Application of asparagine synthetase activator
A technology of asparagine and synthetase, which is applied in the field of medicine, can solve the problems of patient burden, operation cost, large number of people, and unsatisfactory effect, so as to improve cell activity, broaden prevention and treatment ideas, and avoid lens turbidity. Effect
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Embodiment 1
[0031] This embodiment is used to verify that the expression level of ASNS in the lens of naphthalene cataract rats increases, specifically including:
[0032] After the cataract rat lens was treated with naphthalene for 2 weeks, the lens appeared cloudy in different degrees. The present invention classifies according to any naphthalene cataract grading system ( figure 1 A-D) of different degrees of cataract classification. The turbidity of lens treated with naphthalene reached 2 points in 2 weeks (n=6, as figure 1 Shown in F), and reached 4 points in 4 weeks (n=6, such as figure 1 Shown in G), the lens turbidity of the non-treatment group (normal) and the mineral oil treatment group (control) was lower than 2 points. In order to explore the mechanism of naphthalene-induced lens turbidity, the lenses of the naphthalene cataract group and the control group were collected after 1 week of treatment, and RNA was prepared for gene array analysis. Control group increased (n=6, p...
Embodiment 2
[0034] This example is used to verify the oxidation of 1,2-DHN and the increased expression of ASNS under the action of naphthalene metabolite 1,2-DHN, specifically including:
[0035] Using the NADP / NADPH kit, it was found that when different substrates were co-incubated with lens homogenates, the NADPH production of the 1,2-DHN mixture increased over time, but not the N ADPH mixture, which indicated that The lens can promote the oxidation of 1,2-DHN, and it is an autoxidative process (n=3, such as figure 2 shown in A), remove the lens homogenate in the 1,2-DHN oxidation reaction system (n=3, such as figure 2 As shown in D), it was found that the amount of NADPH generated in the lens homogenate system was higher than that in the system without homogenate. NADPH time effect (n=3, such as figure 2 Shown in B and C), it was confirmed that the lens was incubated with 50 μM 1.2-DHN or 50 μM NQ solution for 6 hours, and the lens was collected for western blot detection. Compar...
Embodiment 3
[0037] This embodiment is used to verify that ASNS reduces the decline caused by 1,2-DHN on cell viability and slows down its oxidation rate, specifically including:
[0038] like image 3 As shown, ASNS was overexpressed and knocked down in the SRA01 / 04 cell line with lentivirus respectively, wherein, such as image 3 As shown in A, the relative expression of ASNS at the mRNA level in the SRA01 / 04 cell line overexpressing ASNS and knocking down ASNS was detected by real-time fluorescent quantitative PCR experiment; image 3 B and image 3 As shown in C, the relative expression of ASNS at the protein level in the SRA01 / 04 cell lines overexpressing ASNS and knocking down ASNS was detected by Western blot. In addition, the cell viability of ASNS overexpression (OE), ASNS knockdown (shASNS) and control group (CTL) was detected by MTT method. Compared with the control group, the cell viability of ASNS knockdown group was significantly decreased (n=3, p Figure 4 As shown in A), t...
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