CN116077562A - Application of rose in preparation of products for resisting prostatic hyperplasia/hypertrophy - Google Patents
Application of rose in preparation of products for resisting prostatic hyperplasia/hypertrophy Download PDFInfo
- Publication number
- CN116077562A CN116077562A CN202310078877.1A CN202310078877A CN116077562A CN 116077562 A CN116077562 A CN 116077562A CN 202310078877 A CN202310078877 A CN 202310078877A CN 116077562 A CN116077562 A CN 116077562A
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- rose
- prostatic hyperplasia
- hypertrophy
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/15—Preparation or pretreatment of starting material involving mechanical treatment, e.g. chopping up, cutting or grinding
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/17—Preparation or pretreatment of starting material involving drying, e.g. sun-drying or wilting
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/10—General cosmetic use
Abstract
The invention relates to the technical field of natural medicines for treating prostatic hyperplasia/hypertrophy, in particular to application of rose in preparation of products for resisting prostatic hyperplasia/hypertrophy. Experiments prove that: the rose pollen can obviously reduce the prostate index of a prostate hyperplasia model rat, the levels of dihydrotestosterone and testosterone in the prostate homogenate and the activity of 5alpha-reductase, and reduce the estrogen level in tissues and the ratio of rebound estrogen to androgen in different degrees; can reduce the activities of E2, DHT, E2/T and PACP in serum of a model animal and the MDA content, increase the activity of SOD and have a trend of increasing T; can reduce the number and height of prostate epithelial cells of a model animal, reduce finger protrusion, reduce proliferation of prostate tissue cells and accelerate apoptosis of the cells so as to improve and treat prostatic hyperplasia; and the rose has higher safety. The rose has good application prospect and development potential in preventing and treating the prostatic hyperplasia (hypertrophy) caused by various factors.
Description
Technical Field
The invention relates to the technical field of natural medicines for treating prostatic hyperplasia/hypertrophy, in particular to application of rose in preparation of products for resisting prostatic hyperplasia/hypertrophy.
Background
Benign Prostatic Hyperplasia (BPH), a common disorder of middle-aged and elderly men characterized by progressive frequent urination and difficulty in urination due to a significant increase in the prostate. Epidemiological studies have shown that prostate hyperplasia has a very low incidence before age 40, 40% after age 50, and nearly 90% after age 80, and by 90, nearly 100% of prostate hyperplasia is found when a histological examination of the prostate is performed. With the aging of the population and the increasing deterioration of environmental factors in China, the incidence rate of the traditional Chinese medicine is on the rising trend in recent years.
The treatment modes of the prostatic hyperplasia are mainly divided into operation treatment and drug treatment. Surgical excision was once considered the first method to radically cure BPH, but the patient's advanced age often has a certain limitation in surgical treatment, which is prone to cause various complications, and the statistical surgical complications are about 15% and the death rate is about 1%. It has been shown that the prostate gland produces a variety of immunoglobulins, can synthesize zinc-containing polypeptides with a variety of antimicrobial effects, and has a local immune function of protecting the reproductive system from bacteria and other pathogenic microorganisms, and should therefore be preserved as much as possible. In recent years, the treatment of BPH with drugs has become the main treatment means, mainly including 5alpha-reductase inhibitors, alpha-adrenoceptor antagonists, botanicals and Chinese patent medicines. The preparation can recover the urination function of the patients with prostatic hyperplasia to different degrees and improve the life quality of the patients with prostatic hyperplasia no matter the symptomatic treatment, the cause treatment or the combined treatment, but the chemical drugs have obvious adverse reactions after long-term use, and the Chinese patent medicine preparation has the defects of unknown substance basis and active ingredients, unclear action targets, difficult quality control, difficult guarantee of curative effect, increasing hepatorenal toxicity and the like. Therefore, there is an urgent need for safer and more effective healthy products for the prevention and treatment of BPH to effectively block the disease process and improve the quality of life.
Disclosure of Invention
The invention aims to provide application of rose in preparing a product for resisting prostatic hyperplasia/hypertrophy, so as to solve the technical problem that a natural medicine capable of effectively treating prostatic hyperplasia/hypertrophy is lacking in the prior art.
In order to achieve the above purpose, the invention adopts the following technical scheme:
the application of flos Rosae Rugosae in preparing product for resisting prostatic hyperplasia/hypertrophy is provided.
The principle and the advantages of the scheme are as follows:
flos Rosae Rugosae (Rosa rugosa) is a flower for appreciation and eating in daily life, and is also named as fructus Rosae Davuricae, flos Rosae Rugosae, loiter flower, etc., and is an upright shrub of Rosa of Rosaceae, and is needled with dense clusters and branches and fluffed. The original place of rose is mainly concentrated in north China, japan and Korea, and is cultivated in large areas in China at present, and particularly in Yunnan and Guizhou, the rose becomes an important edible and ornamental flower. The rose has various activities such as anti-inflammatory, antioxidant, anti-tumor, anti-fatigue, blood sugar reducing and blood pressure reducing, and the effect and the application of the rose in resisting prostatic hyperplasia are not reported, and the rose has the effect of preventing and treating prostatic hyperplasia (hypertrophy) for the first time and has great social benefit and economic benefit.
The invention researches the effect of rose on resisting prostatic hyperplasia in vivo, and the result shows that: continuous gastric lavage of rats with prostate hyperplasia model at doses of 0.5, 1.0 and 2.0g/kg for 4 weeks can obviously reduce prostate index and Dihydrotestosterone (DHT), testosterone (T) level and 5α -reductase activity of the model animals, and reduce estrogen (E2) level and rebound estrogen/androgen ratio (E2/T) in tissues to different degrees; can obviously reduce the activities of E2, DHT, E2/T and PACP in serum of a model animal and MDA content, increase SOD activity and have a tendency of increasing T. Histopathological examination showed that roses were able to reduce the number and height of prostate epithelial cells in model animals to varying degrees, reducing finger-like protrusions, i.e. reducing proliferation of prostate tissue cells and accelerating apoptosis to improve prostate hyperplasia.
Experiments prove that the roses do not show lethal toxicity in the acute toxicity experiments of mice, and the maximum tolerance of the roses to the intragastric administration within one day exceeds 24g/kg, which is equivalent to 48 times of the pharmacodynamic effective dose of animals of the same species. The mice are continuously drenched with the rose pollen for 30 days at doses of 1.0, 2.0 and 4.0g/kg, the appearance behavior, liver and kidney functions and general anatomy of the mice are not changed in toxicology significance, no toxic reaction is indicated when the rose pollen is repeatedly dosed to the mice for 30 days, and the result shows that the oral rose has higher safety.
In conclusion, the rose can obviously reduce the prostate index of animals with prostatic hyperplasia, reduce the number and height of prostatic epithelial cells, reduce finger-like protrusions, improve the prostatic hyperplasia by inhibiting the proliferation of prostatic tissue cells and accelerating the apoptosis of the prostatic tissue cells, has the action mechanism related to inhibiting the activity of 5alpha-reductase and regulating the hormone level, and can be used for preventing and treating the prostatic hyperplasia and hypertrophy caused by endocrine factors, immune factors, environmental factors, urethral factors, genetic factors and the like clinically. The invention has the beneficial effects that: the invention discovers that the rose has the effects of reducing proliferation of prostate tissue cells and accelerating apoptosis of the cells to improve the prostate hyperplasia for the first time, and can be applied to medicaments, foods, health care products and cosmetics for preventing and treating the prostate hyperplasia (hypertrophy).
Further, the products include medicines, foods, health products and cosmetics. The rose is an edible plant, and can be prepared into medicines, foods, health-care products and cosmetics, and the aim of preventing and treating prostatic hyperplasia (hypertrophy) is fulfilled.
Further, the product was prepared using petals of rose. The petals of the rose are used as main efficacy raw materials, and are used as medicines.
Further, the petals are first prepared as a powder or extract, and then the powder or extract is used to prepare the product. The bioavailability of the effective components can be increased by pulverizing petals of flos Rosae Rugosae or preparing extract. And particularly, the purpose of fully enriching the functional components can be achieved through the form of the extract.
Further, the extract is prepared by the following method: squeezing the petals of the rose to obtain rose juice, and freeze-drying to obtain the freeze-dried powder extract. The rose juice is obtained by squeezing juice, and then the raw materials are more portable and further enriched with functional components through freeze-drying treatment.
Further, the raw materials of the product also comprise other functional components or auxiliary materials. For enhancing the therapeutic effect and adjusting the dosage form, other functional components or auxiliary materials can be added into the rose.
Further, the auxiliary materials comprise plant starch, microcrystalline cellulose and magnesium stearate. The rose is prepared into a tablet form by taking plant starch, microcrystalline cellulose and magnesium stearate as auxiliary materials, so that the rose is convenient to carry and take.
Further, the other functional components include male silkworm moth and Cordyceps sinensis mycelia. The rose can be singly or in combination with one or more other substances to achieve the effects of strengthening or synergistically preventing and treating the prostatic hyperplasia and the hypertrophy.
Further, the product is tablets, capsules, medicinal granules and oral liquid. The product for resisting the prostatic hyperplasia/hypertrophy can be prepared into various dosage forms according to the actual application requirement, and the aim of treating or preventing the prostatic hyperplasia/hypertrophy can be realized.
Further, the dosage range of the rose is 10 mg-10000 mg per kilogram of body weight. The rose petals are applicable food, and the results of the acute toxicity test of the oral gastric lavage way of the mice and the subacute toxicity test of the oral gastric lavage way of the mice show that the rose petals are high in administration safety, and 10 mg-10000 mg of rose petals can not cause toxic reaction when being taken per kilogram of body weight. The amount of rose used can be adjusted within the above range according to the therapeutic or prophylactic requirements. When the rose is used on a therapeutic or prophylactic basis, it is generally meant that the daily dose is in the range of 10mg to 10000mg per kg of body weight, and may be administered in divided doses if necessary.
Drawings
FIG. 1 is a high performance liquid chromatogram of a blank tube under the final reaction system of Experimental example 1.
FIG. 2 is a high performance liquid chromatogram of the reaction tube under the final reaction system of Experimental example 1.
FIG. 3 is a graph showing the inhibition of 5α -reductase by finasteride of experimental example 1.
Detailed Description
The invention discloses a rose which has good effect of preventing and treating prostatic hyperplasia and has great economic value when being applied to medicines, foods, health care products and cosmetics. It should be noted that all such similar substitutions and modifications will be apparent to those skilled in the art from the disclosure herein that compositions based on rose (or extract, refined) as a core material may be used to develop medicaments, health products, foods and cosmetics for preventing and treating prostatic hyperplasia, and are intended to be encompassed by the present invention. While the invention has been described with reference to preferred embodiments, it will be apparent to those skilled in the relevant art that variations and modifications can be made in the methods and applications described herein, or in appropriate changes and combinations, without departing from the spirit and scope of the invention.
The present invention will be further described in detail with reference to specific examples, which are not intended to limit the scope of the invention, in order to provide a better understanding of the technical aspects of the present invention to those skilled in the art.
Example 1: preparation of Rose pollen
Fresh rose flower 20kg is taken, petals are picked, dried at 45 ℃ and crushed by a crusher (sieving by a 120-mesh sieve) to obtain 1.72kg of rose pollen.
Example 2: preparation of rose freeze-dried powder
Taking 20kg of fresh rose, picking petals, putting into a juicer (extrusion type) to obtain 17.6kg of rose juice, freezing to-40 ℃, putting into a freeze dryer for drying to obtain 1.22kg of freeze-dried powder, and sub-packaging (1 g per bag).
Example 3: preparation of compound rose capsule
Taking 1kg of rose freeze-dried powder, 1kg of male silkworm moth powder, 1kg of cordyceps sinensis mycelium powder (fermented product), uniformly mixing, granulating, drying, finishing, and filling capsules (each granule is 0.5 g).
Example 4: compound rose tablet
Taking 12kg of rose pollen, adding 25kg of sweet potato powder and 3kg of microcrystalline cellulose, mixing, granulating, drying, finishing, adding 0.6kg of magnesium stearate, and tabletting (about 0.5g of each tablet).
Example 5: preparation of superfine rose powder
Taking 20kg of fresh rose, picking petals, drying at 45 ℃ to obtain less than 7% of water, crushing by a crusher (sieving with a 120-mesh sieve), and crushing into wall-broken powder (500 meshes) by a wall-broken crusher to obtain 1.65kg.
Experimental example 1: research on inhibition of 5alpha-reductase in vitro by roses
1. Experimental materials
SPF-grade female SD rats, provided by Kunming medical university; testosterone, shanghai leaf Biotechnology Co., ltd., lot number 20130409; NADPH, sigma company (Lot SLBC 6718V); finasteride, hubei Shubang pharmaceutical Co., ltd (20121001); tris-base, BIOSHARP company (Amresco 0497); sodium chloride, biological company (Lot 3004B 220); DTT, sigma company (Lot BCBK 8875V); absolute ethanol, yunnan pharmaceutical company (LotLN 70N 21); methylene chloride, tianjin Fengsha chemical reagent technology Co., ltd (20100926); chromatographic methanol, beijing Baoling technologies Co., ltd (116481).
2. Experimental method
2.1 architecture establishment
5α -reductase is a well-established target for prostatic hyperplasia. The method of the body system reference Wu Wenzhe (Wu Wenzhe, cheng Zhigong, in light, etc. the 5alpha-reductase inhibitor for treating androgenic alopecia [ J ] is screened from plant extracts by using an in vitro model, modern Chinese medicine research and practice, 2010,24 (2): 35.) is adopted, the variation of the peak area of testosterone in a reaction system is measured by adopting a high performance liquid chromatography, the residual amount of testosterone after reaction is obtained by a standard curve of testosterone concentration and peak area, and the residual testosterone amount after reaction is subtracted by the total testosterone amount participating in the reaction, so that the testosterone amount consumed in the reaction system is obtained, and the activity of enzyme is calculated.
5alpha-reductase was prepared from female SD rat liver homogenate and stored in a refrigerator at-20 ℃. Protein content was measured by the modified BCA method, and the concentration of 5. Alpha. -reductase was measured so as to be 8.67mg/mL in the rat liver microsomes. The stability of the detection system is determined by optimizing the conditions such as the concentration of T, 5alpha-reductase, NADPH, the reaction temperature and the like.
The final reaction conditions were: in a Tris-HCl buffer system with pH of 7.0, the enzyme extract addition concentration is 5.045mg/mL, the T addition concentration is 2mg/mL, the NADPH addition concentration is 5mg/mL, the reaction temperature is 37 ℃, the reaction time is 30min, and the liquid chromatograms of a blank tube and a reaction tube are shown in figures 1 and 2.
Chromatographic conditions: luna C18 (2) column (4.6X250 mm,59 m); column temperature: 40 ℃; sample injection amount: 10uL; flow rate: lmL/min; mobile phase: 70% methanol; detection wavelength: 242nm.
2.2 determination of the Activity of finasteride on 5alpha-reductase
Finasteride is a common drug for clinical treatment of prostatic hyperplasia and is an effective 5alpha-reductase inhibitor. Under the determined reaction conditions, 100% ethanol was replaced with finasteride, the inhibition activity of finasteride on 5α -reductase was measured at each concentration, and one 100% ethanol reaction well and one blank well (dichloromethane was added before enzyme addition to terminate the reaction) were made. The IC50 value of finasteride for 5 a-reductase inhibition was calculated from the inhibition activity = (T finasteride-T reaction)/(T blank-T reaction) ×100%, and the inhibition curve of finasteride for 5 a-reductase is shown in fig. 3.
The IC50 of finasteride on 5alpha-reductase inhibition was calculated to be 0.275 mu mol/L by GraphPad Prism software, and was similar to the results of 0.39 mu mol/L (thaptana K, chaiyavat C, vandee R, eta1.Screening of the three 5alpha-reductase inhibitory activity and total phenolic contentof Thai plants [ J ]. Journal of Medicinal Plants research.2011,5 (7): 1265-1271 ]) measured by thaptana K et al with a liquid phase system, and the results of 0.38.+ -. 0.06 mu mol/L (Park WS, son ED, nam GW, et a1. Torrlin from Torilisjaponica, as a new inhibitor of testosterone 5 alpha-reduction [ J ]. Planta Med,2003,69 (5)) measured by Park WS et al, indicating that the system can be used for drug screening.
3. Determination of the Activity of Rose on inhibition of 5α -reductase
The inhibition of 5α -reductase by using ultrapure water to prepare rose suspensions of different concentrations (prepared by the method of example 5) was measured by the above system, and the result was calculated as 5α -reductase inhibition activity= (T sample-T reaction)/(T blank-T reaction) ×100%, and shown in table 1.
Table 1: inhibition of 5alpha-reductase by roses
From Table 1, IC for rose to inhibit 5alpha-reductase was calculated 50 Is 2.17mg/mL and shows a certain concentration reaction relationship.
Experimental example 2: effect of roses on testosterone propionate replication prostate hyperplasia model mice
1. Test materials
1.1 test article
Rose ultra-fine powder (RO), purple powder, lot number 20160501 (prepared by the method of example 5); finasteride tablets (national drug standard H20030951), 5 mg/tablet, hubei Shubang pharmaceutical Co., ltd., 0161001; uroschesis capsule (national medicine standard Z10960007), 0.3 g/granule, shijia Kedi pharmaceutical company, inc., 161113; preparing suspension with different concentrations by pure water for standby.
1.2 animals
SPF-class Male Kunming mice, 20-24 g, offered by the Kunming medical university laboratory animal center, produced license: SCXK (yunnan) K2015-0002, certification authority: the Kunming scientific and technical office.
2. Experimental method
Methods of reference Li et al (Li, y.f., tang, l.p., he, r.r., xu, z., he, q.q., xiang, f.j., su, w.w., kurihara, h.,2013.Anthocyanins extract from bilberry enhances thetherapeutic effect of pollen of Brassica napus L.on stress-provoked benign prostatichyperplasia in restrained mice.j.functions 5, 1357-1365.), male kunming mice were selected and randomly divided into 7 groups according to body weight: normal control group; model control group; finasteride 1mg/kg group; dysuria capsule 0.45g/kg group; roses 0.5, 1.0, 2.0g/kg dose group; each group of 12. In addition to the normal control group, each model animal group is subcutaneously injected with 5mg/kg testosterone propionate (dissolved in peanut oil) for molding every morning, and is respectively administrated by stomach infusion according to dosage in afternoon; the administration was continued for 21 days, and the body weight was weighed 1 time a week. Taking blood after weighing, and measuring T, E2 level in the separated serum according to an ELLISA kit method; mice were sacrificed by cervical vertebrae removal after blood collection, prostate tissues were dissected and weighed, and prostate indexes (prostate index=wet prostate weight (mg)/weight of mice (g) ×10) were calculated for each group of mice, and 5 α -reductase activity in liver tissue homogenates was determined.
3. Experimental results
3.1 Effect on body weight of mice with prostatic hyperplasia
Table 2: influence on the body weight of mice with prostatic hyperplasia (P >0.05 compared with model group)
As can be seen from table 2, the weight of each group of mice continuously increased during the experiment, and there was no significant difference in weight increase between the model group and the normal control group, and between the administration group and the model group.
3.2 Effect on prostate index in mice with prostatic hyperplasia
As can be seen from table 3, the prostate index of the mice in the model group was significantly increased, and the difference was very significant (P < 0.01) compared with the normal control group, indicating that the modeling was successful; compared with the model group, the prostate index (P < 0.05/0.01) of the model mice can be obviously reduced by the groups except that the effect of the urine retention capsule and the low dosage of the rose is close to the statistical difference (P > 0.05).
Table 3: effect on prostate index in model mice
(as compared to the normal group: ▲▲ P<0.01; compared with the model group: */** P<0.05/0.01)
3.3 Effect on 5α -reductase Activity in liver tissue of mice with prostatic hyperplasia
As can be seen from table 4, the activity of 5 a-reductase in liver homogenate of mice in the model group is significantly higher than that of normal control group, indicating that a large amount of exogenous androgens induce the increase of the activity of 5 a-reductase; except that the effect of the capsule for treating uroschesis is close to the statistical difference (P is more than 0.05), the activity of 5alpha-reductase in animal liver homogenate of three dose groups of positive control finasteride and rose is obviously lower than that of a model group (P is less than 0.01), and the results are consistent with the results of in vitro experiments.
Table 4: effect on 5 a-reductase activity in liver tissue of model mice (compared to normal group: ▲ p<0.05; compared with the model group: ** p<0.01)
3.4 effects on the level of estrogen/androgen in serum of mice with prostatic hyperplasia
As can be seen from Table 5, the serum levels of estrogen and androgen in mice of the model group are significantly higher than those in mice of the normal control group (P < 0.01), indicating that the mice with prostatic hyperplasia are accompanied by endocrine level imbalance. Each of the dosing groups significantly reduced serum E2 levels (P < 0.05/0.01) in the model animals except for the low dose group of finasteride and rose; except for the urine retention comfort and the low-dosage rose groups, each administration group can obviously reduce the serum T level of the model animal (P < 0.05/0.01).
Table 5: effects on hormone levels in serum of model mice
(in contrast to the normal group, ▲▲ p<0.01; in contrast to the set of models, */** p<0.05/0.01)
experimental example 3: effect of roses on castration in combination with testosterone propionate replication in prostate hyperplasia model rats
1. Test materials
1.1 test article
Roses (purple powder), lot number 20160501 (prepared using the method of example 5); finasteride tablets (national drug standard H20030951), 5 mg/tablet, hubei Shubang pharmaceutical Co., ltd., 0161001; uroschesis capsule (national medicine standard Z10960007), 0.3 g/granule, shijia Kedi pharmaceutical company, inc., 161113; preparing suspension with different concentrations by pure water for standby.
1.2 animals
SPF-grade male SD rats, 8-10 weeks old, weighing 250-280 g, provided by the university of Kunming medical laboratory animal center, production license number: SCXK (yunnan) K2015-0002, certification authority: the Kunming scientific and technical office.
1.3 mould-making reagent
Testosterone propionate injection, national drug standard H31030524, specification 1mL/25mg, available from Shanghai general pharmaceutical Co., ltd., batch number 151202; peanut oil, an injectable adjuvant, supplied by the Kunming pharmaceutical group preparation laboratory. For clinical use, the testosterone propionate injection is prepared into 10mg/mL with peanut oil for subcutaneous injection in rats.
2 Experimental methods
2.1 model preparation and group administration
Methods of reference (Qi, p., li, z, chen, m., sun, z, huang, c, 2013.Metabolismand tissue distribution study of Vaccaria seeds (Wang-Bu-Liu-Xing) in benignprostatic hyperplasia model rat: toward an in-depth study for its bioactive components.j Pharm Biomed Anal, 218-230.; hu Lou, li Peng, gu Shufang. Effects of fenugreek on experimental prostatic hyperplasia in rats [ J ]. Pharmacological and clinical chinese medicine, 2011,27 (4): 52-54.; 80 male SD rats were selected, after 7 days of adaptive feeding, 10 were all but a sham group were anesthetized by intraperitoneal injection with 10% chloral hydrate, the extremities and head were fixed on the operating table, and the scrotum and its surrounding skin were sterilized by wiping with iodine and alcohol cotton. Bilateral testis was removed from scrotum under aseptic conditions. The sham operation group was the same as the castrated animals except that testes were not resected. After the castrated rats are recovered to feed for one week, the rats with larger weight deviation or poor conditions are removed, and the rats are randomly divided into 6 groups according to weight: model control group; a chemical positive control finasteride 1.0mg/kg group; 300mg/kg of Chinese medicine positive control uroschesis capsule; low, medium and high dose (0.5, 1.0, 2.0 g/kg) of roses, 11 per group. The sham group rejects 1 animal, with the remaining 9 animals included in the experiment. In addition to the normal control group, each model animal group was subcutaneously injected with testosterone propionate 5mg/kg (dissolved in peanut oil) daily in the morning, and each model animal group and the sham animal group were respectively administered by intragastric administration at the afternoon according to the dosage, and the administration volumes of the model control group and the sham animal group were 10mL/kg for 28 days.
2.2 observations and measurements
2.2.1 general Condition observations
Rats were observed daily for feeding and activity during the experiment, and body weights were measured weekly.
2.2.2 blood-related index detection
After 28 days of continuous administration, the animals are anesthetized by 10% chloral hydrate intraperitoneal injection without water inhibition for 12 hours, 4mL of femoral artery blood is obtained, the animals are centrifuged for 10 minutes at 4000rpm at 4 ℃ in a refrigerated centrifuge, serum is separated and packaged into a centrifuge tube of 1.5mL, and the centrifuge tube is put into a refrigerator at-20 ℃ for preservation. The level of hormones such as E2 and T, DHT in serum and the like and the index such as SOD, MDA, GSH, PACP are measured according to the instruction of the kit, and the operation method is strictly carried out according to the instruction.
2.2.3 organ coefficient measurement
After blood collection, the rats are sacrificed by cervical vertebra, the abdominal cavity is opened rapidly, the prostate, the kidney, the thymus and the spleen are picked up for weighing, and the coefficients of all organs are calculated.
Organ coefficient = organ mass (mg)/animal body weight (g)
2.2.4 detection of hormone levels and 5alpha-reductase Activity in prostate tissue homogenates
After the prostate is weighed, except for reserving tissues at two sides of the urethra for pathological examination, the rest prostate tissues are weighed, pre-cooled PBS is added according to the ratio of 1:4 to prepare 20% tissue homogenate, the tissue homogenate is centrifuged at 4000rpm for 10min at 4 ℃ of a refrigerated centrifuge, the supernatant is taken and split-packed into a centrifuge tube of 1.5mL, and the centrifuge tube is put into a refrigerator of-20 ℃ for preservation. The hormone levels such as E2 and T, DHT and the activity of 5α -reductase in the homogenates were measured according to the kit instructions, and the methods of operation were performed according to the instructions.
2.2.5 histopathological observations
Prostate tissues on two sides of the urethra are taken from each mouse, fixed in 10% formaldehyde solution, dehydrated by gradient alcohol, embedded by paraffin, sliced, and subjected to HE staining, and the pathological changes of the prostate tissues are observed under a lens. Reference (Tian Shaopeng, liang Haiqing, ni Yidong) the inhibition of prostatic hyperplasia by Qianlietong tablet [ J ]. Chinese herbal medicine 2005,36 (10): 1538-1539.) is classified into 4 classes of 0, I, II, III based on the degree of prostatic tissue pathological hyperplasia, and is respectively marked as 0, 1,2, 3.
3. Experimental results
3.1 Effect on testosterone propionate replication in prostate hyperplasia model rats
3.1.1 effects on the general condition of model animals
During the molding and administration period, the sham operation group animals have normal fur, spirit, appetite and activity; the appearance behavior of the model animal is not obviously different from that of the false operation group except for part of vertical hair; animals in the rose group eat and drink normally, the fur is glossy, the state is obviously improved compared with the model group, and no animal death occurs until the experiment is finished.
3.1.2 Effect on model animal body weight
As can be seen from table 6, there was no significant difference in the basal body weight of animals for each model group prior to dosing compared to sham operated groups; during the dosing period, the body weight of each group of animals increased, wherein the increase amplitude of the sham operated group was larger, but there was no statistical difference (P > 0.05) compared to each model animal group; the body weight of each of the animals on days 7, 14, 21, 28 was also not significantly different from that of the model control group (P > 0.05).
Table 6: influence on model rat body weight (p >0.05 compared to model group)
3.2 Effect on visceral index in model animals
As can be seen from table 7, the prostate and kidney indices were significantly increased (P < 0.01) and thymus index was significantly decreased (P < 0.01) in the model group compared to the sham group; compared with the model control group, the prostate index (P < 0.05/0.01) can be obviously reduced by each administration group except for the urine retention comfort and the rose low-dose group, but the prostate index, spleen index and kidney index are not obviously influenced (P > 0.05).
Table 7: influence on organ index of model rat
(in contrast to the sham group, ▲▲ p<0.01; in contrast to the set of models, */** p<0.05/0.01)
3.3 Effect on serum detection index for model animals
3.3.1 effects on serum hormone levels
As can be seen from Table 8, the serum levels of DHT and E2 were significantly increased in rats in the model group compared to the sham group, the T decrease was close to the statistical difference, and E2/T tended to increase. Serum DHT was reduced to a different extent for each dosing group compared to the model group, with the finasteride and rose high dose group being significant (P < 0.01/0.05); except for the uroschesis and the low dosage of the roses, the E2 of each administration group is reduced to different degrees, and the obvious P is less than 0.01/0.05 in the finasteride and the high dosage of the roses; serum T was increased to a different extent for each dosing group, and E2/T tended to decrease (P > 0.05).
Table 8: effects on serum hormone levels in model rats
(in contrast to the sham group, ▲ p<0.05; in contrast to the set of models, */** p<0.05/0.01)
3.3.2 Effect on serum antioxidant index
As seen from table 9, the serum SOD activity was significantly reduced (P < 0.01) and MDA content was significantly increased (P < 0.01) in animals of the model control group compared to the sham group; compared with the model group, serum SOD level of each administration group is increased to different extent, MDA is reduced to different extent, wherein except for the effects of SOD (MDA and MDA) in roses (P > 0.05), the activity of SOD can be obviously improved and the MDA content can be reduced (P < 0.05/0.01) in the rest administration groups.
Table 9: influence on model rat serum antioxidant index
(in contrast to the sham group, ▲/▲▲ p<0.05/0.01; in contrast to the set of models, */** p<0.05/0.01)
3.3.3 Effect on serum PACP
As can be seen from Table 10, the prostate acid phosphatase (PACP) activity was significantly increased in the serum of animals from the model group compared to the sham group (P < 0.01); each dosing group was able to reduce PACP activity in serum of model animals to varying degrees, except that the effect of the low dose rose group was close to statistical difference, all other dosing groups were significant compared to the model group (p < 0.05/0.01).
Table 10: effects on PACP in model rat serum
(in contrast to the sham group, ▲▲ p<0.01; in contrast to the set of models, */** p<0.05/0.01)/>
3.4 Effect on detection index of prostate tissue homogenate in model animals
3.4.1 Effect on 5alpha-reductase Activity in tissue homogenates
As can be seen from Table 11, the 5. Alpha. -reductase activity was significantly increased (P < 0.05) in the prostate homogenate of animals in the model group compared to the sham group; each of the dosing groups reduced 5α -reductase activity in prostate tissue of model animals to a different extent than the model group, with all but the low dose rose groups had a near statistical difference in effect (p < 0.05/0.01).
Table 11: effect on 5α -reductase in model rat prostate tissue
(in contrast to the sham group, ▲ p<0.05; in contrast to the set of models, */** p<0.05/0.01)
3.4.2 effects on hormone levels in tissue homogenates
As can be seen from Table 12, the levels of DHT, E2 and T were significantly increased in the prostate homogenates of the model group compared to the sham-operated group (P < 0.05/0.01). Each of the dosing groups significantly reduced the levels of DHT and T in the prostate homogenate (P < 0.05/0.01) except for the low (DHT) and medium (T) rose dose groups compared to the model control group; all dosing groups had a trend to decrease E2 levels (P > 0.05) but had no significant effect on the E2/T ratio.
Table 12: effects on hormone levels in prostate tissue of model rats
(in contrast to the sham group, ▲/▲▲ p<0.05/0.01; in contrast to the set of models, */** p<0.05/0.01)
3.5 plain mirror inspection
Group of sham operations: no obvious abnormalities were seen in the prostate tissue. The glands of the prostate are orderly arranged, the gland cavities are not expanded, the gland epithelium is in a single-layer column shape, the interstitium is visible among the glands, and a small number of finger-shaped protrusions are occasionally seen at the individual edges.
Model control group: the glandular tissue is obviously increased. The number and height of glandular epithelium are increased, fingers and even cauliflower are formed to protrude into glandular cavity, the interstitium also has hyperplasia with different degrees, partial glandular body is expanded, and the endocrine in the cavity is increased.
Finasteride group: the tissue morphology of the model animal is obviously improved. The expansion of the gland cavity is not obvious, the number of the proliferated glandular epithelial cells is obviously less than that of the model group, and the epithelium is in a low column shape, and part of the epithelium tends to return to normal.
Dysuria-relieving capsule group: the gland arrangement is relatively regular, the epithelial cells are not proliferated or proliferated obviously, the finger-like protrusions are reduced compared with the model group, the cells are in a low column shape or flat shape, and the improvement degree is weaker than that of the finasteride group.
Rose: the three-dose group showed a reduction in glandular epithelial cell height, number and finger structure compared to the model group, but still exhibited a somewhat proliferative state compared to the finasteride group and the sham group, wherein the high-dose group improved glandular proliferation to a better extent than the low-dose group. The specific scores are shown in Table 13.
As can be seen from table 13, animals in the model group had significant differences in prostate tissue pathology scores (P < 0.01) compared to the sham group; compared with the model group, the finasteride and rose three-dose groups can obviously lighten the pathological changes of the prostate tissues of the model rats (P < 0.01/0.05).
Table 13: effects on model rat prostate tissue pathology
Experimental example 4: acute toxicity test of oral gastric lavage route of mice
The pre-test shows that the oral rose micropowder of the mice can not detect the LD50, so the acute toxicity condition of the mice is reflected by the maximum dosage of 3 times of stomach infusion in one day. Taking 40 mice of 18-20g, randomly dividing the mice into a control group and a rose pollen group according to weight and sex, wherein each group comprises 20 mice, and male and female halves. After 8 hours of no water feeding, mice were gavaged 3 times within 1 day at a maximum concentration of 0.2g/mL and a maximum volume of 40mL/kg, each time at 6 hour intervals, and the control group was given the same volume of purified water. The 14d continuous observation recorded the symptoms of poisoning and death. Animals are killed after dislocation at the end of the observation period, dissected, and the volume, color, texture and the like of each tissue and viscera are observed with naked eyes to see whether obvious abnormalities exist. The results are shown in Table 14.
Table 14: rose pollen effect on mice maximum tolerance test body weight (P >0.05 compared to control group)
As can be seen from Table 14, the mice were drenched 3 times with rose pollen at a maximum concentration of 0.2g/mL and a maximum volume of 40mL/kg within one day, no significant toxicity or death of the animals was observed during the observation period of 14 consecutive days, and no significant difference in weight gain was observed between the two groups; no obvious abnormalities in tissues or organs were found in necropsy of the sacrificed animals at the end of the observation period. The maximum tolerance of the rose micropowder orally infused by mice is more than 24g/kg.
Experimental example 5: subacute toxicity test of oral gastric lavage route of mice
Taking 48 mice of 19-21 g, dividing the mice into 4 groups according to sex and weight: vehicle control group; 1.0g/kg of rose pollen, 2.0g/kg of rose pollen and 12 rose pollen per group. Each group of animals was dosed once daily by stomach irrigation, and the control group was dosed with pure water, all at a volume of 20mL/kg for 30 consecutive days. Animals were observed daily for general behavior, toxic response, and mortality, and body weights were weighed weekly to adjust dosing. Taking the weight of the animal, taking the eyeball to separate blood and serum, and measuring ALT, AST, cre, BUN, UA; the heart, liver, spleen, lung, kidney and thymus were dissected and weighed on an analytical balance to calculate the organ coefficients. The results are shown in tables 15-17.
Table 15: influence of continuous infusion of Rose pollen on mouse body weight for 30 days
(in comparison with the control group, ** P<0.01)
| group of | Administration is carried out for 0 day | Administration is carried out for 7 days | Administration is carried out for 14 days | Administration is carried out for 21 days | Administration is carried out for 28 days | Administration is carried out for 30 days |
| Pure water control | 19.7±0.4 | 28.3±2.3 | 30.4±3.2 | 30.3±3.6 | 36.8±4.3 | 36.3±4.5 |
| Rose 1.0g/kg | 19.9±0.9 | 27.1±1.6 | 29.1±3.3 | 28.4±3.0 | 30.5±3.2** | 31.0±2.9** |
| 2.0g/kg of rose | 19.9±0.9 | 27.4±2.4 | 30.1±4.4 | 28.1±4.5 | 29.0±3.3** | 28.1±3.8** |
| 4.0g/kg of rose | 19.7±0.5 | 26.4±2.3 | 26.5±3.3** | 24.0±3.0** | 27.3±3.1** | 26.8±3.6** |
As can be seen from Table 15, the animals in each group were free to move and had a glossy coat during the test, and generally had good conditions, but the weight gain of the rose 3 dose groups was significantly lower than that of the control group, and had a significant dose-effect and a definite time-effect trend. None of the remaining groups died except 4.0g/kg of mice died (anatomic confirmation) due to misuse of the lavage at day 20 of administration. Dissected animals were sacrificed at the end of the test, and visceral tissues such as heart, liver, spleen, lung, kidney were visually observed without significant abnormalities.
As can be seen from Table 16, the mice were continuously perfused with roses at 1.0, 2.0, 4.0g/kg doses for 30 days, and the liver and kidney function indexes were not significantly different from those of the control group except for the significant decrease of serum BUN in the high dose group.
Table 16: effect of continuous gastric lavage of rose on liver and kidney functions of mice for 30 days
(in comparison with the control group, * P<0.05)
as can be seen from Table 17, the mice were continuously perfused with roses at 1.0, 2.0, 4.0g/kg for 30 days, with the exception that the thymus and spleen coefficients were significantly lower in the high dose group than in the control group, and the rest of the organ coefficients were not statistically different from the control group.
Table 17: influence of continuous gastric lavage of flos Rosae Rugosae on organ coefficients of mice for 30 days
(in comparison with the control group, * P<0.05)
the foregoing is merely exemplary of the present invention, and specific technical solutions and/or features that are well known in the art have not been described in detail herein. It should be noted that, for those skilled in the art, several variations and modifications can be made without departing from the technical solution of the present invention, and these should also be regarded as the protection scope of the present invention, which does not affect the effect of the implementation of the present invention and the practical applicability of the patent. The protection scope of the present application shall be subject to the content of the claims, and the description of the specific embodiments and the like in the specification can be used for explaining the content of the claims.
Claims (10)
1. The application of flos Rosae Rugosae in preparing product for resisting prostatic hyperplasia/hypertrophy is provided.
2. Use of rose according to claim 1 for the preparation of a product against prostatic hyperplasia/hypertrophy, characterized in that: the product comprises medicines, foods, health products and cosmetics.
3. Use of rose according to claim 1 for the preparation of a product against prostatic hyperplasia/hypertrophy, characterized in that: the product was prepared using petals of rose.
4. Use of rose according to claim 3 for the preparation of a product against prostatic hyperplasia/hypertrophy, characterized in that: the petals are first prepared as a powder or extract and then the powder or extract is used to prepare the product.
5. Use of rose according to claim 4 for the preparation of a product against prostatic hyperplasia/hypertrophy, characterized in that: the extract is prepared by the following method: squeezing the petals of the rose to obtain rose juice, and freeze-drying to obtain the freeze-dried powder extract.
6. Use of rose according to claim 1 for the preparation of a product against prostatic hyperplasia/hypertrophy, characterized in that: the raw materials of the product also comprise other functional components or auxiliary materials.
7. Use of rose according to claim 6 for the preparation of a product against prostatic hyperplasia/hypertrophy, characterized in that: the auxiliary materials comprise plant starch, microcrystalline cellulose and magnesium stearate.
8. Use of rose according to claim 6 for the preparation of a product against prostatic hyperplasia/hypertrophy, characterized in that: the other functional components comprise male silkworm moth and Cordyceps sinensis mycelia.
9. Use of rose according to claim 1 for the preparation of a product against prostatic hyperplasia/hypertrophy, characterized in that: the product is tablet, capsule, granule or oral liquid.
10. Use of rose according to claim 1 for the preparation of a product against prostatic hyperplasia/hypertrophy, characterized in that: the dosage range of the rose is 10 mg-10000 mg per kilogram of body weight.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0517365A (en) * | 1991-07-03 | 1993-01-26 | Nonogawa Shoji Kk | Testosterone 5alpha-reductase inhibitor |
| KR20200037537A (en) * | 2018-10-01 | 2020-04-09 | 중앙대학교 산학협력단 | Composition for preventing or treating benign prostatic hyperplasia comprising rose flower extract or fraction thereof |
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- 2023-01-17 CN CN202310078877.1A patent/CN116077562A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0517365A (en) * | 1991-07-03 | 1993-01-26 | Nonogawa Shoji Kk | Testosterone 5alpha-reductase inhibitor |
| KR20200037537A (en) * | 2018-10-01 | 2020-04-09 | 중앙대학교 산학협력단 | Composition for preventing or treating benign prostatic hyperplasia comprising rose flower extract or fraction thereof |
Non-Patent Citations (1)
| Title |
|---|
| 张仁荣: "崔学教辨治前列腺增生症用药经验撷萃", 时珍国医国药, vol. 29, no. 01, pages 208 - 209 * |
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