表没食子儿茶素没食子酸酯对哮喘小鼠肺组织中PTEN基因表达的影响
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摘要
目的建立哮喘小鼠急性模型,探讨给予不同剂量的表没食子儿茶素没食子酸酯(EGCG)腹腔注射干预治疗,对小鼠气道炎症及肺组织中PTEN基因表达的影响。方法 32只SPF级BALB/c雌性小鼠随机分为4组,正常对照组、哮喘组、EGCG(5 mg/kg)治疗组、EGCG(50 mg/kg)治疗组,应用卵清蛋白(OVA)致敏、激发建立哮喘模型。采用苏木素-伊红染色观察小鼠气道炎症情况,无创通气方法检测小鼠气道反应性,ELISA方法检测小鼠血清中OVA特异性IgE的水平,Real-time PCR及免疫组化方法检测小鼠肺组织中人类第10号染色体缺失的磷酸酶和张力蛋白同源物基因(PTEN)mRNA及蛋白表达,以及不同剂量EGCG干预治疗对其表达的影响。结果与正常组相比,哮喘组小鼠气道炎性细胞浸润明显增加,气道反应性增高,肺组织中PTEN表达下降,差异有统计学意义(P<0.05),应用高低剂量的EGCG干预治疗后均可减轻气道炎性细胞浸润及降低气道高反应性,同时上调肺组织中PTEN基因的表达,与哮喘组相比差异有统计学意义(P<0.05),且呈剂量依赖性。结论哮喘组小鼠肺组织中PTEN基因表达下降,EGCG干预治疗减轻小鼠气道炎症的,其可能通过上调PTEN基因的表达而发挥作用。
Objective To investigate the effect of epigallo-catechin gallate(EGCG)on the expression of PTEN in the lung tissues in acute asthmatic mice model. Methods Thirty two healthy female BALB/c mice(Specific-pathogen-free grade,18-22 g,6-8 weeks)were randomly divided into normal control group,asthma group,EGCG(5 mg/kg)group and EGCG(50 mg/kg)group.The asthma models were established in BALB/c mice by sensitizing and challenging with ovalbumin. Airway hyperresponsiveness was detected,and lung tissues were examined for inflammatory cell infiltration by the hematoxylin and eosin staining. Serum OVA-specific Ig E level in the serum were evaluated by enzyme-linked immunosorbent assay. The expression of PTEN gene and protein in the lung tissue was measured by real-time PCR and immunohistochemistry respectively. Results Compared with normal control group,the asthma groups showed more pathological changes in the airway,higher level of OVA-s Ig E in serum and airway hyperresponsiveness to aerosolized methacholine,but lower expression of PTEN in the ling tissue(P<0.05). Low or high doses EGCG administration also significantly ameliorated inflammatory cell infiltration,reduced airway hyperresponsiveness,decreased Ig E level in the serum and incresesd the expression of PTEN in the lung tissue compared with asthma group(P<0.05). Conclusion The expression of PTEN was decre Ased in the lung tissue of the asthmatic mice,EGCG administration may play a vital role in ameliorating airway inflammation by upregulating the expression of PTEN gene.
Objective To investigate the effect of epigallo-catechin gallate(EGCG)on the expression of PTEN in the lung tissues in acute asthmatic mice model. Methods Thirty two healthy female BALB/c mice(Specific-pathogen-free grade,18-22 g,6-8 weeks)were randomly divided into normal control group,asthma group,EGCG(5 mg/kg)group and EGCG(50 mg/kg)group.The asthma models were established in BALB/c mice by sensitizing and challenging with ovalbumin. Airway hyperresponsiveness was detected,and lung tissues were examined for inflammatory cell infiltration by the hematoxylin and eosin staining. Serum OVA-specific Ig E level in the serum were evaluated by enzyme-linked immunosorbent assay. The expression of PTEN gene and protein in the lung tissue was measured by real-time PCR and immunohistochemistry respectively. Results Compared with normal control group,the asthma groups showed more pathological changes in the airway,higher level of OVA-s Ig E in serum and airway hyperresponsiveness to aerosolized methacholine,but lower expression of PTEN in the ling tissue(P<0.05). Low or high doses EGCG administration also significantly ameliorated inflammatory cell infiltration,reduced airway hyperresponsiveness,decreased Ig E level in the serum and incresesd the expression of PTEN in the lung tissue compared with asthma group(P<0.05). Conclusion The expression of PTEN was decre Ased in the lung tissue of the asthmatic mice,EGCG administration may play a vital role in ameliorating airway inflammation by upregulating the expression of PTEN gene.
引文
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11 Moon SK,Kim HM,Kim CH.PTEN induces G cell cycle expression of P21 m RNA in HASMCs arrest and inhibits MMP-9 expression via the regulation of NF-B and AP-1 in vascular smooth muscle cells[J].Arch Biochem Biophys,2004,421(2):267-276.
12 Dong X,Yu LG,Sun R et al.Inhibition of PTEN expression and activity by angiotensinⅡinduces proliferation and migration of vascular smooth muscle cells[J].J Cell Biochem,2013,114(1):174-182.
13 Lan H,Zhong H,Gao Y,et al.The PTEN tumor suppressor inhibits human airway smooth muscle cell migration[J].Int J Mol Med,2010,26(6):893-899.
14 Wu D,Wang J,Pae M,et al.Green tea EGCG,T cells,and T cell-mediated autoimmune diseases[J].Mol Aspects Med,2012,33(1):107-18.
15 Kim SH,Park HJ,Lee CM,et al.Epigallocatechin-3-gallate protects toluene diisocyanate-in airway inflammation in a murine model of asthma[J].FEBS Lett,2006,580(7):1883-1890.
2 Liu S,Wang XJ,Liu Y,et al.PI3K/AKT/m TOR signaling is involved in(-)-epigallocatechin-3-gallate-induced apoptosis of human pancreatic carcinoma cells[J].Am J Chin Med,2013,41(3):629-642.
3 Zhang QY,Li MR.Effect of EGCG on the proliferation and apoptosis of human uterine leiomyoma cells in vitro[J].National Medical Frontiers of China,2011,6(1):14-15.
4 Bian EB,Huang C,Ma TT,et al.DNMT1-mediated PTEN hypermethylation confers hepatic stellate cell activation and liver fibrogenesis in rats[J].Toxicol Appl Pharmacol,2012,264(1):13-22.
5 Yamada KM,Araki M.Tumor suppressor PTEN:modulator of cell signaling,growth,migration and apoptosis[J].J Cell Sci,2001,114(13):2375-2382.
6 Cantley LC,Neel BG.New insights into tumor suppression:PTEN suppresses tumor formation by restraining the phosphoinositide 3-kinase/AKT pathway[J].Proc Natl Acad Sci U S A,1999,96(8):4240-4245.
7 Maehama T,Dixon JE.The tumor suppressor,PTEN/MMAC1,dephosphorylates the lipid second messenger,phosphatidylinositol 3,4,5-trisphosphate[J].J Biol Chem,1998,273(22):13375-13378.
8 Sun H,Lesche R,Li DM,et al.PTEN modulates cell cycle progression and cell survival by regulating phosphatidylinositol 3,4,5-trisphosphate and Akt/protein kinase B signaling pathway[J].Proc Natl Acad Sci U S A,1999,96(11):6199-6204.
9 Myers MP,Pass I,Batty IH,et al.The lipid phosphatase activity of PTEN is critical for its tumor suppressor function[J].Proc Natl Acad Sci U S A,1998,95(23):13513-13518.
10 Huang J,Kontos CD.Inhibition of vascular smooth muscle cell proliferation,migration,and survival by the tumor suppressor protein PTEN[J].Arterioscler Thromb Vasc Biol,2002,22(5):745-51.
11 Moon SK,Kim HM,Kim CH.PTEN induces G cell cycle expression of P21 m RNA in HASMCs arrest and inhibits MMP-9 expression via the regulation of NF-B and AP-1 in vascular smooth muscle cells[J].Arch Biochem Biophys,2004,421(2):267-276.
12 Dong X,Yu LG,Sun R et al.Inhibition of PTEN expression and activity by angiotensinⅡinduces proliferation and migration of vascular smooth muscle cells[J].J Cell Biochem,2013,114(1):174-182.
13 Lan H,Zhong H,Gao Y,et al.The PTEN tumor suppressor inhibits human airway smooth muscle cell migration[J].Int J Mol Med,2010,26(6):893-899.
14 Wu D,Wang J,Pae M,et al.Green tea EGCG,T cells,and T cell-mediated autoimmune diseases[J].Mol Aspects Med,2012,33(1):107-18.
15 Kim SH,Park HJ,Lee CM,et al.Epigallocatechin-3-gallate protects toluene diisocyanate-in airway inflammation in a murine model of asthma[J].FEBS Lett,2006,580(7):1883-1890.