Effect of Danggui Buxuetang on Podocyte Pyroptosis in Diabetic Kidney Disease Rats： An Exploration Based on TXNIP/NLRP3/GSDMD Signaling Pathway

He-chao JIN; Sheng-ran LIANG; Guan-wen ZHANG; Jia-wei QIANG; Deng-zhou GUO

doi:10.13422/j.cnki.syfjx.20212441

您当前的位置：

首页 >

文章列表页 >

Effect of Danggui Buxuetang on Podocyte Pyroptosis in Diabetic Kidney Disease Rats： An Exploration Based on TXNIP/NLRP3/GSDMD Signaling Pathway

更新时间：2021-12-31

- Effect of Danggui Buxuetang on Podocyte Pyroptosis in Diabetic Kidney Disease Rats： An Exploration Based on TXNIP/NLRP3/GSDMD Signaling Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 28, Issue 3, Pages: 49-57(2022)
- 作者机构：
  
  1.河北中医学院，石家庄 050200
  2.河北省中医院，石家庄 050011
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20212441
  CLC： R2-0;R289;R33;R587.1
- Received：22 July 2021，
  
  Published Online：08 November 2021，
  
  Published：05 February 2022
- 稿件说明：
移动端阅览
靳贺超,梁胜然,张冠文等.基于TXNIP/NLRP3/GSDMD信号通路探讨当归补血汤对糖尿病肾病大鼠足细胞焦亡的影响[J].中国实验方剂学杂志,2022,28(03):49-57.

JIN He-chao,LIANG Sheng-ran,ZHANG Guan-wen,et al.Effect of Danggui Buxuetang on Podocyte Pyroptosis in Diabetic Kidney Disease Rats： An Exploration Based on TXNIP/NLRP3/GSDMD Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(03):49-57.
靳贺超,梁胜然,张冠文等.基于TXNIP/NLRP3/GSDMD信号通路探讨当归补血汤对糖尿病肾病大鼠足细胞焦亡的影响[J].中国实验方剂学杂志,2022,28(03):49-57. DOI： 10.13422/j.cnki.syfjx.20212441.

JIN He-chao,LIANG Sheng-ran,ZHANG Guan-wen,et al.Effect of Danggui Buxuetang on Podocyte Pyroptosis in Diabetic Kidney Disease Rats： An Exploration Based on TXNIP/NLRP3/GSDMD Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(03):49-57. DOI： 10.13422/j.cnki.syfjx.20212441.

摘要

目的

观察当归补血汤对糖尿病肾病（DKD）大鼠足细胞焦亡的影响，探讨其防治DKD及足细胞焦亡的可能作用机制。

方法

将50只雄性SD大鼠随机分为造模组42只和正常组8只，造模组大鼠高糖高脂饮食6周结合链脲佐菌素（STZ）35 mg·kg

-1

一次性腹腔注射制备2型糖尿病模型。造模成功后随机分为模型组、当归补血汤低剂量（0.72 g·kg

-1

）组、当归补血汤高剂量（1.44 g·kg

-1

）组、厄贝沙坦（0.017 g·kg

-1

）组进行灌胃，正常组及模型组给予等体积生理盐水灌胃，每日1次，连续干预20周。给药期间定期检测各组大鼠空腹血糖（FBG），24 h尿蛋白（24 h-UTP）；给药结束后采用过碘酸六胺银（PASM）染色观察肾组织病理形态学变化；透射电镜（TEM）观察足细胞超微结构变化；酶联免疫吸附测定法（ELISA）检测大鼠血清白细胞介素-1

（IL-1

），白细胞介素-18（IL-18）水平；原位末端标记法（TUNEL）检测大鼠肾组织细胞DNA损伤情况；免疫组化法（IHC）检测大鼠肾组织硫氧还蛋白相互作用蛋白（TXNIP），半胱氨酸天冬氨酸蛋白水解酶-1（Caspase-1），消皮素D（GSDMD）蛋白表达水平；免疫荧光（IF）三标法检测核苷酸结合域样受体蛋白3（NLRP3）及肿瘤蛋白-1（WT-1）在足细胞的表达水平；蛋白免疫印迹法（Western blot）检测大鼠肾组织TXNIP/NLRP3/Caspase-1/GSDMD通路蛋白及足细胞突触足蛋白（Synaptopodin）表达水平。

结果

与正常组比较，模型组大鼠FBG，24 h-UTP显著升高，肾小球肥大，系膜增生，系膜外基质增加，基底膜增厚，出现K-W结节，肾小管上皮细胞空泡变性，足突融合或丢失，血清中IL-1

，IL-18含量明显升高，肾组织TUNEL阳性细胞明显增多，NLRP3在足细胞表达增多且WT-1在足细胞表达减少，Synaptopodin蛋白表达水平显著降低，TXNIP/NLRP3/Caspase-1/GSDMD蛋白表达水平显著升高（

0.01）；与模型组比较，当归补血汤高剂量组明显降低FBG，24 h-UTP水平，明显改善肾组织病理学，改善足细胞的损伤和丢失，减少肾组织TUNEL阳性细胞数，NLRP3在足细胞表达减少且WT-1在足细胞表达增多，升高Synaptopodin蛋白表达水平，降低TXNIP/NLRP3/Caspase-1/GSDMD 蛋白表达水平（

0.05，

0.01）。

结论

当归补血汤可能通过调控TXNIP/NLRP3/GSDMD信号通路，抑制足细胞焦亡以减少蛋白尿，从而延缓DKD的发展进程。

Abstract

Objective

To observe the effect of Danggui Buxuetang on podocyte pyroptosis in diabetic kidney disease （DKD） rats and to explore the possible mechanism of its prevention and treatment of DKD and podocyte pyroptosis.

Method

Eight of the 50 male SD rats were randomly classified into a normal group， and the remaining 42 were fed a high-glucose and high-fat diet for six weeks and then intraperitoneally injected with 35 mg·kg

-1

streptozotocin （STZ） for inducing type 2 diabetes. After successful modeling， they were randomized into the model group， low- （0.72 g·kg

-1

） and high-dose （1.44 g·kg

-1

） Danggui Buxuetang group， and irbesartan （0.017 g·kg

-1

） group and gavaged with the corresponding drugs， while those in the normal group and model group with an equal volume of normal saline， once per day， for 20 weeks. During the medication， the fasting blood glucose （FBG） and 24 h urine protein （24 h-UTP） were measured regularly. After administration， the pathological changes in renal tissues were observed by periodic acid-silver metheramine （PASM） staining， followed by the observation of ultrastructural changes in podocytes under the transmission electron microscope （TEM）. Serum levels of interleukin-1

（IL-1

） and interleukin-18 （IL-18） were determined by enzyme-linked immunosorbent assay （ELISA）. The DNA damage in renal tissue cells of rats was detected by

in situ

nick end-labeling （TUNEL） assay. The protein expression levels of thioredoxin interacting protein （TXNIP）， cysteine-dependent aspartate-directed protease-1 （Caspase-1）， and gasdermin D （GSDMD） in renal tissues of rats were detected by immunohistochemistry （IHC）， the expression levels of nucleotide binding domain like receptor protein 3 （NLRP3） and Wilms tumor protein-1 （WT-1） in podocytes by immunofluorescent （IF） staining， and the expression levels of TXNIP/NLRP3/Caspase-1/GSDMD pathway proteins and Synaptopodin in renal podocytes by Western blot.

Result

Compared with the normal group， the model group exhibited increased FBG and 24 h UTP， glomerular hypertrophy， mesangial hyperplasia， increased extracellular matrix， thickened basement membrane， K-W nodules， vacuolar degeneration in renal tubular epithelial cells， foot process fusion or loss， elevated serum IL-1

and IL-18 levels and TUNEL-positive cells in renal tissue， enhanced NLRP3 but diminished WT-1 expression in podocytes， down-regulated Synaptopodin protein expression， and up-regulated TXNIP/NLRP3/Caspase-1/GSDMD protein expression （

0.01）. Compared with the model group， Danggui Buxuetang high-dose group remarkably lowered FBG， 24-h UTP， and TUNEL-positive cells in renal tissue， improved renal histopathology and podocyte injury and loss， down-regulated NLRP3 expression in podocytes and TXNIP/NLRP3/Caspase-1/GSDMD protein expression levels， and up-regulated WT-1 expression in podocytes and Synaptopodin protein expression （

0.05，

0.01）.

Conclusion

Danggui Buxuetang inhibits podocyte pyroptosis to reduce proteinuria and delays the development of DKD possibly by regulating the TXNIP/NLRP3/GSDMD signaling pathway.

关键词

Keywords

references

FORBES J M ， THORBURN D R . Mitochondrial dysfunction in diabetic kidney disease ［J］. Nature Rev Nephrol ， 2018 ， 14 （ 5 ）： 291 - 312 .

SUN L ， LI W ， LI W ， et al . Astragaloside Ⅳ prevents damage to human mesangial cells through the inhibition of the NADPH oxidase/ROS/Akt/NFkappaB pathway under high glucose conditions ［J］. Int J Mol Med ， 2014 ， 34 （ 1 ）： 167 - 176 .

AN X ， ZHANG Y ， CAO Y ， et al . Punicalagin protects diabetic nephropathy by inhibiting pyroptosis based on TXNIP/NLRP3 pathway ［J］. Nutrients ， 2020 ， 12 （ 5 ）： 1516 .

MANGAN M S J ， OLHAVA E J ， ROUSH W R ， et al . Targeting the NLRP3 inflammasome in inflammatory diseases ［J］. Nat Rev Drug Discov ， 2018 ， 17 （ 9 ）： 688 .

LI J J ， KWAK S J ， JUNG D S ， et al . Podocyte biology in diabetic nephropathy ［J］. Kidney Int Suppl ， 2007 ， 106 ： S36 - S42 .

杜世拔，李国政，卢微嫒，等 . 健脾固肾方联合常规治疗对脾肾亏虚型早期糖尿病肾病患者的临床疗效［J］. 中成药， 2021 ， 43 （ 3 ）： 649 - 653 .

张亚丽，李廷振，汪伟环，等 . 杞菊地黄丸加味联合艾塞那肽对早期糖尿病肾病肝肾阴虚夹瘀证的临床疗效［J］. 辽宁中医杂志， 2020 ， 47 （ 7 ）： 95 - 98 .

KE H L ， ZHANG Y W ， ZHOU B F ， et al . Effects of Danggui Buxue Tang，a traditional Chinese herbal decoction，on high glucose-induced proliferation and expression of extracellular matrix proteins in glomerular mesangial cells ［J］. Nat Prod Res ， 2012 ， 26 （ 11 ）： 1022 - 1026 .

YE T S ， ZHANG Y W ， ZHANG X M . Protective effects of Danggui Buxue Tang on renal function，renal glomerular mesangium and heparanase expression in rats with streptozotocin-induced diabetes mellitus ［J］. Exp Ther Med ， 2016 ， 11 （ 6 ）： 2477 - 2483 .

WANG L ， MA J ， GUO C ， et al . Danggui Buxue Tang attenuates tubulointerstitial fibrosis via suppressing NLRP3 inflammasome in a rat model of unilateral ureteral obstruction ［J］. Biomed Res Int ， 2016 ， doi： 10.1155/2016/9368483 http://dx.doi.org/10.1155/2016/9368483 .

KE R ， WANG Y ， HONG S ， et al . Endoplasmic reticulum stress related factor IRE1alpha regulates TXNIP/NLRP3-mediated pyroptosis in diabetic nephropathy ［J］. Exp Cell Res ， 2020 ， 396 （ 2 ）： 112293 .

靳贺超，于文涛，刘晓，等 . 补肾活血方对血管性痴呆大鼠脑海马细胞凋亡及ERK2，CREB表达的影响［J］. 中国实验方剂学杂志， 2018 ， 24 （ 12 ）： 129 - 135 .

TANG S C W ， YIU W H . Innate immunity in diabetic kidney disease ［J］. Nat Revs Nephrol ， 2020 ， 16 （ 4 ）： 206 - 222 .

DING X ， JING N ， SHEN A ， et al . MiR-21-5p in macrophage-derived extracellular vesicles affects podocyte pyroptosis in diabetic nephropathy by regulating A20 ［J］. J Endocrinol Invest ， 2021 ， 44 （ 6 ）： 1175 - 1184 .

杨凤文，高飞，陈素枝，等 . 益气养阴活血通络方抑制JAK2/STAT3信号通路减轻糖尿病肾病大鼠肾组织细胞凋亡［J］. 中国实验方剂学杂志， 2021 ， 27 （ 11 ）： 89 - 97 .

王美子，岳丽军，万毅刚，等 . 葛根芩连汤加味方改善糖尿病肾病模型鼠足细胞焦亡和胰岛素抵抗的作用和机制［J］. 中国中药杂志， 2021 ， 46 （ 17 ）： 4471 - 4479 .

胡艳红，杨静，修成奎，等 . 人参-三七-川芎提取物延缓高糖诱导的小鼠血管钙化的机制［J］. 中国实验方剂学杂志， 2020 ， 26 （ 8 ）： 13 - 20 .

孙玉霞，李屹，李林，等 . 益气活血汤对糖尿病肾病患者血清CRP，IL-8的影响［J］. 中国实验方剂学杂志， 2017 ， 23 （ 13 ）： 164 - 168 .

LEI X ， ZHANG B D ， REN J G ， et al . Astragaloside suppresses apoptosis of the podocytes in rats with diabetic nephropathy via miR-378/TRAF5 signaling pathway ［J］. Life Sci ， 2018 ， 206 ： 77 - 83 .

WEN W C ， BI F L . Bioactivities of major constituents isolated fromAngelica sinensis ［J］. Chin Med ， 2011 ， 6 ： 29 .

CHEN Y ， LIU Q ， SHAN Z ， et al . Catalpol ameliorates podocyte injury by stabilizing cytoskeleton and enhancing autophagy in diabetic nephropathy ［J］. Front Pharmacol ， 2019 ， doi： 10.3389/fphar.2019.01477 http://dx.doi.org/10.3389/fphar.2019.01477 .

ZHAI R ， JIAN G ， CHEN T ， et al . Astragalus membranaceus and Panax notoginseng ，the novel renoprotective compound，synergistically protect against podocyte injury in streptozotocin-induced diabetic rats ［J］. J Diabetes Res ， 2019 ， doi： 10.1155/2019/1602892 http://dx.doi.org/10.1155/2019/1602892 .

ZHANG C ， ZHU X ， LI L ， et al . A small molecule inhibitor MCC950 ameliorates kidney injury in diabetic nephropathy by inhibiting NLRP3 inflammasome activation ［J］. Diabetes Metab Syndr Obes ， 2019 ， 12 ： 1297 - 1309 .

DU N ， XU Z ， GAO M ， et al . Combination of Ginsenoside Rg 1 and Astragaloside Ⅳ reduces oxidative stress and inhibits TGF-beta1/Smads signaling cascade on renal fibrosis in rats with diabetic nephropathy ［J］. Drug Des Devel Ther ， 2018 ， 12 ： 3517 - 3524 .

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Danggui Buxuetang-containing Serum Alleviates Endoplasmic Reticulum Stress in Diabetic Kidney Disease to Inhibit Podocyte Apoptosis via PERK/ATF4/CHOP Pathway

Danggui Buxuetang Inhibits Mitochondrial Fission and Apoptosis of Podocytes in Rat of Diabetic Kidney Disease via AKAP1/Drp1 Signaling Pathway

Effect of Danggui Buxuetang on Podocyte Injury in Diabetic Kidney Disease Rats： An Exploration Based on RIPK1/RIPK3/MLKL Signaling Pathway

Danggui Buxuetang Alleviates Oxidative Stress and Inflammation in Diabetic Kidney Disease Rats by Improving Mitochondrial Dysfunction of Podocytes

Related Author

GU Yue

SHEN Yuhang

DING Xin

WANG Yifan

ZHANG Yuanyuan

ZHENG Linlin

GUO Dengzhou

ZHENG Linlin

Related Institution

The First Affiliated Hospital of Hebei University of Chinese Medicine，Hebei Provincial Hospital of Traditional Chinese Medicine

Lucheng Community Health Center of Tongzhou District

Hebei University of Chinese Medicine

Key Laboratory of High Incidence Disease Rasearch in Xinjiang（Xinjiang Medical University）， Ministry of Education

Peking University Third Hospital

AI问答

Postal code：100700
Tel：010-84076882 Email：syfjx_2010@188.com
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备11006657号-10 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰