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Application of hydrogen sulfide releasing agent in preparation of medicament for treating renal fibrosis disease

A release agent, hydrogen sulfide technology, applied in drug combinations, urinary system diseases, active ingredients of sulfur/selenium/tellurium, etc., can solve problems such as deterioration, exacerbation of renal function deterioration, and elevated serum creatinine

Inactive Publication Date: 2013-04-17
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the ability of ACEI to expand the glomerular efferent arteriole is greater than that of the afferent arteriole, it will theoretically reduce the glomerular filtration rate, especially when the patient has renal insufficiency or hypovolemia, ACEI may cause Renal function deteriorates sharply, manifested as elevated serum creatinine and hyperkalemia, which aggravates the deterioration of renal function and increases the risk of cardiac arrest, making it impossible to continue to use it. Therefore, there is currently no specific drug for such patients clinically. Dialysis and Kidney transplantation becomes the last treatment strategy

Method used

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  • Application of hydrogen sulfide releasing agent in preparation of medicament for treating renal fibrosis disease
  • Application of hydrogen sulfide releasing agent in preparation of medicament for treating renal fibrosis disease
  • Application of hydrogen sulfide releasing agent in preparation of medicament for treating renal fibrosis disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1 MTT method for measuring the proliferation of renal interstitial fibroblasts

[0069] Renal interstitial fibroblasts were placed in DMEM medium containing 10% fetal bovine serum at 37°C, 5% CO 2 cultured in an incubator. The medium was changed every other day, digested with 0.25% trypsin, and passaged at a ratio of 1:3 to obtain renal interstitial fibroblasts (NRK-49F).

[0070] Take 2×10 3 Renal interstitial fibroblasts (NRK-49F) were seeded in a 96-well plate and cultured. Serum (FBS) was used to simulate the effects of growth factors such as EGF and PDGF. Group experimental cells according to FBS concentration gradient: 1%FBS, 3%FBS, 5%FBS, 10%FBS, 1%FBS+Na 2 S, 3%FBS+Na 2 S, 5%FBS+Na 2 S, 10%FBS+Na 2 S, where Na 2 The concentration of S was 100 μM. Na 2 S pretreatment for 30 minutes. After 24 hours of cell treatment, 20 μL of MTT (5 mg / mL) was added to each well, the medium was discarded after 4 hours, 150 μL of DMSO was added to each well to di...

Embodiment 2

[0072] Example 2 MTT method for measuring the proliferation of renal interstitial fibroblasts

[0073] Take the 2 × 10 prepared in Example 1 3 Renal interstitial fibroblasts (NRK-49F) were seeded in a 96-well plate and cultured. Serum (FBS) was used to simulate the effects of growth factors such as EGF and PDGF. According to Na 2 S concentration gradient for experimental cell grouping: blank control group, FBS group, 1 μM Na 2 S+FBS, 10 μM Na 2 S+FBS, 50 μM Na 2 S+FBS, 100 μM Na 2 S+FBS, 500 μM Na 2 S+FBS, where FBS uses 10% FBS. Na 2 S pretreatment for 30 minutes. After 24 hours of cell treatment, 20 μL of MTT (5 mg / mL) was added to each well, the medium was discarded after 4 hours, 150 μL of DMSO was added to each well to dissolve formazan, and the OD value was read with a microplate reader at a wavelength of 490 nm. For test results, see figure 2 .

[0074] Depend on figure 2 It can be seen that Na 2 When the S concentration was 10-100 μM, the antiproliferat...

Embodiment 3

[0075] Example 3 BrdU infiltration method to measure the proliferation of renal interstitial fibroblasts

[0076] Take the 4 × 10 prepared in Example 1 4 Cells (NRK-49F) were seeded in 24-well plates and cultured overnight in DMEM medium containing 0.5% fetal calf serum. Experimental grouping after cell attachment: blank control group, 10% FBS group, Na 2 Group S (100μM), 10%FBS+Na 2 S (100 μM) group. Cell proliferation was stimulated by 10% FBS for 5 hours, and Brdu (10 μM) was added to continue incubation for 5 hours. After the cells were treated, they were fixed with 4% paraformaldehyde for 20 minutes, followed by 2N HCL DNA denaturation at 37°C for 20 minutes. After blocking with 5% BSA for 1 hour, anti-Brdu primary antibody (1:100) was added overnight at 4°C, Alexa Fluor555 Donkey Anti-Mouse IgG secondary antibody (1:500) was added at room temperature for 1 hour, and the slides were mounted using Vector mounting media containing DAPI. Observe 10 non-overlapping field...

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Abstract

The invention relates to the field of renal fibrosis disease treatment medicaments, in particular to an application of a hydrogen sulfide releasing agent in preparation of a medicament for treating renal fibrosis disease. According to the application of the hydrogen sulfide releasing agent in the preparation of the medicament for treating the renal fibrosis disease, the hydrogen sulfide releasing agent is sodium hydrosulfide or sodium sulfide. The sodium hydrosulfide or the sodium sulfide acts on renal fibroblasts, so that proliferation and differentiation of the renal interstitial fibroblasts are inhibited, deposition of renal interstitial collagen fibers is inhibited, the renal function is improved, further rise of serum creatinine and serum potassium levels is avoided, and the releasing agent has an effect of resisting renal fibrosis.

Description

technical field [0001] The invention relates to the field of drugs for treating renal fibrosis, in particular to the use of a hydrogen sulfide releasing agent in the preparation of drugs for treating renal fibrosis. Background technique [0002] Chronic kidney disease is a public health problem worldwide, and it is a disease that threatens human health with an incidence rate second only to cardiovascular and cerebrovascular diseases, malignant tumors and diabetes. Chronic kidney disease caused by various reasons will eventually develop into end-stage renal failure and require renal replacement therapy, which seriously threatens human life and health and causes a heavy burden on society and families. In the United States and Australia, the prevalence rate is about 11-16%. A recent report pointed out that the incidence rate of chronic kidney disease in the adult population in Beijing, China is 13.0%, and the incidence rate in various regions of the country is also showing an i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K33/04A61P13/12
Inventor 胡丽芳宋锴刘春风
Owner SUZHOU UNIV
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