Proteinase inhibitor nervous cell protecting function and application
A protease inhibitor and excipient technology, applied in the field of Kunitz-type protease inhibitors
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Embodiment 1
[0051] Example 1: Preparation of rat model of acute cerebral infarction, animal grouping, experimental method and result evaluation method
[0052] 1. Preparation of animal (rat) cerebral ischemia-reperfusion injury model: refer to Zealonga suture method (LongaEZ, Weinstein PR, Carlson S, et al. Reversible middle cerebral arteryocclusion without craniectomy in rats. Stroke, 1989, 20 (1) : 8491.) To prepare the focal cerebral ischemia-reperfusion model of right middle cerebral artery occlusion (MCAO) in rats. A 6.0 cardiac surgery single-strand nylon thread (length 4cm, diameter 0.235mm) was used as the tethered thread. One end of the thread was blunted with fire to make the diameter of the top 0.26-0.32mm, and a mark was made at a distance of 18mm from the end. After cleaning with alcohol, put it in 1% sodium heparin for later use.
[0053] Rats were fasted without food or water for 12 hours before the operation. Anesthetized by intraperitoneal injection of 10% chloral hydra...
Embodiment 2
[0076] Example 2: Effects of rhKD / APP on neurological function in rats with cerebral ischemia-reperfusion injury:
[0077] Rat cerebral ischemia model was prepared by MCAO method. After 3 hours of ischemia and 24 hours of reperfusion, obvious neurological deficits were observed in the model group, and the mortality rate was as high as 26.5%. Compared with the model group, the neurological deficits of animals given the neuroprotective drug Nimodi pine (positive control group) were significantly improved (P<0.05), but the mortality rate was still high (P<0.05). Compared with the model group, the neurological deficits of animals in the rhKD / APP low, middle and high dose groups were also significantly improved (P<0.01); and with the increase of the dose, the neurological function score improved significantly. However, there was no statistically significant difference among the three dose groups. Compared with the model group, the mortality rate of animals treated with low, mediu...
Embodiment 3
[0081] Example 3: Effects of rhKD / APP on cerebral infarction size in acute cerebral infarction model:
[0082]TTC is a water-soluble salt, which can react with the dehydrogenase in the mitochondria of cells to produce dark red fat-soluble substances. Dead cells do not develop color due to the inactivation of mitochondrial dehydrogenase. method. In this study, the TTC staining method was used to observe and calculate the cerebral infarction range of each group after 3 hours of cerebral ischemia and 24 hours of reperfusion (see Table 2). Compared with the model group, the cerebral infarction area in the middle and high dose rhKD / APP group and Nimodipine group was significantly reduced (p<0.05). The size of cerebral infarction in the high-dose rhKD / APP group was significantly lower than that in the model group (P<0.01), and there was an obvious dose-related relationship (see Figure 3).
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