冬凌草甲素抑制NF-KB p65活化对糖尿病肾病幼龄大鼠肾损伤和免疫紊乱的影响

Effect of oridonin on renal injury and immune disorder in diabetic nephropathy young rats by inhibiting activation of NF-KB p65

目的探讨冬凌草甲素(Ori)对糖尿病肾病(DN)幼龄大鼠的保护作用。方法将50只SPF级大鼠随机分为对照组、模型组、DN+Ori (5mg/kg)组、DN+Ori (10mg/kg)组及DN+Ori (20mg/kg)组5组,每组10只,采用高脂喂食结合化学诱导法建立糖尿病大鼠模型。留取24h尿液检测尿蛋白含量,取血检测血清肌酸酐(SCr)和尿素氮(UN);苏木精-伊红(HE)和末端标记法(TUNEL)染色观察肾脏病理情况;Western blot检测半胱氨酸天冬氨酸蛋白酶3(Caspase-3)、Caspase-9、B细胞淋巴瘤因子2(Bcl-2)和Bcl-2相关X蛋白(Bax)及核因子κB p65(NF-κB p65)、巨噬细胞炎性蛋白2(MIP-2)的表达;ELISA检测血清中炎症相关因子白介素4(IL-4)、IL-6、IL-10和肿瘤坏死因子(TNF-α)的表达水平。结果与对照组相比,冬凌草甲素能降低模型大鼠尿蛋白、血清肌酐和尿素氮的含量(P<0.05);改善肾组织病理损伤、降低细胞凋亡率(P<0.05);下调Caspase-3、Caspase-9和Bax表达,上调Bcl-2表达(P<0.05);抑制促炎因子IL-6和TNF-α表达,增强抗炎因子IL-4和IL-10表达(P<0.05);抑制炎症通路蛋白NF-κB p65和MIP-2的蛋白表达(P<0.05)。结论冬凌草甲素通过抑制NF-κB p65的活化而缓解糖尿病肾病大鼠肾损伤和免疫紊乱。

Objective To study the protective effect of oridonin(Ori) on diabetic nephropathy(DN) rats. Methods Rats were randomly divided into Control group, DN model group, DN+Ori(5 mg/kg), DN+Ori(10 mg/kg) and DN+Ori(20 mg/kg) group. The diabetic rat model was established by high-fat diet combined with chemical induction. 24 hours urine was collected to detect urinary protein content, blood was taken to detect serum creatinine(SCr) and urea nitrogen(UN). Hematoxylin-eosin(HE) and TUNEL staining were used to observe renal pathology. Western blot was used to detect the expression of apoptosis-related proteins Caspase-3, Caspase-9, B cell lymphoma factor-2(Bcl-2), Bcl-2-related X protein(Bax) and inflammatory pathway NF-kappa B p65, MIP-2. ELISA was used to detect the expression of IL-1 beta, IL-6 and TNF-alpha in serum. Results Comparing with the control group, Ori could reduce the content of urinary protein, serum creatinine and urea nitrogen in DN model mouse(P<0.05), improve the pathological damage of kidney tissue and reduce the apoptotic rate(P<0.05), down-regulate the expression of Caspase-3, Caspase-9 and Bcl-2, up-regulate the expression of Bax(P<0.05), inhibit the expression of inflammatory factors IL-6 and TNF-alpha, increase IL-4 and IL-10(P<0.05). At the same time, Ori could inhibit the protein expression of NF-kappa B 65 and MIP-2(P<0.05). Conclusion Ori can alleviate renal injury and immune disorder in diabetic nephropathy rats by inhibiting the activation of NF-kappa B p65.