海南医学
海南醫學
해남의학
HAINAN MEDICAL JOURNAL
2015年
1期
11-14,15
,共5页
彭锦%李铁岭%韩丽娜%林晓明%何爽
彭錦%李鐵嶺%韓麗娜%林曉明%何爽
팽금%리철령%한려나%림효명%하상
心肌炎%L-NAME%一氧化氮%基质金属蛋白酶
心肌炎%L-NAME%一氧化氮%基質金屬蛋白酶
심기염%L-NAME%일양화담%기질금속단백매
Myocarditis%L-NAME%Nitric oxide%Matrix metalloproteinase
目的：观察NG-硝基-L-精氨酸甲酯(L-NAME)对实验性自身免疫性心肌炎(EAM)Lewis大鼠模型的治疗效果，并探索可能的治疗机理。方法20只Lewis大鼠建立EAM动物模型：双足底注射心肌C蛋白片段和完全弗氏辅佐剂的油状混合物，腹腔注射百日咳毒素。大鼠随机等分为治疗组和模型对照组，每组各10只。治疗组腹腔注射5 mg·kg-1·d-1 L-NAME，从免疫注射术后第1天开始连续20 d，1次/d。对照组相同时间内给予相同剂量生理盐水腹腔注射。治疗结束后第1天处死动物，心脏取材，进行系列检测。其中组织病理学石蜡切片苏木素-伊红(HE)染色检测心肌炎症分级，免疫组织化学染色检测T淋巴细胞浸润，天狼星红染色检测心肌胶原纤维含量，硝酸还原酶法检测NO水平，明胶酶谱法检测胶原酶活性。结果与对照组比较，L-NAME治疗组心肌炎症级别下降[(3.42±0.31) vs (2.51±0.22)，P<0.01]、T淋巴细胞浸润数目减少[(28.2±4.6) vs (13.2±1.9)，P<0.01]、心肌间质纤维化级别下降[(2.33±0.26) vs (1.14±0.17)，P<0.01]、血清NO水平降低[(68.34±8.61)μmol/L vs (45.71±6.53)μmol/L， P<0.01]，明胶酶活性降低[(254526±4729) vs (184712±3869)，P<0.01]。结论 L-NAME抑制EAM病理发展过程，其机制可能与通过降低NO水平和明胶酶活性，从而降低心肌炎症细胞浸润，延缓心肌间质纤维化有关。
目的：觀察NG-硝基-L-精氨痠甲酯(L-NAME)對實驗性自身免疫性心肌炎(EAM)Lewis大鼠模型的治療效果，併探索可能的治療機理。方法20隻Lewis大鼠建立EAM動物模型：雙足底註射心肌C蛋白片段和完全弗氏輔佐劑的油狀混閤物，腹腔註射百日咳毒素。大鼠隨機等分為治療組和模型對照組，每組各10隻。治療組腹腔註射5 mg·kg-1·d-1 L-NAME，從免疫註射術後第1天開始連續20 d，1次/d。對照組相同時間內給予相同劑量生理鹽水腹腔註射。治療結束後第1天處死動物，心髒取材，進行繫列檢測。其中組織病理學石蠟切片囌木素-伊紅(HE)染色檢測心肌炎癥分級，免疫組織化學染色檢測T淋巴細胞浸潤，天狼星紅染色檢測心肌膠原纖維含量，硝痠還原酶法檢測NO水平，明膠酶譜法檢測膠原酶活性。結果與對照組比較，L-NAME治療組心肌炎癥級彆下降[(3.42±0.31) vs (2.51±0.22)，P<0.01]、T淋巴細胞浸潤數目減少[(28.2±4.6) vs (13.2±1.9)，P<0.01]、心肌間質纖維化級彆下降[(2.33±0.26) vs (1.14±0.17)，P<0.01]、血清NO水平降低[(68.34±8.61)μmol/L vs (45.71±6.53)μmol/L， P<0.01]，明膠酶活性降低[(254526±4729) vs (184712±3869)，P<0.01]。結論 L-NAME抑製EAM病理髮展過程，其機製可能與通過降低NO水平和明膠酶活性，從而降低心肌炎癥細胞浸潤，延緩心肌間質纖維化有關。
목적：관찰NG-초기-L-정안산갑지(L-NAME)대실험성자신면역성심기염(EAM)Lewis대서모형적치료효과，병탐색가능적치료궤리。방법20지Lewis대서건립EAM동물모형：쌍족저주사심기C단백편단화완전불씨보좌제적유상혼합물，복강주사백일해독소。대서수궤등분위치료조화모형대조조，매조각10지。치료조복강주사5 mg·kg-1·d-1 L-NAME，종면역주사술후제1천개시련속20 d，1차/d。대조조상동시간내급여상동제량생리염수복강주사。치료결속후제1천처사동물，심장취재，진행계렬검측。기중조직병이학석사절편소목소-이홍(HE)염색검측심기염증분급，면역조직화학염색검측T림파세포침윤，천랑성홍염색검측심기효원섬유함량，초산환원매법검측NO수평，명효매보법검측효원매활성。결과여대조조비교，L-NAME치료조심기염증급별하강[(3.42±0.31) vs (2.51±0.22)，P<0.01]、T림파세포침윤수목감소[(28.2±4.6) vs (13.2±1.9)，P<0.01]、심기간질섬유화급별하강[(2.33±0.26) vs (1.14±0.17)，P<0.01]、혈청NO수평강저[(68.34±8.61)μmol/L vs (45.71±6.53)μmol/L， P<0.01]，명효매활성강저[(254526±4729) vs (184712±3869)，P<0.01]。결론 L-NAME억제EAM병리발전과정，기궤제가능여통과강저NO수평화명효매활성，종이강저심기염증세포침윤，연완심기간질섬유화유관。
Objective To investigate the therapeutic effect of NG-nitro-L-arginine methyl ester (L-NAME) on experimental autoimmune myocarditis (EAM) in Lewis rats, and to explore the possible therapeutic mechanism. Methods Twenty EAM Lewis rats were treated by injection of myocardial C protein emulsified in completed Freund adjuvant in double footpad and intraperitoneal injection of pertussis toxin. EAM Lewis rats were divided into the treat-ment group and control group (each with 10 cases). In the treatment group, the treatment protocol was intraperitoneal ad-ministration of L-NAME (5 mg·kg-1·d-1) after immunization for 20 days. While in control group the same dose of normal saline was intraperitoneally injected in the same period of time. After experiment at the designate time point, the rats were euthanatized, and their hearts were harvested and tested. Paraffin sections were used for hematoxylin and eosin (HE) stain to determine the inflammation score, for immunohistological stain to determine the infiltration of T lymphocytes, and for picrosirius stain to determine fibrosis score and collagen content. Nitrate reductase method was used to detect serum NO level and gelatin zymography assay to detect the activity of gelatinase. Results The inflammation score in cardiac paraf-fin slides [(3.42±0.31) vs (2.51±0.22), P<0.01], infiltration of T lymphocytes [(28.2±4.6) vs (13.2±1.9), P<0.01], myo-cardial interstitial fibrosis score [(2.33 ± 0.26) vs (1.14 ± 0.17), P<0.01], serum NO level [(68.34 ± 8.61) μmol/L vs (45.71 ± 6.53)μmol/L, P<0.01] and activity of gelatinase [(254 526 ± 4 729) vs (184 712 ± 3 869), P<0.01] in treatment group were all significantly lower than in control group. Conclusion L-NAME plays an important role in the patho-genesis of autoimmune myocarditis, and its mechanism may be related to the decrease of serum NO level and gelatin-ase activity in decreasing myocardial inflammatory cells infiltration and delaying myocardial interstitial fibrosis.
