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Preparation for detecting male reproductive toxicity of environmental chemicals and modeling method

A technology for detecting the environment and male reproduction, applied in the biological field, can solve the problems of time-consuming, rough observation end point, unable to reflect the hidden damage of sperm genetic material, etc., and achieve the effect of shortening the experimental period and accurate evaluation.

Inactive Publication Date: 2010-08-18
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above strategy takes a long time (one month to several months) and the observation end point is relatively rough, which cannot reflect the recessive damage of sperm genetic material

Method used

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  • Preparation for detecting male reproductive toxicity of environmental chemicals and modeling method
  • Preparation for detecting male reproductive toxicity of environmental chemicals and modeling method
  • Preparation for detecting male reproductive toxicity of environmental chemicals and modeling method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] (1) Design and synthesis of the interference sequence of the specific double-stranded siRNA targeting the ogg1 gene.

[0023] ①Design and synthesize specific siRNA interference sequence targeting ogg1 gene. The double-stranded sequence of siRNA-ogg1 sequence design is shown below, and the control group is an irrelevant sequence siRNA double-stranded sequence. The above-mentioned siRNA interference fragments are all designed as a double-stranded combination and chemically modified (mainly with TT modification at the end), which can be stable for at least 3 weeks without being degraded.

[0024] ② siRNA double-stranded sequence targeting ogg1 (probably all or part of the following):

[0025] Candidate duplex 1:

[0026] (RNA)-UUGGGAAGCCAUGAUAAGUGACAUC

[0027] (RNA)-GAUGUCACUUAUCAUGGCUUCCCAA

[0028] Candidate double chain 2:

[0029] (RNA)-AGCUGAAUGAGUCGAGGUCCAAAGG

[0030] (RNA)-CCUUUGGACCUCGACUCAUUCAGCU

[0031] Candidate double chain 3:

[0032] (RNA)-AAACCAAG...

Embodiment 2

[0041] The animal model involved in the present invention was taken, and different concentrations of positive drugs (busulfan 1 mg / kg, 5 mg / kg) were orally poisoned for 4 weeks. Sperm count and sperm DNA damage (TUNEL value) results showed (Table 1, image 3 ): When exposed to a lower dose concentration (1mg / kg), DNA damage was more likely to be detected in the left ogg1 knockdown testis compared with normal exposure (ie, the right control testis). Testicular pathological HE staining ( Figure 4 ) and transmission electron microscope ( Figure 5 ) also showed that: at the same exposure dose (busulfan 1mg / kg), compared with the control group (right testis) in the ogg1 knockdown group (left testis), the knockdown group can more sensitively reflect exogenous compounds testicular damage.

[0042] Table 1: Changes in semen parameters and sperm DNA damage in ogg1 siRNA interference group and normal control group after mice were treated with different concentrations of busulfan fo...

Embodiment 3

[0046] Get the animal model involved in the present invention (left side testis ogg1 knockdown, right side testis control sequence), the same concentration of busulfan (5mg / kg) was orally poisoned, and the reproduction was observed at 0, 2, and 4 weeks respectively. Toxic effects (mainly sperm count, DNA damage). The results show that: compared with the traditional method, the ogg1 knockdown group can detect the DNA damage effect in a shorter time, (Table 2, Figure 6 ).

[0047] Table 2: Changes in the number of sperm and DNA damage in the interference group and the control group for different time periods of busulfan exposure (5mg / kg) in mice

[0048]

[0049] *Compared with 0 weeks (before exposure), P<0.05; #Compared with 0 weeks (before exposure), P<0.01

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Abstract

The invention relates to a double-chain siRNA and application thereof, in particular to a double-chain siRNA of DNA damage repair oggl gene, which is applied to mouse testicle microenvironment knockdown gene expression for improving the sensitivity of a male reproductive system to the damage effect of environmental chemicals so as to provide a method for quickly and sensitively detecting male reproductive toxicity of environmental chemicals. As a new method for detecting the reproductive toxicity of the environmental chemicals, the method can realize sensitive and quick reflection of male reproductive hazard caused by the chemicals, and is favorable for more accurately evaluating the safety of the chemicals, and for shortening the period of pre-clinical reproductive toxicity experiments of medicaments.

Description

1. Technical field [0001] The invention belongs to the field of biotechnology, in particular to a preparation and a model building method for detecting the male reproductive toxicity of environmental chemicals. 2. Background technology [0002] With the rapid development of economy and society, especially modern industry, human beings are exposed to exogenous chemicals such as medicines, pesticides, food additives and various pollutants in the environment. Many of these substances can damage reproductive cells after repeated exposure, causing Reproductive toxicity. In males, the specific manifestations are: damage to the spermatogenic cells at all levels of the testis, which in turn leads to a decline in semen quality and DNA damage in sperm, because sperm DNA damage is not only an important cause of infertility, but also an important factor in the decline in the success rate of assisted reproductive therapy , Moreover, the surviving sperm carrying DNA damage may escape the...

Claims

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Application Information

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IPC IPC(8): C12N15/89A61K49/00A01K67/027
Inventor 王心如顾爱华吉贵祥赵鹏龙燕
Owner NANJING MEDICAL UNIV
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