Effect Of Rolipram On NF-κB In The Lungs Of Adjuvant Arthritis Rats

Objective: To study the dynamic expression of NF-κB and the effect ofrolipram on NF-κB in the lungs of adjuvant arthritis rats, to explore theinfluence of phosphodiesterase in pathology of rheumatoid lung disease, andoffer therapy support for phosphodiesterase inhibitor to treat the disease.Method:54clean healthy male Wistar rats were provided by theExperimental Animal Center of Hebei Medical University, weight160g～180g, were received food and water in a week for acclimation, then wererandomly divided into three group：group A（model group）:18rats, werehypodermic injected0.1ml of complete Freund’s adjuvant(CFA)(containingthe BCG vaccine7.5mg/ml) in right rare paw to establish adjuvant arthritis,from the day to the day them be killed, the model group were intraperitonealinjected0.5ml of0.9%NaCl solution everyday.(2)group B(the rolipramgroup):18rats, were used the same way as the model group to establishadjuvant arthritis, and intervened by intraperitoneal injected0.5ml of rolipramsolution（which was diluted by saline to0.1mg/ml in proportion according tothe0.25mg/kg） everyday from the day to when them be killed.(3)group C(normal control group):18rats, were hypodermic injected0.1ml of0.9%NaCl solution in right rare paw in accordance with the model group means,were intraperitoneal injected0.5ml of0.9%NaCl solution everyday from thefirst day after injection to when them be killed. Rats were evaluated jointswelling by foot measure instrument on the1thday,7thday,14thday,28thday,and they were killed respectively on the7thday,14thday,28thday. Then the ratsleft upper lobe and right pulmonary lower lobe were removed out carefully,and fixed them in the4%paraformaldehyde solution for histologicalexamination by using HE staining and Masson staining method. Thedevelopment degree and classification according to Szapiel method were as followed, they can be divided into4category. Using immunohistochemicalmethod to detect the expression of the NF-κB in lung tissue. Using SPSS20.0statistical software to analysis each set of data.Results:1Joint general observationOn1stday, the joint were wild swelling, there was no statisticallysignificant among three group. The joints swelling degree of right joint of Agroup was31.54±3.26%,49.66±4.03%,32.78±3.26%on7thday,14thday,28thday, B group was24.74±2.27,35.54±2.09,23.87±2.76and C group was9.98±1.64%,11.16±2.01%,11.57±1.09%, A group were aggravated over time,marked increase compared with those in C group（P<0.05）, B group werelighter than A group on7thday,14thday,28thday（P<0.05）, heavier thancontrol group at the same time（P<0.05）. Left side joint swelling of modelgroup appears on12thday, the swelling degree was25.29±3.72%,38.86±3.72%on14thday,28thday, B group was17.74±1.77%,24.51±3.84%,and C group was10.99±1.38%,11.76±1.31%, A group were changed moreserious than C group（P<0.05）, B group were lighter than A group on14thday,28thday（P<0.05）, heavier than control group at the same time（P<0.05）.2Lung histological pathology resultsHE staining: A group: On7thday of the experiment, HE dyeing revealedthat the pleura got thicker, the alveolar epithelial were necrosis, alveolar weredeformation, and the alveolar interval were edema, inflammatory cellsinfiltration and inflammatory substances effusion occasionally, on14thday, theinfiltration was more obvious than on7thday, loss of normal alveolar structure,some myofibroblast proliferation, on28thday, the infiltration of inflammatorycell and normal alveolar structure became less than before, alveolitis score ofA group was2.8±0.837,3.67±0.516,2.17±0.753on7thday,14thday,28thday,and C group was1.0±0.0,1.17±0.408,1.0±0.0, the degree of alveolitis of Agroup was more serious than C group（P<0.05）, compared with A group, therewere no remarkable changing in the C group. The B group showed that itschange were between A group and C group. The alveolitis score of B group was1.83±0.753,2.5±0.548, the degree of alveolitis was more serious than Cgroup on7thday and14thday（P<0.05）, and lower than A group（P<0.05）.Masson staining: There was little green collagen in A group on7thday,more manifested on14thday, and collagen was increased obviously. The lungparenchyma had many collagen fibers formation on the28thday. There wereonly a little green collagen in control group. The fibrosis score of A group, Bgroup, C group were1.2±0.447,1.0±0.0,1.17±0.408on7thday. The degree offibrosis was no significant difference among the three group（P>0.05）. Thefibrosis score of A group was2.5±0.548,3.33±0.817on14thand28thday, andB group was2±0.632,1.8±0.837on14thand28thday, C group was1.0±0.0,1.17±0.408on14thand28thday, The degree of fibrosis of A group and Bgroup was higher than C group on14thand28thday（P<0.05）, B group waslower than A group on28thday（P<0.05）.3The expression of NF-κB in lung tissueImmunohistochemical showed that the positive expression of NF-κB in Agroup were much higher than that in C group all the experimental time, all ofthem had statistical significance (P <0.05). IOD of NF-κB of C group was18285.34±1168.74,17704.03±888.01,17425.58±648.09on7thday,14thday,28thday, and A group was123337.58±2590.75,74002.79±3731.36on day14,day28. The level was highest on day7, IOD of NF-κB of A group was170657.84±2422.13on day7, the expression of NF-κB was mainly in thenuclear of lung substance cell. IOD of NF-κB of B group was79452.24±2566.52,47241.56±2105.12,26183.67±1616.49on7thday,14thday,28thday, the positive expression of NF-κB in B group were much lower thanthat in A group and higher than control group all the experimental time, and allof them had statistical significance (P <0.05).Conclusion: Rolipram might inhibit the degree of lung injury in adjuvantarthritis rat lung tissue by lowering the expression of the NF-κB.