Qian Hong, Liu Lijing
Department of Clinical Medicine, Huaihua Medical College, Huaihua 418000, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2014 Jul;30(7):708-12.
To explore the protective effect of hydrogen sulfide (H2S) on viral myocarditis (VMC) mice and its mechanism.
Seventy BALB/c mice were randomly divided into 4 groups: normal control group (n=10), myocarditis group (n=20), DL-propargylglycine (PAG) group (n=20) and sodium hydrosulfide (NaHS) group (n=20). Mice in the latter three groups were inoculated with 0.1 mL Eagle's solution containing Coxsackievirus B3 (CVB3) intraperitoneally; whereas those in normal control group were treated with 0.1 mL Eagle's solution. On the day of inoculation, mice in PAG and NaHS groups received intraperitoneal administration with 40 mg/kg PAG and 50 μmol/kg NaHS, respectively; however, those in the other two groups were treated with PBS instead. PAG, NaHS or PBS were given one time for one day and persisted for 14 days. On day 15, all mice were killed after weighing body mass (BM). The mortality was compared among groups. Serum was separated and serum cardiac troponin I (cTnI) levels were detected using ELISA. The heart was removed and weighed to calculate heart mass (HM). Histological cross sections of heart were stained with hematoxylin-eosin, and myocardial histopathologic scores were counted under optical microscope. The activities of malondialdehyde (MDA), superoxide dismutase (SOD), glutathione-peroxidase (GSH-Px) and catalase (CAT) in the homogenate of myocardium were examined. Myocardial interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α) contents were detected by ELISA. The expression level of myocardial nuclear factor-κB (NF-κB) p65 in the nucleus was examined using Western blotting.
Compared with normal control group, the HM/BM, serum cTnI levels, myocardial MDA activity, NF-κB p65 expression level in the nucleus and myocardial contents of IL-1β, IL-6 and TNF-α were elevated, but myocardial SOD, GSH-Px and CAT activities were reduced in myocarditis group (P<0.05 or P<0.01). In comparison with myocarditis group, HM/BM, serum cTnI levels, myocardial histopathologic scores, myocardial MDA activity, NF-κB p65 expression level in the nucleus and myocardial contents of IL-1β, IL-6 and TNF-α significantly increased while myocardial SOD, GSH-Px and CAT activities significantly decreased in PAG groups (P<0.05). There was no significant difference in the mortality between PAG group and myocarditis group (P>0.05). The mortality, HM/BM, serum cTnI levels, myocardial histopathologic scores, myocardial MDA activity, NF-κB p65 expression level in the nucleus and myocardial contents of IL-1β, IL-6 and TNF-α were lower, while myocardial SOD, GSH-Px and CAT activities were higher in NaHS group than in myocarditis group (P<0.05).
H2S can produce effective protection against VMC in mice. The mechanism may be associated with enhancing antioxidative ability and inhibiting inflammatory response.
探讨硫化氢（H₂S）对病毒性心肌炎（VMC）小鼠的保护作用及其机制。
将70只BALB/c小鼠随机分为4组：正常对照组（n = 10）、心肌炎组（n = 20）、DL-炔丙基甘氨酸（PAG）组（n = 20）和硫氢化钠（NaHS）组（n = 20）。后三组小鼠腹腔注射0.1 mL含柯萨奇病毒B3（CVB3）的伊格尔氏液；而正常对照组小鼠则注射0.1 mL伊格尔氏液。接种当天，PAG组和NaHS组小鼠分别腹腔注射40 mg/kg PAG和50 μmol/kg NaHS；然而，其他两组小鼠则用PBS代替。PAG、NaHS或PBS每天给药1次，持续14天。第15天，称量所有小鼠体重（BM）后处死。比较各组小鼠的死亡率。分离血清，采用酶联免疫吸附测定（ELISA）法检测血清心肌肌钙蛋白I（cTnI）水平。取出心脏并称重，计算心脏重量（HM）。心脏组织学切片进行苏木精-伊红染色，在光学显微镜下计数心肌组织病理学评分。检测心肌匀浆中丙二醛（MDA）、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）和过氧化氢酶（CAT）的活性。采用ELISA法检测心肌白细胞介素（IL）-1β、IL-6和肿瘤坏死因子-α（TNF-α）含量。采用蛋白质免疫印迹法检测心肌细胞核因子-κB（NF-κB）p65的表达水平。
与正常对照组相比，心肌炎组小鼠的HM/BM、血清cTnI水平、心肌MDA活性、细胞核NF-κB p65表达水平以及心肌IL-1β、IL-6和TNF-α含量升高，但心肌SOD、GSH-Px和CAT活性降低（P < 0.05或P < 0.01）。与心肌炎组相比，PAG组小鼠的HM/BM、血清cTnI水平、心肌组织病理学评分、心肌MDA活性、细胞核NF-κB p65表达水平以及心肌IL-1β、IL-6和TNF-α含量显著升高，而心肌SOD、GSH-Px和CAT活性显著降低（P < 0.05）。PAG组与心肌炎组小鼠的死亡率无显著差异（P > 0.05）。NaHS组小鼠的死亡率、HM/BM、血清cTnI水平、心肌组织病理学评分、心肌MDA活性、细胞核NF-κB p65表达水平以及心肌IL-1β、IL-6和TNF-α含量均低于心肌炎组，而心肌SOD、GSH-Px和CAT活性高于心肌炎组（P < 0.05）。
H₂S可对小鼠VMC产生有效保护作用。其机制可能与增强抗氧化能力和抑制炎症反应有关。
