Purpose of kinsenoside to preparation of medicines for preventing and treating neurogenic inflammation
A technique of clematis and neuroinflammation, applied in the field of use of clematis in the preparation of medicines for preventing and treating neuroinflammation
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Embodiment 1
[0038]Example 1 Experiment of auroside inhibiting the expression of inflammatory factor TNFα
[0039] The experimental steps are as follows: ① Seed BV2 microglial cells in a 12-well cell culture plate, 100,000 cells / well; ② After 12 hours, pretreat with auroside with a final concentration of 20 μM (the molar concentration of the stock solution is 80 mM) 1h, the cells that were not pretreated with auroglutinin were used as the unstimulated control group; ③The cells pretreated with auroglutinin for 1h were divided into two groups, and lipopolysaccharide (LPS) was added (final concentration: 1 μg / ml). One group was stimulated with LPS for 12 hours, and the other group was stimulated with LPS for 24 hours; ④ When LPS was stimulated to the predetermined time point, the cells were lysed with Trizol reagent, and RNA was extracted; ⑤ After RNA was extracted, 1 μg of total RNA was taken from each sample, and random The primers were reverse-transcribed to obtain cDNA. The cDNA was quan...
Embodiment 2
[0044] Example 2 Experiment of auroside inhibiting the expression of inflammatory factor IL6
[0045] The experimental steps are as follows: ① Seed BV2 microglial cells in a 12-well cell culture plate, 100,000 cells / well; ② After 12 hours, pretreat with auroside with a final concentration of 20 μM (the molar concentration of the stock solution is 80 mM) 1h, the cells that were not pretreated with auroglutin were used as the unstimulated control group; ③The cells that were pretreated with auroglutin for 1h were divided into two groups, and LPS was added respectively (final concentration: 1 μg / ml), and one group LPS was stimulated for 12 hours, and the other group was stimulated with LPS for 24 hours; ④When LPS was stimulated to the predetermined time point, the cells were lysed with Trizol reagent, and RNA was extracted; Reverse transcription to obtain cDNA. The cDNA was quantified by real-time fluorescent quantitative PCR and sybergreen dye method to detect the expression o...
Embodiment 3
[0050] Example 3-Auroside inhibits the expression of the inflammatory factor iNOS
[0051] The experimental steps are as follows: ① seed BV2 microglial cells in a 12-well cell culture plate, 100,000 cells / well; ② 12 hours later, use auroside with a final concentration of 20 μM (dilute with a stock solution with a molar concentration of 80 mM) The cells were pretreated for 1 hour, and the cells that were not pretreated with auroglutin were used as the unstimulated control group; ③ The cells that were pretreated for 1 hour were divided into two groups, and LPS (final concentration: 1 μg / ml) was added respectively. One group was stimulated with LPS for 12 hours, and the other group was stimulated with LPS for 24 hours; ④ When LPS was stimulated to the predetermined time point, the cells were lysed with Trizol reagent and RNA was extracted; Random primers were used for reverse transcription to obtain cDNA. The cDNA was carried out by real-time fluorescence quantitative PCR, and...
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