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人参皂苷 Rg1 通过调控缝隙连接拮抗铅致大鼠心肌细胞损伤的作用机制 |
李艺,王琼,冯斌,何珍
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1.山东省职业卫生与职业病防治研究院,山东 济南 250062;2.山东第一医科大学/山东省医学科学院;3.济南市疾病预防控制中心
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摘 要: 目的 观察醋酸铅染毒大鼠经人参皂苷 Rg1(ginsenoside Rg1,以下简称Rg1)干预前后心肌缝隙连接蛋白Cx43(connexin 43,Cx43)及其磷酸化蛋白(p-Cx43)的表达变化,探究Rg1通过缝隙连接对铅致心肌细胞损伤的保护作用。方法 将无特定病原体级的Sprague Dawley雄性大鼠按随机数字表法分为对照组,以二甲基亚砜(DMSO)溶解辛醇,作为辛醇对照组,辛醇为Cx43特异性阻滞剂,设置DMSO组及辛醇组,且不做干预;低、中、高剂量铅染毒组(90、180、360 mg/kg),Rg1干预组分为不同剂量铅染毒+Rg1组、辛醇+Rg1组、高剂量铅染毒+辛醇+Rg1 组(Pb360 mg/kg+辛醇+Rg1),共11组,每组8只。对照组给予等量蒸馏水,DMSO组、辛醇组按照体质量进行腹腔注射染毒(5 mmol/kg),不同剂量铅染毒组和铅染毒+Rg1 组经口灌胃醋酸铅溶液,每天染毒1次,每周连续5d,造模4周后,各Rg1(20 mg/kg)干预组每天给药1次,每周连续 5 d,共4周;对照组及不同剂量铅染毒组每天经口灌胃蒸馏水。4 周后处死大鼠收集心脏组织,计算心脏脏器系数,观察心肌组织病理学变化,检测全血血铅浓度,Cx43、p-Cx43、Bcl-2、Bax、Cleaved caspase-3、LC3-Ⅰ和 LC3-Ⅱ蛋白表达水平。结果 与对照组相比,染毒组血铅水平显著升高(P<0.05);高剂量铅染毒组心脏脏器系数显著降低,Rg1干预后脏器系数升高(P<0.05)。HE染色显示,对照组、DMSO组及辛醇组心肌结构无明显变化;低、中剂量铅染毒组出现心肌纤维排列紊乱;高剂量铅染毒组心肌纤维断裂。与铅染毒组、辛醇组相比,Rg1 干预组心肌组织结构明显改善,心肌纤维排列相对整齐紧密,无明显炎症。Western blot检测结果显示,铅染毒组Cx43、p-Cx43、Bcl-2及LC3-Ⅰ蛋白表达低于对照组(P<0.05),Bax、Cleaved caspase-3 及 LC3-Ⅱ蛋白表达高于对照组(P<0. 05);Rg1干预组Cx43、p-Cx43、Bcl-2及LC3-Ⅰ蛋白表达高于各剂量铅染毒组(P<0.05),Bax、Cleaved caspase-3及LC3-Ⅱ蛋白表达低于各剂量铅染毒组(P<0.05)。辛醇组与对照组、辛醇+Rg1 组相比,Cx43蛋白表达差异无统计学意义。结论 人参皂苷Rg1对铅致心肌细胞损伤的保护作用可能与其升高Cx43、p-Cx43 蛋白表达有关。 |
关键词: 人参皂苷 Rg1(Rg1) 醋酸铅 缝隙连接蛋白 43(Cx43) |
中图分类号: R135. 11; R994. 3
文献标识码: A
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基金项目: 山东省中医药科技项目(编号:2021Z038) |
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Mechanism of ginsenoside Rg1 antagonising lead-induced cardiomyocyte injury in rats by modulating gap junctions |
LI Yi,WANG Qiong,FENG Bin,HE Zhen
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Shandong Provincial Academy of Occupational Health and Occupational Medicine;Shandong First Medical University/Shandong Provincial Academy of Medical Sciences, Jinan, Shandong 250062,China
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Abstract: Objective To observe the changes in the expression of myocardial gap junction protein Cx43 (Cx43) and its phosphorylated protein (p-Cx43) before and after the intervention of ginsenoside Rg1 (Rg1) in lead acetate-contaminated rats,and to investigate the protective effect of Rg1 on lead-induced cardiomyocyte injury through gap junctions.Methods Sprague Dawley male rats without specific pathogen grade were divided into control groups by random number table method,and octanol was dissolved in DMSO as a control group of octanol,which is a Cx43-specific blocker,setting DMSO group,octanol (Oct) group, low,medium and high dose of Pb trained and poisoned group (90,180 and 360 mg/kg) and Rg1 intervention group as low,medium and high dose of Pb poisoning+Rg1 intervention group,Oct+Rg1 intervention group,and high dose lead poisoning+ Oct+Rg1 intervention group (Pb 360 mg/kg+Oct+Rg1),totalling 11 groups of 8 animals each.The control group was given an equal amount of distilled water,the DMSO group and the octanol group (5 mmol/kg) were stained by peritoneal injection according to their body mass,and different doses of lead-stained groups (90,180,360 mg/kg) and each dose of lead-stained+ Rg1 group were stained by oral gavage of the lead acetate solution once a day for 5 d consecutive per week.After 4 weeks of modeling, Rg1 (20 mg/kg) intervention was administered once daily for 5 d consecutive per week for 4 weeks in each group;distilled water was orally gavaged daily in the control group and the low,medium,high dose lead-stained groups.Cardiac tissues were collected from rats after 4 weeks of execution, cardiac apparatus coefficients were calculated, myocardial histopathological changes were observed, and whole blood lead concentration, Cx43, p-Cx43, Bcl-2,Bax,Cleaved caspase-3,LC3-Ⅰ
and LC3-Ⅱprotein expression levels were detected. Results Blood lead levels were significantly higher in all tainted groups compared to the control group (P<0.05);cardiac apparatus coefficient levels were significantly lower in the high-dose tainted group (360 mg/kg) and higher after Rg1 intervention (P<0.05).HE staining showed no significant change in myocardial structure in the control,DMSO,and octanol groups;myocardial fiber arrangement disorders were observed in the low and mediumdose lead-stained groups;and myocardial fibers were broken in the high-dose lead-stained group.Compared with the leadstained and octanol groups,the myocardial tissue structure of the Rg1 intervention group was significantly improved, and the myocardial fibers were arranged relatively neatly and tightly without obvious inflammation.The results of Western blot showed that the protein expression of Cx43,p-Cx43,Bcl-2,and LC3-Ⅰ in the lead-tainted group was lower than that in the control group (P<0.05),and that the protein expression of Bax,Cleaved caspase-3 and LC3-Ⅱ was higher than that in the control group (P<0.05);in the Rg1 intervention group, the protein expression of Cx43, p-Cx43, Bcl-2, and LC3-Ⅰ protein expression was higher than that in the different dose lead-tainted group (P<0.05),and Bax,Cleaved caspase-3 and LC3-Ⅱ protein expression were lower than that with the different dose lead-tainted group (P<0.05).There were no statistically significant difference in Cx43 protein expressed in the octanol group compared to the control group and the Oct+Rg1 group.Conclusion Ginsenoside Rg1 effectively protects against lead-induced cardiomyocyte injury,possibly related to its elevated Cx43 and p-Cx43 protein secretion. |
Keywords: ginsenoside Rg1 (Rg1) lead acetate gap junction protein 43 (Cx43) |
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