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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2022, Vol. 42 Issue (11): 109-116    DOI: 10.13523/j.cb.2209026
生物质资源     
黄芩汤对糖尿病肾病大鼠肾脏NF-κB/NLRP3/Caspase-1细胞焦亡通路的影响*
刘嘉1,严宝飞1,张景正1,马宇霆1,田朝晖2,曾庆琪1,**()
1 江苏卫生健康职业学院 南京 211800
2 恩施土家族苗族自治州中心医院 恩施 445000
Effect of Huangqin Decoction on NF-κB/NLRP3/Caspase-1 Signaling Pathway and Pyroptosis in Kidney of Diabetic Nephropathy Rats
LIU Jia1,YAN Bao-fei1,ZHANG Jing-zheng1,MA Yu-ting1,TIAN Zhao-hui2,ZENG Qing-qi1,**()
1 Jiangsu Health Vocational College, Nanjing 211800, China
2 The Central Hospital of Enshi Tujia and Miao Autonomous Prefecture, Enshi 445000, China
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摘要：

目的:研究黄芩汤对糖尿病肾病(diabetic nephropathy,DN)大鼠肾组织核因子κB(nuclear factor kappa-B,NF-κB)/NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)/胱天蛋白酶-1(cysteinyl aspartate specific proteinase-1,Caspase-1)细胞焦亡通路的影响。方法: 将SD大鼠随机分为空白组、模型组、厄贝沙坦组(27 mg/kg)和黄芩汤低、高剂量组(5 g/kg和20 g/kg),高脂饲料喂养6周联合一次性腹腔注射链脲佐菌素(35 mg/kg)诱导DN大鼠模型,每组9只。灌胃给药6周后检测大鼠血清空腹血糖(fasting blood glucose,FBG)、总胆固醇(total cholesterol,TC)、甘油三酯(triacylglycerol,TG)、尿蛋白(urine protein,UP)、尿素氮(blood urea nitrogen,BUN)、血肌酐(serum creatinine,Scr)、白介素-1β(interleukin 1β,IL-1β)和IL-18水平;HE染色和Masson染色观察大鼠肾脏病理变化;Western blot和免疫组化检测肾脏NF-κB/NLRP3/Caspase-1细胞焦亡通路相关蛋白及阳性细胞表达。结果: 与空白组比较,模型组大鼠FBG、TC、TG、UP、BUN、Scr、IL-1β和IL-18水平明显升高(P<0.01);肾脏出现肾小球体积增大及基底膜增厚,肾小管管腔扩张,炎性浸润及纤维化明显等病理变化;肾脏组织NF-κB的磷酸化水平,以及NLRP3、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing a CARD,ASC)、Caspase-1、IL-1β和消皮素D(gasdermin D,GSDMD)的蛋白质表达明显升高(P<0.01);肾脏组织NLRP3和GSDMD阳性细胞表达水平明显升高(P<0.01)。与模型组比较,黄芩汤组大鼠上述血糖、血脂、肾功能及炎性因子水平均得到明显改善(P<0.05,P<0.01);肾脏肾小球及肾小管结构趋于正常,炎性浸润及纤维化程度得到改善;肾脏组织NF-κB的磷酸化水平,以及NLRP3、ASC、Caspase-1、IL-1β和GSDMD的蛋白质表达水平明显降低(P<0.05,P<0.01);肾脏组织NLRP3和GSDMD阳性细胞表达明显降低(P<0.05,P<0.01)。结论: 黄芩汤对DN大鼠具有确切的疗效,机制可能与抑制NF-κB/NLRP3/Caspase-1细胞焦亡通路有关。
关键词： 黄芩汤;  糖尿病肾病;  细胞焦亡;  核因子κB;  NOD样受体热蛋白结构域相关蛋白3;  胱天蛋白酶-1    
Abstract:

To study the effect of Huangqin Decoction on nuclear factor kappa B (NF-κB)/NOD-like receptor thermal protein domain associated protein 3 (NLRP3)/cysteinyl aspartate specific proteinase-1 (Caspase-1)-dependent pyroptosis pathway in diabetic nephropathy (DN) rat model. Methods: Rats were randomly divided into control group, model group, irbesartan group (27 mg/kg), and Huangqin Decoction low- and high-dose groups (5 g/kg and 20 g/kg), and the DN rat model was induced by high-fat diet for 6 weeks combined with a single intraperitoneal injection of streptozotocin (35 mg/kg). Serum fasting blood glucose (FBG), total cholesterol (TC), triacylglycerol (TG), urine protein (UP), blood urea nitrogen (BUN), serum creatinine (Scr), interleukin 1β (IL-1β) and IL-18 levels were measured after 6 weeks of gavage administration; renal pathological changes were observed by HE and Masson staining; renal NF-κB/NLRP3/Caspase-1/pyroptosis pathway-related proteins and positive cell expression were detected by Western blot and immunohistochemistry. Results: Compared with the control group, the levels of FBG, TC, TG, UP, BUN, Scr, IL-1β and IL-18 were significantly increased in the model group (P<0.01); the kidneys showed pathological changes such as increased glomerular volume and basement membrane thickening, tubular lumen expansion, inflammatory infiltration and fibrosis; the phosphorylation levels of NF-κB in kidney tissues and NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), Caspase-1, IL-1β and gasdermin D (GSDMD) protein expression were significantly increasedc (P<0.01); NLRP3 and GSDMD positive cells expression in renal tissue were significantly increased P<0.01). Compared with the model group, the above-mentioned blood glucose, lipids, renal function and inflammatory factor levels were significantly improved in the Huangqin Decoction groups (P<0.05, P<0.01); renal glomerular and tubular structures were normalized, and the degree of inflammatory infiltration and fibrosis was improved; the phosphorylation levels of NF-κB in renal tissue as well as NLRP3, ASC, Caspase-1, IL-1β and GSDMD protein expressions were significantly reduced (P<0.05, P<0.01); renal tissue NLRP3 and GSDMD positive cells expression were significantly reduced (P<0.05, P<0.01). Conclusion: Huangqin Decoction exerts a definite effect on DN rats, at least partly, through inhibition of the NF-κB/NLRP3/Caspase-1-dependent pyroptosis signaling pathway.
Key words: Huangqin Decoction    Diabetic nephropathy    Pyroptosis    NF-κB    NLRP3    Caspase-1
收稿日期: 2022-09-13 出版日期: 2022-12-07
ZTFLH:  Q819  
基金资助: *江苏省高校“青蓝工程”优秀教学团队培养对象(苏教师[2020]42号);江苏省中医药科技发展计划(YB2020100)
通讯作者: **电子信箱:zengqq111@126.com   
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刘嘉
严宝飞
张景正
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田朝晖
曾庆琪

引用本文:

刘嘉, 严宝飞, 张景正, 马宇霆, 田朝晖, 曾庆琪. 黄芩汤对糖尿病肾病大鼠肾脏NF-κB/NLRP3/Caspase-1细胞焦亡通路的影响*[J]. 中国生物工程杂志, 2022, 42(11): 109-116.

LIU Jia, YAN Bao-fei, ZHANG Jing-zheng, MA Yu-ting, TIAN Zhao-hui, ZENG Qing-qi. Effect of Huangqin Decoction on NF-κB/NLRP3/Caspase-1 Signaling Pathway and Pyroptosis in Kidney of Diabetic Nephropathy Rats. China Biotechnology, 2022, 42(11): 109-116.

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https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.2209026    或    https://manu60.magtech.com.cn/biotech/CN/Y2022/V42/I11/109

图1  黄芩汤对糖尿病肾病大鼠生化指标的影响( x -± s,n=6)
图2  黄芩汤对糖尿病肾病大鼠IL-1β和IL-18水平的影响( x -± s,n=6)
图3  黄芩汤对糖尿病肾病大鼠肾脏病理的影响(HE染色和Masson染色,×20)
图4  黄芩汤对糖尿病肾病大鼠肾脏NF-κB/NLRP3/Caspase-1细胞焦亡通路相关蛋白表达的影响( x -± s,n=6)
图5  黄芩汤对糖尿病肾病大鼠肾脏NLRP3和GSDMD阳性细胞表达的影响( x -± s,n=6)
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