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Application of luteolin to preparation of medicine for treating oxidative stress injury of testicular tissue

A technology of oxidative stress and luteolin, applied in the field of male reproductive system, to achieve good protection effect and broad application prospects

Inactive Publication Date: 2017-09-26
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Few drugs are currently used to treat oxidative stress injury in testicular tissue

Method used

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  • Application of luteolin to preparation of medicine for treating oxidative stress injury of testicular tissue
  • Application of luteolin to preparation of medicine for treating oxidative stress injury of testicular tissue
  • Application of luteolin to preparation of medicine for treating oxidative stress injury of testicular tissue

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Protective effect of luteolin on Sertoli cell damage induced by hydrogen peroxide

[0032] The logarithmic phase TM4 cells were inoculated into 96-well culture plates (5000 cells / well), and after 24 hours of adherent culture, they were treated with luteolin (2.5, 5 and 10 μmol / L) or positive drug quercetin (5 μmol / L) respectively. ) After 12 and 24 hours of intervention and protection, add H 2 o 2 (600mM) the culture medium was discarded after 2 hours of injury, and luteolin and quercetin were not added to the model group, only H 2 o 2 ; In the normal control group, neither luteolin, quercetin nor H 2 o 2 . The cell viability of normal control group, model group and experimental group with drug intervention was detected by MTT method. Specific operation: Add 10 μL of 0.5 g / L MTT (3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide) solution to each well, and continue to culture in the incubator 4h. Discard the cell supernatant, add DMSO (150 μL / ...

Embodiment 2

[0035] Example 2 Protective effect of luteolin on Sertoli cell damage induced by triptolide

[0036] TM4 cells in the logarithmic phase were seeded into 96-well culture plates (5000 cells / well), and cultured for 24 hours. The experimental group, positive drug control group, model group and normal group were set up, and the treatment methods of each group were as follows: the experimental group was protected with luteolin (2.5, 5 and 10 μmol / L) for 12 and 24 hours, and then triptolide was added. (300mM) was injured for 24h and the medium was discarded; the positive drug control group was treated with quercetin (5μmol / L) for 12 and 24h after the intervention and protection, and triptolide (300mM) was added for 24h after the injury was discarded; the model group was not added For drug intervention protection, only triptolide was added; no reagent was added to the normal group. The cell viability of the normal group, the model group and the experimental group with drug interventio...

Embodiment 3

[0039] Example 3 Effect of luteolin on the apoptosis rate of Sertoli cells damaged by hydrogen peroxide

[0040] The effects of different concentrations of luteolin on the apoptosis rate of TM4 cells were detected by Annexin V-FITC / PI cell apoptosis detection kit (KGI, China). The kit includes Binding buffer, AnnexinV-FITC and PropidiμmIodide staining solution.

[0041] 2mL (1×10 5 per mL) TM4 cells were seeded into 6-well culture plates and cultured for 24 h. The experimental group, positive drug control group, model group and normal group were set up, and the treatment methods of each group were as follows: the experimental group was protected with luteolin (2.5, 5 and 10 μmol / L) for 24 hours, and then added H 2 o 2 (600mM) injury for 2h; the positive drug control group was intervened and protected by the positive drug quercetin (5μmol / L) for 24h, and then added H 2 o 2 (600mM) injury for 2h; the model group did not add drug intervention protection, only added H 2 o 2...

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Abstract

The invention provides application of luteolin to the preparation of a medicine for treating an oxidative stress injury of testicular tissue, and relates to the field of male reproduction and endocrinic medicines. An active component of a medicinal composition is the luteolin. Discovered through an experiment, the injury, which is caused by hydrogen peroxide and triptolide, of a testicular sustentacular cell can be reversed through the intervention treatment of the luteolin; the apoptosis of the testicular sustentacular cell induced by the hydrogen peroxide is inhibited; the accumulation of ROS (Reactive Oxygen Species) is inhibited; the collapse of a cellular mitochondrial membrane potential caused by the hydrogen peroxide is reversed; the expression of a downstream antioxidase gene can be activated; the autologous oxidation-resistance capacity of the sustentacular cell is improved; the injury to the testicular tissue of a mouse induced by the triptolide is effectively reversed. Besides, the luteolin can be also used for inhibiting the increment of the permeability of a blood-testis barrier, which is caused by the triptolide-induced oxidative stress injury, through regulating and controlling gap junction, occlusion junction and tight junction protein; the completeness of the blood-testis barrier is protected. Therefore, the luteolin can be applied to the preparation of the medicine for treating the oxidative stress injury of the testicular tissue.

Description

technical field [0001] The invention relates to the field of the male reproductive system, in particular to the application of luteolin in the preparation of medicines for treating testicular tissue damage. Background technique [0002] Male infertility is one of the common and frequently-occurring diseases in andrology. According to the WHO survey, 15% of couples of childbearing age have infertility problems, and in some areas of developing countries, it can be as high as 30%. The reasons for both men and women account for 50% each. Epidemiological survey data show that: In the past 50 years, the sperm density of men has almost doubled, and the sperm count of American men has dropped by almost 1.5% every year. It can be seen that testicular spermatogenesis dysfunction is one of the important causes of male infertility. According to reports: more than 80% of male infertile patients have seminal plasma reactive oxygen species (ROS, reactive oxygen species) levels that exceed...

Claims

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Application Information

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IPC IPC(8): A61K31/352A61P15/00
CPCA61K31/352
Inventor 马博张琪王超赵昂张杰朱志铭
Owner NANJING UNIV OF TECH
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