人参皂苷Rb1对川崎病小鼠心肌损伤及血管炎症严重程度的影响

目的探讨人参皂苷Rb1对川崎病小鼠心肌损伤及血管炎症严重程度的影响。方法选择40只5周龄无特定病原体级C57BL/6小鼠,按照随机数字表分为空白对照组、川崎病模型组、Rb1低剂量组和Rb1高剂量组,每组各10只。采用单次腹腔注射干酪乳酸杆菌细胞壁提取物(LCWE)建立川崎病模型,空白对照组和川崎病模型组进行等量0.9%氯化钠溶液灌胃,Rb1低剂量组和Rb1高剂量组分别灌胃30 mg/kg和60 mg/kg人参皂苷Rb1,1次/d,连续21 d。分别于干预前1 d以及干预后7、14、21 d对各组小鼠进行称重并记录。干预结束后,苏木精-伊红染色观察各组小鼠心肌组织病理情况;采用酶联免疫吸附试验(ELISA)法检测心肌组织匀浆中内皮素-1(ET-1)水平,总一氧化氮(NO)检测试剂盒检测心肌组织匀浆中NO水平;采用ELISA法检测小鼠血清中促炎因子白细胞介素(IL)-6和肿瘤坏死因子α(TNF-α)以及抗炎因子IL-10的水平;采用全自动凝血仪检测各组小鼠血浆纤维蛋白原水平,D-二聚体检测试剂盒(免疫荧光法)检测D-二聚体水平,小鼠ELISA试剂盒检测小鼠血浆AT-Ⅲ水平。结果干预后14 d,与空白对照组相比,其余3组不同时间点小鼠体重均显著减少,差异均有统计学意义(P<0.05);与川崎病模型组相比,Rb1高剂量组小鼠体重显著增加,差异有统计学意义(P<0.05);干预后21 d,与空白对照组相比,其余3组不同时间点小鼠体重均显著减少,差异均有统计学意义(P<0.05);与川崎病模型组相比,Rb1低、高剂量组小鼠体重显著增加,差异均有统计学意义(P<0.05)。与空白对照组相比,其余3组心肌组织病理评分、ET-1和NO水平均显著升高,差异均有统计学意义(P<0.05);与川崎病模型组相比,Rb1低、高剂量组小鼠以上指标则显著降低,且Rb1高剂量组也显著低于Rb1低剂量组,差异均有统计学意义(P<0.05)。与空白对照组相比,其余3组小鼠血清IL-6和TNF-α水平均显著升高,IL-10水平则显著降低,差异均有统计学意义(P<0.05);与川崎病模型组相比,Rb1低、高剂量组小鼠血清IL-6和TNF-α水平均显著降低,IL-10水平则显著升高,且Rb1高剂量组以上指标也较Rb1低剂量组显著改善,差异均有统计学意义(P<0.05)。与空白对照组相比,其余3组小鼠血清纤维蛋白原和AT-Ⅲ水平均显著降低,D-二聚体水平则显著升高,差异均有统计学意义(P<0.05);与川崎病模型组相比,Rb1低、高剂量组小鼠血清纤维蛋白原和AT-Ⅲ水平均显著升高,D-二聚体水平则显著降低,且Rb1高剂量组以上指标也较Rb1低剂量组显著改善,差异均有统计学意义(P<0.05)。结论人参皂苷Rb1治疗川崎病可显著促进小鼠体重恢复,从而降低川崎病对小鼠生长发育的影响,其还可减轻小鼠心脏冠状动脉损伤,并进一步改善血管炎症和凝血功能。

Effect of ginsenoside Rb1 on the severity of myocardial injury and vascular inflammation in mice with Kawasaki disease

Objective To investigate the effect and mechanism of ginsenoside Rb1 on the severity of myocardial injury and vascular inflammation in mice with Kawasaki disease(KD).Methods Forty 5-week-old SPF C57BL/6 mice were selected and divided into blank control group,KD model group,low-dose Rb1 group and high-dose Rb1 group according to random number table method,each with 10 mice.A single intraperitoneal injection of lactobacillus casei cell wall extract(LCWE)was used to establish a KD model.The blank control group and the KD model group were given the same amount of 0.9%sodium chloride solution intragastrically,the low-dose Rb1 group and the high-dose Rb1 group were intragastrically administered 30 mg/kg and 60 mg/kg ginsenoside Rb1,once a day for 21 consecutive days.The mice in each group were weighed and recorded before intervention 1 d,after the intervention 7,14 and 21 d.After the intervention,the pathological conditions of the myocardial tissues of the mice in each group were observed by hematoxylin-eosin staining.The level of endothelin-1(ET-1)in the myocardial tissue homogenate was detected by the enzyme-linked immunosorbent assay(ELISA)method,and the nitric oxide(NO)level in the myocardial tissue homogenate was detected by the total NO detection kit.The levels of the pro-inflammatory factors interleukin(IL)-6 and tumor necrosis factor alpha(TNF-α)and the anti-inflammatory factor IL-10 in the serum of mice were detected by ELISA.The plasma FIB level of the mice in each group was detected by the automatic coagulation method,the D-Dimer detection kit(immunofluorescence method),and he plasma AT-Ⅲ level of the mice was detected by ELISA.Results Intervention for 14 days,compared with the blank control group,the weight of the mice in the other three groups were significantly reduced at different time points,the differences were statistically significant(P<0.05).Compared with the KD model group,the weight of the mice in the high-dose Rb1 group was increased significantly,the difference was statistically significant(P<0.05);Intervention for 21 days,compared with the blank control group,the weight of the mice in the other three groups at different time points were significantly reduced,the differences were statistically significant(P<0.05).Compared with the KD model group,the weight of the mice in the low and high-dose Rb1 groups were significantly increased,the differences were statistically significant(P<0.05).Compared with the blank control group,the myocardial histopathological scores,ET-1 and NO levels of the other three groups were significantly increased,the differences were statistically significant(P<0.05).Compared with the KD model group,the indicators in the low and high dose Rb1 group were significantly reduced,and the high-dose Rb1 group was also significantly lower than the low-dose Rb1 group,the differences were statistically significant(P<0.05).Compared with the blank control group,the levels of serum IL-6 and TNF-αin the other three groups were increased significantly,while the levels of IL-10 were significantly decreased,the differences were statistically significant(P<0.05).Compared with the KD model group,the levels of serum IL-6 and TNF-αin the low and high dose Rb1groups were significantly reduced,while the levels of IL-10 were significantly increased,and compared with the Rb1 low-dose group,the above indicators of the high-dose Rb1 group were also significantly improved,the differences were statistically significant(P<0.05).Compared with the blank control group,the levels of serum FIB and AT-Ⅲin the other three groups were significantly reduced,while the level of D-Dimer was significantly increased,the differences were statistically significant(P<0.05).Compared with the KD model group,the levels of serum FIB and AT-Ⅲin the low,high-dose group were significantly increased,while the level of D-Dimer was significantly reduced,and compared with the Rb1 low-dose group,the above indicators of the high-dose Rb1 group were also significantly improved,the differences were statistically significant(P<0.05).Conclusion Ginsenoside Rb1 treatment of Kawasaki disease can significantly promote the weight recovery of mice,thereby reducing the influence of Kawasaki disease on the growth and development of mice.It can also reduce the damage to the coronary arteries of the mouse heart and further improve the vascular inflammation and blood coagulation function.