【摘要】：目的:討論P(yáng)yrin重組蛋白對(duì)博萊霉素誘導(dǎo)的大鼠肺纖維化是否有作用,Pyrin重組蛋白是否通過VEGF/VEGFR2/MMP-9信號(hào)通路來影響肺纖維化。實(shí)驗(yàn)方法:隨機(jī)將60只SD大鼠分成4組,正常對(duì)照組、博來霉素模型組、Pyrin重組蛋白組、SU5416陽性對(duì)照組。所有組于第14天及第28天分兩批進(jìn)行取材,用HE染色和MASSON染色分別觀察肺泡炎程度及肺纖維化程度,免疫組化及RT-PCR分別檢鋇VEGF、VEGFR-2、MMP-9蛋白和mmRNA表達(dá)情況。結(jié)果:1.各組大鼠肺組織病理變化:HE染色分級(jí)示:肺泡炎程度的比較中博來霉素模型組最為嚴(yán)重,而正常對(duì)照組無明顯肺泡炎改變(P0.05)。Pyrin重組蛋白組在第14天及第28天時(shí)肺泡炎程度較正常對(duì)照組明顯加重,而較博來霉素模型組減輕(P0.05)。Masson染色分級(jí)示:博來霉素模型組在纖維化程度最為嚴(yán)重,而正常對(duì)照組未見明顯肺纖維化(P0.05)。而在第14天及第28天時(shí)Pyrin重組蛋白組的肺纖維化程度均較博來霉素模型組的肺纖維化程度減輕(P0.05)。2肺組織免疫組化染色(VEGF、VEGFR2、MMP-9):在第14天時(shí),正常對(duì)照組肺組織中的VEGF、VEGFR2、MMP-9有少量表達(dá),而博來霉素模型組大鼠肺組織的VEGF、VEGFR2、MMP-9表達(dá)明顯增多(P0.05);SU5416陽性對(duì)照組和Pyrin重組蛋白組大鼠肺組織VEGF、VEGFR2、MMP-9表達(dá)較正常對(duì)照組明顯增多,但較博來霉素模型組有所減弱(P0.05)。當(dāng)?shù)?8天時(shí),SU5416陽性對(duì)照組和Pyrin重組蛋白治療組VEGF、VEGFR2、MMP-9表達(dá)水平仍較正常對(duì)照組明顯增強(qiáng)(P0.05),但較博來霉素模型組VEGF、VEGFR2、MMP-9表達(dá)有所減弱(P0.05)。3.mRNA表達(dá)(VEGF、VEGFR2、MMP-9):博來霉素模型組、SU5416陽性對(duì)照組和Pyrin重組蛋白組的VEGF、VEGFR2、MMP-9表達(dá)水平均較正常對(duì)照組相比顯著增高(P0.05)。其中第28天VEGF、VEGFR2、MMP-9表達(dá)水平與第14天相比有升高趨勢(shì),但未見顯著性差異(P0.05)。而正常對(duì)照組肺組織的VEGF、VEGFR2、MMP-9 mRNA表達(dá)水平相對(duì)穩(wěn)定,Pyrin重組蛋白組的VEGF、VEGFR2、MMP-9表達(dá)水平較博來霉素模型組相比在第14天及第28天均明顯降低(P0.05)。結(jié)論:1.Pyrin重組蛋白可能通過下調(diào)VEGF/VEGFR2/MMP-9信號(hào)通路而抑制肺纖維化新生血管的生成。2.Pyrin重組蛋白可能通過下調(diào)VEGF/VEGFR2/MMP-9信號(hào)通路而抑制肺纖維化ECM的沉積。3.Pyrin重組蛋白可能通過下調(diào)VEGF/VEGFR2/MMP-9信號(hào)通路而發(fā)揮抗肺纖維化作用。
[Abstract]:Aim: to investigate the effects of Pyrin recombinant protein on bleomycin induced pulmonary fibrosis in rats and whether Pyrin recombinant protein affects pulmonary fibrosis through VEGF/VEGFR2/MMP-9 signaling pathway. Methods: sixty SD rats were randomly divided into 4 groups: normal control group, bleomycin model group, Pyrin recombinant protein group and SU5416 positive control group. The alveolitis degree and pulmonary fibrosis degree were observed by HE staining and MASSON staining. The expression of barium VEGF,VEGFR-2,MMP-9 protein and mmRNA were detected by immunohistochemistry and RT-PCR, respectively. The result is 1: 1. Pathological changes of lung tissue of rats in each group: HE staining grade showed that the degree of alveolitis was the most serious in bleomycin model group. But there was no obvious change of alveolitis in normal control group (P0.05) the degree of alveolitis in). Pyrin recombinant protein group was significantly worse than that in normal control group on the 14th and 28th day. But compared with bleomycin model group (P0.05). Masson staining grade: bleomycin model group in the fibrosis degree is the most serious, but the normal control group did not see significant pulmonary fibrosis (P0.05). On the 14th and 28th day, the degree of pulmonary fibrosis in the Pyrin recombinant protein group was lower than that in the bleomycin model group (P0.05). 2 the lung tissue immunohistochemical staining (VEGF,VEGFR2,MMP-9): on the 14th day, the pulmonary fibrosis degree of the Pyrin recombinant protein group was lower than that of the bleomycin model group (P0.05). There was a little expression of VEGF,VEGFR2,MMP-9 in lung tissue of normal control group, while the expression of VEGF,VEGFR2,MMP-9 in lung tissue of bleomycin model group was significantly increased (P0.05). The expression of VEGF,VEGFR2,MMP-9 in lung tissue of SU5416 positive control group and Pyrin recombinant protein group was significantly higher than that of normal control group, but decreased compared with bleomycin model group (P0.05). On the 28th day, the expression of VEGF,VEGFR2,MMP-9 in SU5416 positive control group and Pyrin recombinant protein treatment group was significantly higher than that in normal control group (P0.05), but it was higher than that in bleomycin model group (P0.05). MMP-9 expression decreased (P0.05). 3.mRNA expression (VEGF,VEGFR2,MMP-9): VEGF,VEGFR2, in bleomycin model group, SU5416 positive control group and Pyrin recombinant protein group The expression of MMP-9 was significantly higher than that of normal control group (P0.05). The expression of VEGF,VEGFR2,MMP-9 on the 28th day was higher than that on the 14th day, but there was no significant difference (P0.05). The expression of VEGF,VEGFR2,MMP-9 mRNA in lung tissue of normal control group was relatively stable, and the expression of VEGF,VEGFR2,MMP-9 in Pyrin recombinant protein group was significantly lower than that in bleomycin model group on the 14th and 28th day (P0.05). Conclusion: 1.Pyrin recombinant protein may inhibit angiogenesis of pulmonary fibrosis by down-regulating VEGF/VEGFR2/MMP-9 signaling pathway, and 2.Pyrin recombinant protein may inhibit pulmonary fibrosis by down-regulating VEGF/VEGFR2/MMP-9 signaling pathway. The deposition of fibrotic ECM. 3.Pyrin recombinant protein may play an anti-pulmonary fibrosis effect by down-regulating VEGF/VEGFR2/MMP-9 signaling pathway.
【學(xué)位授予單位】：延邊大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R563
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本文編號(hào)：2304359