CN116211916A - Application of grass mulberry chrysanthemum extract in preparing medicament for treating hypertonic xerophthalmia - Google Patents
Application of grass mulberry chrysanthemum extract in preparing medicament for treating hypertonic xerophthalmia Download PDFInfo
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- CN116211916A CN116211916A CN202310495599.XA CN202310495599A CN116211916A CN 116211916 A CN116211916 A CN 116211916A CN 202310495599 A CN202310495599 A CN 202310495599A CN 116211916 A CN116211916 A CN 116211916A
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- chrysanthemum
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- mulberry
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- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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Abstract
The invention provides application of a mulberry chrysanthemum extract in preparing a medicament for treating hypertonic xerophthalmia, and belongs to the technical field of traditional Chinese medicine pharmacy. The grass-leaved sweetflag chrysanthemum extract comprises water extraction alcohol immersion paste of selfheal, mulberry leaf and wild chrysanthemum flower and ethanol immersion liquid with a certain concentration of wild chrysanthemum flower. The invention proves that the grass mulberry chrysanthemum extract has good effect on xerophthalmia caused by hyperpermeability through an in vitro human cornea epithelial cell hypertonic model and an in vivo xerophthalmia rat model, can remarkably promote tear secretion, has the effect of improving hypertonic xerophthalmia, and is superior to grass mulberry chrysanthemum water extraction alcohol immersion paste. The preparation method of the grass mulberry chrysanthemum extract provided by the invention is simple, the raw materials are easy to obtain, and the grass mulberry chrysanthemum extract has no cytotoxicity and has good practical application prospect.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicine pharmacy, and particularly relates to application of a Xiasangju extract in preparation of a medicine for treating hypertonic xerophthalmia.
Background
Dry eye is a disease that is associated with tears and ocular surfaces, where tear quality or quantity and kinetics are abnormal, resulting in unstable tear films and/or abnormalities of ocular surfaces, producing chronic inflammation, with symptoms of ocular discomfort. Dry eye is a common frequently occurring disease and has a significant impact on the quality of life of patients. Decreased tear flow and/or increased tear evaporation lead to tear hyperosmosis, a key factor in the malignant circulation of dry eye pathology. The existing method for treating hypertonic xerophthalmia mainly comprises hypotonic artificial tears, glucocorticoid and immunosuppressant, the duration of the artificial tears in eyes is relatively short, the symptoms and the root causes are not treated, and secondary adverse reactions are easily caused by long-term administration of the glucocorticoid and the immunosuppressant. The hypertonic dry eye disease causes a heavy economic burden on families of patients and even society, and a need exists for searching new, effective and safe medicines, and the traditional Chinese medicine has the overall regulation effects of multiple components and multiple targets, and is an ideal medicine for treating the hypertonic dry eye disease.
"Huangdi's Nei Jing" is: the five zang-organs transform into liquid, the liver being tears, indicating that tear production and homeostasis are closely related to the liver. "the disease sources and the theory of disease" is: the symptoms of the liver, the essence of viscera, the focus of the pulse and the ascending of fluid, the bleeding of the fluid, the astringency of the eyes, suggest that the deficiency of the liver has a close relationship with the occurrence of dry eye. The liver is induced to open eyes, liver qi rises and liver blood can nourish eyes, and dry eyes can be seen in red eyes with swelling and pain of excessive eyes or dim eyes with deficiency symptoms, the occurrence of the dry eyes is always related to the liver, and the treatment is usually from liver clearing. The internal treatment of traditional Chinese medicine is the main method for treating xerophthalmia, mainly tonifying deficiency and clearing heat, and commonly used herbs mainly enter liver meridian, such as Ju Hua and Sheng Di Huang.
Patent (CN 105106318A) discloses a traditional Chinese medicine composition and a preparation method thereof, wherein the traditional Chinese medicine composition is prepared from the following raw materials: pulverizing folium Mori, flos Chrysanthemi Indici, and herba Menthae respectively, making into powder, and mixing. The obtained traditional Chinese medicine composition is used for preparing common fumigation bags, and the preparation process is simple, and only the raw materials of the formula are crushed into coarse powder, split-packed into fumigation bags and sterilized. The Chinese medicinal composition can be used for treating xerophthalmia, especially xerophthalmia caused by meibomian gland dysfunction, and has simple and easy treatment process and good effect.
Patent (CN 107432909 a) discloses a traditional Chinese medicine composition for preventing and treating xerophthalmia, which is prepared from the following raw materials in parts by weight: 10-20 parts of medlar; 10-15 parts of dwarf lilyturf tuber; 10-15 parts of radix scrophulariae; 10-15 parts of radix rehmanniae; 5-10 parts of peppermint; 5-10 parts of liquorice; 5-10 parts of mulberry leaves; 5-10 parts of radix scutellariae; 5-10 parts of chrysanthemum/wild chrysanthemum; 5-10 parts of ginseng. According to the effects of nourishing liver and kidney of medlar, heat-clearing and fluid-producing of dwarf lilyturf tuber, nourishing yin and reducing fire, heat-clearing and fluid-producing of rehmannia root, sang Sheliang blood and improving eyesight, and wind-dispelling and heat-clearing of baical skullcap root, chrysanthemum/wild chrysanthemum flower are taken as the medicines for harmonizing the medicines, and according to the damage and apoptosis of corneal epithelial cells existing in the pathogenesis of xerophthalmia, the medlar is purposefully reused, ginseng is added, and the optimal dosage ratio of medlar and ginseng is screened, so that the obtained traditional Chinese medicine composition can effectively enhance the stability of tear film, increase tear secretion, has obvious protection effect of corneal epithelial cells, can effectively relieve the clinical symptoms of xerophthalmia, and reduce the proportion of secondary conjunctivitis, corneal ulcer and blindness of xerophthalmia patients.
The patent (CN 114081927A) provides a traditional Chinese medicine eye drop, which comprises, by weight, 10-20 parts of climbing groundsel herb, 8-18 parts of garden balsam stem, 5-15 parts of lycopodium clavatum, 7-16 parts of divaricate saposhnikovia root, 10-20 parts of suberect spatholobus stem, 5-14 parts of szechuan lovage rhizome, 12-22 parts of cassia seed, 10-18 parts of sea-ear shell, 15-25 parts of dried dendrobium and 3-16 parts of night-time luminous sand. The obtained traditional Chinese medicine eye drop can treat teenager myopia, amblyopia and astigmatism, can also treat cataract of middle-aged and elderly people, and has good effect without causing other eye problems.
As can be seen, various traditional Chinese medicine preparations have been used in the prior art for treating eye diseases. Prunella vulgaris, mulberry leaf and wild chrysanthemum are three common traditional Chinese medicinal materials, and the Chinese patent medicine of the wild chrysanthemum has the effects of clearing liver and improving vision, dispelling wind and cooling, removing dampness and removing sore toxin, and can be used for treating wind-heat type common cold, conjunctival congestion and headache, hypertension, dizziness and tinnitus, sore throat and furuncle and pyogenic infections. At present, no patent or report on the treatment of xerophthalmia by using the chrysanthemum morifolium ramat has been made, and the chrysanthemum morifolium ramat extract provides a new idea for the treatment of xerophthalmia patients.
Disclosure of Invention
Aiming at the problems existing in the prior art, the invention provides application of a mulberry chrysanthemum extract in preparing a medicament for treating hypertonic xerophthalmia. The said extract contains extract of herba Desmodii Styracifolii and flos Chrysanthemi Indici. According to the invention, through an in vitro human cornea epithelial cell hypertonic model and an in vivo xerophthalmia rat model caused by hypertonic, the grass-mulberry chrysanthemum extract has the effect of improving hypertonic xerophthalmia, and is superior to grass-mulberry chrysanthemum water extraction alcohol immersion paste. The preparation method of the grass mulberry chrysanthemum extract provided by the invention is simple, the raw materials are easy to obtain, the grass mulberry chrysanthemum extract has no cytotoxicity, and the grass mulberry chrysanthemum extract has good effects of resisting oxidization and promoting tear secretion, and is suitable for popularization.
In order to achieve the above object, in a first aspect, the present invention provides an application of a summer mulberry chrysanthemum extract in preparing a medicament for treating hypertonic dry eye, wherein the summer mulberry chrysanthemum extract is an extract prepared from raw materials of selfheal, sang She and wild chrysanthemum in a weight ratio of 500:175:80; the grass-mulberry chrysanthemum extract comprises grass-mulberry chrysanthemum extract and wild chrysanthemum impregnating solution.
In a preferred embodiment, the preparation method of the wild chrysanthemum flower impregnating solution comprises the following steps: taking 8% -20% of the weight of the wild chrysanthemum flower in the raw materials, adding an ethanol solution with the volume concentration of 50% -95% which is 1.5-10 times of the weight of the wild chrysanthemum flower, and extracting at 15-40 ℃ by an impregnation method or a percolation method to obtain the wild chrysanthemum flower extract;
the preparation method of the grass mulberry chrysanthemum extract comprises the following steps: decocting the rest flos Chrysanthemi Indici, all Prunellae Spica and folium Mori in water for 1-3 times each for 1-2 hr, mixing decoctions, filtering, concentrating, adding ethanol to concentrate until the volume concentration of ethanol content reaches 63%, standing overnight, filtering to obtain supernatant, recovering ethanol from the filtrate, and concentrating under reduced pressure to obtain extract with relative density of 1.20-1.30.
In a preferred embodiment, the preparation method of the wild chrysanthemum flower impregnating solution is an impregnating method or a percolating method;
the dipping method comprises the following steps: taking wild chrysanthemum flower, dividing the wild chrysanthemum flower into three parts, respectively filling the three parts into a container according to the weight ratio of 4-6:2-4:1-3, soaking the wild chrysanthemum flower in 95% ethanol which is 1.5-3 times of the weight of the wild chrysanthemum flower, adding the wild chrysanthemum flower into the 1 st part, soaking for 1-3 days, transferring the discharged liquid into the 2 nd part, soaking for 1-3 days, transferring the discharged liquid into the 3 rd part, soaking for 1-3 days, filtering the leaching solution, and obtaining the wild chrysanthemum flower soaking liquid;
the percolation method comprises the following steps: placing flos Chrysanthemi Indici into a percolating cylinder, adding 2-3 times of 95% ethanol solution by volume concentration of flos Chrysanthemi Indici, soaking for 12-24 hr, adding 7-8 times of 95% ethanol by volume concentration of flos Chrysanthemi Indici, percolating at a flow rate of 1-4mL per minute, and collecting ethanol solution to obtain flos Chrysanthemi Indici soaking solution.
In a preferred embodiment, the drug content of the grass-leaved chrysanthemum extract is 3-30g crude drug/g, and the drug content of the wild chrysanthemum impregnation liquid is 0.3-5g crude drug/mL.
In a preferred embodiment, the said extract of the said chrysanthemum is prepared from extract of the chrysanthemum and the impregnating solution of the wild chrysanthemum according to the extract of the chrysanthemum: the wild chrysanthemum impregnation liquid= (5-25) g/1 mL ratio, more preferably, the grass-mulberry chrysanthemum extract is obtained by mixing grass-mulberry chrysanthemum extract and wild chrysanthemum impregnation liquid according to the grass-mulberry chrysanthemum extract/wild chrysanthemum impregnation liquid=16 g/1 mL ratio.
In a preferred embodiment, the effect concentration of said Xiasangju extract is between 0.5 and 1000 μg/mL, more preferably between 0.8 and 625 μg/mL.
In a preferred embodiment, the drug for treating hypertonic dry eye has a good antioxidant effect on hypertonic dry eye and is capable of significantly promoting tear secretion.
In a second aspect, the present invention provides the use of the aforementioned grass-leaved chrysanthemum extract in the manufacture of a medical device or facility for the treatment of hypertonic dry eye.
Compared with the prior art, the invention has the following beneficial effects:
1. the invention provides application of a summer mulberry chrysanthemum extract in preparing a medicine for treating hypertonic xerophthalmia, wherein the summer mulberry chrysanthemum extract comprises a water extraction alcohol immersion paste (namely the summer mulberry chrysanthemum extract) of selfheal, mulberry leaves and wild chrysanthemum and a wild chrysanthemum ethanol immersion liquid (namely the wild chrysanthemum immersion liquid) with a certain concentration. The invention proves that the grass mulberry chrysanthemum extract has good effect on xerophthalmia caused by hyperpermeability through an in vitro human cornea epithelial cell hypertonic model and an in vivo xerophthalmia rat model, can remarkably promote tear secretion, has the effect of improving hypertonic xerophthalmia, and is superior to grass mulberry chrysanthemum water extraction alcohol immersion paste.
2. The Xiasangju extract provided by the invention has no cytotoxicity when treating hypertonic xerophthalmia, has obvious antioxidation effect, and can obviously promote tear secretion.
3. The preparation method of the grass mulberry chrysanthemum extract provided by the invention is simple, raw materials are easy to obtain, and the grass mulberry chrysanthemum extract has good practical application prospect.
Drawings
FIG. 1 effects of different osmotic pressures on human corneal epithelial cell activity;
FIG. 2 effect of Xiasangju extract on human corneal epithelial cell activity;
FIG. 3 protection of hypertonically induced human corneal epithelial cells by Xiasangju extract;
FIG. 4 effect of Xiasangju extract on ROS levels in human corneal epithelial cells under hyperosmotic induction;
FIG. 5 effect of Xiasangju extract on human corneal epithelial cell secretion IL-6 levels under hyperosmotic induction;
FIG. 6 is a comparison of the effect of a Xiasangju extract and a single herbal extract on ROS levels in human corneal epithelial cells under hyperosmotic induction;
FIG. 7 comparison of the protective effect of a Xiasangju extract on hypertonically induced human corneal epithelial cells with a single herbal extract.
Detailed Description
The present invention will be described in further detail with reference to the following examples, which are not intended to limit the present invention, but are merely illustrative of the present invention. The experimental methods used in the following examples are not specifically described, but the experimental methods in which specific conditions are not specified in the examples are generally carried out under conventional conditions, and the materials, reagents, etc. used in the following examples are commercially available unless otherwise specified.
EXAMPLE 1 preparation of Chamomile extract
The Xiasangju extract is from Guangzhou white cloud mountain star (pharmaceutical industry) stock company, the batch number is B200208, and the medicine content is 7.78g crude drug/g; the flos Chrysanthemi Indici soaking solution is from Guangzhou white cloud mountain star (pharmaceutical industry) Co., ltd., batch number of B200202, and medicine liquid concentration of 1.33g crude drug/mL.
In the embodiment, the preparation method of the raw drug of the grass-mulberry chrysanthemum extract is that the grass-mulberry chrysanthemum extract and the wild chrysanthemum impregnating solution are compounded, and the preparation method is as follows: the extract (g) of the mulberry chrysanthemum in summer is as follows: wild chrysanthemum infusion (mL) =16:1. The concentration of the obtained grass-mulberry chrysanthemum extract in the raw material of the grass-mulberry chrysanthemum extract is 1.08g extract/mL, namely 8.40g crude drug/mL; the concentration of the wild chrysanthemum flower soaking liquid is 67.16 mu L soaking liquid/mL, namely 89.32mg crude drug/mL.
The preparation method of the grass mulberry chrysanthemum extract and wild chrysanthemum impregnating solution comprises the following steps:
the extract of the chrysanthemum is prepared from the raw materials of selfheal, sang She and wild chrysanthemum in a weight ratio of 500:175:80, and comprises the extract of the chrysanthemum and the wild chrysanthemum impregnating solution.
The concrete preparation method of the grass mulberry chrysanthemum extract comprises the following steps: decocting 88% of flos Chrysanthemi Indici and all Prunellae Spica and folium Mori in water (10 times of the weight of the materials each time) for 1.5 times, each time for 2 hr, mixing decoctions, filtering, concentrating the filtrate to about 1/2 of the weight of the materials to obtain concentrated solution, adding ethanol to the concentrated solution to reach volume concentration of ethanol content of 63%, standing for 24 hr, filtering to obtain supernatant, recovering ethanol from the filtrate, and concentrating under reduced pressure to obtain extract with relative density of 1.25-1.26 at 30deg.C.
The specific preparation method of the wild chrysanthemum impregnating solution comprises the following steps: taking 12% of the weight of the wild chrysanthemum in the raw materials, filling the wild chrysanthemum into a percolating cylinder, adding an ethanol solution with the volume concentration of 95% which is 3 times of the weight of the wild chrysanthemum, soaking for 20 hours, adding ethanol with the volume concentration of 8 times of the weight of the wild chrysanthemum, percolating at the flow rate of 3mL per minute, and collecting ethanol solution to obtain the wild chrysanthemum impregnating solution.
EXAMPLE 2 preparation of Xiasangju extract
Taking 6.4g of wild chrysanthemum, dividing the wild chrysanthemum into 1, 2 and 3 parts according to the proportion of 5:3:2, respectively filling the three parts into a container, adding 9.6mL of 80% ethanol as a solvent into the 1 st part, soaking for 1 day, transferring the discharged liquid into the 2 nd part, soaking for 1 day, transferring the discharged liquid into the 3 rd part, soaking for 1 day, and filtering the leaching solution to obtain 6.5mL of wild chrysanthemum soaking liquid. 500g of selfheal, 175g of mulberry leaf and 73.6g of wild chrysanthemum are taken, water is added for decoction twice, each time for 1.5 hours, the decoction is combined, filtered, the filtrate is concentrated to 373ml, 95 percent ethanol is added until the ethanol content reaches 63 percent, the mixture is stirred with the addition, the mixture is stood overnight for 24 hours, the supernatant is filtered, the ethanol is recovered from the filtrate, and the filtrate is concentrated under reduced pressure to 91g of the grass mulberry chrysanthemum extract with the relative density of 1.25 (30 ℃). The preparation method of the original drug of the grass-mulberry chrysanthemum extract is characterized by compounding the grass-mulberry chrysanthemum extract and the wild chrysanthemum impregnating solution, and comprises the following steps: the extract (g) of the mulberry chrysanthemum in summer is as follows: wild chrysanthemum infusion (mL) =14:1. Wherein the content of the drug in the grass mulberry chrysanthemum extract is 7.61g crude drug/g; the concentration of the liquid medicine of the wild chrysanthemum impregnating solution is 1.39g crude drug/mL.
Comparative example 1 preparation of ethanol precipitation extractum extracted from summer Sang Jushui
The conventional grass-mulberry chrysanthemum extract is an extract used by common grass-mulberry chrysanthemum particles in the market, and the product is prepared by extracting three medicinal materials of selfheal, sang She and wild chrysanthemum in a weight ratio of 500:175:80. The specific process comprises the following steps: taking selfheal, mulberry leaf and wild chrysanthemum as raw materials, adding 10 times of water, decocting for 1.5 hours, decocting for 2 times, filtering, combining filtrates, concentrating the filtrate to fluid extract with the relative density of 1.06-1.10 (80 ℃), adding 95% ethanol until the ethanol concentration is 63%, precipitating with ethanol for 24 hours, filtering, recovering ethanol from the filtrate, concentrating under reduced pressure to obtain extract with the relative density of 1.25-1.26 (30 ℃) (namely the water-extracted alcohol immersed paste of the mulberry leaf and the wild chrysanthemum).
Comparative example 2 preparation of Prunella Spica extract (XKC), wild chrysanthemum flower extract (YJH) and mulberry leaf extract (SY)
Respectively taking 500g of selfheal and mulberry leaf, decocting and extracting twice, adding 10 times of water each time, decocting for 1.5 hours, merging decoction, filtering, concentrating filtrate to obtain clear paste with the relative density of 1.06-1.10 (80 ℃), adding 95% ethanol to ensure that the ethanol content reaches 63%, fully stirring, standing overnight, filtering, recovering ethanol from the filtrate, concentrating under reduced pressure to a proper amount, and respectively obtaining the selfheal extract (XKC, 1g is equivalent to the crude drug amount of 3.56 g) and the mulberry leaf extract (SY, 1g is equivalent to the crude drug amount of 3.24 g).
Taking 500g of wild chrysanthemum flower, wherein 50g of the wild chrysanthemum flower is soaked in ethanol to obtain wild chrysanthemum flower soaking liquid for standby; decocting the rest 450g of flos Chrysanthemi Indici with water twice, each time with 10 times of water for 1.5 hr, mixing decoctions, filtering, concentrating the filtrate to obtain fluid extract with relative density of 1.06-1.10 (80deg.C), adding 95% ethanol to reach alcohol content of 63%, stirring thoroughly, standing overnight, filtering, mixing the filtrates with the above flos Chrysanthemi Indici soaking solution, recovering ethanol, and concentrating under reduced pressure to appropriate amount to obtain flos Chrysanthemi Indici extract (YJH, 1g corresponds to crude drug content 2.89 g).
Application example 1 Effect of different osmotic pressures on human corneal epithelial cell Activity
The effect of different concentrations of NaCl on the activity of human corneal epithelial cells was explored to select the appropriate modeling concentration for the experiment.
MTS is a new generation tetrazolium blue salt compound, and the Formazan product formed by reduction is higher in water solubility and stability and faster in color development. The application principle of MTT is the same as that of MTT, namely, the MTT is reduced into various colored formazan products by various dehydrogenases in mitochondria of living cells, and the color depth of MTT is highly related to the number of living cells of certain sensitive cell strains in a certain range.
The cells were treated with different concentrations (0, 30, 50, 70, 90, 110, 130 mM) of NaCl for 24h, the medium was changed, after 2h incubation with MTS, absorbance (OD) values at 490nm were measured with a microplate reader, and the cell viability of each group was calculated according to the formula: cell viability = experimental OD/control OD x 100%.
At 24h of treatment with different osmotics, 350, 400 and 450mOsm/L osmotics have no significant effect on the activity of human corneal epithelial cells, and 500-550mOsm/L osmotics reduce the activity of human corneal epithelial cells by not more than 50%. The 600mOsm/L osmotic pressure reduced its activity below 50%. The results are shown in FIG. 1.
Selecting 600mOsm/L osmotic pressure treatment to observe the protective effect of the mulberry chrysanthemum on cells; 450mOsm/L osmotic pressure molding is selected to observe the anti-inflammatory and antioxidant effects of the Xiasangju.
Application example 2 Effect of the Chamomile extract of example 1 on human corneal epithelial cell Activity
The effect of different concentrations of the grass mulberry chrysanthemum extract on the activity of human cornea epithelial cells was investigated. The study was performed with the appropriate dosing concentrations selected.
MTS is a new generation tetrazolium blue salt compound, and the Formazan product formed by reduction is higher in water solubility and stability and faster in color development. The application principle of MTT is the same as that of MTT, namely, the MTT is reduced into various colored formazan products by various dehydrogenases in mitochondria of living cells, and the color depth of MTT is highly related to the number of living cells of certain sensitive cell strains in a certain range.
The cells were treated with different concentrations (100 mg/ml, 10mg/ml, 1mg/ml, 100. Mu.g/ml, 10. Mu.g/ml, 1. Mu.g/ml) of the Xiasangju extract of example 1 for 24 hours, the medium was changed, MTS was added for 2 hours incubation, absorbance (OD) values at 490nm were measured using an ELISA reader, and cell viability of each group was calculated according to the formula: cell viability = experimental OD/control OD x 100%.
As shown in figure 2, when the concentration of the extract of the grass mulberry chrysanthemum is more than or equal to 10mg/ml, the extract has remarkable inhibition effect on the cell activity; the concentration of the grass mulberry chrysanthemum extract is less than or equal to 5mg/ml, and the grass mulberry chrysanthemum extract has no obvious influence on the activity of cells. The efficacy of XSJ was evaluated by setting a plurality of concentrations at a maximum concentration of 5 mg/ml.
Application example 3 protection of hypertonically induced human corneal epithelial cells by the extract of Chamomile of example 1
One of the causes of xerophthalmia is corneal epithelial oxidative stress and inflammatory infiltration caused by hyperfiltration, and the experiment explores the protection effect of different concentrations of the grass mulberry chrysanthemum extract on human corneal epithelial cells under the condition of hyperfiltration.
Human corneal epithelial cells in the logarithmic growth phase were inoculated into 96-well cell culture plates at a density of 5000 cells/dish after digestion with 0.25% pancreatin, and placed in an incubator at 37℃for overnight culture.
Preparing NaCl solution and a Xiasangju extract: accurately weighing a proper amount of sodium chloride solid and the extract of the mulberry chrysanthemum in the embodiment 1, placing the extract in an ultra-clean bench, irradiating ultraviolet for 30min for sterilization, adding a DMEM culture medium, and carrying out ultrasonic treatment at 37 ℃ and 100% power for 30min after vortex mixing uniformly to prepare corresponding concentration. It is ready for use.
And (3) grouping setting: the control group, 600mosm/L group, 600 mosm/L+5mg/mL group, 600 mosm/L+2.5mg/mL group, 600 mosm/L+1.25mg/mL group, 600 mosm/L+625. Mu.g/mL group, 600 mosm/L+312.5. Mu.g/mL group, 600 mosm/L+156. Mu.g/mL group, 600 mosm/L+78. Mu.g/mL group, plating the next day of the primary culture supernatant, adding PBS for 2 times, adding 100. Mu.L of solutions with different concentrations corresponding to the groups into each culture dish according to the group setting, and placing the culture dishes in a 37 ℃ incubator for continuous culture; 3 replicates were set and incubated for 24h.
MTS detection: the crude culture supernatant was discarded, washed 2 times with PBS, 100. Mu.l of DMEM medium and 20. Mu.l of MTS were added, and incubated at 37℃for 1-4 hours. The absorbance of each well at 490nm was measured while blank wells (medium and MTS solution, cell-free) were set.
The average absorbed light intensity was calculated using GraphPad prism8.0.1 software counts, from which the protective effect of the extract of shasangju and single medicinal material on hypertonically induced human corneal epithelial cells was investigated. The results are shown in FIG. 3.
Compared with the model group, 0.625mg/ml and 1.25mg/ml XSJ can significantly improve the activity of human corneal epithelial cells, and the cell activity is improved from 45% to 67.3% and 64.9% of the model group. The extract of the mulberry leaf chrysanthemum has obvious inhibition effect on the cell activity reduction caused by high osmotic pressure.
Application example 4 Effect of the extract of Chamomile of example 1 on ROS level in human corneal epithelial cells under hyperosmotic Induction
Under the induction of hyperosmosis, ROS of human cornea epithelial cells can be increased, and the experiment explores the influence of different concentrations of the grass mulberry chrysanthemum extract on ROS levels of human cornea epithelial cells under the condition of hyperosmosis.
The reactive oxygen species (ROS Assay Kit) is a Kit for detecting reactive oxygen species by using a fluorescent probe DCFH-DA. DCFH-DA itself has no fluorescence, can freely pass through cell membrane, and can be hydrolyzed by intracellular esterase to generate DCFH after entering into cells. Whereas DCFH cannot penetrate the cell membrane, thus making the probe easily loaded into the cell. Intracellular reactive oxygen species can oxidize non-fluorescent DCFH to produce fluorescent DCF. The level of intracellular active oxygen can be known by detecting the fluorescence of DCF.
Human cornea epithelial cell line, xiasangju extract, naCl, HCE-T special culture medium, DMEM culture medium, phosphate buffer (phosphate buffer solution, PBS), 0.25% pancreatin-EDTA, ROS detection kit, flow cytometry, ultra-clean bench, 6-hole cell culture plate, carbon dioxide incubator, electrothermal constant temperature water bath pot, etc.
Well-grown human cornea epithelial cells are inoculated into a 6-well plate after 0.25% trypsin digestion, centrifugation and resuspension, and then are continuously cultured. Ensure that the cell confluency reaches 50-70% during detection.
Preparing NaCl solution and a Xiasangju extract: accurately weighing a proper amount of sodium chloride solid and the extract of the mulberry chrysanthemum in the embodiment 1, placing the extract in an ultra-clean bench, irradiating ultraviolet for 30min for sterilization, adding a DMEM culture medium, and carrying out ultrasonic treatment at 37 ℃ and 100% power for 30min after vortex mixing uniformly to prepare corresponding concentration. It is used for each time.
Modeling and drug administration grouping setting: control, 450mosm/L, 450 mosm/L+1.25mg/mL, 450 mosm/L+625. Mu.g/mL, 450 mosm/L+312.5. Mu.g/m, positive drug (NAC) group. Ensuring that the cell confluency reaches 50-70%, adding PBS, cleaning for 2 times, respectively adding 1mL blank culture medium or solutions with different concentrations into each culture dish according to the grouping arrangement, placing into a 37 ℃ incubator for continuous culture, setting 3 repetitions, and culturing for 24 hours.
Thawing DCFH-DA, and directly adding the thawed DCFH-DA into serum-free DMEM medium according to a ratio of 1:1000, and shading with tinfoil for later use. Mu.l pancreatin was added to each well of the 6-well plate, incubated for 5min, the digestion was stopped by adding an equal volume of HCE-T complete medium, cells in the wells were properly blown off, centrifuged for 5min (500 rcf,5 min) in each set of EP tubes, and the supernatant was discarded, taking care not to aspirate the pellet. 1ml PBS was added for resuspension, washing, centrifugation for 5min, and the supernatant was discarded. The EP tubes were put together by resuspension (1 ml per group) with the DCFH-DA solution prepared previously, taking care of shading with tinfoil, incubating for 20-30min, centrifuging for 5min,500rcf, and discarding the supernatant. 1ml of PBS was added for resuspension, washing, centrifugation for 5min, and the supernatant was discarded. 1ml of PBS was added again to resuspend, wash, centrifuge for 5min, and discard the supernatant. 600 μl PBS was added for resuspension and ready for up-flow. Care was taken to avoid light.
The mean fluorescence intensities were calculated by counting using GraphPad prism8.0.1 software, respectively, from which the effect of the shasangju extract on ROS levels in human corneal epithelial cells under hypertonic induction was investigated.
ROS were detected after staining, and the results are shown in FIG. 4. The 450mOsm/L group showed significantly increased ROS production compared to the normal group; the 450mOsm/L+625 μg/ml group showed significantly reduced ROS production compared to the 450mOsm/L group.
Application example 5 Effect of the extract of Chamomile of example 1 on the secretion of IL-6 by human corneal epithelial cells under hypertonic Induction
Under the induction of hyperosmosis, IL-6 of human cornea epithelial cells can be increased, and the experiment explores the influence of different concentrations of the grass mulberry chrysanthemum extract on ROS level of human cornea epithelial cells under the condition of hyperosmosis.
Coating IL-6 antibody in 96-well plate to form solid phase carrier, adding standard substance or specimen to the micro-pore, combining IL-6 with antibody connected to the solid phase carrier, adding biotinylated IL-6 antibody, washing unbound biotinylated antibody, adding HRP, and washing thoroughly again, adding TMB substrate for color development. TMB is converted to blue under the catalysis of peroxidase and to final yellow under the action of acid, the shade of the color and the IL-6 concentration in the sample being positively correlated. The absorbance was measured at 450m wavelength using a microplate reader, and the sample concentration was calculated.
Instrument and material: human corneal epithelial cell line, the extract of Mulberry leaf in example 1, naCl, HCE-T special medium, DMEM medium, phosphate Buffer (PBS), 0.25% pancreatin-EDTA, ELISA detection kit, ultra clean bench, 12-well cell culture plate, carbon dioxide incubator, electrothermal thermostatic water bath pot, etc.
Modeling with 450mOsm/L, treating with different concentrations of XIASANGJU extract for 24 hr, collecting culture supernatant, and detecting with ELISA kit.
(1) Sample adding: and respectively arranging a standard hole, a sample hole to be tested and a blank hole. Setting a standard hole 7 holes, sequentially adding 100 mu L of standard blank holes with different concentrations and 100 mu L of standard diluent, yu Kongjia to-be-detected sample 100 mu L, and adding a coating film on an ELISA plate, and incubating for 1 hour at 37 ℃.
(2) Discard the liquid, spin-dry, and do not need washing.
(3) 100 mu L of the working solution of the detection solution A (prepared before 15 min) is added to each well, and the ELISA plate is covered and incubated for 1 hour at 37 ℃.
(4) The wells were discarded, each well was washed with 350 μl of wash solution, soaked for 1-2 minutes, and the elisa plate was tapped on absorbent paper to remove all the liquid from the wells. The plate was washed repeatedly 3 times. After the last washing, the residual washing buffer solution is sucked or poured out, the ELISA plate is reversely buckled on the water absorbing paper, and all the liquid remained in the hole is absorbed.
(5) 100 mu L of working solution (prepared before use) of detection solution B is added to each well, and the ELISA plate is covered and incubated for 30 minutes at 37 ℃.
(6) Discarding the liquid in the holes, spin-drying, washing the plate for 5 times, and the method is the same as the step (4).
(7) Adding 90 mu L of TMB substrate solution into each well, coating an ELISA plate, and developing at 37 ℃ in a dark place (the reaction time is controlled to be 10-20 minutes and not more than 30 minutes, and stopping when the gradient of the front 3-4 wells of the standard well is obviously blue and the gradient of the rear 3-4 wells is not obvious).
(8) The reaction was stopped by adding 50. Mu.L of stop solution to each well, at which time the blue color turned yellow immediately. The order of addition of the stop solution should be as similar as possible to the order of addition of the substrate solution. If the color is uneven, the ELISA plate is gently shaken to mix the solution uniformly.
(9) After ensuring that the bottom of the ELISA plate has no water drop and no bubble in the hole, the optical density value (OD value) of each hole is measured by an ELISA plate at 450nm wavelength
The results are shown in FIG. 5, where the IL-6 secretion levels were significantly reduced in the three different concentrations of the XiasangJu extract groups compared to the model group (450 mosm/L). The extract of the mulberry leaf chrysanthemum has an inhibiting effect on the generation of the cell IL-6 caused by high osmotic pressure.
Application example 6 comparison of the effect of the extract of Chamomile of example 1 on the ROS level of human corneal epithelial cells under hypertonic induction with the extract of a single medicinal material
According to the prescription proportion, proper concentration is given to cells, the influence of the extract of the mulberry chrysanthemum and the extract of the single medicinal material on the ROS level of the human cornea epithelial cells under the hypertonic condition is compared, and whether the three medicinal materials have a synergistic effect or not is examined.
Reactive oxygen species detection was performed using a reactive oxygen species detection Kit (ROS Assay Kit) using the fluorescent probe DCFH-DA.
The instrument and the material comprise a human cornea epithelial cell line, a grass mulberry chrysanthemum extract (example 1), a single medicinal material extract (comparative example 2), naCl, a special culture medium for HCE-T, a DMEM culture medium, phosphate Buffer (PBS), 0.25% pancreatin-EDTA, a ROS detection kit, a flow cytometry instrument, an ultra-clean bench, a 6-hole cell culture plate, a carbon dioxide incubator, an electric heating constant temperature water bath pot and the like.
Well-grown human cornea epithelial cells are inoculated into a 6-well plate after 0.25% trypsin digestion, centrifugation and resuspension, and then are continuously cultured. Ensure that the cell confluency reaches 50-70% during detection.
Preparing NaCl solution, xiasangju extract and single medicinal material extract: proper amounts of sodium chloride solid, a Xiasangju extract (XSJ, example 1), a Prunella vulgaris extract (XKC, comparative example 2), a wild chrysanthemum extract (YJH, comparative example 2) and a Mulberry leaf extract (SY, comparative example 2) are accurately weighed, placed in an ultra-clean bench, irradiated with ultraviolet for 30min for sterilization, then added with DMEM culture medium, and subjected to vortex mixing, and subjected to ultrasonic treatment with 100% power for 30min at 37 ℃ to prepare corresponding concentrations. It is ready for use.
And (3) grouping setting: control, 450mosm/L, 450 mosm/L+625. Mu.g/mL XSJ, 450 mosm/L+414. Mu.g/mL XKC, 450 mosm/L+145. Mu.g/mL SY, 450 mosm/L+66. Mu.g/mL YJH.
Ensuring that the cell confluency reaches 50-70%, adding PBS, cleaning for 2 times, respectively adding 1mL blank culture medium or solutions with different concentrations into each culture dish according to the grouping arrangement, placing into a 37 ℃ incubator for continuous culture, setting 3 repetitions, and culturing for 24 hours.
Thawing DCFH-DA, and directly adding the thawed DCFH-DA into serum-free DMEM medium according to a ratio of 1:1000, and shading with tinfoil for later use. Mu.l pancreatin was added to each well of the 6-well plate, incubated for 5min, the digestion was stopped by adding an equal volume of HCE-T complete medium, cells in the wells were properly blown off, centrifuged for 5min (500 rcf,5 min) in each set of EP tubes, and the supernatant was discarded, taking care not to aspirate the pellet. 1ml PBS was added for resuspension, washing, centrifugation for 5min, and the supernatant was discarded. The EP tubes were put together by resuspension (1 ml per group) with the DCFH-DA solution prepared previously, taking care of shading with tinfoil, incubating for 20-30min, centrifuging for 5min,500rcf, and discarding the supernatant. 1ml of PBS was added for resuspension, washing, centrifugation for 5min, and the supernatant was discarded. 1ml of PBS was added again for resuspension, washing, centrifugation for 5min, and the supernatant was discarded. 600 μl PBS was added for resuspension and ready for up-flow. Care was taken to avoid light.
The average fluorescence intensity was counted and calculated by using GraphPad prism8.0.1 software, and the effect of the grass mulberry chrysanthemum extract and single medicinal material on the ROS level of human corneal epithelial cells under the induction of hyperosmosis was explored, and the result is shown in figure 6.
Compared to the model group, the cellular ROS levels of 625 μg/ml and 414 μg/ml Prunella vulgaris (XKC) groups were significantly reduced from 380% to 177% and 215%. 145. The cellular ROS levels of the Mulberry leaf (SY) group and the wild chrysanthemum flower (YJH) group with the concentration of 66 mu g/ml are not significantly different, so that the inhibition effect of the grass mulberry chrysanthemum extract (XSJ) on the generation of cellular ROS caused by high osmotic pressure is better than that of a single medicinal material, and the antioxidation effect of the grass mulberry chrysanthemum extract is better than that of the grass mulberry chrysanthemum extract.
Application example 7 comparison of the protective effect of the extract of Chamomile and the extract of Single medicinal material on hypertonically induced human corneal epithelial cells of example 1
MTS is a new generation tetrazolium blue salt compound, and the Formazan product formed by reduction is higher in water solubility and stability and faster in color development. The application principle of MTT is the same as that of MTT, namely, the MTT is reduced into various colored formazan products by various dehydrogenases in mitochondria of living cells, and the color depth of MTT is highly related to the number of living cells of certain sensitive cell strains in a certain range.
Instrument and material: human corneal epithelial cell line, xiasangju extract (example 1), single medicinal material extract (comparative example 2), naCl, HCE-T dedicated medium, DMEM medium, phosphate Buffer (PBS), 0.25% pancreatin-EDTA, MTS kit, fluorescence microplate reader, ultra clean bench, 96-well cell culture plate, carbon dioxide incubator, electrothermal thermostat water bath, etc.
Human corneal epithelial cells in the logarithmic growth phase were inoculated into 96-well cell culture plates at a density of 5000 cells/dish after digestion with 0.25% pancreatin, and placed in an incubator at 37℃for overnight culture.
Preparing NaCl solution, xiasangju extract and single medicinal material extract: accurately weighing appropriate amount of sodium chloride solid, XIASANGJU extract (XSJ, example 1), prunellae Spica extract (XKC, comparative example 2), flos Chrysanthemi Indici extract (YJH, comparative example 2), and folium Mori extract (SY, comparative example 2), placing in an ultra-clean bench, irradiating with ultraviolet for 30min for sterilization, adding DMEM culture medium, mixing under vortex, and performing ultrasonic treatment at 37deg.C under 100% power for 30min to obtain corresponding concentration. It is ready for use.
And (3) grouping setting: the control group, 600mosm/L group, 600 mosm/L+625 mug/mL XSJ group, 600 mosm/L+414 mug/mL XKC group, 600 mosm/L+145 mug/mL SY group, 600 mosm/L+66 mug/mL YJH group, plating the next day, discarding the original culture supernatant, adding PBS for 2 times, adding 100 mug of blank culture medium or solutions with different concentrations into each culture dish according to the grouping arrangement, and placing the culture dishes into a 37 ℃ incubator for continuous culture; 3 replicates were set and incubated for 24h.
The crude culture supernatant was discarded, washed 2 times with PBS, 100. Mu.l of DMEM medium and 20. Mu.l of MTS were added, and incubated at 37℃for 1-4 hours. The absorbance of each well at 490nm was measured while blank wells (medium and MTS solution, cell-free) were set.
The average absorbed light intensity was calculated using GraphPad prism8.0.1 software counts and the protective effect of the extract of shasangju and the extract of single medicinal material on hypertonically induced human corneal epithelial cells was investigated, and the results are shown in figure 7.
Only 0.625mg/ml of Xiasangju extract (XSJ) in the group significantly increased human corneal epithelial cell activity compared to the model group, from 48% to 75% in the model group. Whereas the cell activities of the 414. Mu.g/ml Prunella vulgaris (XKC) group, the 145. Mu.g/ml mulberry leaf (SY) group and the 66. Mu.g/ml wild chrysanthemum flower (YJH) group were not significantly affected. The method shows that the extract (XSJ) of the mulberry leaf and chrysanthemum has a remarkable inhibiting effect on the cell activity reduction caused by high osmotic pressure compared with the extract of a single medicinal material.
Application example 8 test of Effect on hypertonic solution induced rat Dry eye
Selecting a plurality of rats qualified in adaptability observation to perform binocular cornea fluorescence staining scoring and tear secretion amount measurement, eliminating animals with cornea problematic fluorescence staining abnormality and large binocular tear secretion amount difference, taking the measurement data as basic data, and screening the current diary as D0. The 24 animals qualified in the screening are selected and divided into a negative control group, a model control group, a summer mulberry chrysanthemum extract group (example 1) and a summer mulberry chrysanthemum water extraction alcohol immersion paste group (comparative example 1) according to the average value of the lacrimal secretion of two eyes, and each group of 6 animals, 12 eyes and both male and female are provided.
After screening, D1 was used to draw 20. Mu.L of the hypertonic solution and drop the solution into conjunctival sac of eyes of a molding animal, the interval is about 2 hours for 5 times/day, the animal eyelid was passively closed for about 90s after dropping for molding, the molding was performed without additional administration group as a model control group, and the molding was performed with the addition of the Xiasangju extract (example 1) and the Xiasangju aqueous alcohol extract paste (comparative example 1) as the Xiasangju extract group and the Xiasangju aqueous alcohol extract paste group respectively. Animals in the administration group were administered by intragastric administration of D1 (converted by body surface area equivalent dose method, approximately 4 times the clinical dose), 1 time daily, 28 days continuously, and 3 times on the day of D28. The negative control animals were instilled with an equal volume of physiological saline alone. The animals of each group were subjected to measurement of the amount of lacrimal secretion of both eyes at D14 and D28, respectively.
The results show that the extract of the summer mulberry and the extract of the summer Sang Jushui extract of the comparative example 1, which are respectively (converted by a body surface area equivalent dose method, and the dose is about 4 times of the clinical dosage), show the effect of remarkably increasing the tear secretion of rats with dry eye symptoms after being continuously administered for 28 days, wherein the extract of the summer mulberry has more remarkable improving effect than the extract of the summer mulberry which is extracted with water. Specific results are as follows (Table 1).
TABLE 1 influence on the lacrimal secretion of rats in dry eye model
±S，n=12）/>
Finally, it should be noted that the above description is only for illustrating the technical solution of the present invention, and not for limiting the scope of the present invention, and that the simple modification and equivalent substitution of the technical solution of the present invention can be made by those skilled in the art without departing from the spirit and scope of the technical solution of the present invention.
Claims (8)
1. The application of the grass-leaved sweetflag chrysanthemum extract in preparing the medicine for treating the hypertonic xerophthalmia is characterized in that the grass-leaved sweetflag chrysanthemum extract is an extract taking selfheal to Sang She wild chrysanthemum with the weight ratio of 500:175:80 as raw materials; the grass-mulberry chrysanthemum extract comprises grass-mulberry chrysanthemum extract and wild chrysanthemum impregnating solution.
2. The use according to claim 1, wherein the preparation method of the wild chrysanthemum impregnating solution comprises the following steps: taking 8% -20% of the weight of the wild chrysanthemum flower in the raw materials, adding an ethanol solution with the volume concentration of 50% -95% which is 1.5-10 times of the weight of the wild chrysanthemum flower, and extracting at 15-40 ℃ by an impregnation method or a percolation method to obtain the wild chrysanthemum flower extract; the preparation method of the grass mulberry chrysanthemum extract comprises the following steps: decocting the rest flos Chrysanthemi Indici, all Prunellae Spica and folium Mori in water for 1-3 times each for 1-2 hr, mixing decoctions, filtering, concentrating, adding ethanol to concentrate until the volume concentration of ethanol content reaches 63%, standing overnight, filtering to obtain supernatant, recovering ethanol from the filtrate, and concentrating under reduced pressure to obtain extract with relative density of 1.20-1.30.
3. The use according to claim 2, wherein the preparation method of the wild chrysanthemum impregnating solution is an impregnation method or a percolation method; wherein, the dipping method is as follows: taking wild chrysanthemum flower, dividing the wild chrysanthemum flower into three parts, respectively filling the three parts into a container according to the weight ratio of 4-6:2-4:1-3, soaking the wild chrysanthemum flower in 95% ethanol which is 1.5-3 times of the weight of the wild chrysanthemum flower, adding the wild chrysanthemum flower into the 1 st part, soaking for 1-3 days, transferring the discharged liquid into the 2 nd part, soaking for 1-3 days, transferring the discharged liquid into the 3 rd part, soaking for 1-3 days, filtering the leaching solution, and obtaining the wild chrysanthemum flower soaking liquid; the percolation method comprises the following steps: placing flos Chrysanthemi Indici into a percolating cylinder, soaking in 2-3 times of 95% ethanol solution for 12-24 hr, adding 7-8 times of 95% ethanol, percolating at a flow rate of 1-4mL per minute, and collecting ethanol solution to obtain flos Chrysanthemi Indici soaking solution.
4. The use according to claim 1, wherein the drug content of the grass-leaved chrysanthemum extract is 3-30g crude drug/g, and the drug content of the wild chrysanthemum impregnation liquid is 0.3-5g crude drug/mL.
5. The use according to claim 1, wherein said extract of sumac chrysanthemum is obtained by mixing a sumac chrysanthemum extract and a wild chrysanthemum flower extract in a ratio of sumac chrysanthemum extract to wild chrysanthemum flower extract= (5-25) g to 1 mL.
6. The use according to claim 1, wherein the active concentration of said extract of songaria chamomile is between 0.5 and 1000 μg/mL.
7. The use according to claim 1, wherein the medicament for treating hypertonic dry eye has a good effect on hypertonic dry eye and is capable of significantly promoting tear secretion.
8. The application of the grass-mulberry chrysanthemum extract in preparing medical equipment or medical facilities for treating hypertonic xerophthalmia is characterized in that the grass-mulberry chrysanthemum extract comprises grass-mulberry chrysanthemum extract and wild chrysanthemum impregnating solution.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101297864A (en) * | 2008-06-27 | 2008-11-05 | 广州星群（药业）股份有限公司 | Preparation of Xia sang ju formulation |
| CN105106318A (en) * | 2015-09-15 | 2015-12-02 | 北京市眼科研究所 | Traditional Chinese medicine composition for treating xerophthalmia and preparation method of traditional Chinese medicine composition |
| CN109125401A (en) * | 2018-08-15 | 2019-01-04 | 康美保宁（四川）制药有限公司 | A kind of preparation method of mulberry and chrysanthemum granules thick paste |
| CN109432338A (en) * | 2019-01-18 | 2019-03-08 | 河南德盟医药科技有限公司 | It is a kind of for the Chinese medicine composition of eyeshield health care, the preparation method of Chinese medicine composition and eye sticker |
| CN111888390A (en) * | 2020-08-27 | 2020-11-06 | 广州白云山星群(药业)股份有限公司 | Application of Xiasangju extract in inhibiting human coronavirus |
-
2023
- 2023-05-05 CN CN202310495599.XA patent/CN116211916B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101297864A (en) * | 2008-06-27 | 2008-11-05 | 广州星群（药业）股份有限公司 | Preparation of Xia sang ju formulation |
| CN105106318A (en) * | 2015-09-15 | 2015-12-02 | 北京市眼科研究所 | Traditional Chinese medicine composition for treating xerophthalmia and preparation method of traditional Chinese medicine composition |
| CN109125401A (en) * | 2018-08-15 | 2019-01-04 | 康美保宁（四川）制药有限公司 | A kind of preparation method of mulberry and chrysanthemum granules thick paste |
| CN109432338A (en) * | 2019-01-18 | 2019-03-08 | 河南德盟医药科技有限公司 | It is a kind of for the Chinese medicine composition of eyeshield health care, the preparation method of Chinese medicine composition and eye sticker |
| CN111888390A (en) * | 2020-08-27 | 2020-11-06 | 广州白云山星群(药业)股份有限公司 | Application of Xiasangju extract in inhibiting human coronavirus |
Non-Patent Citations (2)
| Title |
|---|
| 李青青等: "中医药治疗干眼症研究进展", 河南中医, vol. 37, no. 11, pages 2051 - 2054 * |
| 金明主编: "《眼科疾病分册 中成药临床应用指南》", 中国中医药出版社, pages: 41 * |
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