丹蛭降糖胶囊治疗肥胖引起的慢性肾病的机制研究

The preliminary mechanism of Danzhi Jiangtang Capsule for improving high fat diet-induced kidney damages

目的探究丹蛭降糖胶囊(DJC)改善肥胖引起的大鼠慢性肾病的初步机制。方法♂SD大鼠随机分为基础饲料组(CON)、高脂饲料组(HFD)、高脂饲料+丹蛭降糖胶囊低剂量组(HFD+DJCL,500 mg·kg^(-1)·d^(-1))、高脂饲料+丹蛭降糖胶囊高剂量组(HFD+DJCH,1 000 mg·kg^(-1)·d^(-1)),除CON组外,均给予高脂饮食12周,治疗组再灌胃治疗8周。检测血清血脂和肾代谢指标,HE、油红O、PAS染色观察肾脏的病理学变化,10%肾匀浆测定肾组织TG、TSOD、Cu Zn-SOD、CAT、GSH-Px、TNOS、MDA含量,免疫组化法检测CD36的表达变化,RT-PCR分析SREBP1c及FASN的表达,Western blot分析DJC对AMPK、磷酸化AMPK、PPARα、PPARγ表达的影响。结果对比CON组,HFD组大鼠体质量增加,血清TC、TG、UA、BUN、Scr升高,HDL-C下降,肾小球出现明显的损伤及脂质沉积,GSH-Px、MDA升高,TSOD、CAT、Cu Zn-SOD、TNOS水平降低,CD36、SREBP1c、FASN、PPARγ的表达上升,PPARα、AMPK、磷酸化AMPK表达下降;对比HFD组,HFD+DJCL和HFD+DJCH组均明显逆转上述指标,且有一定的剂量效应。结论 DJC可有效降低高脂饮食引起的肾内脂质沉积和氧化应激水平,其机制可能是通过激活AMPK-PPARα通路,抑制CD36和脂肪生成基因PPARγ、SREBP1c、FASN的表达,并有一定的剂量效应。

Aim To study the preliminary mechanism of Danzhi Jiangtang Capsule（ DJC） on the high-fat diet-induced injury in kidney of rats. Methods Male SD rats were randomly divided into control diet group（ CON）,high-fat diet group（ HFD）,HFD and DJC low dose treatment group（ HFD ＋ DJCL）,HFD and DJC high dose treatment group（ HFD ＋ DJCH）. The latter three groups were fed HFD for 12 weeks,then the latter two groups were given DJC 500 mg·kg-1·d-1 and 1 000 mg·kg-1·d-1 via gavage for 8 weeks respectively. The levels of serum lipid and kidney metabolism indices were detected. HE,Oil red O and PAS staining were used to observe the pathological changes of kidney. 10% kidney homogenate was used to detect triglyceride（ TG）,total superoxide dismutase（ TSOD）,Cu Zn superoxide dismutase（ Cu Zn-SOD）,catalase（ CAT）,glutathione peroxidase（ GSH-Px）,total nitric oxide synthase（ TNOS）,malonaldehyde（ MDA）. Expression and distribution of FAT/CD36 were detected using immunohistochemistry. Expression of SREBP1 c and FASN were detected using RT-PCR.Expression of AMPK, phosphorylation of AMPK,PPARα,PPARγ were analyzed using Western blot.Results Compared with CON group rats,rats in HFD group showed greater weight,higher serum levels of TC,TG,UA,Scr,BUN,and lower serum levels of HDL-C. Obvious injury and lipid deposition were observed in the kidney,GSH-Px and MDA content increased,meanwhile TSOD,Cu Zn-SOD,CAT,TNOS levels decreased. Expression of FAT/CD36,SREBP1 c,FASN and PPARγ increased,while that of PPARα,AMPK and phosphorylation of AMPK decreased. Compared with HFD group,the HFD ＋ DJC groups showed significantly improved above indices in a somewhat dose-dependent manner. Conclusions DJC effectively reduces HFD-induced lipid deposition and oxidative stress in kidney,and the mechanisms are possibly related with activated AMPK-PPARα signaling and decreased expression of CD36 and lipogenic genes PPARγ,SREBP1 c,FASN,exhibiting a dose-dependent effect.