| Objective: In order to lay a foundation for further study of rheumatoidlung, we establish the model of adjuvant arthritis rats, observe the morphologycharacteristics of pulmonary lesions in AA rat, investigate whether the AA ratswith lung injury can simulate the morphology of rheumatoid lung, observe thebest time of pulmonary alveolar inflammation and fibrosis lesions, and throughdifferent concentrations of Complete freund’s adjuvant (CFA) in particulartime point to observe the effect of lung injury in AA rats.Method:The experimental design:Part one: Dynamic observation in the lung injury of AA rats over the time.48clean healthy male Wistar rats were provided by the ExperimentalAnimal Center of Hebei Medical University, weight(170±10)g, received foodand water in a week of acclimation, and then randomly divided them into twogroups: Control group (n=24) and Model group (n=24). Model group:24rats,which were hypodermic injected0.1ml about CFA (containing BCG vaccine7.5mg/ml) in right rare paw and established adjuvant arthritis. Control group:24rats, which were hypodermic injected0.1ml about0.9%NaCl solution inright rare paw in accordance with the model group means. the animal werekilled respectively on the7th day,14th day,28th day and56th day.Part two: On the basis of the first part of study, to choose differentconcentrations of CFA in particular time point to observe the effect of lunginjury in AA rats.48clean healthy male Wistar rats were provided by the ExperimentalAnimal Center of Hebei Medical University, weight (170±10)g, received foodand water in a week of acclimation, and then randomly divided them into fourgroups: group CFA5mg, group CFA7.5mg, group CFA10mg and Control group. which were hypodermic injected0.1ml about CFA in differentconcentrations in right rare paw and established adjuvant arthritis. Controlgroup were hypodermic injected0.1ml about0.9%NaCl solution in right rarepaw in accordance with the model group means. the animal were killedrespectively on the14th day and the28th day.Method about experment:To monitor the degree of rat paw swelling and mark the arthritis index inthe process of experiment. Kill the rats and collect their blood at the same time,to detect rheumatoid factor (RF) content in serum of rats with ELISA. Gatherthe right pulmonary lower lobe of rats, then put them in the4%paraformaldehyde solution to fix, use the method of HE staining to observe thechange of lung tissue morphology. Use the method of Masson staining toobserve the collagen fibers deposition in rat lung tissue. And take the left lowerlung tissue alkaline solution determination of lung tissue hydroxyproline (HYP)content. Take the secondary inflammatory edema of rats ankle, put them in the4%paraformaldehyde solution to fix, take off the calcium treatment, use themethod of HE staining to observe the change of joint histopathological. Tomark the histopathology of lung tissue and joint with method of reference.Using SPSS20.0statistical software to analysis each set of data.Results:Part one: Dynamic observation of the lung injury in AA rats over the time.1The level of alveolitis of modle group was more serious than that incontrol group on each time point (P<0.05), the degree of alveolar inflammationin modle group14th days is the highest; the level of fibrosis in modle groupwas had no significant difference than that in control group on the7th day(P>0.05), the level of fibrosis in modle group was more serious than that incontrol group on the14th day, the28th day and the56th day (P<0.05), thedegree of fibrosis in modle group28th days is the most obvious.2The contents of HYP in model group shown an increase tendencyaccompany with the time and reached a peak on the28th day, compare withcontrol group, there were significant difference on each time point (P<0.05). 3、The levels of RF in model group of rats serum obviously increasedafter14days,28days,56days always maintain a high level,compare withcontrol group, there were significant difference on each time point (P<0.05).4、 The joints swelling degree of right joint in model group wassignificant higher on each time point, than control group (P<0.05), the mostobvious on the14th day; The swelling degree of left joint in model wassignificant higher than control group on the14th day,28th day and56th day(P<0.05), the peak subsided after the28th day; The score of AI in model groupwere significant higher on each time point, compare with control group(P<0.05). The score of joint pathologic in model group had no significantdifference on the7th day, but had significant difference on the14th day, the28th day and the56th day compare with control group (P<0.05), and was thehighest on the56th day.Part two: To select the optimum concentration of BCG in CFA to establishthe most typical lung injury in AA rats.1The level of alveolitis of modle group was more serious than that incontrol group on the14th day (P<0.05); The level of alveolitis in7.5mg and10mg group is higher than that in5mg group (P<0.05), but there was nosignificant difference between7.5mg and10mg group (P>0.05). The level offibrosis in modle group was more serious than that in control group on the28thday (P<0.05). The level of fibrosis in7.5mg and10mg group is higher than thatin5mg group, but there was no significant difference between7.5mg and10mggroup (P>0.05).2The contents of HYP on each time point in model group were all higherthan that in control group （P<0.05）. The contents of HYP in7.5mg and10mg group is higher than that in5mg group (P<0.05), but there was nosignificant difference between the two groups (P>0.05).3The levels of RF on each time point in model group were all higher thanthat in control group （P<0.05）. The levels of RF in7.5mg and10mg groupwas higher than that in5mg group (P<0.05), but there was no significantdifference between the two groups (P>0.05). 4The joints swelling degree in7.5mg and10mg group is higher than thatin5mg group on the14th day (P<0.05), but there was no significant differencebetween the two groups (P>0.05). The scroe of AI and pathologic in modelgroup had no significant difference compared with control group (P<0.05). Thescroe of AI in7.5mg and10mg group were higher than that in5mg group(P<0.05), but there was no significant difference between the twogroups(P>0.05). The joints swelling degree in model group was higher thanthat in control group（P<0.05）. The swelling degree in7.5mg and10mggroup were higher than that in5mg group（P<0.05）, but there was nosignificant difference between the two groups (P>0.05). The scroe of AI in7.5mg and10mg group is higher than that in control and5mg group, but therewas no significant difference between7.5mg and10mg group (P>0.05). Thepathologic score in7.5mg and10mg group was higher than control group（P<0.05）,the pathologic score in10mg group was significant higher thanthat in5mg group（P<0.05）, there was no significant difference between7.5mg and5mg group (P>0.05).Conclusion:1The model of AA rat can be observed the morphology characteristics ofpulmonary lesions over the time,the degree of alveolar inflammation in modlegroup14th days is obvious, visible thickening of alveoar interval, alveolarepithelial necrosis, alveolar edema deformation, within the pulmonaryinterstitial cell infiltration. the degree of fibrosis on the28th day is obvious, alarge amount of collagen fiber was dyed green. It prove the animal model in theevent of pathological change and pulmonary fibrosis change at the same time,that AA rats can be used to study the animal model of rheumatoid lung injury.2To observe the optimal concentration of CFA with BCG is7.5or10mg/ml in a particular time,which influences lung injury of AA rats mostobviously and there is no obvious difference between them. |
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