三七皂苷R1对心房颤动大鼠心肌炎症相关因子和金属基质蛋白酶表达的影响

【目的】研究三七皂苷R1对心房颤动大鼠细胞间黏附分子-1（ICAM-1）、肿瘤坏死因子-α（TNF-α）、金属基质蛋白酶-2（MMP-2）和金属机制蛋白酶-2抑制因子（TIMP-2）的影响,探讨三七皂苷R1防治心房颤动的机制。【方法】将102只大鼠根据随机数字法分为对照组、心房颤动组和三七皂苷R1组,每组34只。采用尾静脉注射乙酰胆碱-氯化钙建立心房颤动大鼠模型。三七皂苷R1组大鼠腹腔注射2mL三七皂苷R1水溶液。心电图测定心房颤动持续时间,Masson染色观察心肌纤维化程度,免疫组化法测定心房组织MMP-2和TIMP-2蛋白表达,酶联免疫吸附实验（ELISA）测定血清ICAM-1、TNF-α、MMP-2和TIMP-2水平,免疫印迹法（Western blotting）测定心房组织ICAM-1、TNF-α和Ⅰ型胶原蛋白水平。【结果】三七皂苷R1治疗前,两组心房颤动持续时间比较差异无统计学意义（P>0.05）;治疗后,三七皂苷R1组心房颤动持续时间（6.37±2.02）s,低于治疗前和心房颤动组（P<0.05）。Masson染色显示：对照组大鼠心房肌间质胶原纤维量正常;心房颤动组大鼠心房肌间质见大量胶原纤维;三七皂苷R1组大鼠心房肌间质见片、点状胶原纤维。与对照组比较,心房颤动组和三七皂苷R1组大鼠血清ICAM-1、TNF-α、MMP-2水平升高［（137.52±16.59）10-6g/L、（14.25±1.08）10-6g/L、（435.26±17.63）10-9g/L;（109.25±14.62）10-6g/L、（12.31±1.27）10-6g/L、（288.47±15.52）10-9g/L］（P<0.05）,TIMP-2水平降低［（3541.27±331.24）10-9g/L;（3975.46±313.24）10-9g/L］（P<0.05）,心房组织ICAM-1、TNF-α和Ⅰ型胶原蛋白水平（0.23±0.07、0.51±0.09、0.63±0.14;0.15±0.06、0.22±0.07、0.27±0.12）升高（P<0.05）,心房组织MMP-2蛋白光密度值（0.35±0.07;0.18±0.06）升高（P<0.05）,TIMP-2蛋白光密度值（0.11±0.04;0.18±0.03）降低（P<0.05）;与心房颤动组比较,三七皂苷R1组大鼠血清ICAM-1、TNF-α、MMP-2水平降低（P<0.05）,TIMP-2水平升高（P<0.05）,心房组织ICAM-1、TNF-α和Ⅰ型胶原蛋白水平降低（P<0.05）,心房组织MMP-2蛋白光密度值降低（P<0.05）,TIMP-2蛋白光密度值升高（P<0.05）。【结论】三七皂苷R1通过降低心房颤动大鼠血清和心房组织ICAM-1、TNF-α、MMP-2水平,升高TIMP-2水平发挥对心房颤动的防治作用。

Effects of Notoginsenoside R1 on Expression of Myocardial Inflammation-related Factors and Metalloproteinase in Atrial Fibrillation Rats

【Objective】To study the effects of notoginsenoside R1 on the levels of intercellular adhesion molecule- 1（ICAM- 1）,tumor necrosis factor- α （TNF- α）,metalloproteinase- 2 （MMP- 2） and metalloproteinase- 2 inhibitor （TIMP- 2）in rats with atrial fibrillation in order to explore the mechanism of notoginsenoside R1 on the preventing and treating atrial fibrillation. 【Methods】 102 rats were randomly divided into control group ,atrial fibrillation group and notoginsenoside R1 group,with 34 rats in each group. The rat model of atrial fibrillation was established by injection of acetylcholine-calcium chloride into the tail vein. The rats in the notoginsenoside R1 group were intraperitoneally injected with 2 mL of notoginsenoside R1. ECG was used to measure the duration of atrial fibrillation. Masson staining was used to observe the degree of myocardial fibrosis. Immunohistochemistry was used to detect the expression of MMP-2 and TIMP-2 in atrial tissue. The serum ICAM-1,TNF-α,MMP-2 and TIMP-2 levels were determined by enzyme-linked immunosorbent assay（ELISA）. The levels of ICAM-1,TNF-α and type I collagen in atrial tissue were determined by Western blotting.【Results】Before the treatment of notoginsenoside R1 ,there was no significant difference in the duration of atrial fibrillation between the two groups（P > 0.05）. After treatment,the duration of atrial fibrillation in the notoginsenoside R1 group［（6.37±2.02）s］was lower than that in the pre-treatment and the atrial fibrillation groups（P < 0.05）. Masson staining showed：the amount of atrial fibrillar collagen fibers in control group was normal;a large number of collagen fibers were seen in the atrial myocytes of atrial fibrillation group;the patchy and punctate collagen fibers were seen in the atrial myocytes of notoginsenoside R1 group. Compared with control group,the serum levels of ICAM-1,TNF- α and MMP-2 ［（137.52±16.59）10-6 g/L,（14.25±1.08）10-6 g/L,（435.26±17.63）10-9 g/L;（109.25±14.62）10-6 g/L ,（12.31±1.27）10-6 g/L, （288.47±15.52）10-9 g/L］were increased（P < 0.05）,the serum levels of TIMP-2 levels［（3 541.27±331.24）10-9 g/L ; （3 975.46 ± 313.24）10- 9 g/L］was decreased（P < 0.05）,the atrial tissue ICAM- 1,TNF- α and type I collagen levels （0.23±0.07 ,0.51±0.09 、0.63±0.14 ;0.15±0.06 ,0.22±0.07 ,0.27±0.12）were increased（P < 0.05）,the atrial tissue MMP-2 protein optical density（0.35±0.07;0.18±0.06）was increased（P < 0.05）,the atrial tissue TIMP-2 protein optical density（0.11±0.04;0.18±0.03）was decreased（P < 0.05）in atrial fibrillation group and the notoginsenoside R1 group; Compared with atrial fibrillation group,the levels of serum ICAM-1,TNF- α and MMP-2 were decreased（P < 0.05）, the levels of serum TIMP-2 was increased（P < 0.05）,the atrial tissue ICAM- 1 ,TNF- α and type I collagen levels were decreased（P < 0.05）,the density of MMP-2 protein in atrial tissue was decreased（P < 0.05）,and the optical density of TIMP-2 protein in atrial tissue was increased（P < 0.05）in the rats in notoginsenoside R1 group.【Conclusion】 Notoginsenoside R1 can prevent and treat atrial fibrillation by reducing the levels of ICAM- 1,TNF- α and MMP-2 and increasing the levels of TIMP-2 in serum and atrial tissue of rats with atrial fibrillation.