基于《伤寒论》六经病“欲解时”探讨昼夜节律对脓毒症大鼠急性肾损伤的影响

Study on effects of circadian rhythm on sepsis induced acute kidney injury based on six meridians disease tending to be cured from Shanghan Lun

目的:基于《伤寒论》六经病“欲解时”探讨昼夜节律对脓毒症大鼠急性肾损伤的影响。方法:36只SD大鼠随机分为3组,对照组、申时(ZT10)组和寅时(ZT22)组,每组12只。通过腹腔内注射脂多糖(LPS,10 mg/kg)构建脓毒症大鼠模型,6 h后检测血清肌酐、尿素氮、肿瘤坏死因子(TNF)-α和白细胞介素(IL)-6水平,肾HE染色并计算肾损伤评分,肾脏称重计算湿/干比和肾脏指数,检测时间节律基因Bmal-1的mRNA和蛋白表达差异。结果:与ZT22组比较,ZT10组平均动脉压显著降低(P<0.05),TNF-α和IL-6水平显著升高(P<0.01),肾损伤积分显著升高(P<0.05)。ZT10组大鼠肾组织ZT16的Bmal-1 mRNA和蛋白水平显著低于对照组(P<0.01)。结论:在少阴病欲解时(寅时)构建的脓毒症AKI大鼠肾损伤程度相对较轻,而欲作/剧时(申时)则相对较重,可能与Bmal-1水平波动相关。

Objective:To evaluate the effects of circadian rhythm on sepsis induced acute kidney injury based on six meridians disease tending to be cured from Shanghan Lun.Methods:Thirty-six SD rats were randomly divided into three groups:control group,Shen-point(the period of the day from 3 p.m.to 5 p.m.) group(ZT10 group) and Yin-point(the period of the day from 3 a.m.to 5 a.m.) group(ZT22 group),12 rats in each group.Intraperitoneal injection of 10 mg/kg LPS was used to build sepsis rat model and we measured the levels of serum creatinine,urea nitrogen,TNF-α and IL-6,performed HE staining and rated renal injury scores,calculated wet/dry rate and kidney index,and evaluated mRNA and protein expression levels of time rhythm gene Bmal-1 in each group after 6 h.Results:Compared with ZT22 group,ZT10 group showed lower mean arterial pressure(P<0.05),significantly higher levels of TNF-α and IL-6(P<0.01) and higher renal injury score(P<0.05).The levels of Bmal-1 mRNA and protein in ZT10 group were lower than control group after 6 h of LPS injection(P<0.01).Conclusion:In comparison of Yinpoint,it seems to aggravate kidney injury at Shen-point of LPS injection in sepsis induced AKI rat model,which may contribute to the fluctuation of Bmal-1.