摘要
病毒性心肌炎(viral myocarditis,VMC)是小儿心血管系统常见疾病,其发病机制尚未完全阐明,病情轻重不一,虽然大多数急性VMC患者心功能可完全恢复,但部分患者可能出现反应性心肌纤维化,心室重塑,发展成为慢性VMC、扩张性心肌病(DCM),心肌纤维化的病理过程伴随着VMC的始终,至今国内外对其尚无特效的治疗手段。Periostin是日本学者发现的一种具有调节成骨细胞黏附和分化的功能骨黏附分子,当时被命名为成骨细胞特异性因子-2(osteoblast-specific factor-2,OSF-2),现已归类于细胞外基质(extracellular matrix,ECM)蛋白。近年来研究表明periostin在心梗时作为TGF-β1的效应因子具有趋化心肌成纤维细胞,促进胶原合成和成熟,形成稳定胶原网,参与心室重构的作用。有体外实验表明,periostin与心肌成纤维细胞相互作用,并可调节胶原-I的交联和成熟,与心梗后心肌纤维化密切相关。但目前periostin与病毒性心肌炎心肌纤维化的关系报道尚少。本研究通过构建柯萨奇B_3型病毒(CVB3)BALB/C小鼠VMC模型,运用半定量RT-PCR方法、免疫组织化学方法确定了periostin在VMC发病过程中的表达及可能作用,证实了periostin的表达与心肌纤维化程度、血清血管紧张素II水平,TGF-β1表达呈正相关,提示periostin可能作为血管紧张素II,TGF-β1的下游因子参与VMC心肌纤维化过程。冬虫夏草是我国传统中药,具有调节免疫、清除自由基、抗心律失常及直接抑制病毒复制等作用,本研究主要探讨冬虫夏草是否有抗病毒性心肌炎小鼠心肌纤维化的作用,同时以血管紧张素受体拮抗剂(AT1)厄倍沙坦作为疗效对比,结果表明二者均可通过对periostin的抑制,改善心肌纤维化,同时小剂量冬虫夏草效果与厄倍沙坦相当,大剂量冬虫夏草则优于厄倍沙坦,冬虫夏草的抗心肌纤维化作用,呈量效依赖性,本研究为病毒性心肌炎的临床治疗提供理论和实验依据。
本研究创新点:1. periostin在病毒性心肌炎的表达少有报道,本研究探讨periostin在病毒性心肌炎各期的表达及可能作用。2.本研究探讨了冬虫夏草对periostin表达的影响,及其抗病毒性心肌炎心肌纤维化作用和可能机制。
Viral myocarditis (VMC) is one of the most common infective diseases in cardiovascular diseases. Most of the patients are annoyed with acute VMC, but someone develops into the chronic VMC, dilated cardiomyopathy with survival rate about five years is around 50%. VMC has been threatening children health severely. Myocardial fibrosis refers to the excessive accumulation of collagen fibers in the normal tissues and structures, marked elevation of collagen concentration, or the changes of collagen constituents. This pathological change exists in every stage of VMC. Now, it is believed that this change is associated tightly with cardiac arrhythmia, cardiac dysfunction and dilated cardiomyopathy. The periostin gene was initially cloned from a mouse calvarial cell line (MC3T3-E1) and originally named osteoblast specific factor-2 (OSF-2). Periostin is a member of a growing family of matricellular proteins. Periostin has been shown to play a crucial role during tissue morphogenesis. Periostin is attended as a novel factor responsible for ventricular dilation. Perostin has been shown to be involved in the progress of Myocardial fibrosis after myocardial infarction It is believed that the abnormal expression of periostin may be related with the occurrence and development of VMC.
(1) The expression and function of periostin in mice model with viral myocarditis
Materials and methods : In this study we first established VMC models through BALB/c mice infected with coxsackie virus B3. 100 pure bred male Balb/c mice aged 4 weeks and weighted 18±1g, were divided into control group(n=40) and VMC group (n=60) randomly. VMC model was founded by injecting intraperitoneally 0.1ml CVB3 Nancy solution one time a day for three days, while the control group was injected with 0.1ml Engeal’s fluid excluding virus. The model mice were sacrificed by extirpating eyeballs on day 7, day 14,day 28, and day 56 respectively, after injecting CVB3 ,0.8-1.0ml blood were extracted by burettes. The serums were separated and stored at -70℃, preparing for the test of Ang II and CK-MB. Then all mice were killed by cutting necks, obtaining hearts and putting them partly into 10% formaldehyde solution for biopsy, the other part was stored at -70℃for RT-PCR. The mice of control group were treated by the same methods. After virus inoculation until day 0,day 7,day14,day28,day56, we examined the contents of periostin in the myocardium by SP and RT-PCR . Meanwhile, we examined the score for myocardial necrosis, the level of CK-MB and AngII in serum, CVF and the contents of TGF-β1 mRNA in the myocardium. Based on the results, we further studied the relationship between periostin and other index. Our study would provide new theoretic base and experimental evidence for prevention and therapy of VMC.
Results: We found that the model mice displayed tarnishable furs, reduced movements, been weary spirits and poor appetites ,vaulted backs, showed indifference to stimulation or tended to irritation and been lower temperature since the third day. Death occured on day 4, and reached the top from day 7 to day 14. Among 60 mice in model group, 21 died. The death rate was 35.0%. None of the control mice had any symptom and death. The serum CK-MB concentrations of the VMC group were much higher than that of the control mice at each time spot(P<0.01).The peak is in day 14 , after that day the serum CK-MB concentrations decreased. The other indexs include serum AngII concentrations ,CVF, TGF-β1 increased gradually in the course of disease. The expression of periostin was detected in myocardial tissue at the day0, day7, day14, day28, and day56 after CVB3 infection quantitatively by semiquantitative analysis RT-PCR. The results showed that the expression of periostin increased slightly at the 7 day after infection, then the expression was increased continuously, and the expression of periostin was highest at the 56th day after infection. Linear correlation analysis showed the contents of periostin in the myocardium with the level of AngII in serum, CVF and the contents of TGF-β1 in the myocardium is positive correlation remarkably, but score for myocardial necrosis and level of CK-MB in serum have no correlation with the contents of periostin in the myocardium.
Conclusions: It suggested that periostin may be involved in the myocardial fibrosis in myocardium of VMC. Ang II and TGF-β1 maybe increase periostin expression, and eventually contributed to the myocardial fibrosis in VMC mice model. Periostin may be a new target in the therapy of VMC.
(2): The therapeutical effect and possible mechanism of [Cordyceps sinensis (Berk.) Sacc.] (CS) and Irbesartan in the VMC mice models in chronic phase.
Materials and methods :70 pure bred male Balb/c mice aged 4 weeks and weighted 18±1g, were divided into control group(n=10) and VMC group (n=60) randomly. VMC model was founded by injecting intraperitoneally 0.1ml CVB3 Nancy solution one time a month for three monthes , while the control group was injected with 0.1ml Engeal’s fluid excluding virus.Then we successfully established the models of viral myocarditis in chronic phase through BALB/C mice infected with CVB3 .42 models of viral myocarditis in chronic phase were divided into 4 groups. They were VMC-control group (n=12), Irbesartan group (n=10), CS-large group (n=10) and CS-small group(n=10). VMC-control group was treated intragastric with water 1ml/d . Irbesartan group was treated intragastric with Irbesartan at a dose of 10mg/kg.d . CS-large group was treated intragastric with CS at a dose of 7.5g/kg.d CS-small group was treated intragastric with CS at a dose of 2.5g/kg.d .After feeding drugs for 60 days, we weighed the mice, examined the heart function changes, observed Myocardial ultrastructural changes and myocardial collagen fiber distribution,measured CVF and the level of AngII in blood serum. At the same time, the contents of collagen I and collagen III were detected by immunohistochemical techniques. TGF-β1and periostin were detected by reverse transeription polymerase chain reaction Then the therapeutic effect of CS and Irbesartan were discussed.
Results:Compared with VMC-control group, the mortality matched Irbesartan group, CS-large group showed significant decrease, while the parameters of heart function were improved (P<0.01), but the parameters is not normal yet (P<0.01). Compared with VMC-control group, the level of Ang II in serum was significantly inhibited by CS (P<0.01). But Irbesartan seemed not act(P>0.05). Compared with VMC-control group, Irbesartan group and CS-large group have lower parameters of CVF(p<0.05). The effect of CS large group is better than the effects of Irbesartan group and CS-smallgroup (p<0.05). Compared with VMC-control group, the expressions of TGF-β1and periostin were attenuated in Irbesartan group and CS-large group (p<0.05). There was no evident difference between CS-small group and VMC-control group(P>0.05). Compared with VMC-control group ,the expression of collagen I was decreased in Irbesartan group, CS-large group and CS-small group (p<0.05). AT the same time, the expression of collagen III was decreased in CS-large group and CS-small group (p<0.05).
Conclusions: periostin play an important role in the process of viral myocarditis. CS can offer some protection to myocardial fibrosis in viral myocarditis mice . The inhibition of the expression of periostin may be one of the mechanisms .
引文
[1] Takeshita S, Kikuno R, Tezuka K, et al. Osteoblast-specific factor 2: cloning of a putative bone adhesion protein with homology with the insect protein fasciclin I[J]. Biochem J, 1993, 294(Pt 1):271-278.
[2] Skonier J, Neubauer M, Madisen L, Bennett K, Plowman GD, Purchio AF cDNA cloning and sequence analysis of beta ig-h3, a novel gene induced in a human adenocarcinoma cell line after treatment with transforming growth factor-beta. DNA Cell Biol 1992, 11:511-522.
[3] Politz O, Gratchev A, McCourt PA, Schledzewski K, Guillot P, Johansson S, Svineng G, Franke P, Kannicht C, Kzhyshkowska J, Longati P, Velten FW, Goerdt SStabilin-1 and -2 constitute a novel family of fasciclin-like hyaluronan receptor homologues. Biochem J, 2002, 362:155–164.
[4] Horiuchi K, Amizuka N, Takeshita S, et al. Identification and characterization of a novel protein, periostin, with restricted expression to periosteum and periodontal ligamentand increased expression by transforming growth factor beta[J]. J Bone Miner Res, 1999, 14(7): 1239-1249.
[5] Rios H, Ko Llshik SV, Wang H, Wang J, Zhou HM, Lindsley A, Rogers R, Chen Z, Maeda M, Kruzynska-Frejtag A, Feng JQ, Conway SJ. Periostin null mice exhibit dwarfism, Ineisor enamel defects,and an early-onset Periodontal disease-like phenotype. Mol Cell Biol. 2005, 25:11131-44.
[6] Kikuchi Y, Kashima T, Nishiyama T, Shimazu K, Morishita Y, Shimazaki M, Kiil, Horie H, Nagai H, Kudo A, Fukayama M. Periostin is expressed in Pericryptal fibroblasts And cancer-assoeiated fibroblasts in the colon. J Histochem Cytochem. 2008,56:753-64.
[7] Hamilton DW. Funetional role of Periostin in development and wound repair: Implications for connective tissue disease. JCell Commun Signal, 2008,2:9-17.
[8] Goetsch SC, Hawke TJ, Gallardo TD, Richardson JA, Garry DJ. Transcriptional profiling and regulation of the extracellular matrix during muscle regeneration. Physiol Genomics 2003,14:261-271.
[9] Kudo Y, Siriwardena BS, Hatano H, Ogawa I, Takata T. Periostin:novel diagnostic and therapeutic target for cancer. Histol Histopathol, 2007, 22:1167–1174.
[10] Li P, Oparil S, Feng W, Chen YF. Hypoxia-responsive growth factors upregulateperiostin and osteopontin expression via distinct signaling pathways in rat pulmonary arterial smooth muscle cells. J Appl Physiol 2004, 97:1550–1558 discussion 1549.
[11] Lindner V, Wang Q, Conley BA, Friesel RE, Vary CP. Vascular injury induces expression of periostin:implications for vascular cell differentiation and migration. Arterioscler Thromb Vasc Biol, 2005, 25:77–83.
[12] Takayama G, Arima K, Kanaji T, Toda S, Tanaka H, Shoji S, et al. Periostin:a novel component of subepithelial fibrosis of bronchial asthma downstream of IL-4 and IL-13 signals. J Allergy Clin Immunol, 2006, 118:98–104.
[13] Oku E, Kanaji T, Takata Y, Oshima K, Seki R, Morishige S, Imamura R, Ohtsubo K, Hashiguchi M, Osaki K, et al. Periostin and bone marrow fibrosis. Int J Hematol 2008, 88(1):57–63.
[14] Ruan K, Bao S, ouyang G. The multifaceted role of Periostin in tumorigenesis. Cell Mol Sci. 2009. Epub ahead of Print.
[15] Sasaki H, Sato Y, Kondo S, et al. Expression of the periostin mRNA level in neuroblastoma[J]. J Pediatr Surg,2002,37(9):1293-1297.
[16] Kudo Y, Ogawa I, Kitajima S, et al. Periostin promotes invasion and anchorage- independent growth in the metastatic process of head and neck cancer[J]. Cancer Res, 2006, 66(7):6928-6935.
[17] Siriwardena BS, Kudo Y, Ogawa I, et al. Periostin is frequently overexpressed and enhances invasion and angiogenesis in oral cancer[J]. Br J Cancer, 2006, 95(10): 1396-1403.
[18] Chang Y, Lee TC, Li JC, et al. Differential expression of osteoblast-specific factor-and polymeric immunoglobulin receptor genes in nasopharyngeal carcinoma[J]. Head Neck, 2005, 27(10):873-882.
[19] Fluge O, Bruland O, Akslen LA, et al. Gene expression in poorly differentiated papillary thyroid carcinomas[J]. Thyroid, 2006, 16(2):161-175.
[20] Shao R, Bao S, Bai X, et al. Acquired expression of periostin by human breast cancers promotes tumor angiogenesis through up-regulation of vascular endothelial growth factor receptor2 expression[J]. Mol Cell Biol, 2004, 24(5):3992-4003.
[21] Grigoriadis A, Mackay A, Reis-Filho JS, et al. Establishment of the epithelial-specific transcriptome of normal and malignant human breast cells based on MPSS and array expressiondata[J]. Breast Cancer Res, 2006, 8(5):R56.
[22] Bao S, Ouyang G, Bai X,et al. Periostin potently promotes metastatic growth of coloncancer by augmenting cell survival via the Akt/PKB pathway[J]. Cancer Cell, 2004, 5(4): 329-339.
[23] Baril P, Gangeswaran R, Mahon PC, et al. Periostin promotes invasiveness and resistance of pancreatic cancer cells to hypoxia induced cell death: role of the beta(4) integrin and the PI3k pathway[J]. Oncogene, 2007, 26(14):2082-2094.
[24] Mert E, J rgk, Andre G, et al. Periostin creates a tumor-supportive microenvironment in the pancreas by sustaining fibrogenic stellate cell activity[J]. Gastroenterology, 2007, 132(4): 1447-1464.
[25] Gillan L, Matei D, Fishman DA, et al. Periostin secreted by epithelial ovarian carcinoma is a ligand for alpha(V)beta(3) and alpha(V)beta(5) integrins and promotes cell motility[J]. Cancer Res, 2002, 62(9):5358-5364.
[26] Norris RA, Moreno-Rodriguez RA, Sugi Y, Hoffman S, Amos J, Hart MM, Potts JD, Goodwin RL, Markwald RR Periostin regulates atrioventricular valve maturation. Developmental Biology 2008, 316:200-213.
[27] Oka T, Xu J, Kaiser RA, Melendez J, Hambleton M, Sargent MA, Lorts A, Brunskill EW, Dorn GW 2nd, Conway SJ, Aronow BJ, Robbins J, Molkentin JD Genetic manipulation of periostin expression reveals a role in cardiac hypertrophy and ventricular remodeling. [see comment]. Circulation Research 2007, 101:313-321.
[28] Shimazaki M, Nakamura K, Kii I, Kashima T, Amizuka N, Li M, et al. Periostin is essential for cardiac healing after acute myocardial infarction J Exp Med. 2008, 18 205(2): 295–303.
[29] Iekushi K, Taniyama Y, Azuma J, Katsuragi N, Dosaka N, Sanada F, et al. Novel mechanisms of Valsartan on the treatment of acute myocardial infarction through inhibition of the antiadhesion molecule periostin. Hypertension, 2007, 49:1409-1414.
[30] Katsuragi N, Morshita R, Nakamura N, Ochiai T, Taniyama Y, Hasegawa Y, Kawashima K, Kaneda Y, Ogihara T, Sugimura K. Periostin as a novel factor responsible for ventricular dilation. Circhlution, 2004, 110:1806-1813.
[31] Norris R, Damon B, Mironov V, Kasyanov V, Ramamurthi R, Moreno-Rodriguez R, Trusk T, Potts J, Goodwin R, Davis J, Hoffman S, Wen X, Sugi Y, Kern K, Mjaatvedt C, Turner D, Oka T, Conway SJ, Molkentin J, Forgacs G, Markwald RR. Periostin regulates collagen fibrillogenesis and biomechanical properties of connective tissues. J Cell Biochem, 2007, 101:695–711.
[32] Sun M, et al. Temporal response and localization of integrins b1 and b3 in the heart aftermyocardial infarction: regulation by cytokines. Circulation 2003, 107:1046–1052.
[33] Snider P, Hinton RB, Moreno-Rodriguez RA, Wang J, Rogers R, Lindsley A, Li F, Ingram DA, Menick D, Field L, Firulli AB, Molkentin JD, Markwald R, Conway SJ Periostin is required for maturation and extracellular matrix stabilization of noncard- iomyocyte lineages of the heart. Circ Res 2008, 102:752–760.
[34] Rios H, Koushik SV, Wang H, Wang J, Zhou HM, Lindsley A, RogersR, Chen Z, Maeda M, Kruzynska-Frejtag A, Feng JQ, Conway SJ.Periostin null mice exhibit dwarfism, incisor enamel defects, and anearly-onset periodontal disease-like phenotype. Mol Cell Biol. 2005, 25:11131–11144.
[35] Kiotsekoglou A, Sutherland GR, Moggridge JC, Nassiri DK, Camm AJ, Child AH. The unravelling of primary myocardial impairment in Marfan syndrome by modern echocardiography. Heart. 2009, 95:1561–1566.
[36] Cohn RD, van Erp C, Habashi JP, Soleimani AA, Klein EC, Lisi MT, Gamradt M, ap Rhys CM, Holm TM, Loeys BL, Ramirez F, Judge DP, Ward CW, Dietz HC. Angiotensin II type 1 receptor blockade attenuates TGF-beta-induced failure of muscle regeneration in multiple myopathicstates. Nat Med. 2007, 13:204–210.
[37] LindsleyA., W.Li, J.Wang, N.Maeda, R.Rogers, and S.J. Conway. Comparison of the four mouse fasciclin-containing genes expression patterns during valvuloseptal morphogenesis. Gene Expr. Patterns 2005, 5:593–600.
[38] Takayama G, Arima K, Kanaji T, Toda S, Tanaka H, Shoji S, McKenzie AN, Nagai H, Hotokebuchi T, Izuhara K. Periostin: a novel component of subepithelial fibrosis of bronchial asthma downstream of IL-4 and IL-13 signals. J. Allergy Clin. Immunol. 2006, 118:98–104.
[39] Norris R, Damon B, Mironov V, Kasyanov V, Ramamurthi A, Moreno-Rodriguez R, Trusk T, Potss J, Goodwin RL, Davis J, Hoffman S, Wen X, Sugi Y, Kern CB, Mjaatvedt CH, Turner D, Oka T, Conway SJ, Molkentin JD, Forgacs G, Markwald R. Periostin regulates collagen fibrillogenesis and the biomechanical properties of connective tissues. J. Cell Biochem. 2007, 101:695–711.
[40] Sugiura T, Takamatsu H, Kudo A, Amann E. Expression and characterization of murine osteoblast-specific factor 2 (OSF-2) in a baculovirus expression system. Protein Expr. Purif. 1995, 6:305–311.
[41] Christiansen DL, Huang EK, Silver FH. Assembly of type I collagen: fusion of fibril subunits and the influence of fibril diameter on mechanical properties. Matrix Biol 2000,19:409–420.
[42] Gillan L, Matei D, Fishman DA, Gerbin CS, Karlan BY, Chang DD. Periostin secreted by epithelial ovarian carcinoma is a ligand for alpha(V)beta(3) and alpha(V)beta(5) integrins and promotes cell motility. Cancer Res. 2002, 62:5358–5364.
[43] Yan W, Shao R. Transduction of a mesenchyme-specific gene periostin into 293T cells induces cell invasive activity through epithelial-mesenchymal transformation. J. Biol. Chem. 2006, 281:19700–19708.
[44] Bornstein P, Sage EH. Matricellular proteins: extracellular modulators of cell function. Curr Opin Cell Biol. 2002, 14:608–616.
[45] Bornstein P. Diversity of function is inherent in matricellular proteins: an appraisal of thrombospondin 1. J Cell Biol. 1995, 130:503–506.
[46] Katsuragi N, Morishita R, Nakamura N, et al. periostin as a novelfactor responsible for ventricular dilation[J]. Circulation, 2004, 110(13): 1806-1813.
[47] Derek S, Wheeler, MD, FAAP, et al. A formidable challenge: the diagnosis and treatment of viral myocarditis in children .Crit Care Clin 2003, 19(3): 365-391.
[48] Kii I, Amizuka N, Minqi L, Kitajima S, Saga Y, Kudo A. Periostin is an extracellular matrix protein required for eruption of incisors in mice. Biochem Biophys Res Commun. 2006, 342:766–772.
[49] Kishimoto C, Crumpacker CS , et al. Ribavirin treatment of coxsackie B3 myocarditis with analysis of lymphocyte subsets. J Am Coll Cardiol 1988, 12:1334–1341.
[50] Burgess ML, Terracio L, Hirozane T, Borg TK. Differential integrin expression by cardiac fibroblasts from hypertensive and exercise-trained rat hearts. Cardiovasc Pathol. 2004, 11:78–87.
[51] Horiuchi K, Amizuka N, Takeshita S, Takamatsu H, Katsuura M, OzawaH, Toyama Y, Bonewald LF, Kudo A. Identification and characterization of a novel protein, Periostin, with restricted expression to periosteumand periodontal ligament and increased expression by transforming growth factor beta. J Bone Miner Res. 1999, 14:1239–1249.
[52] Borg TK, Markwald R. Periostin: more than just an adhesion molecule.Circ Res. 2007, 101:230–231.
[53] Ku¨hn B, del Monte F, Hajjar RJ, Chang YS, Lebeche D, Arab S, Keating MT. Periostin induces proliferation of differentiated cardiomyocytes and promotes cardiac repair. Nat Med. 2007, 13:962–969.
[54] Conway SJ, Molkentin JD. Postn as a heterofunctional regulator of cardiac developmentand disease. Curr Genomics, 2008, 9:548–555.
[55] Lorts A, Schwanekamp JA, Elrod JW, Sargent MA, Molkentin JD.Genetic manipulation of periostin expression in the heart does not affect myocyte content, cell cycle activity, or cardiac repair. Circ Res. 2009, 104:e1–e7.
[56] Shimazaki M, Nakamura K, Kii I, Kashima T, Amizuka N, Li M, Saito M, Fukuda K, Nishiyama T, Kitajima S, Saga Y, Fukayama M, Sata M, Kudo A. Periostin is essential for cardiac healing after acute myocardial infarction. J Exp Med. 2008, 205:295–303.
[57] Norris RA, Damon B, Mironov V, Kasyanov V, Ramamurthi A, Moreno-Rodriguez R, Trusk T, Potts JD, Goodwin RL, Davis J, Hoffman S, Wen X, Sugi Y, Kern CB, Mjaatvedt CH, Turner DK, Oka T, Conway SJ, Molkentin JD, Forgacs G, Markwald RR. Periostin regulates collagen fibrillogenesis and the biomechanical properties of connective tissues. J Cell Biochem. 2007, 101:695–711.
[58] Oka T, Xu J, Kaiser RA, Melendez J, Hambleton M, Sargent MA, Lorts A, Brunskill EW, Dorn GW, Conway SJ, Aronow BJ, Robbins J, Molkentin JD. Genetic manipulation of periostin expression reveals a role in cardiac hypertrophy and ventricular remodeling. Circ Res. 2007, 101:313–321.
[59] Kruzynska-Frejtag A, Machnicki M, Rogers R, Markwald RR, Conway SJ. Periostin is expressed in the embryonic mouse heart during valve formation. Mech Dev. 2001, 103:183–188.
[60] Bao S, Ouyang G, Bai X, Huang Z, Ma C, Liu M, Shao R, Anderson RM, Rich JN, Wang XF. Periostin potently promotes metastatic growth of colon cancer by augmenting cell survival via Akt/PKB pathway. Cancer Cell, 2004, 5:329–339.
[61] Butcher JT, Norris RA, Hoffman S, Mjaatvedt CH, Markwald RR. Periostin promotes atrioventricular mesenchyme matrix invasion and remodeling mediated by integrin signaling through Rho/PI 3-kinase. Dev Biol 2007, 302:256–266.
[62] Litvin J., Zhu S., Norris R. & Markwald, R. Periostin family of proteins:therapeutic targets for heart disease. Anat. Rec. A Discov. Mol. Cell. Evol. Biol. 2005, 287:1205– 1212.
[63] Horiuchi K, Amizuka N, Takeshita S, Takamatsu H, Katsuura M, Ozawa H, Toyama Y, Bonewald LF, Kudo A. Identification and characterization of a novel protein, Periostin, with restricted expression to periosteum and periodontal ligament and increased expression by transforming growth factor beta. J Bone Miner Res. 1999, 14:1239–1249.
[64] Li G, Oparil S, Sanders JM, Zhang L, Dai M, Chen LB, Conway SJ, Mc Namara C,Sarembock IJ. Phosphatidylinositol-3-kinase signaling mediates vascular smooth muscle cell expression of Periostin in vivo and in vitro. Atherosclerosis. 2006, 188:292–300.
[65] Chen YF, Feng JA, Li P, Xing D, Zhang Y, Serra R, Ambalavanan N, Majid-Hassan E, Oparil S. Dominant negative mutation of the TGF-beta receptor blocks hypoxia- induced pulmonary vascular remodeling.J Appl Physiol. 2006, 100:564-571.
[66] Goetsch SC, Hawke TJ, Gallardo TD, Richardson JA, Garry DJ. Transcriptional Profiling and regulation of the extracellular matrix during muscle regeneration. Physiol Genomics. 2003, 14:261–271.
[67] Ouyang G, Liu M, Ruan K, Song G, Mao Y, Bao S. Upregulated expression of periostin by hypoxia in non-small-cell lung cancer cells promotes cell survival.
[68] Snider P, Hinton RB, Moreno-Rodriguez RA, Wang J, Rogers R, Lindsley A, Li F, Ingram DA, Menick D, Field L, Firulli AB, Molkentin JD, Marwald R, Conway SJ. Periostin is required for maturation and extracellular matrix stabilization of noncardio- myocyte lineages of the heart. Circ Res. 2008, 102:752-760.
[69] Iekushi K, Taniyama Y, Azuma J, Katsuragi N, Dosaka N, Sanada F, Koibuchi N, Nagao K, Ogihara T, Morishita R. Novel mechanisms of valsartan on the treatment of acute myocardial infarction through inhibition of the antiadhesion molecule periostin. Hypertension. 2007, 49: 1409-1414.
[70] Chen YF, Feng JA, Li P, Xing D, Zhang Y, Serra R, Ambalavanan N,Majid-Hassan E, Oparil S. Dominant negative mutation of the TGF-beta receptor blocks hypoxia- induced pulmonary vascular remodeling. J Appl Physiol. 2006, 100: 564-571.
[71] Du¨nker N, Krieglstein K. Tgfbeta2 _/_ Tgfbeta3 _/_ double knockout mice display severe midline fusion defects and early embryonic lethality. Anat Embryol (Berl). 2002, 206:73-83.
[72] Azhar M, Schultz Jel J, Grupp I, Dorn GW II, Meneton P, Molin DG, Gittenberger-de Groot AC, Doetschman T. Transforming growth factor beta in c ardiovascular develop- ment and function. Cytokine Growth Factor Rev. 2003, 14:391– 407.
[73] Azhar M, Runyan RB, Gard C, Sanford LP, Miller ML, Andringa A, Pawlowski S, Rajan S, Doetschman T. Ligand-specific function of transforming growth factor beta in epithelial-mesenchymal transition in heart development. Dev Dyn. 2009, 238:431-442.
[74] Foitzik K, Lindner G, Mueller-Roever S, Maurer M, Botchkareva N, Botchkarev V, Handjiski B, Metz M, Hibino T, Soma T, Dotto GP, Paus R. Control of murine hair follicle regression (catagen) by TGF-beta1 in vivo. FASEB J. 2000, 14:752-760.
[75] Proetzel G, Pawlowski SA, Wiles WV, Yin M, Boivin G, Howles PN,Ding J, Ferguson MWJ, Doetschman T. Transforming growth factor-3 is required for secondary palate fusion. Nat Genet. 1995, 11:409–141.
[76] Bartram U, Molin DG, Wisse LJ, Mohamad A, Sanford LP, Doetschman T, Speer CP, Poelmann RE, Gittenberger-de GA. Double-outlet right ventricle and overriding tricuspid valve reflect disturbances of looping, myocardialization, endocardial cushion differentiation, and apoptosis in Tgfb2 knockout mice. Circulation. 2001, 103: 2745–2752.
[77] Wang YX, da Cunha V, et al. Antiviral and myocyte protective effects of murine interferon-beta and -{alpha}2 in coxsackievirus B3-induced myocarditis and epicarditis in Balb/c mice. Am J Physiol Heart Circ Physiol. 2007, 293(1):H69-76.
[78] Li G, Oparil S, Sanders JM, Zhang L, Dai M, Chen LB, Conway SJ, Mc Namara CA, Sarembock IJ. Phosphatidylinositol- 3-kinase signaling mediates VSMC expression of periostin in vivo and in vitro.Atherosclerosis. 2006, 188:292–300.
[79] Kuhl U, Pauschinger M, et al. Interferon-treatment eliminates cardiotropic viruses and improves left ventricular function in patients with myocardial persistence of viral genomes and left ventricular dysfunction. Circulation. 2003, 107:2793-8.
[80] Paige Snider, Robert B. Hinton, Ricardo A. Moreno-Rodriguez, Jian Wang, et al. Periostin Is Required for Maturation and Extracellular Matrix Stabilization of Noncardiomyocyte Lineages of the Heart Circulation Research. 2008, 102:752-760.
[81] Webster AD. Pleconaril-an advance in the treatment of enterovira infection in immuno-compromised patients. J Clin Virol, 2005, 32(1):1-6.
[82] Berk, B.C., K. Fujiwara, and S. Lehoux.ECM remodeling in hypertensive heart disease. J. Clin. Invest. 2007, 117:568–575.
[83] Galvin KM, Donovan MJ, Lynch CA, Meyer RI, Paul RJ, Lorenz JN, Fairc Huntress V, Dixon KL, Dunmore JH, Gimbrone MA Jr, Falb D, Huszar D. A role for Sma development and homeostasis of the cardiovascular system. Nat Genet. 2000, 4:171– 174.
[84] Bauer S, Gottesman G, et al. Severe Coxsackie virus B infection in preterm newborns treated with pleconaril. Eur J Pediatr. 2002 Sep, 161(9):491-493.
[85] Fohlman J, Pauksen K, et al. Antiviral treatment with WIN 54954 reduces mortality in murine coxsackievirus B3 myocarditis. Circulation, 1996, 94(9):2254-2259.
[86] Noutsias M, Pauschinger M, et al. Immunomodulatory treatment strategies in inflammatory cardiomyopathy:current status and future perspectives.Expert RevCardiovasc Ther. 2004, 2:37-51.
[87]马沛然,王述昀.皮质激素对病毒性心肌炎疗效及不良反应预防的实验研究[J].临床儿科杂志, 2004, 22(7): 467-469.
[88] Liu C, Chen J, et al. Immunosuppressive treatment for inflammatory cardiomyopathy: meta-analysis of randomized controlled trials. Int Heart J. 2005, 46(1):113-122.
[89] Martini S, Peters H, et al. Current perspectives on FTY720.Expert Opin Investig Drugs, 2007, 16(4): 505-518.
[90] Miyamoto T, Matsumori A, et al. Therapeutic effects of FTY720, a new immuno- suppressive agent, in a murine model of acute viral myocarditis[J]. JAm Coll Cardiol, 2001, 37(6):1713-1718.
[91] Kishimoto C, Shioji K, et al. Treatment of acute inflammatory cardiomyopathy with intravenous immunoglobulin ameliorates left ventricular function associated with suppression ofinflammatory cytokines and decreased oxidative stress[J]. Int J Cardio, l 2003, 91(2):173-178.
[92] Amabile NF, Fraisse A, et al. Outcome of acute fulminant myocarditis in children. Heart 2006, 92:1269-1273.
[93] Tanaka T, Kanda T, et al. Overexpression of interleukin-6 aggravates viral myocarditis: impaired increase in tumor necrosis factor-alpha. J Mol Cell Cardiol. 2001, 33(9): 1627-1635.
[94] Nishio R, Matsumori A, et al. Treatment of experimental viral myocarditis with interleukin-10 [J]. Circulation. 1999, 100(10):1102-1108.
[95] Nishii M, Inomata T, et al. Serum level of interleukin-10 on admission as a prognostic predictor of human fulminant myocarditis[J]. Science direct, 2004, 13(11):32.
[96] Daniela Cihakova, Jobert G Barin, et al. Interleukin-13 Protects Against Experimental Autoimmune Myocarditis by Regulating Macrophage Differentiation American Journal of Pathology. 2008, 172:1195-1208.
[97] Yoshida A, Kand T, et al. Interleukin-18 reduces expression of cardiac tumor necrosis factor-alpha and atrial natriuretic peptide in a murine model of viral myocarditis [J]. Life Sc, 2002, 70(11):1225-1234.
[98] DeLisa Fairweather, Sylvia Frisancho-Kiss,et al. IL-12 Protects against Coxsackievirus B3-Induced Myocarditis by Increasing IFN-γand Macrophage and Neutrophil Populations in the Heart The Journal of Immunology, 2005, 174: 261-269.
[99]郭富强,于小华,等.巨噬细胞移动抑制因子在病毒性心肌炎小鼠心肌中表达[J].临床儿科杂志, 2007,25(10):818.
[100] Matsui Y, Okamoto H, et al. Blockade of macrophage migration inhibitory factor ameliorates experimental autoimmune myocarditis[J]. J Mol Cell Cardiol, 2004, 37(2): 557-566.
[101]孙景辉,翟淑波,等.核因子κB在病毒性心肌炎小鼠发病中的作用.临床儿科杂志, 2008, 26(3):233-235.
[102] Matsumori A, Nunokawa Y, et al. Suppression ofcytokines and nitric oxide production, and protection against lethal endotoxemia and viral myocarditis by a new NF-κB inhibitor. Eur J Heart Fail 2003, 6:137–144.
[103] Yue-Chun L, Li-Sha G, et al. Protective effects of carvedilol in murine model with the coxsackievirus B3-induced viral myocarditis. J Cardiovasc Pharmacol. 2008, 51(1): 92-98.
[104] Daniels MD, Hyland KV, et al. Treatment of experimental myocarditis via modulation of the renin-angiotensin system. Curr Pharm Des, 2007, 13(13):1299-1305.
[105] Zhang ZC, Li SJ, et al. Effect of astragaloside on cardiomyocyte apoptosis in murine coxsackievirus B3 myocarditis. J Asian Nat Prod Res. 2007 Mar, 9(2):145-151.
[106] Akira matsumori, MD, PhD, et al. French Maritime Pine Bark Extract Inhibits Viral Replication and Prevents Development of Viral Myocarditis Journal of Cardiac Failure 2007, 13(9):785-791.
[107]赵余庆,于明,陈立君,等.冬虫夏草属真菌化学研究概况.中草药, 1999,30(12):950-953.
[108]李祥麟,石玉洁,黄檀溪.略述北冬虫夏草生物活性物质的应用(临床)试验研究.传染病药学, 2003, 13(4):13-16.
[109]张兴辉,石力夫,胡晋红.冬虫夏草化学成分和药理作用研究进展.中药材, 2000, 23(11):722-724.
[110]索志荣,刘效珍.宁心宝治疗室性早搏的临床观察[J].中西医结合脑血管病杂志, 2005, 3(7):643.
[111]龚晓健,季晖,曹琪,等.人工虫草提取物抗心律失常作用的研究[J].中国药科大学学报, 2001, 32(3):221-223.
[112]王赫.冬虫夏草对豚鼠心室肌细胞胞浆钙浓度及L-型钙电流的影响[J].中国病理生理杂志, 2003, 19(11):1581.
[113]俞宙,何建新.冬虫夏草水提液对触发性心律失常的治疗作用[J].中国医药学报, 1999, 14(1):32-34.
[114]吴垠.虫草菌粉有效部位的主要药效学及作用机理研究[D].杭州:浙江中医药大学, 2006.
[115]温元祥.冬虫夏草的药理作用及临床作用[J].天津药学, 1998, 10.(1):47.
[116]崛尾忠正,杨美玲.冬虫夏草菌丝体抗哇巴因所致豚鼠心脏毒性和抗氧自由基作用的研究[J].中国药房, 1997, 8(2):57.
[117]季晖,龚晓健,卢顺高,等.人工虫草菌丝体提取物抗哇巴因所致心脏毒性作用的研究[J].中国药科大学学报, 2000, 31(2):118.
[118]刘凤芝,李延平,黄明莉,等.冬虫夏草醇提物对大鼠缺血再灌注过程心肌保护作用的研究[J].中国病理生理杂志, 1999, 15(3):240-241.
[119]许宏远,郑昕.冬虫夏草对阿霉素性心肌的保护作用[J].中医药学报, 2000,28(3):64.
[120]冯鸣国,周前贵,冯高闳,等.人工培养虫草菌丝体对麻醉犬血管的扩张作用[J].中药通报, 1987, 12(2):41-43.
[121]吴秀香,马克玲,李淑云.冬虫夏草降压作用实验研究[J].锦州医学院学报, 2001, 22(2):10-11.
[122]蔡久英,任旭荣,范仲凯,等.黄芪和冬虫夏草对心脏病左心室舒张功能及血脂的影响[J].中国中西医结合急救杂志, 2002, 9(3):174-175.
[123]赵鹏,杨俊峰,李彬.蛹虫草菌丝体降血脂作用的动物试验研究[J].中国食品卫生杂志, 2004, 16(5):434.
[124] Koh J冬虫夏草菌丝体热水提取物的降胆固醇作用[J].国外医学·中医中药分册, 2004, 26(5):305.
[125]李锋,高兴玉,饶邦复,等.冬虫夏草提取液对小鼠病毒性心肌炎心肌脂质过氧化作用的影响[J].南京中医药大学学报, 2006, 22(1):21-23.
[126]朱照静,李峰,饶邦复,高兴玉冬虫夏草增强病毒性心肌炎小鼠免疫反应.中药药理与临床, 2002, 18(6):22-24.
[127]李峰.冬虫夏草提取液对实验性病毒性心肌炎小鼠免疫功能的影响[J].细胞与分子免疫学杂志, 2006, 22(3).
[128] Sato S, Tsutsumi R, Burke A, et al. Persistence of replicating coxsackievirusB3 in the athymic murine heart is associated with development of myocarditic lesions[J]. J GenVirol, 1994, 75(11):2911-2924.
[129] Kawai C. From myocarditis to cardiomyopathy:mechanisms of inflammation and cell death: learning from the past for future[J]. Circμlation. 1999, 99(8):1091-1100.
[130] Seko Y, Shinka Y, Kawasaki A, et al. Evidence of perforin mediated cardiac myocyte injury in acute marine myocarditis caused by coxsackie virus B3[J]. J Pathol, 1993, 170(1): 53-58.
[131] Young LH, Joas SV, Zheng LM, et al. Perforin-mediated myocardial damage in acute myocarditis[J]. Lancet, 1990, 336(8722):1019-1021.
[132] Stanton, L.W., L.J. Garrard, D.Damm, B.L. Garrick, A. Lam, A.M. Kapoun, Q. Zheng, A.A. Protter, G.F. Schreiner, and R.T. White. Altered patterns of gene expression in response to myocardial infarction. Circ. Res. 2000, 86:939–945.
[133] Chocron S, Alwan W, Toubin G, et al. Effects of myocardial ischemias the release of cardiac troponin I in isolated rat hearts[J]. J Thorac Cardiovasc Surg, 1996, 112(2): 508-513.
[134]李丽,凡栋,王程,等.血管紧张素II促进Periostin表达的信号转导通路研究[J].基础医学与临床, 2009, 29(增刊):35.
[135] Kuhl U, Pauschinger M, Seeberg B, Lassner D, Noutsias M, Poller W, et al. Viral persistence in the myocardium is associated with progressive cardiac dysfunction [J]. Circulation, 2005, 112(13): 1965-1970.
[136]陈淳媛,孙跃女,杨作成,龙燕琼.雷帕霉素对柯萨奇病毒B3诱导的心肌成纤维细胞eIF-4E表达的影响.中国当代儿科杂志[J], 2007, 9(6):587-590.
[137]朱伟旺,赵凤琴.厄贝沙坦对SHR左心室肥厚和心肌纤维化的影响[J].心脏杂志,2009,21(2):190-196.
[138]孙绍骞,张志国,赵学忠.厄贝沙坦对急性心肌梗死大鼠心室重构的影响[J].中国老年学杂志. 2008, 4(28):746-748.
[139] Gray MO, Long CS, Kalinyak JE, et al. AngiotensinII stimulates cardiac myocyte hypertrophy via paracrine release of TGF-beta 1 and endothelia-1 from fibroblasts[J]. Cardiovasc Res, 1998, 40:352-363.
[140]荆志成,程显声,杨英珍.氯沙坦干预病毒性心肌炎恢复期、慢性期心脏胶原表达及心功能的实验研究[J].中华心血管病杂志, 1999, 27(2):140-143.
[141] Nakamura H, Yamamoto T, Yamamura T, et al. Repetitive Coxsackie virus infectioninduces cardiac dilation in post-myocarditic mice. Jpn Circ J 1999;63 (10): 794-802.
[142]李风华,刘平,熊卫国,等.冬虫夏草菌丝对二甲基亚硝胺诱导的大鼠肝纤维化的作用[J].中西医结合学报, 2006, 4(5):514-517.
[143]许伯钧,陈铭鸿.冬虫夏草改善SARS后肺纤维化[J].华夏医药, 2006, 1(3):172-176.
[144]闵亚丽,于黔,肖俊.冬虫夏草在肾间质纤维化大鼠模型中的作用及其对TGF-β1、α-SMA的影响[J].中国中西医结合肾病杂志, 2007, 8(2):92-93,100-102.
[145]窦敬芳,翁孝刚.发酵虫草菌粉对糖尿病大鼠肾脏结构的影响[J].中国临床康复, 2006, 10(15):105-107.
[146]马瑞霞,刘丽秋,赵秀珍,等.虫草制剂对肾小球硬化大鼠细胞外基质的影响[J].中国临床康复, 2006, 10(23):102-104.
[147] Taylor K, Patten RD, Smith J, et al. Divergent effects ofangiotensing-converting enzyme inhibition and angiotensinⅡreceptor antagonismon myocardial cellular proliferation and collagen deposition aftermyocardial infarction in rats. J Cardiovasc Pharmaco,l 1998, 31: 654-667.
[148] Yoshida J, Yamamoto K, Mano T, et al. AT1 receptor blocker added to ACE inhibitor provides benefits at advanced stage of hypertensive diastolic heart failure. Hypertension, 2004, 43: 686-691.
[149] Yamamoto K, Mano T, Yoshida J, et al. ACE inhibitor and angiotensinⅡtype 1 receptor blocker differently regulate ventricularfibrosis in hypertensive diastolic heart failure. JHypertens, 2005, 23: 393-400.
[150] Verrecchia F, Mauviel A. Transforming growth factor-beta signaling through the Smad pathway: role in extracellular matrix gene expression and regulation[J]. J Invest Dermatol, 2002, 118(2):211-215.
[151] Brand T, Schneider MD. The TGF beta superfamily in myocardium: ligands, receptors, transduction, and function[J]. J Mol Cell Cardiol, 1995, 27:5-18.