotty hemorrhage and ecchymosis were seen; lungs on 14th day were gray, on which there were inequality of size nodus, sppotty hemorrhage; on 28th day, lungs were gray, smaller and harder, with nodus and bead-like aerate vacuole on the surface. The lung's appearance of each treatment group was better than that of model group.4.2 Observation under light microscopeSevere injuries were observed in lung tissues of model group, which were characterized by widened septa of alveoli, diffuse infiltration of inflammatory cells accompanied by atrophied or disappeared alveolitis, and compensatory alveolar emphysema. In the late period, midrange or severe pulmonary interstitial fibrosis developed; more collagenous fibers in lung matrix were observed by MASSON dyeing. Compared with model group, alveolitis and pulmonary fibrosis of each treatment group were declined significantly, especially in high-dose group.5 YF's effect on the the expression of iNOS of IPF ratsthe iNOS protein expressed negatively in rate lung tissuse frozen section of the normal control and negative control group, and which is expressed strong positive in the 14 days model group, the positive signals are buffy. The high dose group and the prednisone group show weakly posotive signals in cytoplasm, and the positive signals of low dose group show no significant difference with model group.Conclusion:1 Contents of MDA increased in serum and lung tissue of IPF rats , while SOD contents were decreased. These indicated that the disequilibrium of oxidation-antioxidation was one of the important pathogenesis and free radical played a direct role in the formation of IPF. YQYYF could resist IPF effectively by reducing MDA content or increasing SOD content.2 Generous production of NO aggravated inflammatory reaction and tissue damage in IPF rats by increasing vasopermeability. ET-1, pro-fibrous factor, played the same effect. YQYYF could resist the formation of IPF by improving circulatory condition and reducing the synthesis and releasing of NO and ET-1. 3 Significant iNOS expression was seen in lung tissue of IPF rats.YQYYF could reduce the lung injury done by NO by down-regulating iNOS expression.
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