Application of ouabain in preparation of anti-kidney stone kidney tubule injury drug

A technique for renal tubules and stones, applied in the direction of drug combinations, pharmaceutical formulas, medical preparations containing active ingredients, etc.

Pending Publication Date: 2019-05-28
NANJING UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the damage effect of cardiac glycosides on renal tubular epithelial cells in the case of kidney stones has not been reported so far. In our previous work, we found that Na + / K + - The ATPase inhibitor ouabain can alleviate the injury of renal tubular epithelial cells caused by COM at low doses, and at the same time alleviate the accumulation of calcium oxalate in mouse kidneys caused by glyoxylic acid and the resulting kidney damage

Method used

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  • Application of ouabain in preparation of anti-kidney stone kidney tubule injury drug
  • Application of ouabain in preparation of anti-kidney stone kidney tubule injury drug
  • Application of ouabain in preparation of anti-kidney stone kidney tubule injury drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Oouabain reduces the damage of HK-2 cells caused by calcium oxalate monohydrate

[0033] 1.1 Experimental materials

[0034] Human proximal renal tubular epithelial cells HK-2 were purchased from the Cell Bank of the Type Culture Collection Committee of the Chinese Academy of Sciences (Shanghai Cell Bank); ouabain and PI were purchased from Sigma Corporation of the United States; calcium oxalate monohydrate COM was purchased from Sangon Bioengineering (Shanghai Cell Bank); ) Co., Ltd.; CCK-8 detection kit was purchased from Biyuntian, and LDH detection kit was purchased from Nanjing Jiancheng Bioengineering Research Institute; EGFP-Annexin V was purified and prepared by the State Key Laboratory of Pharmaceutical Biotechnology of Nanjing University; Cleaved-PARP and Cleaved-Caspase3 antibody was purchased from Cell Signaling.

[0035] 1.2 Experimental method

[0036] 1.2.1 Establishment of COM on HK-2 cell injury model and administration of ouabain

[0037] ...

Embodiment 2

[0048] Example 2 Establishment of a mouse model of calculus nephropathy

[0049] 2.1 Experimental materials

[0050] Male C57BL / 6 mice aged 6-8 weeks were obtained from Nanjing University-Institute of Model Animals; glyoxylic acid was purchased from Tokyo Chemical Industry Co., Ltd.; creatinine (Cr) assay kit and urea nitrogen (BUN) assay kit were purchased from From Nanjing Jiancheng Institute of Bioengineering; Von kossa staining kit was purchased from Wuhan Sevier Biotechnology Co., Ltd.

[0051] 2.2 Experimental method

[0052] 2.2.1 Establishment of mouse model of calculous nephropathy

[0053] Male C57BL / 6 mice aged 6-8 weeks were fed adaptively for several days, and were randomly divided into control group and experimental group. The mice in the experimental group were given continuous intraperitoneal injection of glyoxylic acid at a dose of 50 mg / kg / d for 12 days, and the mice in the control group were given intraperitoneal injection of an equal volume of sterile no...

Embodiment 3

[0060] Example 3 Oouabain alleviates weight loss and renal dysfunction in mice caused by glyoxylic acid

[0061] 3.1 Experimental materials

[0062] Ouabain was purchased from Sigma, and other experimental materials were the same as in Example 2.

[0063] 3.2 Experimental method

[0064] Male C57BL / 6 mice were randomly divided into 4 groups with 7-9 mice in each group. The mice in the Control group were given intraperitoneal injection of the same amount of normal saline at 9:00 and 21:00 every day; the mice in the ouabain group were given intraperitoneal injection of 0.1 mg / kg ouabain at 9:00 every day, and the same amount of normal saline was injected intraperitoneally at 21:00; Mice in salt nephropathy group were given intraperitoneal injection of the same amount of normal saline at 9:00 every day, and 50 mg / kg glyoxylic acid was given intraperitoneal injection at 21:00; mice in ouabain treatment group were given 0.1 mg / kg ouabain intraperitoneally at 9:00 every day Injec...

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Abstract

The invention relates to application of ouabain in preparation of an anti-kidney stone kidney tubule injury drug. The experiment shows that the plant-derived ouabain compound adopted can effectively improve the decline in vitality of human proximal kidney tubule epithelial cells caused by calcium oxalate monohydrate, release of lactate dehydrogenase in cells and apoptosis caused thereby. At the same time, the ouabain also can alleviate the mouse weight loss and kidney function decrease caused by glyoxylic acid, reduce the accumulation of calcium oxalate crystals in the kidney caused by glyoxylic acid, and the resulting mouse kidney tubule injury and kidney apoptosis. At present, the drug which effectively resists the kidney tubule injury caused by kidney stone is deficient, especially in the case of small molecule drugs, the ouabain has good development value and application prospects.

Description

technical field [0001] The invention relates to a new application of ouabain, an inhibitor of sodium potassium ATPase, in the treatment of renal tubule damage in calculus nephropathy. Background technique [0002] Kidney stones are one of the common obstructive kidney diseases. Currently, the incidence of kidney stones in the global population is about 5%-15%, and about 80% of the stone crystals are oxalate crystals. The traditional methods for the treatment of complex kidney stones are mainly open surgery and extracorporeal impact lithotripsy, but open surgery has a large trauma, many postoperative complications, and a long recovery time; while extracorporeal shock wave lithotripsy is the most effective treatment for patients with complex kidney stones. The effect is not ideal enough, and it is difficult to remove the stones. In addition, extracorporeal shock wave lithotripsy for patients with complex stones often requires repeated lithotripsy, which has a great impact on...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7048A61P13/12
Inventor 殷武季晓君周日程小迎
Owner NANJING UNIV
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