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Application of Gansu Huangcao compound in the preparation of medicaments for the treatment of atherosclerosis

A technique for atherosclerosis and the extract of C. chinensis, which is applied in the field of treating or relieving diseases of C. chinensis and its compounds, and can solve problems such as untapped application potential

Active Publication Date: 2021-06-22
MACAU UNIV OF SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the yellow grass has been used in folk prescriptions for protecting the liver, its mechanism of action and its application potential in the treatment of other diseases have yet to be explored

Method used

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  • Application of Gansu Huangcao compound in the preparation of medicaments for the treatment of atherosclerosis
  • Application of Gansu Huangcao compound in the preparation of medicaments for the treatment of atherosclerosis
  • Application of Gansu Huangcao compound in the preparation of medicaments for the treatment of atherosclerosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1: Induction of autophagy by the total extract of C. chinensis

[0067] 1.1 Western blotting to detect the expression of autophagy-related proteins in Hela cells

[0068] Add 15-30 μg / mL drugs to the cultured Hela cells and incubate overnight: the extracts of the flowers of C. chinensis, the extracts of the leaves of C. sylvestris and the stems of C. sylvestris. Rapamycin (Rapa) was used as the autophagy positive control . After drug treatment, RIPA lysate (Cell Signaling Technologies Inc. Beverly, MA, USA) was added to each well in an amount of 70 μl / well to lyse HeLa cells, and Bradford reagent (Bio-Rad, Hercules, CA, USA) was used to measure protein concentration . Add an equal amount of protein to each well of the SDS-PAGE electrophoresis gel, electrophoresis at 300mA for 2 hours, and transfer the protein on the SDS-PAGE gel to the PVDF membrane. The PVDF membrane was blocked with 5% skimmed milk powder for 60 minutes and washed three times with TBST (1X)...

Embodiment 2

[0088] Example 2: Compounds TA, PG, TB, PG1 induce autophagy

[0089] 2.1 Determination of GFP-LC3 green fluorescent particles

[0090] The U87 cell line with GFP-LC3 stably transfected was transfected and established as in Example 1. After drug treatment, GFP-LC3 green fluorescent particles were detected.

[0091] 2.2 Western blotting

[0092] As described in Example 1, 4-8 μM compounds TA, PG, TB, and PG1 were used to treat HUVEC cells, and the no-drug group was used as a blank control, and rapamycin (Rapa) was used as a positive control. After drug treatment, the HUVEC cells in each group were lysed by the method described above, SDS-PAGE protein electrophoresis, and the protein was transferred to PVDF membrane. After incubation with antibodies, ECL western blot detection solution (Invitrogen, Paisley, Scotland, UK) was used to expose and develop protein bands.

[0093] Result analysis: The autophagy activity of the above four compounds was analyzed by immunofluorescence...

Embodiment 3

[0132] Example 3: TA can reduce the level of oxidative stress in the aorta of the ApoE gene knockout atherosclerosis mouse model (ApoE-KO mice)

[0133] A 2-month-old ApoE gene knockout atherosclerosis mouse (ApoE-KO) model was established, C57BL / 6J wild-type mice were used as the control group, and each group was given or not given intraperitoneal injection of TA (0.1mg / kg per day) for one month To evaluate the role of TA in the process of early aortic atherosclerosis in ApoE-KO knockout mice.

[0134] 3.1 Measurement of ROS in aortic vessels

[0135] 2',7'-dichlorofluorescin diacetate (2',7'-dichlorofluorescin diacetate, DCFH-DA) was used to measure the intracellular ROS content. Aortic vascular cells of ApoE-KO mice and C57BL / 6J mice were collected by Nylonsieve mesh combined with trypsin method. Add 10 μM DCFH-DA to the above cells, incubate together at 37°C for 30 minutes, and then wash three times with serum-free medium. Intracellular esterase hydrolyzes DCFH-DA into ...

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Abstract

The invention discloses the application of the extract of the traditional Chinese medicine C. chinensis in the preparation of a drug for treating atherosclerosis or alleviating Alzheimer's disease, which works by inducing autophagy, and discloses four key compounds in the extract. The extracts of C. chinensis and its compounds exhibit anti-atherosclerotic oxidative stress damage in vivo and in vitro experiments, and have inhibitory effects on the formation of β-amyloid fibrils and the expression of Tao protein that cause Alzheimer's disease , has potential applications for treating atherosclerosis and alleviating Alzheimer's disease.

Description

technical field [0001] The present invention relates to the application of C. chinensis and its compounds in disease treatment or alleviation, in particular to the application in the preparation of medicines for treating atherosclerosis and alleviating Alzheimer's disease (Alzheimer disease, AD). Background technique [0002] It is a perennial herbaceous plant belonging to the saxifrage family and is a traditional Chinese medicinal material. It is recorded in ancient books that it has the effects of clearing away heat and detoxification, reducing yellowness and dampness, promoting blood circulation and dispelling blood stasis, diuresis and detumescence. figure 1 Shown are the above-ground parts of C. japonica plants, including flowers, leaves and stems (1A), and sun-dried leaves (1B). Modern research through the separation and identification of the effective components of C. chinensis shows that the flavonoids and organic acids contained in it are related to its antioxidant ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/7048A61P9/10A61P25/28
CPCA61K31/7048A61P9/10A61P25/28A61K2300/00
Inventor 黄锦伟罗婉君孙晓磊何延政
Owner MACAU UNIV OF SCI & TECH
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