Application of cytokine activin C in treatment of neuropathic pain
A cytokine and activin technology, applied in the field of biomedicine, can solve problems such as unclear role
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Embodiment 1
[0029] Embodiment one, experiment preparation
[0030] 1. Source of experimental animals:
[0031] In the present invention, animals and animal studies were performed in accordance with the recommendations set forth in the Arrival Guidelines and the British Journal of Pharmacology. All experiments were performed in accordance with the guidelines of the International Association for Pain Research and approved by the Experimental Animal Use Committee of Nantong University. To avoid confounding effects of estrous cycle on pain processing, only male animals were used in this study.
[0032] 1-1. Adult male SD rats (200-250 g), young male SD rats (90-110 g) and 8-12-week-old male C57 / BL mice were provided by Shanghai SLAC Experimental Animal Company.
[0033] 1-2. TRPV1 knockout (KO) mice were purchased from Jackson Laboratory in Bar Harbor, Missouri, USA, and crossed with adult female C57BL / 6J wild-type (WT) mice.
[0034] 1-3. Conduct behavioral tests with male TRPV1 KO and WT...
Embodiment 2
[0045] Embodiment two, experimental method
[0046] 1. Sciatic nerve cut, autotomy scoring and dorsal root ganglion isolation
[0047] The sciatic nerve of adult rats was deeply anesthetized with 3% pentobarbital sodium (80 mg / kg). The left sciatic nerve was transected in the mid-thigh, cutting the nerve (1.0 cm). Experimental animals were anesthetized with 0.1 M phosphate cold buffer solution (PBS, pH 7.4), perfused through the left ventricle, and L4~5DRG were rapidly isolated under ribonuclease-free conditions at 0.5, 1, 2, 7, 14, and 28 days after operation , and served as a complete control on the 0th day after operation (n = 9 / time point).
[0048] For axotomy scoring, each rat was scored daily for the level of autotomy for four weeks, which was modified from the prior art in a double-blind manner. The following rule was used: 0.1 points for the loss of 1 nail or 1 bleeding finger, and 1 point for each distal half-finger attacked. Then, add one point for each proximal...
Embodiment 3
[0079] Example 3. Up-regulation of the expression of activin C in the lumbar dorsal root ganglion of rats with peripheral nerve injury
[0080] In order to determine the dose of activin C injected intrathecally into adult rats, the present inventors firstly studied the expression of this cytokine in the lumbar dorsal root ganglion of chronic neuropathic pain rats.
[0081] In this example, rat sciatic nerve was cut, which is a simple and feasible peripheral nerve injury model. A custom microarray was used (Liu et al., 2012).
[0082] figure 1 In the present invention, after the rat sciatic nerve is cut off, the expression of activin βC subunit and activin C protein in the lumbar 4-5 dorsal root ganglion (DRG) is up-regulated. in, figure 1 a is a graph of custom microarray results, N = 6 separate experiments; figure 1 b is the real-time RT-PCR analysis graph, N = 6 separate experiments; figure 1 c is the western blot display, activin C at 25 kDa is composed of βC and βC su...
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