Application of snow tea extract to preparation of medicine for preventing and treating zymloid protein diseases
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An amyloid protein and extract technology, which can be used in drug combinations, nervous system diseases, pharmaceutical formulations, etc., and can solve problems such as complex active ingredients
Inactive Publication Date: 2018-11-13
LIAONING UNIVERSITY
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Because snow tea is a complex biological organic symbiosis, its active ingredients are more complex
Method used
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Embodiment 1
[0032] Example 1 In vitro culture study of snow tea extract inhibiting chicken lysozyme into fibrosis
[0033] (1) Snow tea extract
[0034] The preparation method of snow tea extract is:
[0035]1) Snow tea medicinal materials are dried and pulverized, cut into pieces with scissors, weighed 5g of the powder, soaked in 100mL of deionized water for 12 hours; then ultrasonically extracted at 60°C and 350W for 30 minutes; Recorded as group A (water extract of snow tea) and filter residue.
[0036] 2) Filter the residue with 100 mL of 75% ethanol, reflux the ethanol at 65°C, repeat twice, each time for 1 hour; filter under reduced pressure; combine the two filtrates as (Group B); mix the filtrates of Group AB, concentrate under reduced pressure, Dry at 65°C. After being dissolved in DMSO to 50mg / mL, the color is reddish-yellow brown. It is simply referred to as XC in the charts. The snow tea medicinal material is Tibetan white snow tea.
[0037] (2) Solution preparation
[...
Embodiment 2
[0057] Example 2 The effect of snow tea water extract on the expression of amyloid protein by Pichia pastoris
[0058] (1) Preparation of Snow Tea Water Extract
[0059] The preparation method of the snow tea extract is a combination of water extraction and ultrasonic extraction. Snow tea medicinal materials were dried and pulverized, cut into pieces with scissors, weighed 5g of the powder, soaked in 100mL of deionized water for 12 hours; then ultrasonically extracted at 60°C and 350W for 30 minutes; filtered under reduced pressure, and the filtrate was concentrated under reduced pressure, 65 ℃ drying. The snow tea medicinal material is Tibetan white snow tea.
[0060] (2) Strain selection
[0061] The original Pichia pastoris strain Pichiapastoris X-33 was purchased from Invitroge Company. The yeast expression strain of cystatin mutant I66Q was constructed by our laboratory by transferring the pPICZαA-cC-I66Q plasmid into X-33.
[0062] (3) Detection of the antibacterial ...
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Abstract
The invention relates to application of a snow tea extract to preparation of a medicine for preventing and treating zymloid protein diseases. The influence on zymloid protein fiber formation by the snow tea extract is mainly explored, a new effective treatment medicine for preventing and treating zymloid deposition diseases is expected to find, and support is provided for medicinal research on thesnow tea. Typical zymloid protein chicken lysozyme is mainly subjected to in-vitro culture to form fiber and secreted protein of a recombinant pichia pastoris strain of chicken cystatin forms fiber.The result indicates that the snow tea extract achieves the effect of inhibiting the zymloid protein from forming the fiber; furthermore, in the water extract group, the effect is more significant andthe IC50 value is about 0.018 mg / mL. The bacterial level protein expression experiment indicates that the snow tea extract can inhibit a chicken cystatin zymloid mutant I66Q from forming aggregate extracellularly in a concentration dependency way; and when the extract concentration reaches to 1.5 mg / mL, a certain inhibition effect can be generated on oligomerization and aggregation of the secreted protein.
Description
technical field [0001] The invention relates to the field of medicine and health products; in particular, it relates to the application of snow tea extract in the preparation of medicines for preventing and treating amyloid diseases. Background technique [0002] According to the report, with the improvement of people's living standards, improvement of medical conditions, and prolongation of life expectancy, the incidence of Alzheimer's disease and other "geriatric diseases" is increasing year by year. One of the pathological manifestations of Alzheimer's disease is the deposition of β-amyloid protein. Most of β-amyloid protein is produced by protein misfolding. It is named for the positive reaction that occurs when it encounters iodine like starch and turns blue. β-Amyloid is rich in β-sheet structures from which amyloid fibrils, the insoluble, more inert, unbranched fibrous substance, can form. Those diseases characterized by abnormal deposition in the body due to protein...
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