IBDV感染对雏鸡法氏囊纤维增生及TGF-β1、MMP-9表达的影响
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摘要
试验通过Masson染色、免疫组织化学染色和实时荧光定量PCR(qRT-PCR)等方法研究雏鸡感染传染性法氏囊病病毒(IBDV)过程中法氏囊内纤维形成、转化生长因子β-1(TGF-β1)和基质金属蛋白酶-9(MMP-9)表达及分布规律,探讨雏鸡IBDV感染后TGF-β1和MMP-9的表达变化与纤维增生之间的相互关系。结果表明:雏鸡IBDV感染后,法氏囊内黏膜下层、滤泡周围的固有层纤维大量增生,在感染后4 d(4 dpi)达到最高,随后逐渐减少。同时,法氏囊内TGF-β1的蛋白表达水平逐渐增高至7 dpi达到最大值,且3～7 dpi显著高于对照组(P<0.05)。TGF-β1基因转录水平呈现先增高后降低趋势,且2～5 dpi均显著高于对照组,4 dpi达到最高。MMP-9的蛋白表达和基因转录均呈现先增高后降低的趋势。蛋白表达水平在4 dpi最高,且在1～4 dpi显著高于对照组(P<0.05),基因转录水平在2～7 dpi显著高于对照组且在5 dpi时最高(P<0.05)。结论:雏鸡IBDV感染导致法氏囊中纤维增生和TGF-β1、MMP-9的表达基本一致,提示TGF-β1和MMP-9可能对IBDV感染雏鸡法氏囊中纤维的发生、发展发挥一定作用。
In this study, fibrous hyperplasia, the expression and distribution of TGF-β1 and MMP-9 in bursa of chicken infected with infectious bursal disease virus(IBDV) were studied by Masson staining, immunohistochemical staining and qRT-PCR. The relationship of the expression of TGF-β1 and MMP-9 with fibrous hyperplasia was also discussed.The results showed that fibers became more in the submucosa and the lamina proper of bursa after chicks infected with IBDV, reaching the highest level on 4 days post-infection(4 dpi), and then gradually decreased. At the same time,the protein expression level of TGF-β1 in bursa continued to increase till to 7 dpi, and it was significantly higher than that of the control during 3 to 7 dpi(P<0.05). The gene transcription level of TGF-β1 increased gradually and reached the highest level on 4 dpi, and then decreased, showing significantly higher than that of the control during 2 to 5 dpi(P<0.05). The protein expression and gene expression of MMP-9 have the same pattern, showing increasing first and then decreasing. Protein expression was the highest on 4 dpi and higher significantly than that of the control during 1 to4 dpi(P<0.05). Gene expression was higher than that of the control on 2 to 7 dpi(P<0.05). Conclusion: The patterns of expression changes of fibrous hyperplasia, TGF-β1 and MMP-9 were similar, which suggested that TGF-β1 and MMP-9may play a role in the occurrence and development of fiber in IBDV-infected chicken.
In this study, fibrous hyperplasia, the expression and distribution of TGF-β1 and MMP-9 in bursa of chicken infected with infectious bursal disease virus(IBDV) were studied by Masson staining, immunohistochemical staining and qRT-PCR. The relationship of the expression of TGF-β1 and MMP-9 with fibrous hyperplasia was also discussed.The results showed that fibers became more in the submucosa and the lamina proper of bursa after chicks infected with IBDV, reaching the highest level on 4 days post-infection(4 dpi), and then gradually decreased. At the same time,the protein expression level of TGF-β1 in bursa continued to increase till to 7 dpi, and it was significantly higher than that of the control during 3 to 7 dpi(P<0.05). The gene transcription level of TGF-β1 increased gradually and reached the highest level on 4 dpi, and then decreased, showing significantly higher than that of the control during 2 to 5 dpi(P<0.05). The protein expression and gene expression of MMP-9 have the same pattern, showing increasing first and then decreasing. Protein expression was the highest on 4 dpi and higher significantly than that of the control during 1 to4 dpi(P<0.05). Gene expression was higher than that of the control on 2 to 7 dpi(P<0.05). Conclusion: The patterns of expression changes of fibrous hyperplasia, TGF-β1 and MMP-9 were similar, which suggested that TGF-β1 and MMP-9may play a role in the occurrence and development of fiber in IBDV-infected chicken.
引文
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[13]亓慧琴,张连莲,邢利平,等.慢性阻塞性肺疾病患者应用异丙托溴铵联合乙酰半胱氨酸治疗前后血清中TGF-β和MMP-9水平的变化及其临床意义[J].吉林大学学报(医学版),2016,42(4):777-782.
[14]SARIYA N S,ATIPHAN P,NEERACHA S,et al.TGF-β1 induced MMP-9 expression in HNSCC cell lines via Smad/MLCK pathway[J].Biochemical and Biophysical Research Communications,2008(371):713-718.
[15]FENG M,WANG H,WANG Q R Z,et al.Matrix metalloprotease 9 promotes liver recovery from ischemia and reperfusion injury[J].Journal of Surgical Research,2013,180(1):156-161.
[2]刘晓晓,余燕,邵营格,等.雏鸡感染IBDV后法氏囊组织结构的动态变化[J].河南科技学院学报(自然科学版),2015,43(4):40-43.
[3]SHARON M WAHL.Transforming growth factorp:innately bipoIar[J].Current Opinion Inimmunology,2007(19):55-62.
[4]张蕊,高雪丽,李博韬,等.传染性法氏囊病毒感染SPF雏鸡哈德尔腺和盲肠扁桃体TGF-β1 m RNA表达变化[J].中国家禽,2013,35(9):15-18.
[5]SUWANABOL P A,KENT K C,LIU B.TGF-βand restenosis revisited:A Smad link[J].Journal of Surgical Research,2011,167(2):287-297.
[6]ANDREA K,ULRIKE K,MICHAELA K,et al.Expression of matrix metalloproteinases(MMPs)in primary human breast cancer and breast cancer cell lines:New findings and review of the literature[J].BMC Cancer,2009(9):188.
[7]ZAOUI P,CANTIN J F,ALIMARDANI B M,et al.Role of metalloproteases and inhibitors in the occurrence and progression of diabetic renal lesions[J].Diabetes Metab,2000,26(4):25-29.
[8]戈宏焱,张仕华.基质金属蛋白酶与肝纤维化关系的研究进展[J].临床肝胆病杂志,2017(3):563-566.
[9]CAMARGO K C,GOMESA J R.,LODDI M M,et al.MT1-MMP and its potential role in the vertebrate intestinal morphogenesis[J].Acta Histochemica,2016,7(9):1-7.
[10]AGNIESZKA L W,ANNA H.Involvement of matrix metalloproteinases(M-MP-2,-7,-9)and their tissue inhibitors(TIMP-2,-3)in the chicken oviductregression and recrudescence[J].Cell Tissue Research,2016(366):443-454.
[11]高红艳,刘芳,谢军,等.TGF-β1及MMP-9在宫腔粘连内膜组织中的表达与意义[J].中国妇幼保健,2014,29(24):3971-3974.
[12]FORBES J M,FUKAMI K,COOPER M E.Diabetic nephropathy:Where hemodynamics meets metabolism[J].Experimental and Clinical Endocrinology,2007,115(2):69-84.
[13]亓慧琴,张连莲,邢利平,等.慢性阻塞性肺疾病患者应用异丙托溴铵联合乙酰半胱氨酸治疗前后血清中TGF-β和MMP-9水平的变化及其临床意义[J].吉林大学学报(医学版),2016,42(4):777-782.
[14]SARIYA N S,ATIPHAN P,NEERACHA S,et al.TGF-β1 induced MMP-9 expression in HNSCC cell lines via Smad/MLCK pathway[J].Biochemical and Biophysical Research Communications,2008(371):713-718.
[15]FENG M,WANG H,WANG Q R Z,et al.Matrix metalloprotease 9 promotes liver recovery from ischemia and reperfusion injury[J].Journal of Surgical Research,2013,180(1):156-161.