|
|
- 2017
重组人松弛素-2对肝星状细胞增殖、Ⅰ型胶原合成及TGF-β1与CTGF mRNA表达的影响
|
Abstract:
摘要：目的 在细胞水平研究松弛素的抗肝纤维化作用，并初步探讨其分子机制，为肝纤维化的治疗提供实验依据。方法 体外培养大鼠肝星状细胞(HSC-T6)，分别用20、50、100ng/mL重组人松弛素-2(recombinant human relaxin-2, RLX-2)干预，采用噻唑蓝(MTT)比色实验检测RLX-2对HSC-T6增殖的影响，双抗体夹心酶联免疫吸附法(ELISA)检测对照组及RLX-2作用48h后的细胞培养基上清液Ⅰ型胶原水平，利用逆转录酶-聚合酶链反应(RT-PCR)技术检测药物干预48h的结缔组织生长因子(CTGF)、转化生长因子-β1(TGF-β1)mRNA表达情况。结果 RLX-2对HSC的增殖具有抑制作用，可降低HSCⅠ型胶原水平，RLX-2抑制HSC的CTGF mRNA表达，对HSC的TGF-β1 mRNA表达无明显影响。结论 RLX-2可能通过抑制体外培养大鼠HSC细胞的增殖、Ⅰ型胶原及CTGF mRNA的表达而发挥抗大鼠肝纤维化作用。
ABSTRACT: Objective To validate that relaxin can resist hepatic fibrosis at the cellular level and explore its molecular mechanism in order to provide experimental basis for the treatment of liver cirrhosis. Methods Cultured HSC-T6s were treated with different concentrations (20, 50 and 100ng/mL) of recombinant human relaxin-2 (RLX-2). The proliferation of HSC-T6 was measured by MTT colorimetric assay. The content of type Ⅰ collagen in the cell culture supernatant of each group was detected by ELISA at 48h of drug intervention; RT-PCR was used to detect the mRNA expressions of CTGF and TGF-β1 in HSC-T6 at 48h of drug intervention. Results RLX-2 inhibited the proliferation of HSC and reduced type Ⅰ collagen content of HSC cells. It also inhibited the CTGF mRNA expression of HSC, but did not have a significant effect on the expression of TGF-β1 mRNA. Conclusion In the experiment of culturing HSC-T6 in vitro, RLX-2 may play a role in rat liver fibrosis by inhibiting cell proliferation and type Ⅰ collagen and CTGF mRNA expressions
| [1] | BATALLER R, BRENNER DA. Liver fibrosis[J]. J Clin Invest, 2005, 115(2):209-218. |
| [2] | BENNETT RG, HEIMANN DG, SINGH S, et al. Relaxin decreases the severity of established hepatic fibrosis in mice[J]. Liver Int, 2014, 34(3):416-426. |
| [3] | WESTON BS, WAHAB NA, MASON RM. CTGF mediates TGF-beta-induced fibronectin matrix deposition by upregulating active alpha5beta1 integrin in human mesangial cells[J]. J Am Soc Nephrol, 2003, 14(3):601-610. |
| [4] | IHN H, YAMANE K, KUBO M, et al. Blockade of endogenous transforming growth factor beta signaling prevents up-regulated collagen synthesis in scleroderma fibroblasts: association with increased expression of transforming growth factor beta receptors[J]. Arthritis Rheum, 2001, 44(2):474-480. |
| [5] | YOKOI H, SUGAWARA A, MUKOYAMA M, et al. Role of connective tissue growth factor in profibrotic action of transforming growth factor-beta: a potential target for preventing renal fibrosis[J]. Am J Kidney Dis, 2001, 38(4 Suppl 1):S134-138. |
| [6] | LEASK A, SA S, HOLMES A, et al. The control of ccn2 (ctgf) gene expression in normal and scleroderma fibroblasts[J]. Mol Pathol, 2001, 54(3):180-183. |
| [7] | ULLOA L, TABIBZADEH S. Lefty inhibits receptor-regulated Smad phosphorylation induced by the activated transforming growth factor-beta receptor[J]. J Biol Chem, 2001, 276(24):21397-21404. |
| [8] | 赵青，陈楠，周同. TGF-β诱导CTGF表达的信号转导途径及与纤维化形成[J]. 生命科学, 2003, 15(3):155-158. |
| [9] | 朱艳艳，张怀勤，周浩. 松弛素抗纤维化作用的研究进展[J]. 医学综述, 2014, 20(15):2714-2716. |
| [10] | FALLOWFIELD JA, HAYDEN AL, SNOWDON VK, et al. Relaxin modulates human and rat hepatic myofibroblast function and ameliorates portal hypertension in vivo[J]. Hepatology, 2014, 59(4):1492-1504. |
| [11] | 周玉平，涂国卿，胡俊义，等. 松弛素对肝纤维化大鼠的作用及对转化生长因子-β1的影响[J]. 中西医结合肝病杂志, 2012, 22(5):284-286. |
| [12] | WILLIAMS EJ, BENYON RC, TRIM N, et al. Relaxin inhibits effective collagen deposition by cultured hepatic stellate cells and decreases rat liver fibrosis in vivo[J]. Gut, 2001, 49(4):577-583. |
| [13] | GUPTA S, CLARKSON MR, DUGGAN J, et al. Connective tissue growth factor: potential role in glomerulosclerosis and tubulointerstitial fibrosis[J]. Kidney Int, 2000, 58(4):1389-1399. |
| [14] | SAFADI R, FRIEDMAN SL. Hepatic fibrosis―role of hepatic stellate cell activation[J]. Med Gen Med, 2002, 4(3):27-27. |
| [15] | CASSIMAN D, LIBBRECHT L, DESMET V, et al. Hepatic stellate cell/myofibroblast subpopulations in fibrotic human and rat livers[J]. J Hepatol, 2002, 36(2):200-209. |
| [16] | FRIEDMAN SL. Hepatic stellate cells: Protean, multifunctional, and enigmatic cells of the liver[J]. Physiol Rev, 2008, 88(1):125-172. |
| [17] | ALCOLADO R, ARTHUR MJ, IREDALE JP. Pathogenesis of liver fibrosis[J]. Clin Sci(Lond), 1997, 92(2):103-112. |
| [18] | GRESSNER AM, WEISKIRCHEN R, BREITKOPF K, et al. Roles of TGF-beta in hepatic fibrosis[J]. Front Biosci, 2002, 7:D793-D807. |
| [19] | 李光明，李定国. 结缔组织生长因子与肝纤维化[J]. 国外医学(消化系疾病分册), 2004, 24(4):198-200. |
| [20] | BENNETT RG, DALTON SR, MAHAN KJ, et al. Relaxin receptors in hepatic stellate cells and cirrhotic liver[J]. Biochem Pharmacol, 2007, 73(7):1033-1040. |
