CN105213493A - The application in alpha-glucosidase inhibitor of Radix Rumicis, Radix Actinidiae Chinensis or Herba Hyperici Japonici extract - Google Patents
The application in alpha-glucosidase inhibitor of Radix Rumicis, Radix Actinidiae Chinensis or Herba Hyperici Japonici extract Download PDFInfo
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- CN105213493A CN105213493A CN201510717112.3A CN201510717112A CN105213493A CN 105213493 A CN105213493 A CN 105213493A CN 201510717112 A CN201510717112 A CN 201510717112A CN 105213493 A CN105213493 A CN 105213493A
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Abstract
The invention discloses the application in alpha-glucosidase inhibitor of Radix Rumicis, Radix Actinidiae Chinensis or Herba Hyperici Japonici extract, can be used for treatment or prevent diabetes and disease of viral infection.The preparation method of extract, after Ethnic crude drugs is suitably pulverized, adds 4-10? times amount water decocts respectively, and every taste medical material decocts 2 times, each 1 hour, the decoction liquor that merging decoction is 2 times; Then add 70-95% ethanol, make alcohol content reach 70%, hold over night, get supernatant concentration dry medical material total extract.
Description
Technical field
The present invention relates to three kinds of medicinal substances extract application in alpha-glucosidase inhibitor, these three kinds of Ethnic crude drugs respectively: Radix Rumicis, Radix Actinidiae Chinensis, Herba Hyperici Japonici, belong to medical art.
Background technology
Alpha-glucosidase (α-glucosidase)--be distributed widely in organism, the polysaccharide in food is degraded to monosaccharide by it, is a kind of enzyme played an important role in body metabolic processes.Alpha-glucosidase--inhibitor (α-glucosidaseinhibitor, α-GI) be the class new oral hypoglycemic drug researched and developed in the seventies later stage, its mechanism of action is: by the activity of competitive inhibition alpha-glucosidase, retardance disaccharidase is hydrolyzed into monosaccharide, delay the absorption of sugar, make blood glucose steadily and maintain certain level lentamente, be widely used for reducing postprandial hyperglycemia, be prevent diabetes, reduce one of diabetic complication and the important measures reducing mortality rate.The AGI of current research can be divided into microbial metabolic products according to its source, the inhibitor of natural plant extracts and artificial chemistry synthesis, one kind new medicine of the treatment diabetes wherein brought into use clinically has the acarbose of nineteen ninety-five Bayer A.G's listing, the miglitol of Japan's listing in 1997 etc. (Nishioka, 1998).Along with going deep into and some synthesis AGI more side effect in use of AGI research work, the application of natural A GI is more and more by people are paid attention to.
Meanwhile, alpha-glucosidase also take part in biosynthesis and the catabolic process of glycolipid and glycoprotein in body, and diabetes, obesity, hyperlipidemia, inflammation, canceration, immunoreation and viral infection etc. all have substantial connection with the compound saccharic of cell surface.Therefore, alpha-glucosidase inhibitor can treat multiple relevant to metabolism and relevant with viral infection disease, has broad application prospects.At present, the kind of alpha-glucosidase--inhibitor is less, and price costly, and has certain side effect.Therefore, find efficient, low toxicity, the alpha-glucosidase that price is comparatively cheap simultaneously--inhibitor becomes one of focus of research both at home and abroad.
Summary of the invention
The technical problem to be solved in the present invention is: providing can the Chinese crude drug of Inhibiting α-glucosidase or its extract, and provides effective site and the extracting method thereof of each medical material, to solve the deficiencies in the prior art.
Technical scheme of the present invention is: the application in alpha-glucosidase inhibitor of Radix Rumicis, Radix Actinidiae Chinensis or Herba Hyperici Japonici extract.
The preparation method of extract is after Ethnic crude drugs is suitably pulverized, and add 4-10 times amount water and decoct respectively, every taste medical material decocts 2 times, each 1 hour, the decoction liquor that merging decoction is 2 times; Then add 70-95% ethanol, make alcohol content reach 70%, hold over night, get supernatant concentration dry medical material total extract.
The effective site of Radix Rumicis is ethyl acetate extract.
The effective site of Radix Actinidiae Chinensis is n-butanol portion.
The effective site of Herba Hyperici Japonici is water position and precipitation position thereof.
The preparation method of medical material effective site is each medical material 70%-95% soak with ethanol 3 times, be scattered in water after supernatant concentration, more namely obtain each medical material effective site after extracting 3 each extraction position concentrate dryings successively respectively with petroleum ether, ethyl acetate, n-butanol solvent respectively.
Its medicinal substances extract can be used for prevention or treatment diabetes.
Its medicinal substances extract can be used for prevention or treatment disease of viral infection.
Beneficial effect of the present invention: the present invention finds that the total extract of the Ethnic crude drugs such as Radix Rumicis, Radix Actinidiae Chinensis, Herba Hyperici Japonici all has stronger inhibit activities to alpha-glucosidase, its IC
50be respectively 1.90 μ g/mL, 1.55 μ g/mL, 1.30 μ g/mL; Its activity is better than positive control drug acarbose (IC50=175.97 μ g/ml).Further, the effective site of the present invention to the stronger Guizhou Ethnic crude drugs extract of three kinds of activity is studied, find the ethyl acetate extract (IC50=0.22 μ g/ml) of Radix Rumicis, the n-butanol portion (IC50=1.94 μ g/ml) of Radix Actinidiae Chinensis and the water position (IC50=1.06 μ g/ml) of Herba Hyperici Japonici and precipitate position (IC50=0.86 μ g/ml) that all there is significant inhibit activities, and to the inhibit activities of alpha-glucosidase, there is concentration-dependent relation within the scope of finite concentration.
Detailed description of the invention
reagent
Alpha-glucosidase (α-glucosidase) is purchased from OSAKABIO company (Japan); 4-Nitrobenzol-α-D-pyranglucoside (4-nitrophenyl-α-D-glucopyranoside, PNPG) available from Sigma; Acarbose (Acarbose, acarbose) is purchased from Bayer A.G; All the other reagent are analytical pure.
instrument
Max-190 type microplate reader (MolecularDevices company, the U.S.), constant incubator (Shanghai Bo Tai experimental facilities company limited, China), Rotary Evaporators (Shanghai Yarong Biochemical Instrument Plant, China), analytical precision balances (upper current chart level instruments and meters company limited, China), 96 porocyte plates purchased from CorningCostar(CambridgeMA company, the U.S.).
the preparation of medicinal substances extract
The each 100g of Ethnic crude drugs, after suitably pulverizing, traditionally decocting method decocts respectively, and every taste medical material decocts 2 times, each 1 hour, and the decoction liquor that merging decoction is 2 times, adds 95% ethanol, make alcohol content reach 70%.Hold over night, get supernatant concentration dry total extract ,-20 DEG C of preservations are stand-by.
Various nationalities' medical material 95% soak with ethanol 3 times, is scattered in water after supernatant concentration, then extracts 3 times respectively successively with petroleum ether, ethyl acetate, n-butyl alcohol equal solvent respectively.After the concentrate drying of each extraction position ,-20 DEG C of preservations are stand-by.
The drying sample at the Ethnic crude drugs extract of concentrate drying and medical material difference extraction position is taken out a certain amount of respectively, is dissolved in pure DMSO(dimethyl sulfoxide) in, be made into the high concentration mother solution that concentration is 100mg/mL; Then be diluted to desired concn with phosphate buffer (PB buffer) before use, the final volume mark of DMSO is 0.1%.
the detection method of alpha-glucosidase activity
This detection employing 96 orifice plate screening system, the method of reaction system reference literature report, reaction system after improving and optimizating is: add alpha-glucosidase (α-glucosidase in 80 μ L kaliumphosphate buffers (pH6.9), 0.1U/ml) 10 μ L, add certain density sample solution 10 μ L, 37 DEG C of constant temperature 10min, then add PNPG (2.5mmol/L) 20 μ L, 37 DEG C of isothermal reaction 15min, then the Na adding 40 μ L
2cO
3solution (1mol/L) cessation reaction, measures absorbance (A value) under 405nm wavelength, and the cumulative volume of this reaction system is 160 μ L.
The positive control that acarbose (Acarbose) is this law, set blank (only with buffer) and negative control (only with buffer and enzyme liquid), inhibition of enzyme activity rate is calculated as follows simultaneously; Adopt Origin7.5 computed in software half-inhibition concentration (IC
50) value.
statistical disposition
All data acquisition mean+SD represent, compare employing to each group of data
tinspection,
p< 0.05 thinks there is statistical significance.
experimental result
three kinds of Ethnic crude drugs extracts are on the impact of alpha-glucosidase activity
In three kinds of Ethnic crude drugs extracts, Radix Rumicis total extract has stronger inhibitory action to alpha-glucosidase, Radix Rumicis total extract in the concentration range of 0.01 ~ 100 μ g/mL can concentration dependent ground Inhibiting α-glucosidase activity, when concentration is 100 μ g/ml, suppression ratio (Inhibitory) can reach 97%, as calculated its IC
50be about 1.90 μ g/mL.The results are shown in Table-1.
In three kinds of Ethnic crude drugs extracts, the activity of Radix Actinidiae Chinensis extract energy Inhibiting α-glucosidase in the concentration range of 0.1 ~ 100 μ g/mL, and have certain concentration-dependent relation, when concentration is 100 μ g/mL, its suppression ratio (Inhibitory) can reach 96.2%; Its IC as calculated
50be about 1.55 μ g/mL; And Herba Hyperici Japonici extract can the activity of Inhibiting α-glucosidase strongly in the concentration range of 0.1 ~ 100 μ g/mL, and there is certain concentration-dependent relation, its IC as calculated
50be about 1.30 μ g/mL, the results are shown in Table-1.
Positive drug acarbose is 1, and the activity of energy concentration dependant ground Inhibiting α-glucosidase in the concentration range of 10,100,1000 μ g/mL, when concentration is 1000 μ g/mL, its suppression ratio is 82.5%, its IC
50be about 175.97 μ g/mL, the results are shown in Table-1.
radix Rumicis the different extracted parts is to the inhibitory action of alpha-glucosidase activity
Radix Rumicis chloroform extraction position, ethyl acetate extract part, n-butanol extraction position, and residue water position is respectively 0.1, can the activity of concentration dependent ground Inhibiting α-glucosidase in the concentration range of 1,10,100 μ g/mL, its IC
50be about 1.57 respectively, 0.22,0.67 μ g/ml and 9.51 μ g/ml, its activity is better than positive drug acarbose (IC
50=175.97 μ g/ml); Wherein, the activity of ethyl acetate extract part is the strongest.Measurement result is in Table-2.
radix Actinidiae Chinensis the different extracted parts is to the inhibitory action of alpha-glucosidase activity
The activity of Petroleum ether extraction position (10,100 μ g/mL) energy Inhibiting α-glucosidase when higher concentration of Radix Actinidiae Chinensis, as calculated, its IC
50be about 13.76 μ g/mL; Radix Actinidiae Chinensis ethyl acetate extract part and n-butanol extraction position, can the activity of Inhibiting α-glucosidase in the concentration range of 0.1,1,10,100 μ g/mL 0.01, and there is certain concentration-dependent relation, its IC
50be about 3.98 μ g/mL and 1.94 μ g/mL respectively; The residue water position of Radix Actinidiae Chinensis, can the activity of Inhibiting α-glucosidase significantly under the concentration of 100 μ g/mL 10, its IC
50be about 3.91 μ g/mL.In each extract part of Radix Actinidiae Chinensis, the activity at n-butanol extraction position is the strongest, the results are shown in Table-3.
herba Hyperici Japonici the different extracted parts is to the inhibitory action of alpha-glucosidase activity
The activity of Petroleum ether extraction position (10,100 μ g/mL) energy Inhibiting α-glucosidase when higher concentration of Herba Hyperici Japonici, its IC
50be about 26.77 μ g/mL; The ethyl acetate extract part of Herba Hyperici Japonici and n-butanol extraction position, can the activity of concentration dependent ground Inhibiting α-glucosidase in the concentration range of 1,10,100 μ g/mL 0.1, its IC
50be about 2.34 μ g/mL and 1.37 μ g/mL respectively; The residue water position of Herba Hyperici Japonici, also can the activity of Inhibiting α-glucosidase in the concentration range of 1,10,100 μ g/mL 0.1, its IC as calculated
50be about 1.06 μ g/mL; The precipitation position of Herba Hyperici Japonici is 0.1, and the activity of energy concentration dependent ground Inhibiting α-glucosidase in the concentration range of 1,10,100 μ g/mL, when concentration is 100 μ g/mL, its suppression ratio can reach 99.9%, as calculated its IC
50be about 0.86 μ g/mL, the results are shown in Table-4.
Claims (8)
1. Radix Rumicis, Radix Actinidiae Chinensis or the Herba Hyperici Japonici extract application in alpha-glucosidase inhibitor.
2. Radix Rumicis according to claim 1, Radix Actinidiae Chinensis or the Herba Hyperici Japonici extract application in alpha-glucosidase inhibitor, it is characterized in that: the preparation method of extract is after Ethnic crude drugs is suitably pulverized, add 4-10 times amount water to decoct respectively, every taste medical material decocts 2 times, each 1 hour, merge decoction liquor; Then add 70-95% ethanol, make alcohol content reach 70%, hold over night, get supernatant concentration dry medical material total extract.
3. Radix Rumicis according to claim 1, Radix Actinidiae Chinensis or the Herba Hyperici Japonici extract application in alpha-glucosidase inhibitor, is characterized in that: the effective site of Radix Rumicis is ethyl acetate extract.
4. Radix Rumicis according to claim 1, Radix Actinidiae Chinensis or the Herba Hyperici Japonici extract application in alpha-glucosidase inhibitor, is characterized in that: the effective site of Radix Actinidiae Chinensis is n-butanol portion.
5. Radix Rumicis according to claim 1, Radix Actinidiae Chinensis or the Herba Hyperici Japonici extract application in alpha-glucosidase inhibitor, is characterized in that: the effective site of Herba Hyperici Japonici is water position and precipitation position thereof.
6. according to the application in alpha-glucosidase inhibitor of the Radix Rumicis one of claim 3-5 Suo Shu, Radix Actinidiae Chinensis or Herba Hyperici Japonici extract, it is characterized in that: the preparation method of medical material effective site is each medical material 70%-95% soak with ethanol 3 times, be scattered in water after supernatant concentration, more namely obtain each medical material effective site after extracting 3 each extraction position concentrate dryings successively respectively with petroleum ether, ethyl acetate, n-butanol solvent respectively.
7. Radix Rumicis according to claim 1, Radix Actinidiae Chinensis or the Herba Hyperici Japonici extract application in alpha-glucosidase inhibitor, is characterized in that: its medicinal substances extract can be used for prevention or treatment diabetes.
8. Radix Rumicis according to claim 1, Radix Actinidiae Chinensis or the Herba Hyperici Japonici extract application in alpha-glucosidase inhibitor, is characterized in that: its medicinal substances extract can be used for prevention or treatment disease of viral infection.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201510717112.3A CN105213493B (en) | 2015-10-30 | 2015-10-30 | The application of Rumex madaio, Chinese actinidia root or Herba Hyperici Japonici extract in alpha-glucosidase restrainer |
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| CN201510717112.3A CN105213493B (en) | 2015-10-30 | 2015-10-30 | The application of Rumex madaio, Chinese actinidia root or Herba Hyperici Japonici extract in alpha-glucosidase restrainer |
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| CN105213493B CN105213493B (en) | 2019-10-29 |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004075638A (en) * | 2002-08-21 | 2004-03-11 | Okinawa Pref Gov | Functional material having action to suppress increase of blood sugar level and suppress increase of blood pressure |
| CN103550200A (en) * | 2013-11-05 | 2014-02-05 | 赵全成 | Application of pharmaceutical composition in preparation of medicament for inhibiting blood sugar rise and medicament for reducing blood fat |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004075638A (en) * | 2002-08-21 | 2004-03-11 | Okinawa Pref Gov | Functional material having action to suppress increase of blood sugar level and suppress increase of blood pressure |
| CN103550200A (en) * | 2013-11-05 | 2014-02-05 | 赵全成 | Application of pharmaceutical composition in preparation of medicament for inhibiting blood sugar rise and medicament for reducing blood fat |
Non-Patent Citations (3)
| Title |
|---|
| 张琳等: "田基黄的化学成分研究", 《中国药学杂志》 * |
| 潘艺井等: "7种贵州民族药材提取物对α-葡萄糖苷酶的抑制作用", 《中华中医药杂志》 * |
| 聂昌平等: "民族药材土大黄中α-葡萄糖苷酶抑制剂的初步研究", 《时珍国医国药》 * |
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