Traditional Chinese medicine composition for treating perimenopausal syndrome as well as preparation method and application of traditional Chinese medicine composition

Abstract

The invention belongs to the technical field of traditional Chinese medicine compositions, and particularly relates to a traditional Chinese medicine composition for treating perimenopausal syndrome as well as a preparation method and application of the traditional Chinese medicine composition. The traditional Chinese medicine composition is prepared from 8-12 parts of rhizoma curculiginis, 8-12 parts of herba epimedii, 8-12 parts of radix morindae officinalis, 8-12 parts of radix angelicae sinensis, 4-6 parts of cortex phellodendri, 4-6 parts of rhizoma anemarrhenae, 8-12 parts of fructus ligustri lucidi, 8-12 parts of herba ecliptae, 13-17 parts of light wheat, 8-12 parts of radix glycyrrhizae, 8-12 parts of fructus ziziphi jujubae, 8-12 parts of fossil fragments and 8-12 parts of concha ostreae. The traditional Chinese medicine composition can improve the effect of treating the perimenopausal syndrome.

Description

Technical Field

[0001] The present invention belongs to the technical field of Chinese medicine compositions, and in particular relates to a Chinese medicine composition for treating perimenopausal syndrome, a preparation method and use thereof. Background Art

[0002] Perimenopausal syndrome, also known as menopausal syndrome, is a common gynecological disease in contemporary middle-aged and elderly women; patients often experience irregular menstruation, hot flashes, night sweats, mood swings, sleep disorders and other symptoms. At present, Western medicine mainly treats this disease with hormone replacement, including oral hormone therapy or transdermal and vulvar administration. Estradiol tablets and estradiol dydrogesterone tablets are common therapeutic hormones, but most Western medicine treatments have the disadvantage of treating symptoms but not the root cause.

[0003] Research progress on the mechanism and related complications of TCM treatment of perimenopausal syndrome, Zheng Xiaojun, Zhang Lei, Wu Xiaozhen, Xu Leqin, Wu Xiaoyun, TCM Information, 2024-08-01 10:45; Shao Yan used Erxian Decoction to treat perimenopausal hypertension patients and found that it can effectively reduce the expression of serum inflammatory factors and blood pressure variability. Fan Shunhua et al. used modified Erxian Decoction to treat perimenopausal hypertension patients caused by Chongren disorder and found that it can not only effectively improve the clinical symptoms of patients, but also regulate the release of neurotransmitters and sex hormone levels. Summary of the invention

[0004] The technical problem to be solved by the present invention is to provide a Chinese medicine composition for treating perimenopausal syndrome, a preparation method and use thereof, so as to improve the effect of treating perimenopausal syndrome.

[0005] The embodiment of the present invention provides a traditional Chinese medicine composition for treating perimenopausal syndrome, comprising the following traditional Chinese medicine components in parts by weight: 8-12 parts of Curculigo, 8-12 parts of Epimedium, 8-12 parts of Morinda officinalis, 8-12 parts of Angelica sinensis, 4-6 parts of Phellodendron, 4-6 parts of Anemarrhena, 8-12 parts of Ligustrum lucidum, 8-12 parts of Eclipta prostrata, 13-17 parts of Triticum tiliaceus, 8-12 parts of Licorice, 8-12 parts of Jujube, 8-12 parts of Dragon Bone, and 8-12 parts of Oyster.

[0006] Preferably, it is composed of the following traditional Chinese medicine components in parts by weight: 8-12 parts of Curculigo, 8-12 parts of Epimedium, 8-12 parts of Morinda officinalis, 8-12 parts of Angelica sinensis, 4-6 parts of Phellodendron chinense, 4-6 parts of Anemarrhena asphodeloides, 8-12 parts of Ligustrum lucidum, 8-12 parts of Eclipta prostrata, 13-17 parts of Triticum tiliaceus, 8-12 parts of Licorice, 8-12 parts of Jujube, 8-12 parts of Dragon Bone, and 8-12 parts of Oyster.

[0007] Preferably, the composition comprises the following Chinese medicinal components in parts by weight: 10 parts of Curculigo, 10 parts of Epimedium, 10 parts of Morinda officinalis, 10 parts of Angelica sinensis, 5 parts of Phellodendron, 5 parts of Anemarrhena, 10 parts of Ligustrum lucidum, 10 parts of Eclipta prostrata, 15 parts of Triticum tiliaceum, 10 parts of Licorice, 10 parts of Jujube, 10 parts of Dragon Bone, and 10 parts of Oyster.

[0008] The embodiment of the present invention provides a method for preparing the Chinese medicine composition, comprising mixing the Chinese medicine components, decocting with water twice, adding 8-12 times the amount of water for the first time and adding 5-7 times the amount of water for the second time, combining the decoctions, filtering and concentrating to obtain the Chinese medicine composition.

[0009] Preferably, 10 times the amount of water is added for the first time and 6 times the amount of water is added for the second time.

[0010] Preferably, the first decoction time is 1-2 hours, and the second decoction time is 0.5-1 hour.

[0011] Preferably, the relative density of the concentrated medicinal solution is 1.3-1.4.

[0012] An embodiment of the present invention provides a use of the traditional Chinese medicine composition, wherein the traditional Chinese medicine composition is used to prepare a medicine for treating perimenopausal syndrome.

[0013] The beneficial effects of the present invention are that curculigo, epimedium, Morinda officinalis, angelica, phellodendron, rhizoma anemarrhenae, fructus ligustris, eclipta prostrata, floating wheat, licorice, jujube, keel and oyster are added to the traditional Chinese medicine prescription of the present invention. The present invention treats perimenopausal syndrome, mainly adds components such as floating wheat, keel and oyster, and also adds components such as phellodendron and rhizoma anemarrhenae. The results of animal experiments show that the traditional Chinese medicine composition of the present invention can significantly improve the effect of treating perimenopausal syndrome, the effects of the middle and low dose groups and the positive drug group are equivalent, the efficacy of some high dose groups is better than that of the positive drug group, and the present invention has a good therapeutic effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 Results of consecutive vaginal smears of rats (200×, scale bar = 100 μm).

[0015] Figure 2 These are the open field movement trajectories of rats in each group, including A: sham operation group, B: model group, C: positive drug group, D: high-dose Chinese medicine group, E: medium-dose Chinese medicine group, and F: low-dose Chinese medicine group.

[0016] Figure 3 These are the results of hypothalamic HE staining (40×), where A: sham operation group, B: model group, C: positive drug group, D: high-dose Chinese medicine group, E: medium-dose Chinese medicine group, and F: low-dose Chinese medicine group.

[0017] Figure 4The results of HE staining of uterus (200×). A: sham operation group, B: model group, C: positive drug group, D: high-dose Chinese medicine group, E: medium-dose Chinese medicine group, F: low-dose Chinese medicine group.

[0018] Figure 5 The protein electrophoresis of kisspeptin, GPR54, GnRH in the hypothalamic ARC region and pituitary GnRH-R in each group was shown. DETAILED DESCRIPTION

[0019] The present invention is described in detail below in conjunction with specific implementation modes.

[0020] Example 1

[0021] A traditional Chinese medicine composition for treating perimenopausal syndrome comprises the following traditional Chinese medicine components in parts by weight: 10 parts of Curculigo, 10 parts of Epimedium, 10 parts of Morinda officinalis, 10 parts of Angelica sinensis, 5 parts of Phellodendron, 5 parts of Rhizoma Anemarrhenae, 10 parts of Fructus Ligustri Lucidi, 10 parts of Eclipta prostrata, 15 parts of Triticum aestivum, 10 parts of Licorice, 10 parts of Jujube, 10 parts of Os Dracaenae, and 10 parts of Ostreae Conchae.

[0022] The preparation method of the Chinese medicine composition for treating perimenopausal syndrome comprises the following steps: mixing the above Chinese medicine components, decocting with water twice, adding 10 times the amount of water for the first time and decocting for 2 hours, and adding 6 times the amount of water for the second time and decocting for 1 hour, combining the decoctions, filtering, and concentrating to a relative density of 1.3-1.4 to obtain the Chinese medicine composition.

[0023] Example 2

[0024] Experimental animals

[0025] 60 SPF female SD rats, 8-10 weeks old, weighing 280-320 g, were provided by Hunan Slake Jingda Experimental Animal Co., Ltd., animal certificate number SCXK (Xiang) 2021-0002, and were routinely raised in the Experimental Animal Center of Hunan University of Chinese Medicine. The breeding conditions were as follows: temperature 22-26°C, daily temperature difference ≤3°C, relative humidity 40%-70%, light-dark alternation time 12 / 12h, and rats were free to eat and drink water. This experiment was approved by the Animal Experiment Ethics Committee of Hunan University of Chinese Medicine (ethics approval number HNUCM21-2408-03).

[0026] Main drugs and reagents

[0027] Ampicillin sodium USP grade (Aladdin, item number: A105483); estradiol valerate tablets (Bayer Healthcare Co., Ltd. (Guangzhou), item number: J20171038); 0.9% sodium chloride injection; GnRH test kit (item number: J3396-A), purchased from Jiangsu Jingmei Biotechnology Co., Ltd.; LH test kit (item number: J3676-A), purchased from Jiangsu Jingmei Biotechnology Co., Ltd.; FSH test kit (item number: J21359-A), purchased from Jiangsu Jingmei Biotechnology Co., Ltd.; E2 test kit (Cat. No.: J3450-A) was purchased from Jiangsu Jingmei Biotechnology Co., Ltd.; IL-1β test kit (Cat. No. JM-01454R1) was purchased from Jiangsu Jingmei Biotechnology Co., Ltd.; IL-18 test kit (Cat. No. JM-01601R1) was purchased from Jiangsu Jingmei Biotechnology Co., Ltd.; TNF-α test kit (Cat. No. JM-01587R1) was purchased from Jiangsu Jingmei Biotechnology Co., Ltd.; RIPA protein lysis buffer (Cell Signaling Technology, Catalog No. 9806), polybutylcyanoacrylate (BCA) protein quantification kit (Cell Signaling Technology, Catalog No. 7780), β-actin (β-actin) antibody (abcam, Catalog No. ab8227), kisspeptin antibody (Bioss, Catalog No. bs-0749R), GPR54 antibody (abcam, Catalog No. ab100896), GnRH antibody (proteintech, Catalog No. 26950-1-AP), GnRH-R antibody (proteintech, Catalog No. 19950-1-AP), hematoxylin-eosin (HE) staining solution set, Beso Gram fast staining solution kit (Zhuhai Beso Biotechnology Co., Ltd., Catalog No. BA4012), horseradish peroxidase (HRP) labeled goat secondary antibody (abcam, Catalog No. ab205719).

[0028] Animal grouping, modeling and drug administration

[0029] ① Animal grouping: 48 SD rats were randomly divided into sham operation group, model group, low-dose Chinese medicine group, medium-dose Chinese medicine group, high-dose Chinese medicine group and positive drug group, with 8 rats in each group.

[0030] ② Modeling: A rat model of perimenopausal syndrome was established with reference to the method for preparing the animal model of perimenopausal syndrome developed by the Chinese Association of Traditional Chinese Medicine Experimental Pharmacology Committee (Miao Mingsan, Tian Shuo, Xin Weiyun, et al., Specification for Preparation of Animal Model of Perimenopausal Syndrome (Draft), Chinese Journal of Traditional Chinese Medicine, 2018, 33(03):996-1000).

[0031] The model was established by incomplete castration: rats were given 1 mL of 0.35% sodium pentobarbital for every 100 g of body weight, and anesthetized by intraperitoneal injection. After anesthesia, the rats were fixed in the abdominal position, and the hair was cut about 2 cm away from the spine on the midaxillary line below the last rib on the back. After disinfection with iodine, the skin and back muscles were cut 1.5-2.0 cm, and a mass of white shiny fat was seen in the field of vision, in which the ovaries were embedded; the fat mass was gently clamped with small tweezers, pulled out of the incision and separated, and a light pink follicular ovary was seen. After ligating the fallopian tube, the ovaries were removed, and the left ovary was completely removed, and the right ovary was only removed by 80%. The uterine horn was returned to the abdominal cavity, and penicillin was given locally in the incision. Suture, and penicillin sodium salt was injected intraperitoneally at 5.0×104U / d after surgery for 3 consecutive days to prevent infection. The sham operation group did not remove the ovaries, and the rest of the method was the same as above. The animals were fed normally and subjected to vaginal smear examination once a day for 5 consecutive days starting from 5 days after surgery. If no estrus reaction occurred, the rat perimenopausal syndrome model was successfully established.

[0032] ③ Drug administration: Start drug administration after successful model establishment, once a day, and continue for 4 weeks.

[0033] Positive control drug: estradiol valerate, 0.09 mg / kg / d;

[0034] Example 1 The concentration was adjusted to a liquid concentration of 0.7614 g / mL (containing crude drugs). The drug dosage was based on the animal surface area-dose conversion method (He Shilin, Wang Jian, Wang Jingjing, Chinese Medicine Research Design and Statistics: Chinese Medicine Research Design and Statistical Methods, Hunan Science and Technology Press, 2001: 64). The liquid concentrations of the low-dose group, medium-dose group, and high-dose group of Chinese medicine were 1.9035 g / kg / d, 3.807 g / kg / d, and 7.614 g / kg / d, respectively. According to the above-mentioned doses, the six groups were gavaged with a dosage volume of 10 mL / kg / d.

[0035] Sampling and index testing

[0036] The open field test and novel object recognition test were performed 24 hours after the last gavage. After the animal behavior experiment, the samples were collected. After anesthesia, blood was collected to separate serum; the uterus was taken, weighed and fixed in 4% paraformaldehyde; the ovary and pituitary were taken and fixed in 4% paraformaldehyde; the hypothalamus of 4 rats were randomly taken from each group of rats and fixed in 4% paraformaldehyde; the remaining hypothalamus of each group was quickly frozen and stored at -80°C.

[0037] ① Determination of animal body weight, uterine wet weight and uterine coefficient.

[0038] ② Determine the total time of rats exploring objects and the discrimination index (DI) in the new object experiment.

[0039] ③ Determination of GnRH, LH, FSH and E2.

[0040] The levels of GnRH, LH, FSH and E2 in serum were determined according to the kit instructions.

[0041] ④ Elisa was used to detect the levels of serum inflammation-related factors IL-1β, IL-18 and TNF-α.

[0042] ⑤Hematoxylin-eosin (HE) staining was used to observe the pathological changes of the hypothalamus, pituitary, uterus and ovarian tissues.

[0043] The hypothalamus, pituitary, uterus and ovarian tissues fixed in 4% paraformaldehyde solution were dehydrated, embedded, sliced ​​and stained according to the conventional HE staining method. After sealing with neutral gum, the pathological changes of the hypothalamus, pituitary, uterus and ovarian tissues were observed under an inverted microscope and photographed.

[0044] Statistical processing

[0045] SPSS 25.0 software was used for statistical analysis. If the data conform to normal distribution and the variance is homogeneous between groups, LSD method is used. If the variance is not homogeneous, Dunnett's T3 method is used. If the data do not conform to normal distribution, non-parametric test is used. P < 0.05 means the difference is statistically significant.

[0046] Results and Conclusions

[0047] The results of rat vaginal smear are shown in Figure 1 As shown in the figure, the incomplete castration group is the model group.

[0048] Open field test

[0049] Compared with the sham operation group, the average speed, total distance and activity time of the rats in the model group decreased significantly (P<0.01); compared with the model group, the average speed, total distance and activity time of the positive drug group (estradiol valerate group) and the Chinese medicine medium dose group and Chinese medicine high dose group increased significantly (P<0.05, P<0.01). See Table 1. Open field movement trajectory is shown in Figure 2 .

[0050] Table 1 Changes in open field test of rats in each group ( n=4)

[0051]

[0052] The total exploration time and discrimination index (DI) of rats in each group are shown in Table 2.

[0053] Table 2 Total exploration time and discrimination index (DI) of rats in each group

[0054]

[0055] The body weight and uterine index of rats in each group are shown in Table 3.

[0056] Table 3 Body weight and uterine index of rats in each group

[0057]

[0058] Note: Compared with the sham operation group 1) P<0.01; compared with the model group 2) P<0.05, 3) P<0.01.

[0059] The comparison of GnRH, LH, FSH and E2 in the serum of rats in each group is shown in Table 4.

[0060] Table 4 Comparison of serum GnRH, LH, FSH and E2 in rats of each group

[0061]

[0062] Comparison of IL-1β, IL-18 and TNF-α in serum of rats in each group See Table 5.

[0063] Table 5 Comparison of IL-1β, IL-18 and TNF-α in serum of rats in each group

[0064]

[0065] The results of hypothalamic HE staining are shown in Figure 3 , HE staining results of uterus are shown in Figure 4 .

[0066] The expressions of kisspeptin, GPR54, GnRH in the hypothalamic ARC region and pituitary GnRH-R protein in each group of rats are shown in Table 6.

[0067] Table 6 Expression of kisspeptin, GPR54, GnRH in hypothalamic ARC region and pituitary GnRH-R protein in each group of rats

[0068]

[0069] From Table 1-6 and Figure 1-5It can be seen that compared with the model group, the Chinese medicine group can reduce the levels of GnRH, FSH, and LH, increase the level of E2, reduce the shrinkage of hypothalamic neurons, slightly increase the number of neurons, thicken the endometrium of uterine tissue, increase the number of glands and improve gland atrophy, significantly reduce the body weight of rats, significantly increase the wet weight and index of uterus, improve the anxiety-like behavior and learning and memory ability of rats, reduce the level of neuroinflammation, and regulate the homeostasis of HPO axis through kisspeptin-GnRH pathway to improve perimenopausal syndrome, among which the high-dose group had the best effect.

[0070] A person skilled in the art should understand that the discussion of any of the above embodiments is merely illustrative and is not intended to imply that the scope of protection of the present application is limited to these examples. In line with the concept of the present application, the technical features in the above embodiments or different embodiments may be combined, the steps may be implemented in any order, and there are many other variations of different aspects of one or more embodiments of the present application as described above, which are not provided in detail for the sake of simplicity.

[0071] One or more embodiments of the present application are intended to cover all such substitutions, modifications and variations that fall within the broad scope of the present application. Therefore, any omissions, modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of one or more embodiments of the present application should be included in the protection scope of the present application.

Claims

1. A Chinese medicine composition for treating perimenopausal syndrome, characterized in that: The invention comprises the following Chinese medicinal components in parts by weight: 8-12 parts of Curculigo, 8-12 parts of Epimedium, 8-12 parts of Morinda officinalis, 8-12 parts of Angelica sinensis, 4-6 parts of Phellodendron amurense, 4-6 parts of Rhizoma Anemarrhenae, 8-12 parts of Fructus Ligustri Lucidi, 8-12 parts of Eclipta prostrata, 13-17 parts of Triticum aestivum, 8-12 parts of Licorice, 8-12 parts of Jujube, 8-12 parts of Dragon Bone and 8-12 parts of Oyster.

2. The Chinese medicine composition according to claim 1, characterized in that: The invention is composed of the following Chinese medicinal components in parts by weight: 8-12 parts of Curculigo, 8-12 parts of Epimedium, 8-12 parts of Morinda officinalis, 8-12 parts of Angelica sinensis, 4-6 parts of Phellodendron chinense, 4-6 parts of Anemarrhena asphodeloides, 8-12 parts of Ligustrum lucidum, 8-12 parts of Eclipta prostrata, 13-17 parts of Triticum tiliaceus, 8-12 parts of Licorice, 8-12 parts of Jujube, 8-12 parts of Dragon Bone and 8-12 parts of Oyster.

3. The Chinese medicine composition according to claim 1, characterized in that: The invention comprises the following Chinese medicinal components in parts by weight: 10 parts of Curculigo orchioides, 10 parts of Epimedium, 10 parts of Morinda officinalis, 10 parts of Angelica sinensis, 5 parts of Phellodendron amurense, 5 parts of Rhizoma Anemarrhenae, 10 parts of Fructus Ligustri Lucidi, 10 parts of Eclipta prostrata, 15 parts of Triticum aestivum, 10 parts of Radix Glycyrrhizae, 10 parts of Jujube, 10 parts of Os Dracaenae, and 10 parts of Ostreae Conchae.

4. A method for preparing the Chinese medicine composition according to any one of claims 1 to 3, characterized in that: The Chinese medicinal components are mixed, and decocted twice with water, with 8-12 times the amount of water added for the first time and 5-7 times the amount of water added for the second time. The decoctions are combined, filtered, and concentrated to obtain the Chinese medicinal composition.

5. The preparation method according to claim 4, characterized in that: Add 10 times the amount of water the first time and 6 times the amount of water the second time.

6. The preparation method according to claim 4, characterized in that: The first decoction time is 1-2h, and the second decoction time is 0.5-1h.

7. The preparation method according to claim 4, characterized in that: The relative density of the concentrated solution is 1.3-1.

4.

8. A use of the Chinese medicine composition according to any one of claims 1 to 3, characterized in that: The traditional Chinese medicine composition is used for preparing medicine for treating perimenopausal syndrome.

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