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Novel applications of exogenous nerve growth factor (NGF)

An exogenous, drug-based technology that is applied in the field of biomedicine to achieve potential therapeutic effects, promote development, and reduce damage

Inactive Publication Date: 2014-07-23
AFFILIATED HOSPITAL OF NANTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether exogenous NGF affects the expression changes of endogenous NGF and HoxA10 in the rat cryptorchidism model has not been reported yet

Method used

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  • Novel applications of exogenous nerve growth factor (NGF)
  • Novel applications of exogenous nerve growth factor (NGF)
  • Novel applications of exogenous nerve growth factor (NGF)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] model creation, such as Image 6 shown.

[0025] Obtain about 30 SD male mice aged 21 days (the testes of SD mice have not yet descended at this time, and the descent of SD mice’s testes usually occurs at the age of 20-30 days), anesthetized by intraperitoneal injection of pentobarbital sodium, and cut open the abdominal cavity in the middle of the lower abdomen , cut off the distal end of the testicular guide band, suture the testicular albuginea, fix the testis on the corresponding peritoneum, close the abdominal cavity, and complete the establishment of the cryptorchidism model. The contralateral testis served as the control group.

[0026] After the model was established, they were divided into 4 groups, 6 rats in each group, high and low injection dose groups of NGF (HN group, LN group), HCG injection group (HC group), and negative control group (NC group). From the 2nd day after operation, the HN group was injected intramuscularly with mouse NGF9000IU each time,...

Embodiment 2

[0027] Embodiment 2 one-step real-time quantitative PCR

[0028] Extraction of total RNA from tissues: 50-100 mg of testicular tissue was taken using a Trizol kit (Invitrogen, USA), total RNA was extracted according to the instructions, and stored in a -70°C refrigerator for later use. To avoid possible genomic DNA contamination, RNase-free DNase (Promega, Madison, WI, USA) was used for all RNA samples. The OD of each sample was measured by UV spectrophotometer 260 / OD 280 (R) value to identify RNA purity and integrity. The R value is shown in Table 1, and 1.8<R<2.0 confirms that the RNA extracted by the Trizol method is of good purity and can be used in subsequent experiments.

[0029] According to the standard protocol, the Rotor-Gene3000 real-time DNA analysis system (Corbett Research, Sydney, Australia) was used, and the SensiMixTM One-Step kit (Quantace, UK) was used to complete one-step fluorescent real-time quantitative PCR (one-step qPCR). Gene real-time analysis s...

Embodiment 3

[0040] Example 3 Digoxigenin-labeled anti-β-NGF mRNA in situ hybridization histochemical method

[0041] Dig-NGF RNA probe preparation: Trizol reagent was used to extract the total RNA in the brain tissue of S-D rats, and the cDNA fragment of β-NGF was extracted by RT-PCR. The upstream and downstream primers used in RT-PCR are

[0042] 5'-GATCGGCGTACAGGCAGAAT-3' and 5-GGCTCGGCACTTGGTCTCAA-3'. Through the T4 ligase reaction, the PCR product was cloned into the pGEM-T vector to construct the NGF / pGEM-T plasmid, and the constructed NGF / pGEM-T plasmid was transformed into DH5a competent cells, and after screening, extraction and purification, Further, after enzyme digestion identification and sequencing verification, the amplified plasmids were digested with restriction endonucleases SalI and ApaI, respectively, to linearize the plasmids, and the digested fragments were recovered respectively as templates for sense or antisense NGF probes. Using Sp6 and T7 transcriptases, accord...

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PUM

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Abstract

The invention discloses novel applications of exogenous nerve growth factor (NGF). Establishment of rat cryptorchism models is realized; control groups are obtained via exogenous injection of NGF; real-time quantitative PCR, in-situ hybridization, immunofluorescence, immunohistochemistry, and transmission electron microscopy are adopted so as to observe morphological change of cryptorchidism tissue, and expression change of NGF, HoxA10mRNA and protein in testicular tissue, and confirm that NGF is capable of promoting formation of testicular sperms. So that, NGF can be widely used for preparation of medicines which are used for promoting sperm generation, promoting cryptorchism model testicular tissue development, reducing cryptorchidism tissue damage, and treating cryptorchidism.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a new application of exogenous NGF. Background technique [0002] Cryptorchidism is undescended testis, which is a malformation of the genitourinary system caused by the failure of one or both testes to enter the scrotum. The incidence rate of newborn boys is 2%-4%, and the incidence rate of premature infants is even higher. Under normal circumstances, before the fetus is born, the testicles first develop in the abdominal cavity, and then gradually descend to the scrotum under the pulling effect of the belt. The unsuccessful descent of the testes into the scrotum was mainly due to the failure to undergo histological changes to the testes. In addition, cryptorchidism is one of the risk factors for male infertility and testicular tumors. Studies have shown that children with cryptorchidism without timely and effective treatment have an irreversible impact on their future fert...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/18A61P15/08A61P15/00
Inventor 咸华黄剑飞咸云刘莉莉
Owner AFFILIATED HOSPITAL OF NANTONG UNIV
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