CN114796113B - Application of blumea balsamifera nanoemulsion in preparation of antitumor drugs and wound repair drugs - Google Patents
Application of blumea balsamifera nanoemulsion in preparation of antitumor drugs and wound repair drugs Download PDFInfo
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Abstract
The invention belongs to the technical field of preparation of medicinal preparations, and in particular relates to application of blumea balsamifera nanoemulsion in preparation of anti-tumor medicaments and wound repair medicaments; the formula of the blumea balsamifera nanoemulsion is screened, and the anti-tumor and wound repair effects are researched, so that the concept that blumea balsamifera with anti-inflammatory and bactericidal effects is widely considered to have good anti-tumor effects is proved, and the anti-tumor effects of the blumea balsamifera nanoemulsion are found to be slightly higher than those of common blumea balsamifera. The combined blumea balsamifera nanoemulsion has the advantages of low manufacturing cost, simple operation process, wound healing and anti-tumor effect. So that the blumea balsamifera nanoemulsion has better development prospect.
Description
Technical Field
The invention belongs to the technical field of preparation of medicinal preparations, and in particular relates to application of blumea balsamifera nanoemulsion in preparation of anti-tumor medicaments and wound repair medicaments.
Background
Nanoemulsions, also known as microemulsions, are thermodynamically stable systems formed from aqueous phase, oil phase, surfactant and cosurfactant in proportions, and have high transparency as emulsions with an average particle size of 10-100nm, and are often used in the pharmaceutical and chemical fields.
The blumea balsamifera oil has good sterilization and anti-inflammation effects, but poor water solubility and volatility limit the wide application of the blumea balsamifera oil, a nano drug carrier is used for wrapping the blumea balsamifera oil, the strong stimulation taste of the blumea balsamifera oil can be covered on the basis of improving the stability of the drug, in addition, in the skin wound repair process (comprising four stages of a hemostatic stage, an inflammatory stage, a value-added stage and a remodelling stage), the inflammatory stage is a necessary stage of skin wound repair, and the blumea balsamifera oil is combined with the sterilization and anti-inflammation effects of the blumea balsamifera oil to the aspects of promoting skin wound repair and the like, and the compound blumea balsamifera granulation promoting gel with the effect of promoting skin ulcer wound repair is disclosed in a patent document with publication number of CN108524570A and a preparation method thereof. It is used for treating diabetic skin ulcer, burn, wound repair, scleroderma, and other skin ulcer. The patent document with publication number of CN110974861A discloses blumea balsamifera liposome which consists of blumea balsamifera, lecithin, cholesterol and PBS solution, can be matched with auxiliary materials acceptable in the fields of medicines and daily chemicals, and further can be prepared into products such as gel, ointment, eye ointment, liniment, aerosol, solution, sol, emulsion, suspension and the like, and has good transdermal absorption effect and anti-inflammatory, analgesic and skin repairing activity. However, the efficacy research of blumea balsamifera is mainly focused on anti-inflammatory and analgesic aspects, and the anti-tumor activity of blumea balsamifera is only in shallow layers, and detailed reports of intensive researches are not seen.
Disclosure of Invention
The invention provides an application of blumea balsamifera nanoemulsion in preparing anti-tumor drugs and wound repair drugs, aiming at solving the problems.
The method is realized by the following technical scheme:
1. the application of blumea balsamifera nanoemulsion in preparing antitumor medicine is that the tumor refers to human non-small cell lung cancer, human malignant embryo rhabdomyoma cell and human tongue squamous carcinoma cell.
Furthermore, the antitumor drug is an oral preparation, and the blumea balsamifera nanoemulsion is combined with pharmaceutically acceptable auxiliary materials to prepare the required preparation.
2. An application of blumea balsamifera nanoemulsion in preparing wound repair medicine is provided.
Further, the application of the blumea balsamifera nanoemulsion in preparing a wound repair medicament is characterized in that the wound repair medicament is an external liniment, and the blumea balsamifera nanoemulsion is combined with pharmaceutically acceptable auxiliary materials to prepare a required preparation.
3. The preparation method of the blumea balsamifera nanoemulsion is characterized by comprising, by mass, 1-10% of blumea balsamifera, 1-5% of isopropyl myristate, 1-10% of a surfactant, 1-7% of a cosurfactant and 100% of water.
Further, the surfactant is PCO40, and the cosurfactant is absolute ethyl alcohol.
Further, the specific preparation method comprises the following steps:
(1) Precisely weighing blumea balsamifera and isopropyl myristate according to the formula proportion, and magnetically stirring and uniformly mixing at room temperature to obtain a mixed oil phase;
(2) Weighing absolute ethyl alcohol, adding the absolute ethyl alcohol into the mixed oil phase obtained in the step (1), and uniformly stirring at the speed of 2000r/min to obtain a mixed solution;
(3) Taking PCO40 according to the formula proportion, adding the PCO40 into the mixed solution obtained in the step (2), and stirring at the rotating speed of 2000-2100r/min for 2-4min to obtain coarse emulsion;
(4) And (3) placing the crude emulsion obtained in the step (3) in an ultrasonic pulverizer, selecting a 2mm ultrasonic amplitude transformer with 400w power, and performing ultrasonic emulsification in an ice-water bath for 1h to obtain blumea balsamifera nanoemulsion, wherein the average particle size is 62.8nm, and the encapsulation rate is more than 98%.
The appearance of blumea balsamifera nanoemulsion and blank nanoemulsion is shown in figure 3; the morphological characteristics of blumea balsamifera nanoemulsion under a transmission electron microscope are shown in figure 4; the particle size distribution of blumea balsamifera nanoemulsion is shown in figure 5.
1. Blumea balsamifera nanoemulsion formula screening
1. Determination of oil phase
The solubility of blumea balsamifera oil in IPM, olive oil and corn oil is observed, and the optimal oil phase suitable for the nanoemulsion system is screened. The detailed results are shown in Table 1.
TABLE 1
IPM was chosen as the mixed oil phase for blumea balsamifera.
2. Determination of cosurfactants
Blumea balsamifera oil is insoluble in water, and is easily dissolved in absolute ethanol, chloroform and diethyl ether, and the solubility of blumea balsamifera oil and organic solvent is determined at room temperature. From the green environmental protection point of view, absolute ethyl alcohol is used as a surfactant.
3. Determination of surfactant
The surfactant plays a key role in the preparation process of the nanoemulsion, the nonionic surfactants TW80, TW20, PCO40 and EL40 with low toxicity are respectively selected as alternative surfactants, absolute ethyl alcohol is used as cosurfactant, IPM is used as blumea balsamifera mixed oil phase to prepare the nanoemulsion, the nanoemulsion is placed for 24 hours at the low temperature of 4 ℃ and the high temperature of 45 ℃, and the stability of the nanoemulsion is examined by comparing the sizes (n=3) of the nanoparticles before and after temperature circulation. The composition of each component is recorded in mass percent. The detailed results of the experiment are shown in Table 2.
TABLE 2
The data show that the nanoemulsion prepared with surfactants TW20, TW80, EL40 had a very large variation in the average particle size before and after warm cycling and poor stability, whereas the nanoemulsion prepared with surfactant PCO40 had a small fluctuation in the average particle size before and after warm cycling, so PCO40 was selected as the optimal surfactant.
4. Determination of Km value (mass ratio of surfactant to cosurfactant)
PCO40 is used as a surfactant, absolute ethyl alcohol is used as a cosurfactant, IPM is used as a mixed oil phase, and the surfactant and the cosurfactant are mixed according to a ratio of 3: 1. 2: 1. 1:1, and then mixing the mixed surfactant and the determined oil phase IPM according to the mass ratio of 1: 1. 1: 2. 3: 5. 5: 3. 7: 2. 8:11, gradually dripping distilled water and continuously stirring until clear and transparent nano emulsion is formed, recording the water adding amount when the critical change occurs in the system, calculating the mass fractions of each component in the system, respectively taking a surfactant/cosurfactant, an oil phase and a water phase as three vertexes, and drawing a pseudo ternary phase diagram by using Origin software as shown in figure 1, wherein the area of a shadow part represents the emulsification rate.
From the graph, it can be found that when the mass ratio of the surfactant to the cosurfactant is 2: since the emulsion ratio of the nanoemulsion was the highest at 1, the km value was determined to be 2.
5. Investigation of surfactant usage
The amount of the surfactant can influence the particle size of the nanoemulsion, PCO40 is used as the surfactant, the Km value is fixed to be 2, the influence of the amount of the surfactant on blumea balsamifera nanoemulsion is examined, and the experimental results are shown in Table 3. The composition of each component is recorded in mass percent.
TABLE 3 Table 3
As can be seen from the results of Table 3, as the amount of the surfactant increases, the particle size of the blumea balsamifera nanoemulsion increases, and when the amount of the surfactant is 2.5% and 5%, the blumea balsamifera nanoemulsion prepared exhibits a milky appearance, but when the amount of the surfactant is increased to 7.5% and 10%, respectively, the blumea balsamifera nanoemulsion prepared exhibits higher transparency and better stability, and from the viewpoint of saving cost, the amount of the surfactant is 7.5% as a standard.
6. Effect of oil Loading on nanoemulsion particle size
The amount of the immobilized surfactant PCO40 was 7.5%, the Km value was 2, and the oil amounts were 3%, 5%, 10%, 15% and 20% respectively to develop blumea balsamifera nanoemulsion, and the data are shown in Table 4.
TABLE 4 Table 4
When the oil consumption is 3% and 5%, the prepared nanoemulsion is poor in stability, respectively shows the micro-cream yellow and cream yellow appearances, when the oil consumption is 10%, 15% and 20%, the blumea balsamifera nanoemulsion is clear and transparent, but when the oil consumption reaches 15% and 20%, the viscosity of the nanoemulsion is too high, three aspects of experiment cost, maximum drug loading capacity of the nanoemulsion and optimal appearance of the nanoemulsion are comprehensively saved, and the oil consumption is 10% as a standard for preparing subsequent nanoemulsions.
7. Determination of the Mixed oil proportion
PCO40 is used as a surfactant, absolute ethyl alcohol is used as a cosurfactant, IPM is used as an oil phase, the fixed Km value is 2:1, and blumea balsamifera oil is used as a main oil phase: IPM is respectively 10: 0. 9: 1. 8: 2. 7: 3. 6: 4. 5:5 develop the preparation experiment and examine its stability, the results are shown in table 5.
TABLE 5
As can be seen from the above table, when blumea balsamifera oil: IPM ratio of 9:1 and 8:2, the stability of the prepared nanoemulsion is best, and the maximum drug loading is considered, so that blumea balsamifera is used in the preparation of nanoemulsion and the process: ipm=9: 1 is used as a standard.
The ratio of blumea balsamifera oil to IPM can affect the particle size and stability of the nanoemulsion, and the stability evaluation is mainly carried out by placing the nanoemulsion in a refrigerator at 4 ℃ and an oven at 45 ℃ for 24 hours respectively, and comparing the particle size change and appearance change of the nanoemulsion before and after temperature cycle, wherein the formula screening process is shown in figure 2. Blumea balsamifera oil: ipm=9: 1, blumea balsamifera nanoemulsion has the best stability.
The raw material components of the nanoemulsion are composed of blumea balsamifera oil, IPM, a surfactant, a cosurfactant and water through formula screening, and the components are recorded as follows by mass percent: blumea balsamifera oil 9%, IPM 1%, surfactant PCO 40.5%, cosurfactant absolute ethanol 3.75%, and water 100%. The nanoemulsion is prepared by a ultrasonic emulsification method.
In summary, the beneficial effects of the invention are as follows: the invention is proved by screening the formula of the blumea balsamifera nanoemulsion and researching the anti-tumor and wound repair effects of the blumea balsamifera nanoemulsion, and the blumea balsamifera nanoemulsion has better anti-tumor effect which is generally considered to have anti-inflammatory and bactericidal effects, and the anti-tumor effect of the blumea balsamifera nanoemulsion is found to be slightly higher than that of common blumea balsamifera. The combined blumea balsamifera nanoemulsion has the advantages of low manufacturing cost, simple operation process, wound healing and anti-tumor effect. So that the blumea balsamifera nanoemulsion has better development prospect.
Blumea balsamifera as the main drug in nanoemulsions, oswald ripening (or oswald ripening) is a phenomenon that can be observed in solid solutions or sols, which describes a change in heterogeneous structure over time, with smaller crystalline or sol particles in the solute dissolving and redepositing onto larger crystalline or sol particles. Eventually leading to the formation of large droplets and even phase separation of the emulsion droplets during storage. The storage stability of blumea balsamifera nanoemulsions is generally improved by incorporating maturation inhibitors into the essential oils to delay droplet growth. The nano emulsion prepared by adopting isopropyl myristate (IPM) as an oswald antagonist has good stability and transparent appearance, and the blumea balsamifera nano emulsion prepared based on the formula has good skin wound healing efficacy and anti-tumor effect. Meets the current market demand. The average particle size of the blumea balsamifera nano emulsion is 62.8nm, and the encapsulation rate is more than 98%.
Drawings
FIG. 1 is a pseudo ternary phase diagram of Km value screening.
FIG. 2 shows screening of blumea balsamifera and IPM ratio.
Fig. 3 shows the appearance of blumea balsamifera nanoemulsion and blank nanoemulsion.
Fig. 4 is a morphological feature of blumea balsamifera nanoemulsion under transmission electron microscopy.
FIG. 5 shows the particle size distribution of blumea balsamifera nanoemulsion.
FIG. 6 is a graph showing the effect of blumea balsamifera nanoemulsion on skin wound repair (scale in the figure indicates 50 mm).
FIG. 7 shows skin wound repair rate and weight change of mice with blumea balsamifera nanoemulsion.
FIG. 8 shows the toxic effects of blumea balsamifera on tumor cells (TCA, RD, A549).
FIG. 9 is a comparison of toxicity effects of blumea balsamifera and blumea balsamifera nanoemulsion on A549 tumor cells.
FIG. 10 shows the toxic effects of PCO40 and EtOH on A549 tumor cells.
FIG. 11 is a graph showing the comparative effect of blumea balsamifera nanoemulsion on skin wound repair (marked ruler in the figure: 50 mm).
FIG. 12 shows skin wound repair rate of blumea balsamifera and weight change of mice.
FIG. 13 is a graph showing the cumulative permeation rate of blumea balsamifera nanoemulsion.
FIG. 14 is a graph showing the cumulative transmittance of blumea balsamifera nanoemulsion.
Detailed Description
The following detailed description of the invention is provided in further detail, but the invention is not limited to these embodiments, any modifications or substitutions in the basic spirit of the present examples, which still fall within the scope of the invention as claimed.
Example 1
Uniformly mixing 2g of blumea balsamifera with 0g of IPM to obtain a mixed oil phase, adding 0.75g of absolute ethyl alcohol into the mixed oil phase to be fully dissolved, then adding 1.5g of PCO40, mixing by a magnetic stirrer until the PCO40 is completely dispersed, dripping 15.75g of distilled water by a rubber head dropper while stirring to obtain a blumea balsamifera crude nanoemulsion, and carrying out ultrasonic treatment by an ultrasonic cell pulverizer for 1h to obtain the blumea balsamifera nanoemulsion (1). The average particle size of the nanoemulsion before and after temperature cycling was 61.1nm and 103.9nm, respectively.
Example 2
Uniformly mixing 1.8g of blumea balsamifera with 0.2g of IPM to obtain a mixed oil phase, adding 0.75g of absolute ethyl alcohol into the mixed oil phase to be fully dissolved, then adding 1.5g of PCO40, mixing by a magnetic stirrer until the PCO40 is completely dispersed, dripping 15.75g of distilled water by a rubber head dropper while stirring to obtain a blumea balsamifera crude nanoemulsion, and carrying out ultrasonic treatment by an ultrasonic cell pulverizer for 1h to obtain the blumea balsamifera nanoemulsion (2). The average particle size of the nanoemulsion before and after temperature cycling was 62.8nm and 62.2nm, respectively.
Example 3
Uniformly mixing 1.6g of blumea balsamifera with 0.4g of IPM to obtain a mixed oil phase, adding 0.75g of absolute ethyl alcohol into the mixed oil phase to be fully dissolved, then adding 1.5g of PCO40, mixing by a magnetic stirrer until the PCO40 is completely dispersed, dripping 15.75g of distilled water by a rubber head dropper while stirring to obtain a blumea balsamifera crude nanoemulsion, and carrying out ultrasonic treatment by an ultrasonic cell pulverizer for 1h to obtain the blumea balsamifera nanoemulsion (3). The average particle size of the nanoemulsion before and after temperature cycling was 87.2nm and 83.4nm, respectively.
Example 4
Uniformly mixing 1.4g of blumea balsamifera with 0.6g of IPM to obtain a mixed oil phase, adding 0.75g of absolute ethyl alcohol into the mixed oil phase to be fully dissolved, then adding 1.5g of PCO40, mixing by a magnetic stirrer until the PCO40 is completely dispersed, dripping 15.75g of distilled water by a rubber head dropper while stirring to obtain a blumea balsamifera crude nanoemulsion, and carrying out ultrasonic treatment by an ultrasonic cell pulverizer for 1h to obtain the blumea balsamifera nanoemulsion (4). The average particle size of the nanoemulsion before and after temperature cycling was 116.6nm and 88.4nm, respectively.
Example 5
Uniformly mixing 1.2g of blumea balsamifera with 0.8g of IPM to obtain a mixed oil phase, adding 0.75g of absolute ethyl alcohol into the mixed oil phase to be fully dissolved, then adding 1.5g of PCO40, mixing by a magnetic stirrer until the PCO40 is completely dispersed, dripping 15.75g of distilled water by a rubber head dropper while stirring to obtain a blumea balsamifera crude nanoemulsion, and carrying out ultrasonic treatment by an ultrasonic cell pulverizer for 1h to obtain the blumea balsamifera nanoemulsion (5). The average particle size of the nanoemulsion before and after temperature cycling was 135.4nm and 100.1nm, respectively.
Example 6
Uniformly mixing 1g of blumea balsamifera with 1g of IPM to obtain a mixed oil phase, adding 0.75g of absolute ethyl alcohol into the mixed oil phase to be fully dissolved, then adding 1.5g of PCO40, mixing by a magnetic stirrer until the PCO40 is completely dispersed, dripping 15.75g of distilled water by a rubber head dropper while stirring to obtain a blumea balsamifera crude nanoemulsion, and carrying out ultrasonic treatment by an ultrasonic cell pulverizer for 1h to obtain the blumea balsamifera nanoemulsion (6). The average particle size of the nanoemulsion before and after temperature cycling was 197.8nm and 197.1nm, respectively.
By comparison, the blumea balsamifera nanoemulsion (2) and the blumea balsamifera nanoemulsion (6) are found to have higher stability, wherein the blumea balsamifera nanoemulsion (2) has larger drug loading, so the blumea balsamifera nanoemulsion (2) is selected as a final formula and is used for carrying out animal experiments
1. Skin wound repair evaluation experiment
Wound repair is a complex and orderly process, each stage being carefully regulated by the body. Wound repair is generally divided into 4 phases: (1) a hemostasis stage; (2) an inflammatory reaction phase; (3) a cell proliferation differentiation stage; (4) and a tissue reconstruction and scar formation stage. It is a complex dynamic process of self-repair by interactions between various growth factors, extracellular matrix and repair cells after injury occurs in tissues. The experiment utilizes the blumea balsamifera nanoemulsion obtained by the invention to compare the skin wound healing effect of a positive group mupirocin and a blank group.
1 Experimental method
1.1. Grouping animals
KM mice, male and female mice are divided into groups randomly, 8 mice in each group are divided into groups: blank (without any treatment), positive (mupirocin ointment), blumea balsamifera nano-drug.
1.2 Experimental methods
The hair on the back of the mice was removed by an epilator, washed with warm water, wiped dry, and anesthetized with 1% chloral hydrate at 0.007 ml/g. Then, the mice are partially disinfected by 5% iodine and 75% alcohol, and circular wounds with the diameter of 10mm are cut at the positions of 10mm on two sides of the back spine of the mice, so that fascia and adipose tissues are not damaged. The wound thickness of the muscle layer was about 3mm, and the wound was rinsed and sterilized with physiological saline. The wound surface is opened, hemostasis is carried out fully, and after waking up, the wound surface is fed and observed for 24 hours in a single cage, and free drinking and eating are carried out in the period. The method takes that the wound has no congestion, swelling and suppuration, the mice have normal feeding and good activity as the judging standard, and the mice are used as the experimental subjects to group experiments. Each group was sterilized with iodine and physiological saline every day, changed once daily (blank group was not treated, positive drug group and blumea balsamifera nanoemulsion drug group were each administered 100 mg), and wound healing was observed by photographing recording on 1 st, 3 rd, 5 th, and 7 th day of wound treatment, respectively, and wound healing rate and weight change were calculated. The experimental results are shown in FIG. 6. It can be found that the wound healing of the positive group and blumea balsamifera nanoemulsion drug group is much higher than that of the blank group. On day 7 after wound treatment, the wound healing of blumea balsamifera nanoemulsion drug group was higher than that of positive drug group.
Healing rate = (original wound area-unhealed wound area)/original wound area x100%
The skin wound repair rate and the change of the body weight of the mice of the blumea balsamifera nanoemulsion are shown in fig. 7.
2. Acute skin toxicity test
2 Experimental methods
2.1 laboratory animals
The KM mice were male and female, and had a body weight of 18-22g, and were divided into A, B groups of 4 samples each, each of which was distilled water (control) and blumea balsamifera nanoemulsion, and each of which was provided with a complete skin group and a broken skin group.
2.2 Experimental methods
The back of the hair was dehaired by electric hair clippers, the area was about 2cm×2cm, and the presence or absence of skin damage was checked before the experiment, and the injured skin was not used for the subsequent experiment. In the experiment, 100mg of the test object is uniformly coated on the dehairing area, wherein the group A is coated with distilled water as a control, and the group B is coated with blumea balsamifera nanoemulsion. After 24 hours of application, the applied medicine was washed away with warm water, and then day 7 was observed day by day, the observation contents including the systemic poisoning manifestations and death conditions of skin, hair, eyes and mucous membrane of the test animal, and respiratory, central nervous system, limb activities, etc.
After 8 mice were further divided into groups by depilating with the above method, the depilating sites were scratched with fine sand paper to make the skin appear dense bleeding points, and the test substances were immediately coated on the scratched sites in the same manner as above for administration, and the observation indexes were the same.
2.3 experimental results
As shown in Table 6, the KM mice were subjected to distilled water and blumea balsamifera nanoemulsion samples, respectively, and had no systemic poisoning or death, and the animals had no weight increase, and had no influence on respiratory cycle, central nervous system, limb activities, etc. The blumea balsamifera nanoemulsion has no irritation to normal skin and damaged skin.
TABLE 6
In conclusion, the blumea balsamifera nanoemulsion provided by the invention has good stability, obvious skin wound repairing effect and no toxicity to skin and mucous membrane.
3. Cytotoxicity test
3 Experimental method
3.1 Experimental cell lines
A549 (human non-small cell lung cancer), RD (human malignant embryo rhabdomyoma cells), TCA (human tongue squamous carcinoma cells)
3.2 Experimental methods
The MTT method is adopted to screen the blumea balsamifera nanoemulsion for anticancer activity. The tumor cells tested were RD, TCA, A549. Preparing 100 mu L of cell suspension in a 96-well plate, and culturing the culture plate for 24 hours at 37 ℃ under the condition of 5% CO 2; then adding 10 mu L of nano-emulsion with different concentrations into the culture plate, and then placing the culture plate in an incubator for incubation for 48 hours; then, 20 mu L MTT solvent is added into each hole, and the culture plate is placed in an incubator for incubation for 1-4 hours; finally, measuring absorbance at 490nm by using an enzyme-labeled instrument, and calculating the cell inhibition rate.
3.4 experimental results
The blumea balsamifera nano emulsion has good inhibition effect on RD, TCA and A549 tumor cells (the concentration gradients are respectively 300 mug/ml, 250 mug/ml, 200 mug/ml, 150 mug/ml and 100 mug/ml), the concentration gradient of the blumea balsamifera nano emulsion is 196 mug/ml, 157 mug/ml, 141 mug/ml and 125 mug/ml, the detailed data result is shown in figure 8, the blumea balsamifera nano emulsion also has obvious inhibition effect on tumor cells, the cytotoxicity effect on the tumor cells caused by the comparison of common blumea balsamifera and blumea balsamifera nano emulsion is found, the same medicine concentration gradient (196 mug/ml, 188 mug/ml, 180 mug/ml and 172 mug/ml) is set, the A549 is taken as a typical tumor cell, the data shows that the antitumor activity of the blumea balsamifera nano emulsion is slightly higher than that of common blumea balsamifera, in addition, PCO4 and EtOH two solvent groups (the concentration and the previous gradient are kept consistent) are set, the effect on the tumor cells is not shown in figure 9, the detailed result is shown in figure, the effect on the growth of the tumor cells is shown in figure 9.
4. Blumea balsamifera oil and blumea balsamifera nanoemulsion for repairing skin wounds
4.1 grouping of animals
KM mice, male and female mice are divided into groups randomly, 8 mice in each group are divided into groups: blank (without any treatment), positive (mupirocin ointment), blumea balsamifera nano-drug and blumea balsamifera drug.
4.2 Experimental methods
The hair on the back of the mice was removed by an epilator, washed with warm water, wiped dry, and anesthetized with 1% chloral hydrate at 0.007 ml/g. Then, the mice are partially disinfected by 5% iodine and 75% alcohol, and circular wounds with the diameter of 10mm are cut at the positions of 10mm on two sides of the back spine of the mice, so that fascia and adipose tissues are not damaged. The wound thickness of the muscle layer was about 3mm, and the wound was rinsed and sterilized with physiological saline. The wound surface is opened, hemostasis is carried out fully, and after waking up, the wound surface is fed and observed for 24 hours in a single cage, and free drinking and eating are carried out in the period. The method takes that the wound has no congestion, swelling and suppuration, the mice have normal feeding and good activity as the judging standard, and the mice are used as the experimental subjects to group experiments. Each group was sterilized with iodine and physiological saline every day, changed once daily (blank group was not treated at all, positive drug group, blumea balsamifera and blumea balsamifera nanoemulsion drug group were all administered 100 mg), and wound healing was observed by photographing recording on 1 st, 3 rd, 5 th and 7 th day of wound treatment, respectively, and wound healing rate and weight change were calculated.
Blumea balsamifera used in this experiment was obtained in this laboratory by steam distillation using blumea balsamifera (purchased from Rodian, guizhou).
The experimental results are shown in FIG. 11. It can be found that the wound healing of blumea balsamifera and blumea balsamifera nanoemulsion drug groups is much higher than that of the blank group. And the wound healing rate of the blumea balsamifera nanoemulsion is higher than that of common blumea balsamifera.
Healing rate = (original wound area-unhealed wound area)/original wound area x100%
The skin wound repair rate (A) and the change in the body weight of the mice (B) are shown in FIG. 12.
5. Transdermal absorption experiments
5.1 Experimental methods
The method comprises the steps of obtaining the isolated skin of a mouse, constructing a skin diffusion cell, wherein the inner diameter of the skin diffusion cell is 1.8cm, the receiving cell is 100ml, and the diffusion cell sequentially comprises a supply cell, the skin of the mouse and the receiving cell from top to bottom. Wherein the adding amount of blumea balsamifera is 0.2g, the adding amount of blumea balsamifera nanoemulsion is 2mL, and 1mL of receiving solution (PH=5.8 PBS buffer solution) of each group is taken out at the temperature of 37 ℃ for 2 h, 4h, 6 h, 8h, 10 h, 12 h and 24h respectively, and the receiving solution is supplemented with the same volume of new receiving solution. The content of l-borneol was measured by GC method, and the cumulative permeation quantity (Qn, ug/cm 2) and the cumulative permeation rate were calculated. The experimental results are shown in fig. 13 and 14.
Blumea balsamifera used in this experiment was obtained in this laboratory by steam distillation using blumea balsamifera (purchased from Rodian, guizhou).
Cumulative transmittance=qn/Q Total (S)
Where Ve represents the sample volume, ci represents the drug concentration at the ith sample, V0 is the total volume of the release medium, cn is the drug concentration at the nth sample, and a represents the receiving pool area.
Cumulative transmittance (%) =qn/(mtotal/a)
Wherein Qn represents the cumulative permeation quantity, mtotal represents the original total drug quantity, and a represents the receiving well area.
5.2 experimental results
The transdermal effect of the blumea balsamifera nanoemulsion is obviously higher than that of common blumea balsamifera, the average maximum cumulative transmittance of the blumea balsamifera nanoemulsion is about 22 percent within 24 hours after administration, and the blumea balsamifera is only about 3.35 percent. Such results might be related to the higher skin wound healing effect of blumea balsamifera nanoemulsions than common blumea balsamifera.
Claims (5)
1. The blumea balsamifera nanoemulsion is characterized by comprising the following raw materials in percentage by mass: blumea balsamifera oil 9%, IPM 1%, surfactant PCO 40.5%, cosurfactant absolute ethanol 3.75%, and water 100%; is prepared by a ultrasonic emulsification method.
2. The use of blumea balsamifera nanoemulsion according to claim 1 for preparing an anti-tumor drug, wherein the tumor is specifically a human non-small cell lung cancer cell, a human malignant embryo rhabdomyoma cell and a human tongue squamous cell carcinoma cell.
3. The use of blumea balsamifera nanoemulsion according to claim 2 in the preparation of an anti-tumor drug, wherein the anti-tumor drug is an oral preparation, and the blumea balsamifera nanoemulsion is combined with pharmaceutically acceptable auxiliary materials to prepare the required preparation.
4. Use of a blumea balsamifera nanoemulsion according to claim 1 for the preparation of a wound repair medicament.
5. The use of blumea balsamifera nanoemulsion according to claim 4 for preparing a wound repair drug, wherein the wound repair drug is an external liniment, and the blumea balsamifera nanoemulsion is combined with pharmaceutically acceptable auxiliary materials to prepare a required preparation.
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