Application of growth differentiation factor 11 in preparation of medicine for treating ischemic stroke
An ischemic stroke, growth differentiation factor technology, applied in cardiovascular system diseases, drug combinations, medical preparations containing active ingredients, etc. Defect score, the effect of promoting cell proliferation in brain region
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Embodiment 1
[0045] Example 1: Therapeutic effect of GDF11 exogenous recombinant protein in cerebral ischemic injury after ischemia
[0046] 1. Establishment of focal cerebral ischemia-reperfusion model in rats with suture method (Middle cerebral arteryocclusion, MCAO model):
[0047] Sprague-Dawley rats with a body weight of 300±5g (purchased from Beijing Weitong Lihua Company) were taken, and after intraperitoneal injection of 10% chloral hydrate (0.3-0.4ml / 100g body weight), they were fixed on the operating table in supine position, and A midline incision was made, the subcutaneous tissue was bluntly dissected, the left common carotid artery (CCA) was dissected and ligated, and the bifurcation of the internal carotid artery (ICA) and external carotid artery (ECA) was dissected upward along the CCA, and the proximal end of the ECA was ligated. Cut a small opening at the intersection of the common carotid artery, and insert a 0.25mm-diameter tethered suture into the ICA at about 20.5±0.5m...
Embodiment 2
[0062] Example 2: The preventive effect of overexpression GDF11 preconditioning in cerebral ischemic injury
[0063] 1. Detection of GDF11 overexpression virus efficiency:
[0064] In order to establish a state of long-term high expression of GDF11 in the brain and study its preventive effect on cerebral ischemic injury, the inventors first constructed a GDF11 overexpression plasmid packaged into a lentivirus with green fluorescent protein (GFP), and The overexpression efficiency of the virus was detected by in vivo and in vitro experiments. 293 cells were recovered and cultured, and when the cell density reached about 80%, 5 μg of negative control virus and GDF11 overexpression virus were added respectively, and fluorescence real-time quantitative PCR was performed 48 hours later. The results showed that the mRNA level of GDF11 overexpression virus was higher than that of the blank virus group 6 times (control group: 1±0; GDF11 overexpression group: 6.08±0.14). The inventor...
Embodiment 3
[0075] Example 3: The molecular mechanism of GDF11 in the treatment and prevention of cerebral ischemic injury
[0076] The cortical ischemic penumbra tissues of cerebral ischemic animals were collected separately, and the protein was extracted for Western Blot. The results showed that after injection of exogenous recombinant GDF11 protein, pSmad2 / 3 in the brain (control group: 1±0.09; GDF11 protein group: 2.0± 0.24; Figure 14 A and B) significantly increased expression. It can be seen from the results that the recombinant GDF11 protein participates in the treatment process of cerebral ischemic injury by regulating Smad2 / 3.
[0077] The tissue protein in the ischemic penumbra of the cortex was extracted and detected by Western Blot. It was found that after injection of the GDF11 overexpression virus, pSmad2 / 3 in the rat brain (control group: 1±0.66; GDF11 overexpression group: 1.57±0.10; Figure 14 The amounts of A and C) are significantly increased. It can be seen from th...
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