Layered bimetal hydroxide / selenium nanocomposite and application thereof
A layered bimetallic, hydroxide technology, applied in the direction of drug combination, sulfur/selenium/tellurium active ingredients, genetic material components, etc., can solve the problems of unsatisfactory cellular absorption effect of nano-selenium, limited clinical application, etc. Promote orderly release, efficient gene silencing effect, inhibit cell proliferation effect
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Embodiment 1
[0031] Example 1 Preparation of layered double hydroxide / selenium nanocomposite SeLDH
[0032] Mg(NO 3 ) 2 (3mmol) and Al(NO 3 ) 3 (1mmol) was dissolved in 10mL deionized water, and quickly added to 40mL NaOH solution (6mmol); the mixture was stirred at room temperature for 10min, and centrifuged (5min, 4500min -1 ) to collect the resulting slurry; wash twice with 40mL deionized water, centrifuge and resuspend in 50mL deionized water; transfer the resulting inhomogeneous suspension to a reactor and heat at 100°C for 16h to obtain a transparent, uniform The layered double metal hydroxide nano sol; the sol contains nearly 4mg / mL uniformly dispersed MgAl-LDH nanoparticles;
[0033] A certain amount of Na 2 SeO 3 The solution (1mL, 1M) was added dropwise to the hydrated solution of MgAl-LDH nanoparticles (2mL, 0.025M), and the stirring was continued for 30min; then, L-cysteine (L-Cys) was added to the mixture, and the Stir until the reaction solution becomes deep red; the...
Embodiment 2
[0034] Example 2 Preparation of layered double hydroxide / selenium nanocomposite SeLDH-siRNA
[0035] Incubate 100nM siRNA and different masses of SeLDH in phosphate buffer (PBS, pH=7.4) for 30min at room temperature (25°C) to obtain SeLDH-siRNA with different mass ratios, wherein the mass ratios of SeLDH and siRNA were respectively 1:1, 2:1, 4:1, 8:1 and 16:1;
[0036] The siRNA is selected from PsiRNA, TsiRNA or MocksiRNA; SeLDH-PsiRNA, SeLDH-TsiRNA or SeLDH-MocksiRNA is obtained.
[0037] The siRNA is selected from the mixture of PsiRNA and TsiRNA to obtain SeLDH-pooled siRNAs, wherein the molar ratio of PsiRNA and TsiRNA is 1:1.
Embodiment 3
[0040] Example 3 SeLDH gene load test
[0041] Agarose gel electrophoresis was used to detect the ability of SeLDH and MgAl-LDH to bind siRNA. Different mass ratios (1:1 to 16:1) of SeLDH / siRNA or LDH / siRNA were prepared in PBS, and electrophoresed by TBE buffer containing 1% agarose for 1 h. The gel was stained with 1 μg / ml ethidium bromide and photographed under a UV imager, see figure 2 . figure 2 A shows that when the mass ratio exceeds 4:1, the migration of LDH-bound siRNA is completely blocked, indicating that LDH can fully bind siRNA and protect it from degradation during electrophoresis. The binding ability of SeLDH is relatively weak, figure 2 It is shown in B that it is not completely blocked until the mass ratio is 8:1. Compared with LDH, SeLDH does not bind siRNA so tightly that it is not easy to release, thus exerting a greater biological effect.
[0042] To detect the release of siRNA, SeLDH / siRNA (16:1) and LDH / siRNA (16:1) were incubated in phosphate bu...
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