人参皂甙Rg3对脓毒症小鼠炎症反应及淋巴细胞内Cleaved Caspase-3蛋白的影响

Effects of ginsenoside Rg3 on inflammatory response and lymphocyte Cleaved Caspase-3 protein expression in septic mice

目的探讨人参皂甙Rg3对脓毒症小鼠炎症反应的保护作用和对淋巴细胞内凋亡相关蛋白裂解的天冬氨酸特异性半胱氨酸蛋白酶3(Cleaved Caspase-3)蛋白表达的影响。方法采用区组随机法将40只小鼠分为对照组、人参皂甙Rg3组、脂多糖(LPS)组、人参皂甙Rg3+LPS组,每组10只。其中LPS组、人参皂甙Rg3+LPS组小鼠通过腹腔注射LPS的方法制备脓毒症模型。对照组用0.3 ml 0.9%氯化钠溶液灌胃,1次/d,连续7 d,第4天腹腔注射0.3 ml 0.9%氯化钠注射液;人参皂甙Rg3组将人参皂甙Rg3按30 mg/kg溶于0.3 ml 0.9%氯化钠溶液后,灌胃,1次/d,连续7 d,第4天腹腔注射0.3 ml 0.9%氯化钠注射液;LPS组用0.3 ml 0.9%氯化钠溶液灌胃,1次/d,连续7 d,第4天腹腔注射0.3 ml LPS溶液(15 mg/kg LPS溶于0.3 ml 0.9%氯化钠注射液);人参皂甙Rg3+LPS组将人参皂甙Rg3按30 mg/kg溶于0.3 ml 0.9%氯化钠溶液后,灌胃,1次/d,连续7 d,第4天腹腔注射0.3 ml LPS溶液(15 mg/kg LPS溶于0.3 ml 0.9%氯化钠注射液)。第7天通过腹主动脉采血,采用ELISA法检测各组小鼠血清炎性因子TNF-α、IL-6、IL-1β水平;并处死小鼠,取小鼠脾脏组织,称重、记录脾脏指数;分离脾脏淋巴细胞采用Western blot法检测淋巴细胞Cleaved Caspase-3蛋白表达。结果与对照组比较,LPS组、人参皂甙Rg3+LPS组小鼠TNF-α、IL-6、IL-1β水平、脾脏指数、Cleaved Caspase-3蛋白表达水平均升高(均P<0.05);与LPS组比较,人参皂甙Rg3组、人参皂甙Rg3+LPS组小鼠TNF-α、IL-6、IL-1β水平、脾脏指数、Cleaved Caspase-3蛋白表达水平均降低(均P<0.05)。结论人参皂甙Rg3具有降低脓毒症小鼠体内炎症反应的作用,并能降低淋巴细胞内Cleaved Caspase-3的表达。

Objective To investigate the effect of ginsenoside Rg3 on inflammatory response and lymphocyte Cleaved Caspase 3 protein expression in septic mice.Methods Forty male BALB/c mice were randomly divided into control group,ginsenoside Rg3 group,lipopolysaccharide(LPS)group and ginsenoside Rg3+LPS group with 10 mice in each group.Mice in control group were given 0.3 ml normal saline by gavage q.d for 7 d,and 0.3 ml normal saline by intraperitoneal injection on d4;mice in ginsenoside Rg3 group were given ginsenoside Rg330 mg/kg by gavage q.d for 7 d,and 0.3 ml normal saline by intraperitoneal injection on d4;mice in LPS group were given normal saline by gavage q.d for 7 d,and 0.3 ml LPS solution by intraperitoneal injection on d4;mice in Rg3+LPS group were given ginsenoside Rg330 mg/kg by gavage q.d for 7 d,then 0.3 ml LPS solution by intraperitoneal injection on d4.On d7,blood samples were collected by abdominal aorta,serum TNFα,IL-6 and IL-1βlevels were detected by ELISA.The mice were sacrificed and the spleen was removed and weighed,the expression of Cleaved Caspase-3 protein in spleen lymphocytes were detected by Western blot.Results Compared with the control group,the levels of TNFα,IL-6,IL-1β,Spleen index and Cleaved Caspase-3 protein expression were significantly increased in LPS group and ginsenoside Rg3+LPS group(all P<0.05).Compared with the LPS group,the levels of TNF-α,IL-6,IL-1β,spleen index and Cleaved Caspase-3 protein expression were significantly decreased in ginsenoside Rg3 group and ginsenoside Rg3+LPS group(all P<0.05).Conclusion Ginsenoside Rg3 can inhibit inflammatory response and down-regulate the Cleaved Caspase-3 expression of lymphocyte in mice with sepsis.