摘要
目的研究大蒜素是否通过TGFβ-Smads信号通路改善压力超负荷大鼠心肌纤维化。方法 40只SD雄性大鼠随机分为假手术组、模型组、大蒜素组和川芎嗪组,每组10只。以腹主动脉缩窄方法制备压力超负荷心肌肥厚大鼠模型。造模后3天,大蒜素组和川芎嗪组分别给予大蒜素注射液5.0mg/kg、盐酸川芎嗪注射液20mg/kg腹腔注射,假手术组和模型组给予生理盐水腹腔注射。给药4周后,天狼猩红染色观察心肌胶原变化,计算心肌胶原容积分数(myocardial collagen volume fraction,CVF)和血管周围胶原面积(perivascular collagen area,PVCA);ELISA法检测血清转化生长因子-β1(transforming growth factor-β1,TGF-β1)水平;免疫组化法观察心肌组织TGF-β1蛋白表达;实时荧光定量PCR检测心肌Smad2和Smad7 mRNA表达量的变化;荧光素酶报告基因检测进一步验证大蒜素对TGF-β信号传导系统的影响。结果与假手术组比较,模型组大鼠心肌CVF、PVCA、血清TGF-β1水平及心肌组织TGF-β1蛋白表达均明显增加(P<0.05,P<0.01);与模型组比较,大蒜素组和川芎嗪组PVCA、血清TGF-β1水平及心肌组织TGF-β1蛋白表达均明显减少(P<0.05,P<0.01)。模型组Smad2 mRNA表达上调而Smad7mRNA表达下调,大蒜素组和川芎嗪组Smad2 mRNA表达明显下调(P<0.05),川芎嗪组Smad7 mRNA表达明显上调(P>0.05)。在2ng/mLTGF-β1干预下,1～2μg/mL大蒜素可以明显抑制相应TGF-β1的荧光素酶活性(P<0.05)。结论大蒜素通过部分阻抑TGFβ1介导的Smad信号减轻压力超负荷大鼠的心肌反应性纤维化。
Objective To observe whether garlicin could ameliorate pressure overload induced myocardial fibrosis in rats through partial inhibiting transforming growth factor β1 （TGF-β1） mediated Smads signal. Methods Forty male SD rats were randomly divided into 4 groups, i.e., the sham-operation group, the model group, the garlicin group, and the Tetramethylpyrazine （TMP） group, 10 in each group. The pressure overload induced my-ocardial fibrosis rat model was prepared using coarctation of aorta. Three days after modeling 5. 0 mg/kg garlicin injection was administered to rats in the garlicin group, 20 mg/kg TMP injection to rats in the TMP group by peri-toneal injection, while normal saline was peritoneally injected to rats in the sham-operation group and the model group. Four weeks after medication, the changes of myocardial collagen were observed by picrosirius red staining. The myocardial collagen volume fraction （CVF） and perivascular collagen areas （PVCA） were calculated. The serum transforming growth factor β1 （TGF-β1） expression was detected using ELISA. The TGF-β1 protein expression in the myocardial tissue was observed using immunohistochemical assay. The changes of myocardial Smad2 and Smad7 mRNA expressions were detected using Real-time RT-PCR. The effects of garlicin on TGF-β1 mediated Smad Signaling through luciferase assay were further verified using Mvl Lu-（CAGA）12-Luc cell line re-sponse to TGF-β1. Results Compared with the sham-operation group, the myocardial levels of CVF and PVCA,the serum TGF-I31 level, and the TGF-[51 protein expression in the myocardial tissue obviously increased in the model group （P〈0.05,P〈0.01）. Compared with the model group, the PVCA level, the serum TGF-~I level, and the TGF-[5~ protein expression in the myocardial tissue of the garlicin group and the TMP group obviously de- creased （P 〈0. 05, P 〈0.01 ）. The Smad2 mRNA expression was up-regulated while Smad7 mRNA expression down-regulated in the model group. The Smad2 mRNA expression was obviously down-regulated in the garlicin group and the TMP group （ P 〈 0. 05）. The Smad7 mRNA expression was obviously up-regulated in the TMP group （ P 〉0.05）. One to 2 iag/mL garlicin could obviously inhibit the luciferase activities of corresponding TGF-β1 under the stimulation of 2 ng/mL TGF-β （ P 〈0.05）. Conclusion Garlicin ameliorated pressure overload in- duced myocardial fibrosis in rats through partial inhibiting TGF[5-Smads signal pathway.
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2012年第5期666-670,共5页
Chinese Journal of Integrated Traditional and Western Medicine
基金
首都医学发展科研基金资助项目(No.03-Ⅲ-02)
中日友好医院科研基金资助项目
