Application of beta-sitosterol in treatment of cerebral arterial thrombosis
A technology for ischemic stroke and ischemic stroke, applied in the field of medicine, can solve problems such as no reports of β-sitosterol, and achieve the effects of improving neurological behavior, inhibiting accumulation, and inhibiting neuron apoptosis
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Embodiment 1
[0018] Example 1: Effects of β-sitosterol on intracellular cholesterol transport and transport receptor NPC1L1 in neurons
[0019] 1. Experimental method
[0020] (1) Primary culture of neurons: Pregnant mice (E14-15d) were killed by excessive anesthesia outside the ultra-clean bench, placed in a dissection tray, sprayed with 75% alcohol for disinfection, and the abdomen was quickly cut open to remove the fetal beads, placed in a container In a glass dish with pre-cooled (ice bath) D-Hank's equilibration solution. Single embryos were isolated under sterile conditions, and the braincases were stripped and the bilateral brains were quickly removed (operated on ice). The vascular membrane was peeled off under a dissecting microscope, and the cerebral cortex tissue was taken, placed in pre-cooled D-Hank's solution, cut into small pieces of 1-2 mm3, transferred to 0.25% trypsin solution, and digested at 37 °C for 20 min. Digestion was terminated by rinsing the tissue 3 times with...
Embodiment 2
[0027] Example 2: Effects of β-sitosterol on apoptosis genes in neurons
[0028] 1. Experimental method:
[0029] (1) Western blot detection of total protein level: same as above
[0030] 2. Experimental results
[0031] like figure 2 As shown, compared with the normal group, the protein expressions of apoptosis genes caspase3, caspase 9 and Bax in the cortex of MCAO model mice were significantly increased; meanwhile, endoplasmic reticulum stress marker proteins GRP78 / Bip and caspase12 were detected. Significantly higher expression; and the β-sitosterol-treated group reversed this effect. This indicates that β-sitosterol can inhibit the apoptosis induced by endoplasmic reticulum stress in ischemic stroke neurons.
Embodiment 3
[0032] Example 3: Effects of β-sitosterol on neurological behavior and cerebral infarction in animal model mice
[0033] 1. Experimental method:
[0034] (1) Determination of cerebral infarct size by triphenyltetrazolium chloride (TTC) staining: 24 hours after reperfusion, animals in each group were intraperitoneally injected with 10% chloral hydrate. Quick-freeze for 15 min, and cut 5 coronal slices with a thickness of 2 mm. Be careful when slicing, as brain tissue tends to stick to the blade, and to prevent loss, the blade is first dipped in a 2% TTC solution. The cut brain slices were quickly placed in TTC solution for staining, and the brain slices were pressed with a cover glass to make the brain tissue flat, incubated in a 7°C incubator in the dark for 30 minutes, fixed with 4% paraformaldehyde, and the color changes were continuously observed. When the surface of the slice begins to turn pink, invert the slice. The stained surface has viable brain tissue; the infarct...
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