Objective To investigate the protective effects of pretreatment of asiatic acid (AA) on acute renal injury of mice with sepsis.Methods Twenty-four male BALB/c mice were randomly divided into the blank control group, lipopolysaccharide (LPS) group, low-dose AA group, and high-dose AA group with 6 mice in each.Mice in the low-dose AA group and high-dose AA group were administered 10 and 30 mg/kg AA for 3 consecutive days, meanwhile, mice in the blank control and LPS group were intragastrically administered equal volume of 0.5% sodium carboxyl methyl cellulose (CMC-Na).At 2 h after the last administration, mice in the blank control group were intraperitoneally injected with PBS, while mice in the other three groups were injected with 10 mg/kg LPS to make the sepsis models.After 4-hour LPS injection on day 3, the level of serum urea nitrogen (BUN) was measured by diacetylmonoxime colorimetric assay.The pathological change of kidney was investigated by HE staining.The mRNA expression levels of cytokines (TNF-α, IL-1β and IL-6) and Notch signaling pathway-related genes (Notch1-4, Dll4, and Jag2) were measured by real-time PCR.Results Compared with the blank control group, the level of BUN in the LPS, low-dose and high-dose AA groups was higher.Meanwhile, the level of BUN in the low-dose AA and high-dose AA groups was lower than that of the LPS group (all P<0.05).The histological examination showed LPS treatment resulted in neutrophil infiltration, tubular necrosis and endothelial cell swelling.Against these changes, the low-dose and high-dose AA groups showed significantly alleviated neutrophil infiltration, endothelial cell swelling and pathological injury as compared with that of the LPS group.The mRNA levels of TNF-α, IL-1β, and IL-6 in the LPS group were higher than those of the blank control group (all P<0.05), at the same time, the mRNA levels of TNF-α, IL-1β, and IL-6 in the low-dose and high-dose AA groups were lower than those of the LPS group (all P<0.05).The mRNA levels of Notch related genes including Notch1, Notch2, Notch3, Notch4, Dll 4 and Jag2 in the LPS group were lower than those of the blank control group (all P<0.05), meanwhile, the mRNA levels in the low-dose and high-dose AA groups were higher than those of the LPS group (all P<0.05).Conclusion AA pretreatment may significantly alleviate the acute renal injury of mice with sepsis by regulating Notch signaling pathway to develop anti-inflammatory effect.%目的 探讨积雪草酸(AA)预处理对脓毒症小鼠急性肾损伤的作用及机制.方法 选择雄性BALB/c小鼠24只,随机分为空白对照组、脂多糖(LPS)组、AA低剂量组、AA高剂量组各6只.AA低剂量组、AA高剂量组每天分别以AA 10、30 mg/kg灌胃预处理,空白对照组、LPS组每天以等量的0.5%羧甲基纤维素钠灌胃.第3天灌胃2 h后,空白对照组腹腔注射PBS,其余三组腹腔注射LPS 10 mg/kg建立小鼠脓毒症模型.各组于第3天腹腔注射4 h后,采用二乙酰肟比色法检测血清尿素氮(BUN)水平;取肾组织,行HE染色观察肾组织病理变化;采用real-time PCR法检测炎症因子(TNF-α、IL-1β、IL-6)及Notch信号通路相关基因(Notch1~4、Dll4 、Jag2)表达.结果 LPS组、AA低剂量组、AA高剂量组血清BUN水平均高于空白对照组,AA低剂量组、AA高剂量组血清BUN水平均低于LPS组(P均<0.05).HE染色可见正常对照组小鼠肾组织正常;LPS组表现为肾小管坏死,内皮细胞肿胀以及炎性细胞浸润;AA低剂量组、AA高剂量组表现为内皮细胞肿胀减轻、炎症细胞浸润减少,病理损伤较LPS组明显减轻.LPS组TNF-α、IL-1β、IL-6 mRNA表达均高于空白对照组(P均<0.05),AA低剂量组、AA高剂量组TNF-α、IL-1β、IL-6 mRNA表达均低于LPS组(P均<0.05).LPS组Notch信号相关基因Notch1、Notch2、Notch3、Notch4、Dll4及 Jag2 mRNA表达均低于空白对照组(P均<0.05),AA低剂量组、AA高剂量组Notch1、Notch2、Notch3、Notch4、Dll4及 Jag2 mRNA表达均高于LPS组(P均<0.05).结论 AA预处理能够明显减轻脓毒症小鼠的肾损伤,其机制可能与调控Notch信号通路发挥抗炎作用有关.
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