CN103566059A - Liver-protecting eyesight-improving folium isatidis and semen cassiae drug pair combined medicament, preparation method and application thereof - Google Patents
Liver-protecting eyesight-improving folium isatidis and semen cassiae drug pair combined medicament, preparation method and application thereof Download PDFInfo
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Abstract
The invention provides a liver-protecting eyesight-improving folium isatidis and semen cassiae drug pair combined medicament. The drug pair combined medicament is an extract by extracting folium isatidis and semen cassiae. The preparation method of the medicament comprises the following steps: extracting a folium isatidis and semen cassiae drug mixture in a refluxing manner by using ethyl acetate which is 7 times that of the weight of the drugs as a solvent twice with 1.0 hour each time. According to the traditional Chinese drug pair combined extract, the drug pair composition has better activity than that of two extracts of single drugs, and has an excellent activity in protecting liver and improving eyesight, even in treating cataract.
Description
Technical field
The present invention relates to a kind of Chinese medicine composition, relate in particular a kind of green grass or young crops and determine liver-nourishing and eyesight-improving medicine to composition of medicine, preparation method and application thereof.
Technical background
Cataract is the primary causes of blindness of China, and its unique and effective Therapeutic Method is operation at present.But the complication that some are serious may appear in cataract operation, this both can increase patient's misery, also can increase patient's financial burden.Therefore, by chemoprophylaxis or treatment cataract, just seem very important in early days.Due to the cataractous slower development of majority, need long-term prescription, and Chinese herbal medicine is pure natural medical, side effect is little, and therefore suitable life-time service has become one of focus of anti-cataract medicine research in recent years to the exploitation of the effective prescription of Chinese herbal medicine.
Summary of the invention
For addressing the above problem, inventor provides a kind of green grass or young crops to determine liver-nourishing and eyesight-improving medicine to composition of medicine, is specially the extract that two kinds of medicinal material extract of Semen Cassiae and Folium Isatidis obtain.
Adopt a kind of mixture obtaining for solvent extraction Semen Cassiae and two kinds of medical materials of Folium Isatidis in ethanol or ethyl acetate.
Employing ethyl acetate is solvent, the mixture that two kinds of medical materials of reflux, extract, Semen Cassiae and Folium Isatidis obtain.
The weight ratio of described Semen Cassiae and Folium Isatidis is 1-10:10-1.
The weight ratio of described Semen Cassiae and Folium Isatidis is 3:1.
The described green grass or young crops preparation method of liver-nourishing and eyesight-improving medicine to composition of medicine of determining, adopting refluxes extracts the mixing medical material of Folium Isatidis and Semen Cassiae, and solvent is ethyl acetate, and the amount of solvent is 7 times of medical material weight, extracts 2 times, each 1.0h.
The green grass or young crops described above liver-nourishing and eyesight-improving medicine application in preparation treatment cataract medicine to composition of medicine of determining.
The green grass or young crops described above liver-nourishing and eyesight-improving medicine application in preparing liver-nourishing and eyesight-improving medicine to composition of medicine of determining.
Useful technique effect of the present invention is: the invention provides the extract of a kind of Chinese medicines to combination, this medicine is active better with respect to the extract of two kinds of single medical materials to compositions, at liver-nourishing and eyesight-improving, and even unexpectedly has good activity in treatment cataract disease.
The specific embodiment
Embodiment 1 Different solution extracts the impact on active component
Take 3 groups of Semen Cassiae that weight ratio is 1:1 and Folium Isatidis medical materials, adopt respectively 70% ethanol, ethyl acetate and water as solvent, with reflow method, extract, the amount of solvent is 7 times of medical material weight, and the number of times of extraction is 2 times, each 1h; Merge to reclaim respectively extracting solution of each group, reclaim solvent and make dry cream, prepare alcohol extraction group, ethyl acetate is put forward group and water extraction group; Detect total solid yield, Determination of chrysophanol and the Content Determination of Indirubin of each group, specifically as shown in table 1.
Table 1 extraction process Orthogonal experiment results
| Group | Total solid yield/(%) | The content of chrysophanol/(mg/g) | Content Determination of Indirubin/(mg/g) |
| Alcohol extraction group | 31.11 | 2.016 | 0.089 |
| Ethyl acetate is put forward group | 20.46 | 4.954 | 0.238 |
| Water extraction group | 14.32 | 0.291 | 0.03 |
The condition that embodiment 2 adopts ethyl acetate to extract Folium Isatidis and Semen Cassiae mixture is investigated
The present embodiment be take and measured the content of chrysophanol in extracting solution and,, the content of indirubin and total solid yield are index, adopt orthogonal experiment, and experimental design is shown in Table 2; The extraction process condition of preferred Semen Cassiae.According to trial test result, choose and affect the factor of extraction effect and level, be shown in Table 3.
Table 2 extraction process factor level table
The preparation of sample solution: take Semen Cassiae and Folium Isatidis medical material that weight ratio is 2:1, by the experimental technique of table 3 test row number, take ethyl acetate as solvent, reflux, extract,, filtration respectively, and be concentratedly settled to 250mL, place standby.
Precision measures each extracting solution 50mL and puts in the evaporating dish of constant weight, and water bath method, puts in 105 ℃ of baking ovens, and dry 3h, takes out and put cooling 0.5h in exsiccator, weighs, and puts in 105 ℃ of baking ovens, and dry 1h, to constant weight, calculates total solid yield.
Table 3 extraction process Orthogonal experiment results
From variance analysis, A, B, C tri-factors have appreciable impact to extraction process.Factor A: level 2 ﹥ 3 ﹥ 1; Factor B: level 2 ﹥ 3 ﹥ 1; Factor C: level 2 ﹥ 1 ﹥ 3.The best of breed of each factor should be A
2b
2c
2: extraction process should be to add 7 times of solvents, extract 2 times, each 1.0h.
The preparation of embodiment 3 laboratory samples
Take Semen Cassiae and Folium Isatidis medical material that weight ratio is 10:1, according to method optimum in embodiment, adopt ethyl acetate as solvent, extract and merge extractive liquid,, reclaim and make dry cream, prepare group I;
Take Semen Cassiae and Folium Isatidis medical material that weight ratio is 1:10, according to method optimum in embodiment, adopt ethyl acetate as solvent, extract and merge extractive liquid,, reclaim and make dry cream, prepare experimental group II;
Take Semen Cassiae and Folium Isatidis medical material that weight ratio is 3:1, according to method optimum in embodiment, adopt ethyl acetate as solvent, extract and merge extractive liquid,, reclaim and make dry cream, prepare experimental group III;
Take Cassia obtusifolia L, according to method optimum in embodiment, adopt ethyl acetate as solvent, extract and merge extractive liquid,, reclaim and make dry cream, prepare Semen Cassiae matched group;
Take Folium Isatidis medical material, according to method optimum in embodiment, adopt ethyl acetate as solvent, extract and merge extractive liquid,, reclaim and make dry cream, prepare Folium Isatidis matched group.
The blue or green certainly liver-nourishing and eyesight-improving medicine of embodiment 4 is treated cataractous activity research to composition of medicine
Wistar rat neonatal rat is divided into 5 groups at random, matched group 1O only, model group 1O only, alcohol extraction group 1O only, 10 of ethyl acetate extraction groups, 10 of water extraction groups.It is identical that each organizes the neonatal rat age, male and female dual-purpose, and body weight is close, with breast milk and plain particles feedstuff, divides cage to feed, and freely drinks water.Give model group, alcohol extraction group, ethyl acetate extraction group, low dose of (3.46mg/kg) sodium selenite of water extraction group rat nape portion subcutaneous injection, the next day 1 time, continuous 3 times.Control rats is left intact.Within after last administration the 3rd day, with slit lamp microscope observation rat lens, change, lenticular opacity is modeling success.
The green grass or young crops liver-nourishing and eyesight-improving medicine of determining is twisted into pieces and with distilled water, is made into suspension afterwards composition of medicine.Preparation different quality concentration, is loaded on respectively in vial, and sealing, puts 4 ℃ of Refrigerator stores of people standby.Give the counter sample preparing in model group, alcohol extraction group, ethyl acetate extraction group, water extraction group administration embodiment 1; The rat respectively embodiment green grass or young crops of gavage 30mg/mL is determined liver-nourishing and eyesight-improving medicine to composition of medicine.Gavage amount is crude drug 3g1kg
-1weight, 1 time/d, continues 30d.Matched group and model group rat need not any medicines.
When experiment finishes, with Tropicamide eye drop, each group experimental rat pupil of both eyes is loose large, with examination with slitlamp microscope and record lenticular opacity situation.The standard grading of lenticular opacity degree reference literature: "-" shows that crystalline lens is transparent without muddy; "+" shows that core is muddy, and muddiness occurs nucleus lentis, but cortex portion is still limpid; " * " shows full muddiness, and nucleus lentis and cortex are all become turbid.
When experiment finishes, with rustless steel eye scissors, put to death and respectively organize experimental rat, take out immediately bilateral eyeball.By after ℃ normal saline flushing of O～4 for eyeball 3 times, peel off crystalline lens, with filter paper, blot, be placed on electronic balance and weigh, and record.Bilateral crystalline lens is put into 5mL homogenate cup, add in proportion 8.6g/L ice normal saline, with temperature control multi-purpose high speed, tissue mashing machine smashs 1.5min to pieces with 10000r/min, makes 3.5% crystalline lens homogenate.Again by homogenate with 2000r/min low-speed centrifugal 8min after, take out supernatant.With xanthine oxidase, measure SOD active, by thiobarbituricacidα-method, measure MDA content, by Coomassie brilliant blue method, measure protein content.
Experimental data with mean ± standard deviation (
± s) represent, between each group, relatively with t check, P<O.05 represents that difference has statistical significance.
Matched group, give after injection sodium selenite 3 times the 3rd day of model group, alcohol extraction group, ethyl acetate extraction group, water extraction group rat, naked eyes and slit lamp microscope observation find that these rat lens of both eyes have all occurred that canescence is in various degree muddy, the rat 100% that shows to inject after sodium selenite has formed cataract in various degree, and control rats occurs lenticular opacity without 1 example.The same day of killing Mus, with examination with slitlamp microscope, respectively organize experimental rat lenticular opacity situation, the results are shown in Table shown in 4.
Table 4 is respectively organized the muddy situation of rat lens
Note: "-" represents that crystalline lens is transparent; "+" represents that nucleus lentis is muddy, but cortex is still limpid; " * " represents that muddiness all appears in nucleus and skin.
Model group rat lens SOD activity is starkly lower than matched group, and MDA content is significantly higher than matched group, and its difference all has significant.Blue or green certainly liver-nourishing and eyesight-improving medicine is to the SOD activity of 2 dosage groups of composition of medicine all higher than model group, and MDA content is all lower than model group, and its difference all has statistical significance, shows that blue or green certainly liver-nourishing and eyesight-improving medicine can improve the oxidation resistance from cataract rat to composition of medicine; Be shown in Table 5.
Table 5 respectively organize rat lens SOD activity or MDA content comparison (
± s)
| Group | Rat quantity/n | SOD(U/mgprot) | MDA(nmol/mgprot) |
| Matched group | 10 | 34.26±3.28 | 0.22±0.27 |
| Model group | 10 | 27.94±3.12 | 1.26±0.26 |
| Alcohol extraction group | 10 | 31.87±3.77 | 0.71±0.35 |
| Ethyl acetate extraction group | 10 | 29.65±4.23 | 0.93±0.45 |
| Water extraction group | 10 | 27.51±4.62 | 1.05±0.65 |
Note:
*with matched group comparison, P ﹤ 0.05;
△with model group comparison, P ﹤ 0.05.
Data show shown in table 5, extract prepared by Folium Isatidis prepared by employing ethyl acetate and Semen Cassiae, treatment cataract best results.
The blue or green certainly liver-nourishing and eyesight-improving medicine of embodiment 5 is treated cataractous activity research 2 to composition of medicine
Wistar rat neonatal rat is divided into 7 groups at random, matched group 1O only, model group 1O only, group I 1O only, 10 of 10 of experimental group II, 10 of experimental group III, 10 of Semen Cassiae matched groups and Folium Isatidis matched groups.It is identical that each organizes the neonatal rat age, male and female dual-purpose, and body weight is close, with breast milk and plain particles feedstuff, divides cage to feed, and freely drinks water.Give model group, group I, experimental group II, experimental group III, Semen Cassiae matched group and Folium Isatidis control rats nape portion subcutaneous injection low dose of (3.46mg/kg) sodium selenite, the next day 1 time, continuous 3 times.Control rats is left intact.Within after last administration the 3rd day, with slit lamp microscope observation rat lens, change, lenticular opacity is modeling success.
The green grass or young crops liver-nourishing and eyesight-improving medicine of determining is twisted into pieces and with distilled water, is made into suspension afterwards composition of medicine.Preparation different quality concentration, is loaded on respectively in vial, and sealing, puts 4 ℃ of Refrigerator stores of people standby.Give the blue or green certainly liver-nourishing and eyesight-improving medicine of embodiment of model group, group I, experimental group II, experimental group III, Semen Cassiae matched group and Folium Isatidis control rats difference gavage 30mg/mL to composition of medicine.Gavage amount is crude drug 3g1kg
-1weight, 1 time/d, continues 30d.Matched group and model group rat need not any medicines.
Method described in employing embodiment 4 detects.
Experimental data with mean ± standard deviation (
± s) represent, between each group, relatively with t check, P<O.05 represents that difference has statistical significance.
After injection sodium selenite 3 times the 3rd day of model group, group I, experimental group II, experimental group III, Semen Cassiae matched group and Folium Isatidis control rats, naked eyes and slit lamp microscope observation find that these rat lens of both eyes have all occurred that canescence is in various degree muddy, the rat 100% that shows to inject after sodium selenite has formed cataract in various degree, and control rats occurs lenticular opacity without 1 example.The same day of killing Mus, with examination with slitlamp microscope, respectively organize experimental rat lenticular opacity situation, the results are shown in Table shown in 6.
Table 6 is respectively organized the muddy situation of rat lens
Note: "-" represents that crystalline lens is transparent; "+" represents that nucleus lentis is muddy, but cortex is still limpid; " * " represents that muddiness all appears in nucleus and skin.
Model group rat lens SOD activity is starkly lower than matched group, and MDA content is significantly higher than matched group, and its difference all has significant.Blue or green certainly liver-nourishing and eyesight-improving medicine is to the SOD activity of 2 dosage groups of composition of medicine all higher than model group, and MDA content is all lower than model group, and its difference all has statistical significance, shows that blue or green certainly liver-nourishing and eyesight-improving medicine can improve the oxidation resistance from cataract rat to composition of medicine; Be shown in Table 7.
Table 7 is respectively organized the comparison (x ± s) of rat lens SOD activity or MDA content
| Group | Rat quantity/n | SOD(U/mgprot) | MDA(nmol/mgprot) |
| Matched group | 10 | 34.26±3.28 | 0.22±0.27 |
| Model group | 10 | 27.94±3.12 | 1.26±0.26 |
| Group I | 10 | 31.87±3.77 | 0.81±0.35 |
| Experimental group II | 10 | 30.61±4.01 | 0.77±0.41 |
| Experimental group III | 10 | 32.59±4.28 | 0.35±0.33 |
| Semen Cassiae matched group | 10 | 29.88±3.63 | 0.81±0.25 |
| Folium Isatidis matched group | 10 | 28.66±3.81 | 1.01±0.32 |
Note:
*with matched group comparison, P ﹤ 0.05;
△with model group comparison, P ﹤ 0.05.
By clinical observation, confirm, blue or green certainly liver-nourishing and eyesight-improving medicine to composition of medicine according to tcm treatment according to syndrome differentiation principle, in side, Folium Isatidis and Semen Cassiae equalization have the effect of liver heat removing and eyesight improving, expelling wind and removing nebula, the senile cataract that both liver and kidney deficiency, asthenia of essence and blood or insufficiency of the spleen mistake fortune and stagnated heat of liver channel etc. the are caused pathogenesis that hits.
The blue or green certainly activity research of liver-nourishing and eyesight-improving medicine to composition of medicine fat-reducing liver-protecting of embodiment 6
72 of Wistar rats, male and female half and half, body weight 150～170g, separate at random 12 as blank group, feed normal diet, all the other equal feed high lipid foods, filling a prescription is: 1% cholesterol, 5% Adeps Sus domestica, 5% Oleum Arachidis hypogaeae semen, 5% yolk powder, 0.02% propylthiouracil, 2% sucrose, 82% normal diet.Feed 10d.
By feed high lipid food rat fasting 12h, after socket of the eye, get blood and survey TC, TG, according to TC, TG value, feed high lipid food rat is divided into 5 groups at random: model control group, blank group, positive controls (1mgkg
-1), blue or green certainly liver-nourishing and eyesight-improving medicine is to composition of medicine low dose group (0.40gkg
-1), middle dosage group (0.80gkg
-1), (high dose group 1.60gkg
-1).Each administration group all gives relative medicine by lmL/100g body weight gavage, and blank group and model control group give normal saline, qd, continuous 5 weeks.During administration, except blank treated animal gives normal diet, all the other each treated animals all give high lipid food and feed.
11h fasting before last administration, 1h after administration, the centrifugal 2400rmin of anesthetized rat abdominal aortic blood
-1extract serum, measure ALT, AST, TCT, TG, LDL-C, HDL-C; Data with mean ± standard deviation (
± s) represent, homogeneity test of variance is checked with F, between group, relatively with t, checks.Blue or green certainly liver-nourishing and eyesight-improving medicine impact on hyperlipidemia rats serum alt, AST, TC, TG, HDL-C and LDL-C on composition of medicine; The results are shown in Table shown in 8.
Table 8 on the impact of hyperlipidemia rat fat (
± s)
Note: with the comparison of blank group matched group,
△p ﹤ 0.05,
△ △p ﹤ 0.01; With model control group comparison,
*p ﹤ 0.05,
*p ﹤ 0.01.
As shown in Table 8, model control group is compared with blank group, Serum ALT value obviously raise (P<0.05); Simvastatin group, blue or green certainly liver-nourishing and eyesight-improving medicine are compared with model control group each group group of composition of medicine, and Serum ALT value has no significant change.Model control group is compared with blank group, serum AST value obviously raise (P<0.05); Simvastatin group, blue or green certainly liver-nourishing and eyesight-improving medicine are compared with model control group each group group of composition of medicine, and serum AST value all obviously reduces (P<0.05).Model control group is compared with blank group, and TC, TG, LDL-C value obviously raise (P<0.01), and HDL-C value obviously reduces (P<0.05); Simvastatin group, blue or green certainly liver-nourishing and eyesight-improving medicine are compared with model control group each group of composition of medicine, TC, TG, LDL-C value obviously reduce (P<0.05), HDL-C value is rising (P<O.05) obviously, in addition, the TC of Semen Cassiae matched group, TG, HDL-C, LDL-C decrease, but not obvious, prove, Folium Isatidis and Semen Cassiae combination are extracted and can be brought into play medicine pairing effect.
Liver plays the role of a nucleus in lipid and Proteometabolism, and liver generation pathological changes, can cause the disorder of lipid metabolism, forms hyperlipemia, shows as blood TC, TG and LDL-C and raises, and HDL-C reduces; When hyperlipemia is serious, can cause again fatty liver to occur, cause hepatocyte function infringement further to increase the weight of, both reciprocal causation relations, thus form vicious cycle.Medicine can be by regulating lipid metabolism extremely bring into play the effect that improves hepatocyte function.
This experiment, by animal feed high lipid food, makes lipid material surpass the needs of body and the ability of removing, thereby has built hyperlipemia model, utilize this model observed blue or green certainly liver-nourishing and eyesight-improving medicine to composition of medicine the preventive and therapeutic effect to hyperlipidemia rats.Result shows that blue or green certainly each group can obviously reduce TC in rat blood serum, TG, LDL-C content, and HDL-C is had to rising effect, and the blue or green certainly liver-nourishing and eyesight-improving medicine of prompting may be brought into play the effect protecting the liver by improving the metabolism of lipid to composition of medicine.Pathological examination shows, there is obvious liver fat degeneration in model group rat, fatty liver is generally with the infringement of liver function clinically, show as the slight rising of serum transaminase, take AST rising as main, the blue or green certainly liver-nourishing and eyesight-improving medicine of this experimental result can obviously reduce serum AST value to composition of medicine, and alleviates liver fat denaturation degrees, proves that blue or green certainly liver-nourishing and eyesight-improving medicine has the effect of alt-reducing and liver-protecting simultaneously to composition of medicine.
Claims (8)
1. green grass or young crops is determined liver-nourishing and eyesight-improving medicine to a composition of medicine, it is characterized in that: the extract obtaining for Semen Cassiae and two kinds of medicinal material extract of Folium Isatidis.
2. green grass or young crops according to claim 1 is determined liver-nourishing and eyesight-improving medicine to composition of medicine, it is characterized in that: adopt a kind of mixture obtaining for solvent extraction Semen Cassiae and two kinds of medical materials of Folium Isatidis in ethanol or ethyl acetate.
3. green grass or young crops according to claim 2 is determined liver-nourishing and eyesight-improving medicine to composition of medicine, it is characterized in that: employing ethyl acetate is solvent, the mixture that two kinds of medical materials of reflux, extract, Semen Cassiae and Folium Isatidis obtain.
4. green grass or young crops according to claim 3 is determined liver-nourishing and eyesight-improving medicine to composition of medicine, it is characterized in that: the weight ratio of described Semen Cassiae and Folium Isatidis is 1-10:10-1.
5. green grass or young crops according to claim 4 is determined liver-nourishing and eyesight-improving medicine to composition of medicine, it is characterized in that: the weight ratio of described Semen Cassiae and Folium Isatidis is 3:1.
6. the green grass or young crops according to claim 5 preparation method of liver-nourishing and eyesight-improving medicine to composition of medicine of determining, it is characterized in that: adopting refluxes extracts the mixing medical material of Folium Isatidis and Semen Cassiae, and solvent is ethyl acetate, and the amount of solvent is 7 times of medical material weight, extract each 1.0h 2 times.
7. the liver-nourishing and eyesight-improving medicine application in preparation treatment cataract medicine to composition of medicine of determining of the green grass or young crops described in claim 1-5.
8. the liver-nourishing and eyesight-improving medicine application in preparing liver-nourishing and eyesight-improving medicine to composition of medicine of determining of the green grass or young crops described in claim 1-5.
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Citations (2)
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| CN101129588A (en) * | 2007-08-20 | 2008-02-27 | 陈江 | Instant powder for conjunctivitis |
| CN101683161A (en) * | 2008-09-26 | 2010-03-31 | 刘泳宏 | Natural tea drink capable of clearing away heat and production method |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101129588A (en) * | 2007-08-20 | 2008-02-27 | 陈江 | Instant powder for conjunctivitis |
| CN101683161A (en) * | 2008-09-26 | 2010-03-31 | 刘泳宏 | Natural tea drink capable of clearing away heat and production method |
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| Title |
|---|
| 邢湘臣: "盛夏话药茶", 《农业考古》, no. 02, 30 June 1999 (1999-06-30), pages 224 - 8 * |
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