化瘀解毒方对梗阻性肾病大鼠肾脏NLRP3、Caspase1、IL-1β表达的影响

Effect of Huayu Jiedu Recipe on the Expressions of NLRP3,Caspase1,and IL-1β in Kidneys of Obstructive Nephropathy Rats

目的观察化瘀解毒方对梗阻性肾病大鼠肾脏核苷酸结合寡聚结构域样受体蛋白3[NOD(nucleotide-binding oligomerization domain)-like receptor protein 3,NLRP3]、天冬氨酸特异性半胱氨酸蛋白酶1(Caspase-1)、IL-1β表达的影响。方法将40只清洁级SD大鼠随机分为假手术组(10只)、造模组(30只),采用单侧输尿管结扎(unilateral ureteral obstruction,UUO)手术复制梗阻性肾病模型,成功后随机分为模型组、西药组和中药组,每组各10只。西药组给予依普利酮100 mg/(kg·d),中药组给予化瘀解毒方13.7 g/(kg·d),假手术组和模型组给予等量生理盐水,连续给药10天。检测各组血清IL-1β含量,检测肾脏NLRP3、Caspase-1、IL-1β蛋白及mRNA表达,检测肾脏TUNEL阳性细胞率。结果假手术组NLRP3表达不明显,Caspase-1及IL-1β呈弱表达,仅见少量TUNEL阳性细胞。与假手术组比较,模型组血清IL-1β含量升高(P<0.01),肾组织NLRP3、Caspase-1及IL-1β蛋白及mRNA表达升高(P<0.01);NLRP3主要表达于肾间质巨噬细胞及肾小管上皮细胞,Caspase-1及IL-1β主要见于肾小管上皮细胞胞浆;TUNEL阳性细胞增多,以间质扩张的远端小管上皮细胞为主(P<0.01)。与模型组比较,两个给药组血清IL-1β含量降低(P<0.01),NLRP3、Caspase-1及IL-1β蛋白及mRNA表达降低,TUNEL阳性细胞率降低(P<0.05,P<0.01)。结论化瘀解毒方可下调NLRP3、Caspase-1、IL-1β蛋白及mRNA表达,减轻肾组织炎症损伤。

Objective To observe the effect of Huayu Jiedu Recipe （H JR） on the expressions of nucleotide-binding oligomerization domain （NOD）-Iike receptor protein 3 （NLRP3）, Caspase-1, IL-1β in kidneys of obstructive nephropathy rats. Methods Totally 40 clean grade SD rats were randomly divided into the sham-operation group （n =10） and the model group （n =30）. The model of obstructive nephropa- thy was established by unilateral ureteral obstruction （UUO）. Totally 30 successfully modeled UUO rats were randomly divided into the model group, the Western medicine group, the Chinese medicine group, 10 in each group. Eplerenone （100 mg ·kg-1· d-1） was administrated to rats in the Western medicine group. H JR （13.7 g ·kg-1 · d-1） was administrated to rats in the Chinese medicine group. Equal volume of normal saline was administered to rats in the sham-operation group and the model group. All medica- tion was performed once daily for 10 successive days. The serum IL-1β level was detected. Protein andmRNA expressions of NLRP3, Caspase-1, and IL-1β in renal tissue were detected. TUNEL positive rate was detected by TUNEL method. Results The expression of NLRP3 was not obviously seen, Caspase-1 and IL-1β were weakly expressed, and only fewer amount of TUNEL positive cells could be seen in the sham-operation group. Compared with the sham-operation group, serum IL-1β level increased （P 〈 0.01 ）, mRNA and protein expression of NLRP3, Caspase-1, and IL-1β were up-regulated in renal tissue of the model group （P 〈0.01）. NLRP3 was mainly expressed in renal interstitial macrophages and renal tubular epithelial cells. Caspase-1 and IL-1β were mainly expressed in the cytoplasm of renal tubular epi- thelial cells. TUNEL positive cells were significantly increased, mainly dominated in interstitial expanded epithelial cells of distal tubules （P 〈0.01 ）. Compared with the model group, serum IL-1β level was signifi- cantly decreased （P 〈0.01 ） ; mRNA and protein expressions of NLRP3, Caspase-1, and IL-1β were ob- viously down-regulated （P 〈0.01 ）, and the TUNEL positive rate was obviously decreased （P 〈0.05, P 〈 0.01 ） in the two medicated groups. Conclusion H JR could down-regulate mRNA and protein expres- sions of NLRP3, Caspase-1, and IL-1β, thus attenuating inflammatory injury of renal tissue.