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New application of piceatannol in preventing and treating human cytomegalovirus infection

A technology of human cytomegalovirus and piceatanol, applied in antiviral agents, active ingredients of hydroxyl compounds, etc.

Inactive Publication Date: 2019-03-01
ZHEJIANG HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But the piceatanol involved in the present invention inhibits the p16 induced by HMCV INK4a And the cell senescence caused by the increase of reactive oxygen species (ROS) and then play an anti-HCMV effect, has not been reported yet

Method used

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  • New application of piceatannol in preventing and treating human cytomegalovirus infection
  • New application of piceatannol in preventing and treating human cytomegalovirus infection
  • New application of piceatannol in preventing and treating human cytomegalovirus infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1: The cytotoxicity of the drug is detected by the MTT method to detect the cell viability. The method is as follows: 3000 fibroblasts are inoculated in a 96-well culture plate per well, and after 24 hours of cultivation, different concentrations of the drug are added, and each concentration is set to 6 Parallel wells were set up with a blank control group without drug and a lysis control group without cells. After culturing for 3 days, add 20 μl of MTT (5 mg / ml, prepared with serum-free DMEM culture solution) to each well, at 37°C, 5% CO 2 Cultivate for 4 hours under conditions, carefully suck up the liquid in the wells, add 150 μl DMSO to each well, and shake gently on the shaker for 10 minutes to fully dissolve the crystals, then measure the light absorbance at 570 nm, and calculate the relative cell viability. The cell viability of the blank group was 100%. See the experimental results figure 1 .

Embodiment 2

[0065] Embodiment 2: HCMV inoculation and piceatanol treatment

[0066] HCMV is strictly parasitic and generally only grows on diploid fibroblasts in vitro. Using 28-35PD human embryonic lung diploid fibroblast WI-38 (from ATCC, USA), press 2×10 with 10% FBS medium 4 / cm 2 Inoculate into a culture dish, replace the medium with 0.2% FBS after 24 hours and continue to culture for 48 hours. Through this method of serum starvation, the cells are synchronized at G0 / G1 at this time, which is more conducive to the infection of HCMV. Afterwards, the HCMV virus (Towne virus strain) was inoculated with an inoculation amount of 0.01 MOI (multiplicity of infection), and the culture was continued until the specified time for relevant detection. When detecting the activity of anti-HCMV drugs, a certain concentration of drugs was added to the medium 2 hours in advance, and then HCMV was inoculated to observe the changes in cell morphology ( figure 2 ) and changes in viral gene expression...

Embodiment 3

[0067] Embodiment 3: Western Blot detects the expression of HCMV-related proteins and senescence-related molecules

[0068] Using human embryonic lung diploid fibroblast WI-38, press 2×10 4 / cm 2 Inoculate into a petri dish, replace the medium with 0.2% FBS after 24 hours and continue to cultivate for 48 hours, then inoculate with HCMV virus (Towne virus strain), the inoculation amount is 0.01 MOI (multiplicity of infection). A certain concentration of drugs was added 2 hours before HCMV inoculation, and the cells were collected with a cell scraper after continuing to culture for a specified time. After the cells were lysed with RIPA lysate, the protein was collected, and the protein concentration was measured with a BCA kit to prepare samples for analysis. Using the corresponding antibody, the HCMV immediate early protein IE1 / 2, early protein UL44 and senescence-related molecule p16 were detected by Western Blot method INK4a protein expression levels. See the experimental ...

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Abstract

The invention provides a new application of piceatannol in preparing a medicament for preventing and treating human cytomegalovirus infection. Experiments show that the single treatment of the piceatannol has no obvious toxicity to HCMV in vitro host cells, namely human embryo lung diploid fibroblast WI-38 in the range of 0-20micrometre, the expression of the immediate early protein IE1 / 2 and theearly protein UL44 of HCMV in WI-38 was inhibited in the range of 10-20micrometre by piceatannol, and the amplification of HCMV was inhibited in the range of 10-20micrometre by piceatannol. Piceatannol exerts anti-HCMV activity by inhibiting the HCMV-induced host cell senescence, and the specific mechanism is to inhibit the HCMV-induced high expression of p16INK4a and the increase of active oxygenfree radical level, thereby exerting the anti-HCMV effect.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular to a new application of piceatanol in preventing and treating human cytomegalovirus infection. Background technique [0002] Human cytomegalovirus (HCMV) belongs to the β-subfamily of herpesviruses, which is common in the population and often carries the virus for life after infection. Patients with cytomegalovirus infection are prone to damage to the nervous system, liver, respiratory system and blood system, and even life-threatening in severe cases (Rev Med Virol, 2010, 20:311-326). Epidemiological data show that in most developed countries, about 50% of the adult population has a serological history of previous HCMV infection; in developing countries, HCMV infection is even as high as 90%-100% in older children and adults. my country is a high-incidence area of ​​HCMV infection. The anti-HCMV positive rate in children is 83.2%-87.2%, and it is as high as 95% in adults (International ...

Claims

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Application Information

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IPC IPC(8): A61K31/05A61P31/22
CPCA61K31/05A61P31/22
Inventor 毛根祥王三应苏慧丽徐小刚万晓青暴一众陈莎莎张婧代继桓
Owner ZHEJIANG HOSPITAL
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