Effect of Jinkui Shenqiwan on Rats with Renal Interstitial Fibrosis Induced by Adenine

YUAN Jin-feng; CHEN Lan-ying; WANG Hui-ling; ZHANG Ni; LI Xue-liang; GUAN Zi-yi; FANG Cong; ZHOU Meng-jing; WANG Ling-ling; YIN Li; SHOU Bin-yao

doi:10.13422/j.cnki.syfjx.20182199

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Effect of Jinkui Shenqiwan on Rats with Renal Interstitial Fibrosis Induced by Adenine

更新时间：2024-09-23

- Effect of Jinkui Shenqiwan on Rats with Renal Interstitial Fibrosis Induced by Adenine
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 21, Pages: 149-156(2018)
- 作者机构：
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20182199
  CLC： R285.5
- Published：2018
- 稿件说明：
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YUAN Jin-feng, CHEN Lan-ying, WANG Hui-ling, et al. Effect of Jinkui Shenqiwan on Rats with Renal Interstitial Fibrosis Induced by Adenine[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(21): 149-156.
DOI：

YUAN Jin-feng, CHEN Lan-ying, WANG Hui-ling, et al. Effect of Jinkui Shenqiwan on Rats with Renal Interstitial Fibrosis Induced by Adenine[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(21): 149-156. DOI： 10.13422/j.cnki.syfjx.20182199.

摘要

目的：研究金匮肾气丸对腺嘌呤致大鼠肾间质纤维化的保护作用，并探讨其作用机制。方法：96只SD大鼠随机分成正常组、模型组、氯沙坦组（5 mg·kg-1）和金匮肾气丸高、中、低剂量组（2.4，1.2，0.6 g·kg-1），采用腺嘌呤灌胃（200 mg·kg-1·d-1）诱导肾间质纤维化大鼠模型，2 h后灌胃给予相应药物，造模连续9周；给药连续16周。采用生化法测定血清肌酐（SCr）和尿素氮（BUN）水平；苏木素-伊红（HE）染色观察大鼠肾脏组织病理改变情况；马松（Masson）染色观察大鼠肾脏胶原沉积情况；以实时荧光定量聚合酶链式反应（Real-time PCR）测定大鼠肾脏组组转化生长因子-β1（TGF-β1），α-平滑肌肌动蛋白（α-SMA），钙黏蛋白（E-cadherin），Ⅰ型胶原（Col-Ⅰ），Ⅲ型胶原（Col-Ⅲ），Ⅳ型胶原（Col-Ⅳ），基质金属蛋白酶-1（MMP-1），基质金属蛋白酶-2（MMP-2），基质金属蛋白酶-9（MMP-9），基质金属蛋白酶抑制剂-1（TIMP-1），基质金属蛋白酶抑制剂-2（TIMP-2）mRNA的表达；以免疫组化法测定大鼠肾脏TGF-β1，α-SMA和E-cadherin的表达。结果：与正常组比较，模型组中SCr和BUN显著升高（P<0.01），给予金匮肾气丸干预后SCr和BUN水平降低（P<0.05）；HE染色和Masson染色结果显示，与正常组比较，模型组肾间质损伤严重且肾间质胶原物质大量沉积，给予金匮肾气丸干预后肾间质小管损伤减轻，肾间质胶原物质沉积减少；Real-time PCR结果显示，与正常组比较，模型组TGF-β1，α-SMA，Col-Ⅰ，Col-Ⅲ，Col-Ⅳ，TIMP-1，TIMP-2 mRNA表达上调（P<0.05），模型组E-cadherin，MMP-1，MMP-2，MMP-9 mRNA表达下调（P<0.05），给予金匮肾气丸干预后E-cadherin，MMP-1，MMP-2，MMP-9 mRNA的表达上调（P<0.05），TGF-β1，α-SMA，Col-Ⅰ，Col-Ⅲ，Col-Ⅳ，TIMP-1，TIMP-2 mRNA的表达明显下调（P<0.05）；免疫组化结果显示，与正常组比较，模型组E-cadherin蛋白表达下调（P<0.01），TGF-β1和α-SMA蛋白表达上调，金匮肾气丸干预后E-cadherin蛋白表达上调（P<0.05），TGF-β1和α-SMA蛋白表达下调（P<0.05）。结论：金匮肾气丸可有效改善肾间质纤维化并对肾脏有一定的保护作用，其改善肾间质纤维化的机制可能与降低基质金属蛋白酶抑制剂（TIMP-1，TIMP-2）活性，TGF-β1蛋白表达，α-SMA蛋白表达，升高基质金属蛋白酶（MMP-1，MMP-2，MMP-9）活性，E-cadherin蛋白表达，从而减少胶原物质的沉积有关。

Abstract

Objective: To study the preventive and therapeutic effects of the Jinkui Shenqiwan on rats with renal interstitial fibrosis induced by adenineand and its mechanism. Method: Totally 96 male rats were randomly divided into 6 groups:the normal group

the model group

the losartan group(5 mg·kg-1) and the Jinkui Shenqiwan high

medium and low dose(2.4

1.2

0.6 g·kg-1). The model was established by ig administration adenine (200 mg·kg-1·d-1) for 9 weeks.In gavaging for 2 h

the drug was esablished for 16 weeks. The levels of serum creatinine (SCr) and blood urea nitrogen (BUN) were measured by biochemical method;the histopathological changes of kidneys in rats were observed by htoxylin and eosin(HE) staining;the collagen deposition in rat kidneyswas observed by Masson's staining;the gene transforming growth factor-β1(TGF-β1)

α-smooth muscle actin(α-SMA)

E-cadherin

collage Ⅰ(Cd-Ⅰ)

collage Ⅲ(Col-Ⅲ)

collage Ⅳ(Col-Ⅳ)

matrix metalloproteinase-1(MMP-1)

MMP-2

MMP-9

tissue inhibitor of metalloproteinase-1(TIMP-1)

TIMP-2 expression in kidney tissue were detected respectively by Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR);the protein TGF-β1

α-SMA and E-cadherin expression in kidney tissue were detected respectively by immunehistochemistry. Result: The results of each experimental group showed that creatinine and urea nitrogen were significantly increased in the model group compared with the normal group (P<0.01)

and creatinine and urea nitrogen levels were decreased after intervention with the Jinkui Shenqiwan (P<0.05); HE staining Compared with the normal group

the model group had severe renal interstitial damage and massive deposition of renal interstitial collagen. The renal interstitial tubule injury was relieved after intervention with the Jinkui Shenqi Wan

and renal interstitial collagen deposition decreased. Real-time PCR results showed that the expression of TGF-β1

α-SMA

Col-Ⅰ

Col-Ⅲ

Col-Ⅳ

TIMP-1

TIMP-2 was up-regulated in the model group compared with the normal group (P<0.05)

and the model group was MMP-1. The expressions of MMP-2 and MMP-9 were down-regulated (P<0.05)

and the expressions of MMP-1

MMP-2 and MMP-9 were up-regulated after intervention with the Jinkui Shenqiwan (P<0.05). The expressions of TGF-β1

α-SMA

Col-Ⅰ

Col-Ⅲ

Col-Ⅳ

TIMP-1 and TIMP-2 were significantly down-regulated (P<0.05). The immunohistochemical results showed that the expression of E-cadherin protein was down-regulated in the model group compared with the normal group (P<0.01). The expression of TGF-β1 and α-SMA protein was up-regulated (P<0.01)

and the expression of E-cadherin protein was up-regulated after the Jinkui Shenqiwan intervention(P<0.05)

TGF-β1 and α-SMA protein expression was down-regulated (P<0.05). Conclusion: The the Jinkui Shenqiwan can effectively improve renal interstitial fibrosis and have a certain protective effect on the kidney. The mechanism ofimproving renal interstitial fibrosis may reduce the activity of matrix metalloproteinase inhibitor (TIMP-1

TIMP-2). TGF-β1 protein expression

α-SMA protein expression

increased matrix metalloproteinase (MMP-1

MMP-2

MMP-9) activity

E-cadherin protein expression

thereby reducing the deposition of collagen.

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