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1.
Article in Chinese | WPRIM | ID: wpr-928157

ABSTRACT

This study aims to explore the effect of icariin(ICA) on mitochondrial dynamics in a rat model of chronic renal failure(CRF) and to investigate the molecular mechanism of ICA against renal interstitial fibrosis(RIF). CRF was induced in male Sprague-Dawley(SD) rats with 5/6(ablation and infarction, A/I) surgery(right kidney ablation and 2/3 infarction of the left kidney). Four weeks after surgery, the model rats were randomized into the following groups: 5/6(A/I) group, 5/6(A/I)+low-dose ICA group, and 5/6(A/I)+high-dose ICA group. Another 12 rats that received sham operation were randomly classified into 2 groups: sham group and sham+ICAH group. Eight weeks after treatment, the expression of collagen-Ⅰ(Col-Ⅰ), collagen-Ⅲ(Col-Ⅲ), mitochondrial dynamics-related proteins(p-Drp1 S616, p-Drp1 S637, Mfn1, Mfn2), and mitochondrial function-related proteins(TFAM, ATP6) in the remnant kidney tissues was detected by Western blot. The expression of α-smooth muscle actin(α-SMA) was examined by immunohistochemical(IHC) staining. The NRK-52 E cells, a rat proximal renal tubular epithelial cell line, were cultured in vitro and treated with ICA of different concentration. Cell viability was detected by CCK-8 assay. In NRK-52 E cells stimulated with 20 ng·mL~(-1) TGF-β1 for 24 h, the effect of ICA on fibronectin(Fn), connective tissue growth factor(CTGF), p-Drp1 S616, p-Drp1 S637, Mfn1, Mfn2, TFAM, and ATP6 was detected by Western blot, and the ATP content and the mitochondrial morphology were determined. The 20 ng·mL~(-1) TGF-β1-stimulated NRK-52 E cells were treated with or without 5 μmol·L~(-1) ICA+10 μmol·L~(-1) mitochondrial fusion promoter M1(MFP-M1) for 24 h and the expression of fibrosis markers Fn and CTGF was detected by Western blot. Western blot result showed that the levels of Col-Ⅰ, Col-Ⅲ, and p-Drp1 S616 were increased and the levels of p-Drp1 S637, Mfn1, Mfn2, TFAM, and ATP6 were decreased in 5/6(A/I) group compared with those in the sham group. The levels of Col-Ⅰ, Col-Ⅲ, and p-Drp1 S616 were significantly lower and the levels of p-Drp1 S637, Mfn1, Mfn2, TFAM, and ATP6 were significantly higher in ICA groups than that in 5/6(A/I) group. IHC staining demonstrated that for the expression of α-SMA in the renal interstitium was higher in the 5/6(A/I) group than in the sham group and that the expression in the ICA groups was significantly lower than that in the 5/6(A/I) group. Furthermore, the improvement in the fibrosis, mitochondrial dynamics, and mitochondrial function were particularly prominent in rats receiving the high dose of ICA. The in vitro experiment revealed that ICA dose-dependently inhibited the increase of Fn, CTGF, and p-Drp1 S616, increased p-Drp1 S637, Mfn1, Mfn2, TFAM, and ATP6, elevated ATP content, and improved mitochondrial morphology of NRK-52 E cells stimulated by TGF-β1. ICA combined with MFP-M1 further down-regulated the expression of Fn and CTGF in NRK-52 E cells stimulated by TGF-β1 compared with ICA alone. In conclusion, ICA attenuated RIF of CRF by improving mitochondrial dynamics.


Subject(s)
Adenosine Triphosphate/pharmacology , Animals , Female , Fibrosis , Flavonoids , Humans , Infarction/pathology , Kidney , Kidney Failure, Chronic , Male , Mitochondrial Dynamics , Rats , Rats, Sprague-Dawley , Renal Insufficiency, Chronic , Transforming Growth Factor beta1/metabolism
2.
Article in Chinese | WPRIM | ID: wpr-943087

ABSTRACT

ObjectiveTo observe the effect of Dahuang Xiezhuo prescription on the changes in renal pathology and reactive oxygen species (ROS)/thioredoxin-interacting protein (TXNIP)/NOD-like receptor protein 3 (NLRP3) pathway expression in the kidney tissues of rats with 5/6 nephrectomy, and to explore the mechanism of Dahuang Xiezhuo prescription in protecting renal function and delaying renal interstitial fibrosis and the possibility. MethodNinety healthy male SD rats were randomly divided into a sham operation group, a model group, low, medium, and high-dose (6.825, 13.65, 27.30 g·kg-1) Dahuang Xiezhuo prescription groups, and a Niaoduqing granule group (2.60 g·kg-1). Except the sham operation group, 5/6 nephrectomy was used to replicate the rat model of chronic renal failure (CRF). After modeling, each administration group was given the corresponding dose of drug suspension by intragastric administration, once a day for consecutive 8 weeks. After administration, serum creatinine (SCr) and urea nitrogen (BUN) levels and 24 h urinary protein quantification (UTP) levels were detected. Western blot assay was used to detect the protein expressions of thioredoxin (TRX), TXNIP, and NLRP3. The protein expressions of TRX, TXNIP, NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), transformation growth factor-β (TGF-β), Collagen Ⅳ, α-smooth muscle actin (α-SMA), and fibronectin (FN) were detected by immunohistochemistry. ResultAs compared with the sham operation group, serum levels of SCr, BUN, and UTP in the model group were increased (P<0.05), TRX, TXNIP, NLRP3, ASC, TGF-β, Collagen Ⅳ, α-SMA, and FN proteins were increased (P<0.01), and renal interstitial fibrosis significantly occurred. As compared with the model group, the levels of SCr, 24 h BUN, and UTP in the low, medium, and high-dose Dahuang Xiezhuo prescription groups and the Niaoduqing granule group were decreased to varying degrees (P<0.05), TRX, TXNIP, NLRP3, ASC, TGF-β, Collagen Ⅳ, α-SMA, and FN were decreased (P<0.01), and renal interstitial fibrosis was improved to varying degrees. ConclusionDahuang Xiezhuo prescription can protect renal function and delay renal interstitial fibrosis in rats with CRF.

3.
Article in Chinese | WPRIM | ID: wpr-940798

ABSTRACT

ObjectiveTo explore the mechanism of Yishen Huoxue prescription in renal interstitial fibrosis (RIF) from the perspective of endothelial cell and cell energy metabolism. MethodThe model was successfully established by unilateral ureteral obstruction (UUO). Seventy-five SPF C57BL/6 mice were randomly divided into a model group, a resveratrol group (50 mg·kg-1·d-1), three Yishen Huoxue prescription low, medium, and high-dose groups (7.1, 14.2, 28.4 g·kg-1·d-1), with 15 mice in each group. In addition, another 15 mice were used to prepare sham operation model. Mice in the sham operation group and the model group were gavaged with equal volume of normal saline. All mice were sacrificed on 7, 14, and 21 d after modeling. The protein expression of platelet endothelial cell adhesion molecule 31 (CD31) was detected by immunohistochemical S-P method. The expression of α-smooth muscle actin (α-SMA), collagen Ⅳ (Col-Ⅳ), angiopoietin 1(Ang-1) and tyrosine kinase receptors 2 (Tie-2), vascular endothelial growth factor (VEGF), vascular endothelial cadherin (VE-cadherin), and occludin in renal tissues was detected by Western blotting. The mRNA expressions of Ang-1/Tie-2, VEGF, VE-cadherin, and occludin in renal tissues were detected by Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR), and the levels of reactive oxygen species (ROS) in mice were detected by enzyme linked immunosorbent assay (ELISA). ResultAs compared with the sham operation group, the expression of CD31 in renal tissues of the model group was significantly decreased and worsened with the extension of modeling time (P<0.05), α-SAM and Col-Ⅳ protein expression levels were significantly increased (P<0.01), but the expression of CD31 was stable in 14-21 d. ROS levels were significantly increased (P<0.01), and the protein and mRNA expressions of Ang-1/Tie-2, VEGF, VE-cadherin, and occludin were significantly down-regulated (P<0.01). As compared with the model group, the expression of CD31 was increased (P<0.05), and α-SAM and Col-Ⅳ in the resveratrol group and the medium and high-dose Yishen Huoxue prescription groups were significantly decreased (P<0.01). The ROS content was significantly decreased (P<0.01), and the protein and mRNA expressions of Ang-1/Tie-2, VEGF, VE-cadherin, and occludin were up-regulated (P<0.01), As compared with the resveratrol group, the protein expressions of Ang-1/Tie-2, VEGF, VE-cadherin, and occludin in the medium and low-dose Yishen Huoxue prescription groups were significantly different (P<0.01). There was no significant difference in the mRNA expressions of CD31 and Ang-1/Tie-2 in the high-dose Yishen Huoxue prescription group, and no significant difference in the ROS level in the medium-dose Yishen Huoxue prescription group. ConclusionThe anti-RIF effect of Yishen Huoxue prescription may be related to promoting vascular endothelial repair, regulating mitochondrial ROS to reduce oxidative stress, protecting the integrity of renal endothelial structure, delaying cell apoptosis, and maintaining cell energy metabolism.

4.
Article in Chinese | WPRIM | ID: wpr-940724

ABSTRACT

ObjectiveTo observe the mechanism of modified Shengjiangsan in necroptosis and renal fibrosis of rats with diabetic nephropathy based on receptor-interacting protein (RIP)1/RIP3/mixed lineage kinase domain-like protein (MLKL) signaling pathway. MethodSeventy-five SD rats were randomly divided into a model group, a normal group, three high, medium, and low-dose modified Shengjiangsan groups (4.365, 8.73, 17.46 g·kg-1), and an irbesartan group (0.013 5 g·kg-1). After 4 weeks of intragastric administration, the levels of 24 h urine protein (UTP), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) of rats in each group were determined, as well as the changes in degree of renal pathology. Real-time quantitative polymerase chain reaction (Real-time PCR) and immunohistochemistry were used to detect the mRNA and protein expression levels of IL-1β, TNF-α, monocyte chemoattractant protein-1 (MCP-1), transforming growth factor-β1 (TGF-β1), and nuclear factor kappa B (NF-κB) in kidney tissues of rats. Western blot assay was used to detect the expression levels of key proteins in the RIP1/RIP3/MLKL signaling pathway. ResultAs compared with the normal group, the renal interstitial fibrosis in the model group was obvious, and the 24 h UTP, IL-1β, TNF-α levels were significantly increased (P<0.05). In the model group, the mRNA and protein expression levels of IL-1β, TNF-α, MCP -1, TGF-β1, and NF-κB in the kidney tissues were significantly increased (P<0.05), and protein expression levels of RIP1, RIP3, p-MLKL, and MLKL were significantly increased (P<0.05). Compared with the model group, all modified Shengjiangsan groups and the irbesartan group improved the levels of renal interstitial fibrosis in rats to varying degrees. As compared with the model group, the 24 h UTP levels in all modified Shengjiangsan groups and the irbesartan group were decreased to varying degrees (P<0.05), the content of IL-1β and TNF-α in the serum were decreased (P<0.05), the mRNA and protein expression levels of IL-1β, TNF-α, MCP-1, TGF-β1, and NF-κB in renal tissues was down-regulated (P<0.05), and the protein expression levels of RIP1, RIP3, p-MLKL, and MLKL were down-regulated (P<0.05). ConclusionModified Shengjiangsan ameliorates renal injury of rats with diabetic nephropathy, and the mechanism may be related to the down-regulation of the RIP1/RIP3/MLKL signaling pathway, the prevention of renal tissue necroptosis, and the inhibition of renal fibrosis.

5.
Article in Chinese | WPRIM | ID: wpr-940579

ABSTRACT

Chronic kidney disease (CKD) has become a public health problem worldwide with renal interstitial fibrosis (RIF) serving as the important pathological feature and pathological outcome of various CKD. Therefore, anti-fibrosis therapy has important practical significance for delaying the progression of CKD and improving the prognosis of CKD patients. The Wnt/β-catenin signaling pathway is a conserved signaling pathway through evolution, which plays a vital role in organ formation, tissue homeostasis, and disease progression during embryonic development. A growing body of research has confirmed that the Wnt/β-catenin signaling pathway is one of the key signaling pathways in a variety of kidney diseases and its activation is closely related to RIF. RIF is aggravated by the specific regulation of the expression of downstream target genes, such as fibroblasts, zinc finger transcription factor 1 (Snail1), M2 macrophages, matrix metalloproteinase-7 (MMP-7), plasminogen activator inhibitor-1 (PAI-1), and renin-angiotensin system(RAS), and relieved by targeting the signaling pathways, such as Klotho, Dickkopf-related protein 1 (DKK1), secreted frizzled-related protein 1 (Sfrp1), and indocyanine green-001. In addition,the pathological view of RIF in traditional Chinese medicine (TCM) coincides with that in western medicine. Based on the etiology and pathogenesis of TCM in the combination of deficiency and excess, TCM regulates fibrosis-promoting mediators by tonifying deficiency, eliminating turbidity, removing the toxin, resolving stasis, and treating both symptoms and root causes in a multi-target, multi-layer, and multi-pathway manner to inhibit the Wnt/β-catenin signaling pathway and play an important role in renal protection. Therefore, this study reviewed the regulatory mechanism of the Wnt/β-catenin signaling pathway in RIF and the protective effect of targeting this signaling pathway on renal function and discussed the potential role of TCM in delaying the progression of RIF, which is expected to provide new targets and strategies for the prevention and treatment of RIF.

6.
Article in Chinese | WPRIM | ID: wpr-940200

ABSTRACT

Renal interstitial fibrosis(RIF)is a common pathway for the progression of chronic kidney disease to renal failure,and its pathogenesis is mainly related to renal inflammatory damage,oxidative stress,apoptosis,and excessive extracellular matrix(ECM) deposition. Transforming growth factor-β1(TGF-β1) signaling pathway,mammalian target of rapamycin(mTOR) signaling pathway and other signaling pathways mediate the occurrence and development of RIF. Because of the complicated mechanism of RIF,there have been no specific prevention and treatment measures in clinical practice. Autophagy is a non-damaging response produced by eukaryotic cells. It maintains the balance of tissue homeostasis through degradation and reabsorption. At present, Chinese medicine has achieved desirable clinical effects with its unique advantages of multiple components,multiple effects,and multiple targets in the treatment of chronic kidney disease to delay the process of RIF. Scholars have found that autophagy is consistent with the Yin-Yang theory and the theory of abdominal mass in traditional Chinese medicine (TCM) to a certain extent,and it is involved in many aspects of RIF. The progression of RIF is closely related to autophagy. The targeted therapy of RIF by intervention in autophagy has become the frontier of research. However,little is known about the role of autophagy in RIF and the regulation of autophagy by Chinese medicine in the treatment of RIF. Therefore,it is necessary to further elucidate the relationship between autophagy and RIF in order to clarify the mechanism of autophagy in RIF and the mechanism of Chinese medicine regulating autophagy in targeted therapy of RIF. This article focused on the correlation between autophagy and RIF based on TCM theory,and systematically summarized the role of autophagy in RIF and the intervention of Chinese medicine by combining the effects of autophagy on inflammation damage,oxidative stress,apoptosis,and excessive ECM deposition in RIF, and the regulation mechanism of autophagy in TGF-β1 and mTOR signaling pathways in RIF. This study was expected to provide a certain reference for the clinical treatment of RIF and the development of new drugs.

7.
Article in Chinese | WPRIM | ID: wpr-940168

ABSTRACT

Renal interstitial fibrosis(RIF)is a common pathway for the progression of chronic kidney disease to renal failure,and its pathogenesis is mainly related to renal inflammatory damage,oxidative stress,apoptosis,and excessive extracellular matrix(ECM) deposition. Transforming growth factor-β1(TGF-β1) signaling pathway,mammalian target of rapamycin(mTOR) signaling pathway and other signaling pathways mediate the occurrence and development of RIF. Because of the complicated mechanism of RIF,there have been no specific prevention and treatment measures in clinical practice. Autophagy is a non-damaging response produced by eukaryotic cells. It maintains the balance of tissue homeostasis through degradation and reabsorption. At present, Chinese medicine has achieved desirable clinical effects with its unique advantages of multiple components,multiple effects,and multiple targets in the treatment of chronic kidney disease to delay the process of RIF. Scholars have found that autophagy is consistent with the Yin-Yang theory and the theory of abdominal mass in traditional Chinese medicine (TCM) to a certain extent,and it is involved in many aspects of RIF. The progression of RIF is closely related to autophagy. The targeted therapy of RIF by intervention in autophagy has become the frontier of research. However,little is known about the role of autophagy in RIF and the regulation of autophagy by Chinese medicine in the treatment of RIF. Therefore,it is necessary to further elucidate the relationship between autophagy and RIF in order to clarify the mechanism of autophagy in RIF and the mechanism of Chinese medicine regulating autophagy in targeted therapy of RIF. This article focused on the correlation between autophagy and RIF based on TCM theory,and systematically summarized the role of autophagy in RIF and the intervention of Chinese medicine by combining the effects of autophagy on inflammation damage,oxidative stress,apoptosis,and excessive ECM deposition in RIF, and the regulation mechanism of autophagy in TGF-β1 and mTOR signaling pathways in RIF. This study was expected to provide a certain reference for the clinical treatment of RIF and the development of new drugs.

8.
Article in Chinese | WPRIM | ID: wpr-909602

ABSTRACT

OBJECTIVE Chronic kidney disease (CKD) has become a global public health problem with 10%-15%incidence rate, and inhibiting the renal interstitial fibrosis is considered to be a potential strategy to delay the progression of CKD. Z-Guggulsterone (Z-GS), an active compound from derived from Commiphora mukul, has been proved to be effective in various diseases. The present study aimes to determine the protective effect and the molecular mechanism of Z-GS on renal fibrosis. METHODS Unilateral ureteral obstruction (UUO) mice and hypoxia-induced HK-2 cells were used to simulate renal fibrosis in vitro and in vivo, respectively. The mice and cells were treated with different doses of Z-GS to observe the pharmacological action. Renal function, including Scr, BUN, and UA, were detected by commercial kits. H&E and Masson staining were performed to observe histopathological changes of kidney. Cell viability and LDH release of HK-2 cells were detected by commercial kits. Cell cycle distribution and apoptosis rate were analyzed by flow cytometry. Fibrosis markers were detected by immunohistochemistry and immunofluorescence analysis. Cell cycle related proteins and Klotho/p53 signaling were analyzed by Western blotting. RESULTS The results showed that Z-GS decreased the rise of Scr, BUN, and UA and lightened renal histopathological injury, which were induced by UUO. Besides, Z-GS administration alleviated renal fibrosis in mice by inhibiting the expressions of α-SMA, TGF-β and colla?genⅣ, and delayed G2/M cell cycle arrest by promoting the expressions of CDK1 and cyclinD1/B1 rate. Experiments in vitro indicated that Z-GS treatment significantly increased the cell viability while decreased the LDH release in hypoxia-induced HK-2 cells. In addition, hypoxia induced fibrosis and G2/M cycle arrest in HK-2 cells were retarded by Z-GS. The study of its possible mechanism exhibited that Z-GS treatment increased the level of Klotho and inhibited P53 level. Nev?ertheless, the effect of Z-GS on Klotho/P53 signaling was reversed by siRNA-Klotho. Moreover, siRNA-Klotho treatment eliminated the effects of Z-GS on G2/M cell cycle arrest and fibrosis. CONCLUSION This study clarified that Z-GS allevi?ated renal fibrosis and G2/M cycle arrest through Klotho/P53 signaling pathway. People who have suffered CKD may potentially benefit from treatment with Z-GS.

9.
Acta Pharmaceutica Sinica B ; (6): 835-847, 2021.
Article in English | WPRIM | ID: wpr-881172

ABSTRACT

Localized delivery, comparing to systemic drug administration, offers a unique alternative to enhance efficacy, lower dosage, and minimize systemic tissue toxicity by releasing therapeutics locally and specifically to the site of interests. Herein, a localized drug delivery platform ("plum‒pudding" structure) with controlled release and long-acting features is developed through an injectable hydrogel ("pudding") crosslinked

10.
Article in Chinese | WPRIM | ID: wpr-906515

ABSTRACT

Objective:To observe the effect of modified Shengjiangsan on renal fibrosis in rats with membranous nephropathy (MN) and to explore the mechanism of its complications of renal fibrosis. Method:Rats were injected with cationized bovine serum albumin(C-BSA)in the tail vein to establish a rat model of membranous nephropathy. The normal group,model group,modified Shengjiangsan group (27.3 g·kg<sup>-1</sup>)and benazepril group(10 mg·kg<sup>-1</sup>)were established in this study. Each group was given corresponding dosage of the drug once a day for 4 weeks of continuous intervention. After the administration,we observed the pathological changes of rat kidneys by the technology of Masson staining, silverhexylamine iodate (PASM) staining, transmission electron microscopy (TEM), immunofluorescence technology (IF) was used to detect immunoglobulin(Ig)G deposition in rat kidneys. The levels of interleukin-1<italic>β</italic> (IL-1<italic>β</italic>), interleukin-6 (IL-6), tumor necrosis factor-<italic>α</italic> (TNF-<italic>α</italic>) in rat serum were detected by enzyme-linked immunosorbent assay (ELISA) method. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and immunohistochemistry (IHC) were used to detect the mRNA and protein expression levels of monocyte chemotactic protein 1 (MCP-1), intercellular adhesion molecule 1 (ICAM-1), nuclear factor kappa B (NF-<italic>κ</italic>B), Toll-like receptor 4 (TLR4), plasminogen activator inhibitor 1 (PAI-1), transforming growth factor-<italic>β</italic><sub>1</sub> (TGF-<italic>β</italic><sub>1</sub>), <italic>α</italic>-smooth muscle actin (<italic>α</italic>-SMΑ) and type Ⅳ Collagen (Collagen Ⅳ) in rat kidney tissues. Result:Compared with normal group, the kidney tissue of the model group was obviously fibrotic, the serum levels of IL-1<italic>β</italic>, IL-6, and TNF-<italic>α</italic> were significantly increased(<italic>P</italic><0.05), and the expressions of MCP-1, ICAM-1, NF-<italic>κ</italic>B, TLR4, PAI-1, TGF-<italic>β</italic><sub>1</sub>, <italic>α</italic>-SMA and Collagen Ⅳ mRNA and protein in kidney tissue were significantly increased(<italic>P</italic><0.05). Compared with model group, modified Shengjiangsan and benazepril significantly improved renal fibrosis in rats, reduced the levels of IL-1<italic>β</italic>, IL-6, and TNF-<italic>α</italic> in the serum of MN rats(<italic>P</italic><0.05), down-regulated MCP-1, ICAM-1, NF-<italic>κ</italic>B, TLR4, PAI-1, TGF-<italic>β</italic><sub>1</sub>, <italic>α</italic>-SMA and Collagen Ⅳ mRNA and protein expression in kidney tissue(<italic>P</italic><0.05). Conclusion:Modified Shengjiangsan can reduce the release and expression of inflammatory factors by down-regulating the TLR4/NF-<italic>κ</italic>B signaling pathway, inhibit renal fibrosis, and reduce renal damage in MN rats.

11.
Article in Chinese | WPRIM | ID: wpr-905831

ABSTRACT

Objective:To investigate the mechanism of Jianzhong Bushen Xiaozheng decoction in regulating the effect of miRNA139 on Wnt/<italic>β</italic>-catenin signaling pathway for renal interstitial fibrosis. Method:The 120 mice were randomly divided into sham operation group, unilateral ureteral obstruction (UUO) group, Jianzhong Bushen Xiaozheng decoction low, middle, high dose group, and Niaoduqing group. The UUO animal model was established to observe the morphological changes in mice. Intragastic administration was started from day 3 after modeling. The sham operation group and UUO group received the same amount of distilled water every day. The low, medium and high-dose groups received Jianzhong Bushen Xiaozheng decoction solution at 6,12,24 g·kg<sup>-1</sup>·d<sup>-1</sup> respectively. The Niaoduqing group received 6.2 g·kg<sup>-1</sup>·d<sup>-1</sup> Niaoduqing granule solution. After 14 d and 21 d, 28 d , the morphological changes, general signs and renal interstitial fibrosis index of the obstructed side were observed, hematoxylin-eosin (HE) staining and Masson staining were used to observe the pathological changes of renal tissue. Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) method was used to detect the miRNA-139 expression in renal tissue volume, Western blot was used to detect expression of beta serial proteins (<italic>β</italic>-catenin) and fibrinolytic enzyme activators inhibitor-1 (PAI-1) in renal tissues, and immunohistochemical assay was used for detection of matrix metalloproteinases-7 (MMP-7) protein expression at the obstruction side. Result:After 14, 21 and 28 days, the expression levels of <italic>β</italic>-catenin and PAI-1 in UUO group were higher than those in sham operation group(<italic>P</italic><0.05),while the expression levels of miRNA139 and MMP-7 protein were lower than those of sham operation group (<italic>P</italic><0.05). The expression levels of <italic>β</italic>-catenin and PAI-1 proteins in mice after treatment in Niaoduqing group and the traditional Chinese medicine groups were lower than those in the UUO group(<italic>P</italic><0.05), the expression of miRNA139 and MMP-7 proteins increased(<italic>P</italic><0.05), and the efficacy of high-dose Jianzhong Bushen Xiaozheng decoction group was better than that of other dosage groups or Niaoduqing group(<italic>P</italic><0.05). Conclusion:Jianzhong Bushen Xiaozheng decoction may regulate miRNA139 to mediate the process of renal interstitial fibrosis through the Wnt/ <italic>β</italic>-catenin pathway and delay the development of renal interstitial fibrosis to improve renal function.

12.
Chinese Journal of Nephrology ; (12): 809-816, 2021.
Article in Chinese | WPRIM | ID: wpr-911903

ABSTRACT

Objective:To explore the mechanism of cisplatin-induced renal interstitial fibrosis and provide a new idea for the prevention and treatment of renal interstitial fibrosis.Methods:Eight-week-old male C57BL/6 mice (specific pathogen-free) were used to carry out the experiment. The mice were divided into cisplatin group (10 mg/kg, n=6) and saline group ( n=6) with intraperitoneal injection on day 0, 7 and 21, and sacrificed on day 28. The kidney tissues were collected for RNA Illumina high-throughput sequencing, real-time PCR, Western blotting, Masson staining and bioinformatics analysis. Results:Through real-time PCR, Western blotting and Masson staining, a mouse model with cisplatin-induced renal interstitial fibrosis was successfully established. Through RNA Illumina high-throughput sequencing, 387 long noncoding RNA (lncRNA) and 2 427 mRNA were differently expressed between cisplatin group and saline group. The expression of the top two lncRNA was confirmed by real-time PCR with the same tendency as RNA sequencing. Complement C3 was found to be at the top among the different expressed mRNA by RNA sequencing. Several terms related to immunity were found to be within the top 20 terms through Gene Ontology (GO) enrichment analysis. Systemic lupus erythematous pathway (ko05322, Q=3.4E-17), including the complement cascade pathway, was found to be the top pathway through Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The mRNA expression levels of C1q, C2, C3 and C4 were up-regulated remarkably in the cisplatin group by RNA sequencing than those in saline group (all P<0.05) and confirmed by real-time PCR. Conclusions:Renal interstitial fibrosis can be induced by intraperitoneal injection of cisplatin periodically in mice, with complement cascade pathway activation in the diseased kidney.

13.
Journal of Integrative Medicine ; (12): 505-513, 2020.
Article in English | WPRIM | ID: wpr-880981

ABSTRACT

OBJECTIVE@#MicroRNAs (miRNAs) may be viable targets for treating renal interstitial fibrosis (RIF). Fuzheng Huayu recipe (FZHY), a traditional Chinese compound herbal medicine, is often used in China to treat fibrosis. This study sought to assess the mechanisms through which FZHY influences miRNAs to treat RIF.@*METHODS@#RIF was induced in rats by mercury chloride and treated with FZHY. Hydroxyproline content, Masson's staining and type I collagen expression were used to evaluate renal collagen deposition. Renal miRNA profiles were evaluated using a miRNA microarray. Those miRNAs that were differentially expressed following FZHY treatment were identified and subjected to bioinformatic analyses. The miR-21 target gene phosphatase and tensin homolog (PTEN) expression and AKT phosphorylation in kidney tissues were assessed via Western blotting. In addition, HK-2 human proximal tubule epithelial cells were treated using angiotensin II (Ang-II) to induce epithelial-to-mesenchymal transition (EMT), followed by FZHY exposure. miR-21 and PTEN expressions were evaluated via quantitative reverse transcription-polymerase chain reaction (qRT-PCR), while E-cadherin and α-smooth muscle actin (α-SMA) expressions were assessed by immunofluorescent staining and qRT-PCR. Western blotting was used to assess PTEN and AKT phosphorylation.@*RESULTS@#FZHY significantly decreased kidney collagen deposition, hydroxyproline content and type I collagen level. The miRNA microarray identified 20 miRNAs that were differentially expressed in response to FZHY treatment. Subsequent bioinformatic analyses found that miR-21 was the key fibrosis-related miRNA regulated by FZHY. FZHY also decreased PTEN expression and AKT phosphorylation in fibrotic kidneys. Results from in vitro tests also suggested that FZHY promoted E-cadherin upregulation and inhibited α-SMA expression in Ang-II-treated HK-2 cells, effectively reversing Ang-II-mediated EMT. We also determined that FZHY reduced miR-21 expression, increased PTEN expression and decreased AKT phosphorylation in these cells.@*CONCLUSION@#miR-21 is the key fibrosis-related miRNA regulated by FZHY. The ability of FZHY to modulate miR-21/PTEN/AKT signaling may be a viable approach for treating RIF.

14.
Article in Chinese | WPRIM | ID: wpr-847566

ABSTRACT

BACKGROUND: Unilateral ureteral obstruction (UUO) is a traditional surgical method that can cause renal interstitial fibrosis in rats in a short period of time (1-2 weeks), but it can develop many postoperative complications and result in a high mortality. OBJECTIVE: To optimize and improve the UUO operation in rats for reducing the incidence of postoperative complications and improving the survival rate of animals, and to detect the pathophysiological indicators of rats modeled by UUO, providing background data for the basic research on functional pharmacology. METHODS: Twenty male Sprague-Dawley rats were randomly divided into four groups, followed by traditional UUO ligation, improved UUO ligation, undissociated ureteral ligation, and opening the abdominal cavity with no ureter ligation (sham group), respectively. At 14 days postoperatively, the successful rate of renal interstitial fibrosis model, mortality rate, and the incidence of postoperative complications were compared between groups. Another 28 Sprague-Dawley rats, half male and half female, were randomly divided into improved UUO group and control group. Physiological indexes, including urine analysis, blood cells analysis, blood biochemical analysis, were tested as background information. Besides, the pathological changes of kidney tissues were compared between groups using hematoxylin-eosin staining and Masson staining at 14 and 21 days postoperatively. RESULTS AND CONCLUSION: Compared with the undissociated ureteral ligation group, the successful rate of modeling was higher in the traditional and improved UUO groups. The improved UUO surgery with double ligation of the middle ureter and no cutting of the middle ureter was characterized by easier operation, smaller surgical wound, faster modeling and higher successful rate of renal interstitial fibrosis model, lower postoperative mortality and lower incidence of complications in animals as compared with the traditional UUO surgery. The improved UUO group showed statistically significant differences from the sham group in blood biochemical indexes such as urea nitrogen, glutamate aminotransferase, and albumin (P < 0.05). Results from hematoxylin-eosin staining and Masson staining of the obstructed kidney revealed typical pathological features of renal interstitial fibrosis.

15.
Article in Chinese | WPRIM | ID: wpr-873225

ABSTRACT

Objective:To observe the effects of Fushengong prescription on p38 mitogen-activated protein kinase(p38 MAPK) signal pathway of rats with chronic renal failure(CRF),and to explore its mechanism of reducing inflammatory reaction of renal tissues and delaying the progress of renal interstitial fibrosis. Method:The 55 male Sprague-Dawley rats were randomly divided into normal group,model group, and low,medium and high dose groups of Fushengong prescription,with 11 rats in each group.The normal group was routinely reared, and the other four groups of rats were fed a diet containing 0.5% adenine to produce a model of CRF, which was continuously molded for 21 days.After successful modeling,all rats switched to conventional feed.Normal group and model group were given normal saline 20 mL·kg-1,and each group of Fushengong prescription was given 4,8,16 g·kg-1 of water prescription once a day for 30 days.After the experiment,Masson staining was used to observe the degree of renal interstitial fibrosis.The expression of monocyte chemotactic protein-1(MCP-1) in renal tissues was detected by immunohistochemistry. The expression of phosphorylated p38 mitogen-activated protein kinase(p-p38 MAPK) and transformed growth factor-β1(TGF-β1) in renal tissues were detected by Western blot. Result:Compared with normal group,the renal interstitial collagen deposition increased significantly,the average optical density value of MCP-1 and the expression levels of p-p38 MAPK and TGF-β1 also increased significantly in model group (P<0.05). Compared with model group,the renal interstitial collagen deposition reduced significantly,the average optical density value of MCP-1 and the protein expression levels of p-p38 MAPK and TGF-β1 also decreased significantly in each dose group of Fushengong prescription(P<0.05). Conclusion:Fushengong prescription can effectively inhibit the expression of related inflammatory factors in the renal tissue of CRF rats,so as to reduce the inflammatory response in the renal tissue and delay the progress of renal interstitial fibrosis,the mechanism of which may be related to inhibit the activation of p38 MAPK signal transduction pathway.

16.
Article in Chinese | WPRIM | ID: wpr-873055

ABSTRACT

Objective:To study the effect of Shenshuai Xiezhuo decoction and its deficiency tonifying and pathogen eliminating components on renal interstitial fibrosis in UUO rats. Method:A rat model of unilateral ureteral obstruction (UUO) was established through ligation of a unilateral ureter. The rats were divided into six groups: sham operation group, model group, benazepril group, Shenshuai Xiezhuo decoction group, Buxufang group, and phlegm group, with 24 rats in each group. On the third day after operation, the rats in the Shenshuai Xiezhuo decoction group, Buxufang group, and phlegm group were given Shenshuai Xiezhuo decoction concentrating agent at a dose of 8.0 g·kg-1·d-1, the rats in the benazepril group were given benazepril 1.5 mg·kg-1·d-1, and the rats in the sham operation group and the model group were given the same volume of saline. On the 7th, 14th and 21st days after operation, the expressions of peripheral cells and relevant signal pathway markers in renal tissue were detected by Western blot and Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) respectively. Result:The stromal damage score and the interstitial collagen accumulation on the 14th and 21st days after UUO were significantly lower in the Shenshuai Xiezhuo decoction group, Buxu prescription group and Qixie prescription group than those in the model group (P<0.05). Except for the sham operation group, the protein expressions of α smooth muscle actin (α-SMA), platelet derived growth factor A (PDGFA), nerve/glial type 2 chondroitin sulfate glycoprotein (NG-2), vascular endothelial growth factor A (VEGFA), vascular endothelial growth factor receptor 1 (VEGFR1), platelet-derived growth factor -β (PDGF-β) and platelet-derived growth factor receptor-β (PDGFR-β) in the kidney tissues of the 14th and 21st days were significantly increased compared with those on the 7th day (P<0.05). On the 21st day after surgery, the expressions of NG-2, VEGFA, VEGFR1 and PDGFR-β in renal tissue of Shenshuai Xiezhuo decoction group were significantly lower than those in the model group (P<0.05). Compared with the model group, relevant expressions of α-SMA, PDGFA, NG-2, VEGFA, VEGFR1, PDGF-β, and PDGFR-β in kidney tissue of Shenshuai Xiezhuo decoction group decreased significantly on the 21st day after operation (P<0.05), and relative expressions of α-SMA, NG-2, VEGFA, VEGFR1 and PDGFR-β in the Shenshuai Xiezhuo decoction group were significantly lower than those in the benazepril group on the 21st day after surgery (P<0.05). Conclusion:Shenshuai Xiezhuo decoction and its deficiency tonifying and pathogen eliminating components have an antagonistic effect on renal interstitial fibrosis in UUO rats. Its mechanism is related to the inhibition of activation of peripheral cells and relevant cell signaling pathways.

17.
Article in Chinese | WPRIM | ID: wpr-872708

ABSTRACT

Objective:To discuss the efficacy and safety of Shenwu Yishenpian on stage 4-5 chronic kidney disease-nondialysis (CKD) with deficiency of spleen and kidney Qi, and the effect on renal interstitial fibrosis (RIF) and microinflammation. Method:One hundred and twenty patients were randomly divided into observation group and control group. A total of 58 patients in control group completed the treatment (including 2 patients falling off or lost). And 58 patients in observation group completed the treatment (including 1 patient was falling off or lost visit, and 1 eliminated). Both groups got comprehensive treatment of western medicine. Patients in control group got simulated medicine of Shenwu Yishenpian, 4 pieces/time, 3 times/day. Patients in observation group got Shenwu Yishenpian, 4 pieces/time, 3 times/day. The treatment lasted for 6 months until the renal replacement therapy, and the 6-month follow-up was recorded. For every month, blood creatinine (SCr) was detected, and glomerular filtration rate (eGFR) were calculated. The 12-month renal replacement (dialysis or kidney transplantation), progress (CKD4 to CKD5) and mitigation (CKD5 to CKD4 or CKD4 to CKD3) were recorded. Before and after treatment, levels of urea nitrogen (BUN), hemoglobin (HB), plasma albumin (ALB), urine protein quantity (24 hUp) and blood uric acid (UA) levels were detected, deficiency of spleen kidney Qi was scored, and transforming growth factor-β1 (TGF-β1), connective tissue growth factor (CTGF), serum Klotho, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1 (Lkn-1) and interleukin-12 (IL-12) were detected. And the safety was evaluated. Result:At the 3th and 6th after treatment, SCr in two groups increased (P<0.01), while eGFR decreased (P<0.01). Compared with control group, SCr was less than that in control group (P<0.01), whereas eGFR was higher than that in control group (P<0.01). During 12 months of observation, the reduction rate of CKD was 13.79% (8/58), which was higher than 1.72% (1/58) in control group. The progress rate of CKD was 11.43% (4/35), which was lower than 31.58% (12/38) in control group (P<0.05). Levels of BUN, 24 hUp and UA were lower than those in control group (P<0.01), while levels of Hb and ALB were higher than those in control group (P<0.01). Effect in observation group was better than that in control group (Z=2.051, P<0.05). And levels of TGF-β1, CTGF, TNF-α, IL-6, Lkn-1 and IL-12 were lower than those in control group (P<0.01), and level of Klotho was higher than that in control group (P<0.01). There was no adverse reaction relating to Shenwu Yishen Pian. Conclusion:Shenwu Yishenpian can delay the progress of renal function and CKD, reverse the progress of renal function in some patients, reduce the risk factors of disease progress, reduce the state of micro inflammation and resist RIF, and protect or improve renal function. Its clinical effect is better than placebo, and it is safe to use.

18.
Article in Chinese | WPRIM | ID: wpr-872648

ABSTRACT

Objective:To explore the effect of Qizhu Zhenwutang on renal interstitial fibrosis in rats ligated with unilateral ureter, transforming growth factor-β1 (TGF-β1)/Smads and oxidative stress. Method:A total of 30 male SD rats were randomly divided into sham operation group, model group, high-dose group, low-dose group and irbesartan group (n=6). The left ureter ligation was performed in the model group and the treatment group. In the sham operation group, the ureter was not ligated, only the ureter was separated, and the abdominal cavity was closed. Rats in each group were given drugs by gavage on the next day after operation. Sham operation group and model group were given aseptic distilled water 10 mL·kg-1 by gavage, high-dose Qizhizhenwu Tang group was given 22.2 g·kg-1 by gavage, low-dose group was given 11.1 g·kg-1 by gavage, and irbesartan group was given 0.02 g·kg-1 by gavage. Rats in each group were sacrificed on the 14th day after operation, 24-hour urine was collected before sampling, and the total amount of 24 hour urine protein (24 h-Upr) was detected. Blood samples were collected from the abdominal aorta to detect serum creatinine(SCr) and blood urea nitrogen (BUN). The tissues were stained with htoxylin eosin (HE) and Masson, and the pathological changes were observed under light microscope, immunohistochemical method was used to detect α-SMA, FN and Col-Ⅰ expressions. Western blot method was used to detect the expressions of TGF-β1, Smad3, p-Smad3 and NOX4. Result:Compared with sham group, SCr, BUN and collagen volume fraction (CVF),24 h-Upr in model group were all increased (P<0.05, P<0.01). The expressions of α-SMA, Col-Ⅰ, FN, TGF-β1, p-Smad3, NOX4 were higher (P<0.05). Compared with the model group, SCr, BUN and CVF were lower in high-dose group and irbesartan group (P<0.05). 24 h-Upr was lower in high-dose group (P<0.05), the expressions of α-SMA, Col-Ⅰ, FN, TGF-β1, Smad3, p-Smad3, NOX4 in traditional Chinese medicine treatment group were less (P<0.05). Conclusion:Qizhi Zhenwutang can reduce the urinary protein of UUO rats, protect the renal function, and inhibit the occurrence and development of renal interstitial fibrosis, the mechanism may be related to the inhibition of TGF-β1/Smads signaling pathway and oxidative stress response.

19.
Article in Chinese | WPRIM | ID: wpr-849684

ABSTRACT

The basic features of chronic kidney disease are persistent decline in renal function, accumulation of extracellular matrix, and progressive tissue fibrosis. Renal interstitial fibrosis and renal anemia are common complications of chronic kidney disease. Renal interstitial fibrosis is a chronic and progressive process. Its severity is closely related to renal dysfunction. Renal anemia may increase the incidence of cardiovascular complications and mortality in patients with chronic kidney disease. Therefore, it will be important to note the diagnosis and treatment of these two complications in the early stage of chronic kidney disease. The key cells mediating renal interstitial fibrosis are activated myofibroblasts, which are a major matrix-secreting cell type, and reactive cells mostly present under injury or pathological conditions; while the important factor mediating renal anemia is the reduction of erythropoietin-producing cells, which resulting in decreased secretion of erythropoietin (which is a glycoprotein hormone that promotes erythropoiesis). In addition, the transformation of erythropoietin-producing cells into myofibroblasts underlies the link between anemia and fibrosis in chronic kidney disease, and the transformation is reversible. Targeting this may open a new way for the treatment of renal anemia and renal interstitial fibrosis. In recent years, researchers have tried to find alternative therapies for the treatment of anemia and fibrosis in chronic kidney disease, and traditional Chinese medicine has been increasingly used to treat chronic kidney disease because of its unique advantages. The relationship has been mainly discussed in present paper between erythropoietin-producing cells and anemia and fibrosis in chronic kidney disease, and the current research status of traditional Chinese medicine is then analyzed and summarized.

20.
Article in Chinese | WPRIM | ID: wpr-752238

ABSTRACT

Objective To study the mechanism of overexpression of retinoic acid receptor alpha( RARα)in attenuating renal interstitial fibrosis(RIP)in rats. Methods Porty 6_week_old male SD rats were randomly divided into 4 groups:sham operation group,model group,negative control group and transfection group,with 10 rats in each group. Rats in model group were separated and double ligated with left ureter;rats in sham operation group were not li_gated with ureter;rats in transfection group and negative control group were transfected with adeno_associated virus and negative control virus carrying RARα gene on the basis of model group,respectively. All rats were sacrificed 2 weeks later. Left kidney tissues were taken for pathological examination and RIP index was calculated. The expression of colla_genⅣ(Col_Ⅳ)and fibronectin(PN)in renal tissue was detected by using immunohistochemistry. The expressions of RARα,prohibitin(DHB)and transforming growth factor_beta 1(TGP_β1)in renal tissue were detected by using real_time fluorescence quantitative polymerase chain reaction( RT _qDCR)and Western blot. Results (1)Com_pared with sham operation group,the RIP index was significantly increased in model group(22. 81 ± 2. 43 vs. 2. 34 ± 0. 55,q﹦24. 94,P〈0. 05);compared with model group,the RIP index was not of significant difference in negative control group(22. 81 ± 0. 43 vs. 22. 26 ± 3. 43,q﹦0. 67,P〉0. 05),however it significantly decreased in transfection group(14. 06 ± 2. 99 vs. 22. 81 ± 2. 43,q﹦10. 66,P〈0. 05).(2)Compared with sham operation group,the mRNA and protein expressions of RARα,DHB significantly decreased in model group,but TGP_β1 mRNA and protein,Col_Ⅳand PN protein expression significantly increased in model group( mRNA:0. 43 ± 0. 17 vs. 1. 00 ± 0. 00,0. 34 ± 0. 08 vs. 1. 00 ± 0. 00,2. 97 ± 0. 54 vs. 1. 00 ± 0. 00,all P〈0. 05;protein:0. 25 ± 0. 10 vs. 0. 51 ± 0. 06,0. 24 ± 0. 07 vs. 0. 58 ± 0. 04,0. 59 ± 0. 09 vs. 0. 33 ± 0. 06,16. 01 ± 0. 87 vs. 8. 79 ± 0. 39,14. 64 ± 0. 32 vs. 9. 36 ± 0. 59,all P〈0. 05);com_pared with model group,the mRNA and protein expressions of RARα,DHB,TGP_β1 and Col_Ⅳand PN protein ex_pression had no significant difference in negative control group(all P〉0. 05);compared with model group,the mRNA and protein expression of RARα,DHB mRNA and protein expression significantly increased,but the TGP_β1 mRNA and protein,Col_Ⅳ and PN protein expression significantly decreased in transfected group( mRNA:0. 86 ± 0. 07 vs. 0. 43 ± 0. 17,0. 89 ± 0. 11 vs. 0. 34 ± 0. 08,1. 65 ± 0. 28 vs. 2. 97 ± 0. 54,all P〈0. 05;protein:0. 40 ± 0. 07 vs. 0. 25 ± 0. 10,0. 45 ± 0. 10 vs. 0. 24 ± 0. 07,0. 43 ± 0. 08 vs. 0. 59 ± 0. 09,11. 57 ± 0. 33 vs. 16. 01 ± 0. 87,11. 67 ± 0. 53 vs. 14. 64 ± 0. 32,all P〈0. 05).(3)Correlation analysis revealed that RARα protein expression was negatively correlated with RIP index,Col_Ⅳ,PN,TGP_β1(r﹦ _0. 78,_0. 78,_0. 76,_0. 76,all P〈0. 05);DHB protein expression was negatively correlated with RIP index,Col_Ⅳ,PN,TGP _β1( r ﹦ _0. 87,_0. 87,_0. 88,_0. 75,all P 〈0. 05);RARα protein was positively correlated with DHB(r﹦0. 85,P〈0. 05). Conclusion Overexpression of RARα could attenuate RIP by enhancing DHB expression in rats subjected to unilateral ureteral obstruction.

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