阅读说明：本技术 一种雷公藤内酯酮减轻cfa诱导的小鼠痛敏反应的方法 (Method for reducing CFA (circulating fluid array) -induced nociceptive response of mice by triptonide ) 是由 董玉林 姜保春 于 2019-11-15 设计创作，主要内容包括：本发明公开了一种雷公藤内酯酮减轻CFA诱导的小鼠痛敏反应的方法,步骤一：小鼠单侧后足注射CFA；步骤二：注射CFA后静脉注射TPN；步骤三：检测CFA注射后背根神经节DRG及内神经元AKT的表达变化；步骤四：检测CFA注射后DRG神经元肿瘤坏死因子α、白细胞介素1β和白细胞介素6mRNA的表达变化；步骤五：使用AKT抑制剂预处理,检测CFA引起的机械性痛觉过敏和热痛觉过敏的变化,以及TNF-α、IL-1β和IL-6等促炎性细胞因子mRNA的表达变化,通过本发明方法,可以得到这些结果为TPN的抗伤害和抗炎作用提供了直接证据,提示TPN在炎症性疼痛治疗中的新应用。(The invention discloses a method for reducing mouse nociceptive response induced by CFA (circulating fluid dynamics) by triptonide, which comprises the following steps: mice were injected with CFA on one hind paw; step two: TPN was injected intravenously after CFA injection; step three: detecting expression changes of DRG and AKT of internal neurons of dorsal root ganglia after CFA injection; step four: detecting the expression changes of DRG neuron tumor necrosis factor alpha, interleukin 1 beta and interleukin 6mRNA after CFA injection; step five: the method of the invention can obtain the results which provide direct evidence for the anti-injury and anti-inflammatory effects of TPN and suggest a new application of TPN in the treatment of inflammatory pain.)

1. A method for reducing CFA-induced nociceptive response of mice by triptonide is characterized in that: the method comprises the following steps:

the method comprises the following steps: mice were injected with CFA on one hind paw, followed by paw swelling and persistent pain hypersensitivity;

step two: injecting TPN intravenously after CFA injection, and detecting the change of paw swelling degree, mechanical hyperalgesia, thermal hyperalgesia and the like;

step three: detecting the expression changes of DRG and AKT of internal neurons of dorsal root ganglia after CFA injection and the change of AKT after TPN whole body treatment;

step four: detecting the expression changes of DRG neuron tumor necrosis factor alpha, interleukin 1 beta and interleukin 6mRNA after CFA injection and the changes of the factors after TPN systemic treatment;

step five: the AKT inhibitor is used for pretreatment, and changes of mechanical hyperalgesia and thermal hyperalgesia caused by CFA and expression changes of proinflammatory cytokine mRNA such as TNF-alpha, IL-1 beta and IL-6 are detected.

2. The method of claim 1, wherein the triptonide reduces the CFA-induced nociceptive response in the mouse comprises: and in the third step, expression changes of DRG (dorsal root ganglion) and AKT (intrinsic killer T) of the internal neurons after CFA (circulating fluid antigen) injection are detected by adopting western blot and immunofluorescence.

3. The method of claim 1, wherein the triptonide reduces the CFA-induced nociceptive response in the mouse comprises: and in the fourth step, expression changes of DRG neuron tumor necrosis factor alpha, interleukin 1 beta and interleukin 6mRNA after CFA injection are detected by adopting western blot and immunofluorescence.

4. The method of claim 1, wherein the triptonide reduces the CFA-induced nociceptive response in the mouse comprises: and step five, detecting the change of mechanical hyperalgesia and thermal hyperalgesia caused by CFA by adopting western blot and immunofluorescence.

Technical Field

The invention relates to the technical field of nociceptive response, in particular to a method for reducing mouse nociceptive response induced by CFA (circulating fluid dynamics) by triptonide.

Background

Triptonide (TPN) is the major component of Tripterygium wilfordii, and recent studies have shown that the PI3K/AKT/mTOR pathway plays an important role in the pathogenesis of chronic pain. Complete Freund's Adjuvant (CFA) is a commonly used inflammatory agent and is currently the most commonly used adjuvant in animal experiments.

Since the nociceptive response provides direct evidence for antinociceptive and anti-inflammatory effects and suggests that TPN plays a role in the treatment of inflammatory pain, there is no experimental approach for TPN in inflammatory pain, and thus an improved technique is needed to solve the problem in the prior art.

Disclosure of Invention

The invention aims to provide a method for reducing CFA (circulating fluid dynamics) -induced nociceptive response of mice by triptonide, which aims to solve the problems in the background technology.

In order to achieve the purpose, the invention provides the following technical scheme: a method for reducing CFA-induced nociceptive response in mice by triptonide comprises the following steps:

the method comprises the following steps: mice were injected with CFA on one hind paw, followed by paw swelling and persistent pain hypersensitivity;

step two: injecting TPN intravenously after CFA injection, and detecting the change of paw swelling degree, mechanical hyperalgesia, thermal hyperalgesia and the like;

step three: detecting the expression changes of DRG and AKT of internal neurons of dorsal root ganglia after CFA injection and the change of AKT after TPN whole body treatment;

step four: detecting the expression changes of DRG neuron tumor necrosis factor alpha, interleukin 1 beta and interleukin 6mRNA after CFA injection and the changes of the factors after TPN systemic treatment;

step five: the AKT inhibitor is used for pretreatment, and changes of mechanical hyperalgesia and thermal hyperalgesia caused by CFA and expression changes of proinflammatory cytokine mRNA such as TNF-alpha, IL-1 beta and IL-6 are detected.

Preferably, in the third step, western blot and immunofluorescence are adopted to detect expression changes of DRG and AKT of dorsal root ganglia after CFA injection.

Preferably, in the fourth step, expression changes of DRG neuron tumor necrosis factor alpha, interleukin 1 beta and interleukin 6mRNA after CFA injection are detected by adopting western blot and immunofluorescence.

Preferably, in the fifth step, the change of mechanical hyperalgesia and thermal hyperalgesia caused by CFA is detected by using western blot and immunofluorescence.

Compared with the prior art, the invention has the beneficial effects that:

these results are obtained by the method of the invention and provide direct evidence for the antinociceptive and anti-inflammatory effects of TPN, suggesting a new use of TPN in the treatment of inflammatory pain.

Drawings

Figure 1 is a graphical representation of the paw swelling (a), mechanical hyperalgesia (B) and thermal hyperalgesia (C) induced by CFA injected intravenously with TPN after CFA injection.

FIG. 2 is a schematic diagram of the expression change of neuronal AKT in the western blot and immunofluorescence detection CFA injection Dorsal Root Ganglion (DRG) and the change of AKT after TPN systemic treatment.

FIG. 3 is a graph showing the expression curves of TNF- α (A), IL-1 β (B) and IL-6 (C) mRNAs in DRG neurons induced by CFA inhibition by TPN.

FIG. 4 is a graphical representation of AKT inhibitor pretreatment to reduce CFA-induced mechanical hyperalgesia (A) and thermal hyperalgesia (B), and to reduce the expression of TNF- α (C), IL-1 β (D) and IL-6 (E).

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The invention provides a technical scheme that: a method for reducing CFA-induced nociceptive response in mice by triptonide comprises the following steps:

the method comprises the following steps: unilateral posterior foot injection of Complete Freund's Adjuvant (CFA) in mice can cause foot swelling and persistent pain hypersensitivity;

step two: injecting TPN intravenously after CFA injection, and detecting the change of paw swelling degree, mechanical hyperalgesia, thermal hyperalgesia and the like;

step three: adopting western blot and immunofluorescence to detect the expression change of DRG and AKT of internal neuron after CFA injection, and the change of AKT after TPN whole body treatment;

step four: detecting the expression changes of mRNA of DRG neuron tumor necrosis factor alpha (TNF-alpha), interleukin 1 beta (IL-1 beta) and interleukin 6 (IL-6) after CFA injection and the changes of the factors after TPN systemic treatment;

step five: the AKT inhibitor is used for pretreatment, and changes of mechanical hyperalgesia and thermal hyperalgesia caused by CFA and expression changes of proinflammatory cytokine mRNA such as TNF-alpha, IL-1 beta and IL-6 are detected.