Notch通路在糖尿病肾病大鼠的表达及化瘀通络中药的干预作用

Expression of Notch pathway in diabetic nephropathy rats and intervention effect of Chinese materia medica for dispersing blood stasis and dredging collateral

目的探讨Notch通路在糖尿病肾病(DN)大鼠肾脏的表达以及化瘀通络中药对其的干预作用。方法 60只SD雄性大鼠,随机取10只为对照组,其余大鼠给予高糖高脂饲料喂养联合ip小剂量链脲佐菌素(STZ)制备DN模型。成模大鼠随机分为模型组、厄贝沙坦组、化瘀通络中药组,厄贝沙坦组、化瘀通络中药组均ig给药,于16周末检测大鼠24 h尿蛋白定量,Real-time PCR法检测Notch1、Jagged1和Hey1 m RNA的表达,免疫组化及Western blotting法检测Notch1、Jagged1和Hey1蛋白的表达。结果与对照组比较,模型组大鼠24 h尿蛋白定量及Notch1、Jagged1、Hey1 m RNA和蛋白的表达量明显升高(P<0.01)。与模型组比较,化瘀通络中药组及厄贝沙坦组大鼠24 h尿蛋白定量及Notch1、Jagged1、Hey1m RNA和蛋白的表达量降低(P<0.01)。结论化瘀通络中药可以减少DN大鼠24 h尿蛋白定量,且能够抑制DN大鼠肾脏Notch通路中Notch1、Jagged1和Hey1的高表达,该作用可能是其减少蛋白尿排泄的主要途径之一。

Objective To investigate the expression of Notch pathway in the kidney of diabetic nephropathy（DN） rats and the intervention effect of Chinese materia medica（CMM） for dispersing blood stasis and dredging collateral. Methods Sixty male Sprague-Dawley rats, in which ten rats were randomly selected as control group（n = 10）, and the other rats were fed with high glucose and high fat diet combined with low-dose streptozotocin（STZ） ip injection as DN models. The model rats were randomly divided into model group, irbesartan group, and Huayu Tongluo Granles（HTG） group. The rats in each group were ig administered with corresponding drugs. At the end of the 16 weeks, the 24 h urinary protein was detected. The expression of Notch1, Jagged1, and Hey1 m RNA and protein in renal tissue was detected by real-time PCR, and immunohistochemical staining and western blotting assay, respectively. Results Compared with the control group, 24 h urinary protein, the m RNA and protein expression of Notch1, Jagged1, and Hey1 in model group was significantly increased（P 〈 0.01）. Compared with the model group, 24 h urinary protein and Notch1, Jagged1, Hey1 m RNA, and protein expression of HTG group and irbesartan group was significantly decreased（P 〈 0.01）. ConclusionCMM for dispersing blood stasis and dredging collateral can reduce 24 h urinary protein in DN rats and inhibit the high expression of Notch1, Jagged1, and Hey1 in the kidney tissue of DN rats, which may be one of the main ways to reduce proteinuria excretion.