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Application of inhibitor SB (Sodium Butyrate) 203580 of p (phosphor) -p38

A technology of p-p38 and inhibitors, applied in the direction of anti-toxic agents, anti-inflammatory agents, non-central analgesics, etc., can solve problems such as unclear activation of microglial cells

Inactive Publication Date: 2012-09-12
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, under the pathological conditions of scorpion sting pain, the relationship between the p38 signaling pathway and the activation of microglia is still unclear.
Combined with the existing research, we can't help but wonder whether the activation of p38 in microglia plays a role in central sensitization

Method used

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  • Application of inhibitor SB (Sodium Butyrate) 203580 of p (phosphor) -p38
  • Application of inhibitor SB (Sodium Butyrate) 203580 of p (phosphor) -p38
  • Application of inhibitor SB (Sodium Butyrate) 203580 of p (phosphor) -p38

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Activation time course of microglial cells in rat L4-L5 spinal cord in BmK I pain model

[0022] Rats were subcutaneously injected with BmK I, then deeply anesthetized with pentobarbital sodium (60mg / kg) and then perfused. First, inject 200ml of normal saline from the ascending aorta, and then replace it with 400ml of 4% paraformaldehyde (dissolved in 0.1M PB buffer, pH=7.4). The lumbar spinal cord at the corresponding time point was fixed with the above formaldehyde solution for 12 hours, and then dehydrated with 20% sucrose PB solution (the tissue should sink to the bottom), and finally repeated tissue dehydration with 30% sucrose PB. Then the tissue was frozen into slices (thickness 14um) and pasted on the plectrum for experimentation. The "avidin-biotin-peroxidase (ABC)" method was used for immunosingle labeling. First, the sections were infiltrated with PBS buffer (0.01M, pH=7.4), and then incubated with the primary antibody (48 h, 4°C ), secondary...

Embodiment 2

[0024] Example 2: Time course of expression of p-p38 in L4-L5 spinal cord in BmK I pain model.

[0025] See the introduction in Example 1 for the slice preparation and result analysis methods. Note: The positive result counted in this implementation case is the number of p-p38 positive cells. The primary antibody used in this example is anti-p-p38 rabbit polyclonal antibody (1:200, CST Company), and the secondary antibody used is biotinylated goat anti-rabbit secondary antibody (1:200). see image 3 with Figure 4 .

[0026] The experimental results showed that in the spinal cord of the L4-L5 segment, the number of p-p38 positive cells was gradually activated with the establishment of the BmK I pain model, and reached a peak at 8 hours. This appears to be highly correlated with the activation time course of microglia. The above results suggest that the activated cellular localization of p-p38 may be associated with microglia.

Embodiment 3

[0027] Example 3: Cellular localization of p-p38 in the spinal cord

[0028] see Figure 5. Slice preparation refers to the method in the implementation case 1. In the fluorescence double-labeling test, the slices were first infiltrated with PBS buffer (0.01M, pH=7.4), and then incubated with the first primary antibody (24 h, 4°C) and secondary antibody (2 h, room temperature) before the second incubation. Two kinds of primary antibody incubation and secondary antibody incubation. Between the above incubation processes, rinse with PBS buffer three times, 10 minutes each time. Sections were observed under a microscope (Leica) after using glycerol slices. Immunofluorescence photos were synthesized with Image J image processing software. The primary antibodies used in this example include: anti-OX-42 mouse monoclonal antibody (CD11b, microglial cell marker, 1:50, Santa Cruz Company), anti-GFAP mouse monoclonal antibody (GFAP, star glial cell marker, 1:300, CST company),...

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Abstract

The invention aims to provide application of an inhibitor SB (Sodium Butyrate) 203580 of p (phosphor)-p38. Activation of phosphor-p38 mediated microglial cells contributes to pain correlated responses of a pain model of BmKI (Biomedical Knowledge Integration); the specific inhibitor SB 203580 of the p-p38 can inhibit the microglial cell and thus relieving the pain correlated responses in the pain model of the BmKI. The invention provides a theoretical basis for pain central sensitization induced by peripheral damages, a study carrier for studying cellular and molecular mechanisms and treatment strategies of nociception and an experimental basis for studying novel drugs for relieving the pain on clinic are provided. The SB 203580 has a prospect of becoming a tool medicine applied in studying correlated pain mechanism of p38 signal path and then has a potential prospect of developing drugs for treating and relieving pathological pain and maintaining corresponding drugs on clinic.

Description

technical field [0001] The present invention relates to the application of p-p38 inhibitor SB203580. technical background [0002] The sting of the East Asian pincer scorpion (Scorpion Buthus martensi Karsch, BmK) releases venom that causes severe pain, skin edema, and a burning sensation. As the main neurotoxic polypeptide component of the venom of the East Asian pincer scorpion, BmK I is considered to be the main factor of BmK venom-induced inflammatory pain behavior. Plantar injection of BmK I induced hind paw edema, spontaneous pain lasting 2 hours, bilateral mechanical allodynia lasting 7 days and thermal hyperalgesia lasting 3 days. [0003] Glial cells were originally thought to play a supportive role in the central nervous system, but accumulating evidence suggests that the communication dialogue between glial cells and neurons plays a role in the development of different types of neurological diseases and the occurrence of pain. important role. Recent studies hav...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/4439A61P29/00A61P39/02
Inventor 吉永华牛青山姜峰华黎明傅晋
Owner SHANGHAI UNIV
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