CN110403926B - Application of nano acidified fatty acid ester in preparation of medicine for preventing and treating contact dermatitis - Google Patents
Application of nano acidified fatty acid ester in preparation of medicine for preventing and treating contact dermatitis Download PDFInfo
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Abstract
The invention discloses an application of nano acidified fatty acid ester in preparing a medicine for preventing and treating contact dermatitis, wherein the nano acidified fatty acid ester is prepared by the following method: firstly, preparing fatty acid ester, wherein the fatty acid ester comprises the following components in parts by weight: 9-15 parts of hexadecanoic acid, 4-8 parts of unsaturated linoleic acid, 63-70 parts of unsaturated oleic acid and 3-5 parts of octadecanoic acid; the fatty acid ester is first nano treated and then ozonized, the grain size is controlled below 100mn, and the acid value is controlled not to be higher than 30mg/kg, so as to obtain the nano acidified fatty acid ester.
Description
Technical Field
The invention relates to fatty acid ester, and particularly discloses application of nano acidified fatty acid ester in preparation of a medicine for preventing and treating primary irritant contact dermatitis and allergic contact dermatitis.
Background
Contact dermatitis is an inflammatory response that occurs at the site of contact, even to an external site, after a single or multiple exposure of the skin or mucosa to an exogenous substance. The contact substance of the primary irritant contact dermatitis has strong irritation to the skin, and the dermatitis can occur after any person contacts the contact substance. 2. Allergic contact dermatitis is basically non-irritating, and a few people who are sensitized by the substance contact with the substance again, and dermatitis occurs at and near the contact site after 12 to 48 hours. Allergic Contact Dermatitis (ACD) is a T cell-mediated immune response to skin contact allergens and has the characteristics of difficult prevention and complex and diverse pathogenesis. ACD has increased dramatically in the past 30 years in populations and especially in children, and its consequences include a potential for serious interference with quality of life, normal physical activities and school educational life, while ACD has a significant potential economic impact on society due to its high prevalence.
At present, statistical survey shows that ACD accounts for 70-90% of common occupational skin diseases, the occupational life of patients is seriously affected, in severe cases, ACD threatens to cause the patients to lose business, and treatment and prevention needed by all ACD patients engaged in related occupations are free from allergens. However, detection of allergens and prevention and treatment thereof is often very difficult to achieve.
The current treatment scheme of ACD patients is mainly limited to avoiding allergen contact and local use of anti-inflammatory drugs such as glucocorticoid, but hormone external drugs have obvious side effects and are not suitable for preventive use and long-term use, and the search for safe and effective drugs, especially drugs capable of being used preventively, is always a research hotspot.
In the case of professional people, the patients cannot be treated with hormone after obtaining ACD, but the hormone can not be used as a preventive medicine for the patients who need to be contacted for work because the long-term use of the hormone can cause side effects such as hormone dependence, skin atrophy, telangiectasia and the like. There are many causes of ACD, and 2,4-Dinitrochlorobenzene (DNCB) is one of the main causes, and the research on a preparation capable of preventing ACD is of great significance to professional population who is easy to cause ACD. For the people who have already suffered from the disease, the research on a preparation which can treat ACD and can replace hormone is also of great significance.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide the application of the nano acidified fatty acid ester in preparing the medicine for preventing and treating the contact dermatitis.
In order to realize the purpose of the invention, the technical scheme of the invention is as follows:
the invention discloses an application of nano acidified fatty acid ester in preparing a medicine for preventing and treating contact dermatitis, wherein the nano acidified fatty acid ester is prepared by the following method: firstly, preparing fatty acid ester, wherein the fatty acid ester comprises the following components in parts by weight: 9-15 parts of hexadecanoic acid, 4-8 parts of unsaturated linoleic acid, 63-70 parts of unsaturated oleic acid and 3-5 parts of octadecanoic acid; the fatty acid ester is first nano treated and then ozonized, the particle size is controlled below 100mn, and the acid value is controlled not to be higher than 30mg/kg, so as to obtain the nano acidified fatty acid ester.
One of the preferred schemes is as follows: the fatty acid ester comprises the following components in parts by weight: hexadecanoic acid: 9.09 parts, linoleic acid: 7.29 parts, oleic acid: 69.36 parts, octadecanoic acid: 3.63 parts.
The second preferred scheme is as follows: the fatty acid ester comprises the following components in parts by weight: hexadecanoic acid: 13.52 parts, linoleic acid: 4.68 parts, oleic acid: 67.25 parts, octadecanoic acid: 4.33 parts.
The third preferred scheme is as follows: the fatty acid ester comprises the following components in parts by weight: hexadecanoic acid: 14.69 parts, linoleic acid: 4.14 parts, oleic acid: 63.83 parts, octadecanoic acid: 4.65 parts.
Further preferred scheme: the acid value ranges from 5 to 10mg/kg.
Further preferred scheme: the control conditions for the ozonation include: the temperature of the system is controlled between 25 ℃ and 30 ℃, the pressure is 0.5 bar to 1.5bar, and the ozone content reaches 80g/L to 120g/L.
Further preferred scheme: the particle size of the nano acidified fatty acid ester is less than 100nm, and the nano acidified fatty acid ester can be obtained by performing nanocrystallization treatment on the fatty acid ester through a nano ultrahigh pressure homogenizer.
The invention takes mometasone as a contrast to observe the protection effect of the nano acidified fatty acid ester on a guinea pig ACD model and searches for a new potential therapeutic drug of ACD.
The invention uses nano acidified fatty acid ester to prevent and treat ACD for preliminary study, and experimental results show that the nano acidified fatty acid ester has a certain protective effect on ACD, and the effect of treating ACD is close to that of the externally applied drug mometasone ointment of intermediate glucocorticoid.
The results in the prevention experiments of the ACD suggest that the nano-acidified fatty acid ester can well prevent and intervene inflammation formation, compared with the non-nano-acidified fatty acid ester, the nano-acidified fatty acid ester prevention group has lighter epidermal hyperplasia and dermal inflammatory cell infiltration, and at present, the main strategy for the prevention research of the ACD is to avoid direct contact with irritative and sensitive products. Meanwhile, the main clinical prevention measure is clinical education and management, and no preventive medicine report aiming at ACD is found in the literature at present. The safety research of the nano acidified fatty acid ester shows that the nano acidified fatty acid ester is safe and effective and does not have the side effect of corticoid. Therefore, the nano acidified fatty acid ester is expected to be an effective preventive medicine for preventing ACD.
Drawings
FIG. 1 is a graph showing the comparison of the change of back skin damage in ACD pre-treatment and anti-dry treatment groups, wherein A is a blank control group; ACD-model group; ACD-conventional ozone oil prevention group; and D, ACD-nano acidified fatty acid ester prevention group.
FIG. 2 is a graph of changes in back skin lesions of groups after ACD treatment, wherein A is ACD-NaCl treatment group; ACD-conventional ozone oil treatment group; ACD-nano acidified fatty acid ester treatment group; and D, ACD-mometasone treatment group.
Fig. 3 is a graph of the histopathological changes of skin (H E staining x 100) following ACD prophylactic intervention: a is blank control group; ACD-model group; ACD-conventional ozone oil prevention group; and D, ACD-nano acidified fatty acid ester prevention group.
FIG. 4 is a graph of the pathological changes in skin tissue (H E staining x 100) following ACD treatment; wherein A is ACD-NaCl treatment group; ACD-conventional ozone oil treatment group; ACD-nano acidified fatty acid ester treatment group; and D, ACD-mometasone treatment group.
Figure 5 is a skin CT detection image after ACD preventative intervention.
Blank control group: the blood vessel in the dermis is not obviously dilated, and no obvious inflammatory cell infiltration is seen around the tube;
ACD model group: the superficial dermis blood vessels expand and are hyperemic, and inflammatory cells infiltrate with unequal amount around the vessels;
ACD conventional ozone oil prevention group: a few capillaries are dilated and congested in part of dermal papilla, and the tube Zhou Shaoxu is infiltrated by inflammatory cells;
the ACD nano acidified fatty acid ester prevention group comprises: the blood vessel in the dermis is not obviously dilated, and no obvious inflammatory cell infiltration is seen in the periphery of the tube.
Figure 6 is a CT scan of skin after ACD treatment, comparing between and within groups, "×", p <0.05 is statistically different: ACD-NaCl group: dermal papilla and superficial blood vessel obviously dilate congestion, and inflammatory cell infiltration is equal in the periphery of the vessel;
ACD conventional ozone oil treatment group: a few capillaries are in the dermal papilla, dilate and engorge, and the tube Zhou Shaoxu is infiltrated by inflammatory cells;
ACD nano acidified fatty acid ester treatment group: a few blood vessels in the dermal papilla are expanded, and no obvious inflammatory cell infiltration is seen around the tubes;
d, ACD mometasone treatment group: the vascular dilatation in individual dermal papilla is hidden, and the infiltration of inflammatory cells around the tube is not obvious D0: before ACD prevention, D12: end of ACD preventive intervention, D19: ACD is treated one week later.
FIG. 7 is a comparison of allergic contact dermatitis in both hands before and after treatment.
FIG. 8 is a comparison graph of the treatment of the strong acid irritant contact dermatitis before and after the treatment.
Detailed Description
Example 1 is further illustrated and explained below with reference to specific experimental data
The purpose is as follows: the effect of the nano acidified fatty acid ester on preventing 2,4-dinitrochlorobenzene (DNC B) from inducing Allergic Contact Dermatitis (ACD) and the treatment effect on the ACD are researched. The method comprises the following steps: the 40 guinea pigs were randomly grouped as: a normal control group, an ACD-model group, an ACD-conventional ozone oil prevention group and an ACD-nano acidified fatty acid ester prevention group; ACD-NaCl treatment group, ACD-conventional ozone oil treatment group, ACD-nano acidified fatty acid ester treatment group, and ACD-mometasone treatment group. And (3) copying an AC D model by using a D N C B model in each group except a normal control group, counting scratching times of the guinea pigs in each period, photographing and recording skin damage conditions of the guinea pigs, observing back skin damage conditions of the guinea pigs by using a skin CT (computed tomography) method, counting the thickness of the skin of each group, taking skin tissues at the back skin damage positions, and dyeing and observing pathological changes of the skin of each group by using a hematoxylin-eosin dyeing method. As a result: nano acidified fatty acid ester prevention experiment: in the statistical comparison of scratching times, compared with a non-nano acidified fatty acid ester intervention group, the scratching times of a nano acidified fatty acid ester prevention group are remarkably reduced (P <0,05); preventing the morphological change of the skin in the experiment, and only red spots are seen in the skin damage prevention group of the nano acidified fatty acid ester; the skin CT result shows the prognosis of dry prevention: the nano acidified fatty acid ester can prevent a small amount of inflammation infiltration in a group, and a large amount of inflammatory cells and expanded blood vessels can be seen in an intervention group; skin thickness measurements showed that the non-prevention group was significantly thicker than the nano-acidified fatty acid ester prevention group, P <0.01; the HE staining result shows that compared with the intervention group of the nano acidified fatty acid ester, the intervention group of the non-nano acidified fatty acid ester has more obvious inflammation expression; nano acidified fatty acid ester treatment experiment: compared with other groups, the scratching times of the nano acidified fatty acid ester treatment group and the mometasone treatment group are obviously reduced (P <0,05), and in comparison of skin loss of each group, the skin loss area of the nano acidified fatty acid ester treatment group and the mometasone treatment group is obviously reduced compared with that of the NaCl and conventional ozone oil treatment group; the skin CT result shows that compared with the pre-treatment inflammatory cells, the nano acidified fatty acid ester treatment group and the mometasone treatment group are greatly reduced, and the visible epidermis thickness is obviously thinned, wherein P is less than 0.01; compared with other treatment groups, the nano acidified fatty acid ester treatment group and the mometasone treatment group in the HE dyeing result obviously reduce the epidermal hyperplasia and inflammatory infiltration degree. And (4) conclusion: the nano acidified fatty acid ester has a certain prevention effect on the formation of ACD, the nano acidified fatty acid ester and mometasone have similar effects of promoting the recovery of inflammation, and the treatment effect on the ACD is close to that of a positive control medicament mometasone.
The formula and preparation of the nano-acidified fatty acid ester are characterized in that fatty acid ester is prepared firstly, see table 1, the components are subjected to nanocrystallization and then ozonization to obtain the nano-acidified fatty acid ester with the particle size controlled below 100mn and the acid value controlled not to be higher than 30 mg/kg.
Table 1 shows 3 specific formulations and control conditions in example
The ozonization control conditions are as follows: the temperature of the system is controlled to be 25-30 ℃, the pressure is 0.5-1.5 bar, and the ozone content reaches 80-120 g/L.
Results of clinical trials
1. Data and method
1.1 data
1.1.1 test animal data:
albino guinea pigs were 40, male and female halves. General grade, weight 300-350 g, provided by Taiping Biotechnology Limited, hunan. SCXK (xiang) 2015-004. The experimental mechanism comprises: the laboratory animal department of the university of China and south China, the temperature is 18-24 ℃, the relative humidity is 55-70 percent,
1.1.2 main reagents and instruments:
conventional ozone oil, commercially available; 0.9% sodium chloride (batch No.: D16091912-1 manufacturer: hunan Kolun pharmaceutical Co., ltd.); mometasone ointment (lot: 20160518, bayer pharmaceuticals (Shanghai) Co., ltd.) chloral hydrate (lot: 20130325, tegaku Mimeou Chemicals Co., ltd.) and the like. 2. 4-Dinitrochlorobenzene (manufacturer: sigma; batch No. 138630-100G). The balance (YP 3001N) is a product of Shunhui scientific instruments, inc. in Shanghai; in-vivo reflective confocal microscopy (VIVASCOPE) is product of Lucid inc.
1.2 methods
1.2.1 Experimental groups and administrations:
40 albino guinea pigs, male and female halves. Guinea pigs were randomly grouped as: ACD-model group, ACD-conventional ozone oil prevention group, ACD-nano acidified fatty acid ester prevention group, ACD-NaCl treatment group, ACD-conventional ozone oil treatment group, ACD-Allolone treatment group, experimental reference [7] ACD model was manufactured: before the experiment, the abdomen of the rat is shaved, about 2cm multiplied by 2cm is selected for standby application, 30 microliter of 7 percent DNCB solution is smeared on the shaved area of the abdomen of the experimental group for exciting sensitization on the 1 st day (sensitization period) of the experiment, 2cm multiplied by 3cm is selected for standby application on the 7 th day, 30 microliter of 1 percent DNCB solution is smeared on the sheared area of the back of the rat on the 8 th day, and the clinical expression of Allergic Contact Dermatitis (ACD) such as redness, swelling, roughness, thickening, lichenification and the like is observed until the back of the rat is observed whether to have redness, swelling, roughness, thickening, lichenification and the like. The ACD is administered with conventional ozone oil and nano acidified fatty acid ester for prevention and intervention in the molding process, and a treatment experiment is carried out after the molding is successful: the components are respectively treated by NaCl, conventional ozone oil, ozone oil and mometasone. In the prevention and treatment experiment, the administration dose of each group except the group treated with the Allolopine was 0.1ml, and the administration dose of the Allolopine group acted on the skin to form a snowflake shape for 1 time/day.
1.2.2 Experimental observations
(1) Taking scratching of front and rear limbs of guinea pigs on the head, the back and the abdomen as main materials, counting scratching times of the head, the abdomen and the back of the guinea pigs in each period within 30min, (2) observing and photographing prevention and treatment experiments before and after the experiments and recording the morphological change of the back skin of each group of guinea pigs, (3) selecting the back skin lesions of mice after the experiments, adopting skin CT detection and HE staining of the histology of the skin lesions, wherein the two detections can simultaneously observe the pathological conditions of the back skin lesions of the mice, and meanwhile, the skin CT counts the thickness of the epidermis of each group of experiments before and after the experiments.
1.2.3 statistical methods
Statistical processing was performed on the data using SPSS 22.0 statistical analysis software. The measurement data adopts mean + -standard deviation
Indicating that t test and F test are performed. P is less than or equal to 0.05, and the difference has statistical significance.
2 results
2.1 statistics of number of scratching of guinea pigs:
compared with the nanometer acidified fatty acid ester, the prevention of the non-nanometer acidified fatty acid ester in the experimental process has the statistical difference that the scratching prevention frequency is increased, and P is less than 0.05; in treatment experiments, compared with the ACD-NaCl group and the conventional ozone oil group, the ACD-mometasone treatment group and the nano acidified fatty acid ester group have obviously reduced scratching times and have statistical difference, and P is less than 0.05.
2.2 morphological changes of the skin
At the end of the ACD prevention experiment, the skin of the model group and the skin of the conventional ozone oil intervention group have obvious erythema exudation and scabbing, and the skin of the nano acidified fatty acid ester group only has erythema (figure 1); in ACD treatment experiments, compared with skin lesions of mometasone treatment groups, skin lesions of a nanometer acidified fatty acid ester treatment group and a normal ozone oil treatment group are obviously improved: the tendency of escharosis and inflammation erythema to disappear (fig. 2)
2.3 pathological changes in the skin lesion tissue
ACD prevention experimental tissue H E was shown under the stained microscope: compared with the intervention group of the nano acidified fatty acid ester, the intervention group of the non-nano acidified fatty acid ester has more obvious inflammation expression: hyperkeratosis of epidermis, thickening of spinous layer, significant inflammatory infiltration of dermal layer, significant collagen proliferation (fig. 3); ACD treatment experimental tissue pathological display: the nano-acidified fatty acid ester treatment group and the mometasone treatment group had a significant reduction in epidermal hyperplasia and dermal inflammatory cell infiltration after treatment compared to the conventional ozone oil treatment group and the NaCl treatment group, but mild hyperkeratosis was still visible (fig. 4).
2.4 skin CT detection prevention experiment skin CT detection: the blank control group can see no obvious inflammatory cells, the model group and the conventional ozone oil prevention group can see obvious large amount of inflammatory cells and dilated blood vessels, the nano acidified fatty acid ester prevention group can see small amount of inflammatory infiltration (figure 5), and the treatment experiment skin CT detection: after treatment, obvious expansion of inflammatory cells and blood vessels can still be seen in the NaCl treatment group and the conventional ozone oil treatment group, the change is not obvious compared with that before treatment, and the inflammatory cells are greatly reduced in the nano acidified fatty acid ester treatment group and the mometasone treatment group compared with that before treatment.
Skin thickness: compared with a blank control group in a prevention experiment, the skins of other groups are thickened, but the thicknesses of a model group and a conventional ozone oil prevention group are obviously thicker, namely the thickness P is less than 0.01; in a treatment test, compared with a Nacl treatment group, the surface skin thickness of a nano acidified fatty acid ester treatment group and a mometasone treatment group is obviously reduced, P is less than 0.01, the thickness of a conventional ozone oil treatment group is not obviously different, and P is more than 0.05.
2.5 clinical trial comparative plot
FIG. 7 is a comparison of allergic contact dermatitis in both hands before and after treatment.
FIG. 8 is a comparison graph of the treatment of the strong acid irritant contact dermatitis before and after the treatment.
From fig. 7 and 8, it can be seen that the nano acidified fatty acid ester of the present invention has a better therapeutic effect on contact dermatitis.
Claims (6)
1. The application of nano acidified fatty acid ester in preparing a medicine for preventing and treating contact dermatitis is disclosed, wherein the nano acidified fatty acid ester is prepared by the following method: firstly, preparing fatty acid ester, wherein the fatty acid ester comprises the following components in parts by weight: 9-15 parts of hexadecanoic acid, 4-8 parts of unsaturated linoleic acid, 63-70 parts of unsaturated oleic acid and 3-5 parts of octadecanoic acid; the fatty acid ester is first nano-treated and then ozonized, the particle size is controlled below 100nm, and the acid value is controlled not to be higher than 30mg/kg to obtain nano-acidified fatty acid ester; the control conditions for the ozonation include: the temperature of the system is controlled to be 25-30 ℃, the pressure is 0.5-1.5 bar, and the ozone content reaches 80-120 g/L.
2. The use of claim 1, wherein the fatty acid ester component comprises the following components in parts by weight: hexadecanoic acid: 9.09 parts, linoleic acid: 7.29 parts of oleic acid: 69.36 parts, octadecanoic acid: 3.63 parts.
3. The use of claim 1, wherein the fatty acid ester component comprises the following components in parts by weight: hexadecanoic acid: 13.52 parts, linoleic acid: 4.68 parts, oleic acid: 67.25 parts, octadecanoic acid: 4.33 parts.
4. The use of claim 1, wherein the fatty acid ester component comprises the following components in parts by weight: hexadecanoic acid: 14.69 parts, linoleic acid: 4.14 parts, oleic acid: 63.83 parts, octadecanoic acid: 4.65 parts.
5. Use according to claim 1, wherein the acid number is in the range of 5-10mg/kg.
6. The use according to any one of claims 1 to 5, wherein the contact dermatitis comprises primary irritant contact dermatitis and allergic contact dermatitis.
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| ITRM20050514A1 (en) * | 2005-10-17 | 2007-04-18 | Gabriele Maietta | OZONIZED OIL, HIGH-STABILITY ACTIVE INGREDIENT BASED ON PEROXIDES USABLE FOR MEDICAL THERAPEUTIC APPLICATIONS AND FOR SIMILAR DOMICILE TREATMENTS TO THOSE OF OZONOTHERAPY, WITHOUT THE USE OF OZONE GASSOUS. |
| CN108057350A (en) * | 2017-12-14 | 2018-05-22 | 长沙思麦食品开发有限公司 | A kind of solubilization method of ozone in tea oil |
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2019
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