Traditional Chinese medicine compound for menopausal syndrome

Abstract

The application provides a traditional Chinese medicine compound for treating menopausal syndrome, and belongs to the technical field of traditional Chinese medicines. The traditional Chinese medicine compound is composed of ten medicinal materials, i.e. radix panacis quinquefolii, herba eriochidis, longan aril, orchid root, radix paeoniae alba, radix alpiniae oxyphyllae, cucumis sativus, rhizoma anemarrhenae, syzygium aromaticum and citrus aurantium. The whole prescription is rigorous in prescription and unique in compatibility, and has the effects of nourishing liver and kidney, clearing heart and soothing nerves, regulating liver and invigorating spleen. In-vivo experiments prove from multiple angles that the compound can significantly inhibit ovary granulosa cell apoptosis, improve ovary reserve function, regulate sex hormone level and neurotransmitter metabolism disorder, and effectively relieve insomnia, emotional abnormalities and other symptoms caused by menopausal syndrome. A plurality of experimental indexes comprehensively show that the traditional Chinese medicine compound is an effective prescription for treating menopausal syndrome, and has a good clinical application prospect.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, specifically to a traditional Chinese medicine compound for menopausal syndrome. Background Technology

[0002] Menopausal syndrome refers to a group of physical and psychological symptoms experienced by women during the perimenopausal period due to the progressive decline in ovarian function, fluctuations in sex hormone levels, and imbalances in neuroendocrine regulation. Its clinical manifestations are complex and diverse, significantly impacting the physical and mental health of patients. Furthermore, fluctuations in hormone levels may increase long-term health risks, such as atrophy of the urogenital system, osteoporosis, and increased susceptibility to cardiovascular disease. Therefore, effectively preventing and treating perimenopausal syndrome and improving the quality of life and subjective well-being of middle-aged and elderly women has become an important clinical issue that urgently needs to be addressed.

[0003] Currently, hormone replacement therapy remains the primary intervention for treating menopausal syndrome, regulating endocrine function and alleviating clinical symptoms through exogenous hormone supplementation. However, this therapy has estrogen and progesterone-related adverse reactions and is subject to numerous precautions and absolute contraindications, which to some extent limits long-term patient adherence. In contrast, Traditional Chinese Medicine (TCM) offers advantages in treating perimenopausal symptoms, including diverse methods, fewer adverse reactions, and higher safety, effectively reducing related symptoms and improving patients' quality of life.

[0004] Traditional Chinese medicine believes that perimenopausal women are in a critical physiological transition period where kidney qi gradually declines, menstrual blood is about to be exhausted, and the Chong and Ren meridians transition from their peak to decline. The fundamental pathogenesis lies in the imbalance of yin and yang in the kidneys. Treatment emphasizes a holistic approach and syndrome differentiation, using comprehensive methods such as regulating qi and blood, unblocking meridians, and balancing yin and yang to regulate bodily functions at multiple targets, thereby stimulating and promoting the body's inherent self-repair capabilities.

[0005] However, the treatment of menopausal syndrome in Traditional Chinese Medicine (TCM) has not yet formed a systematic and standardized theoretical framework, and exploring precise and effective treatment plans remains a key focus of clinical research. Future efforts should focus on strengthening research into the mechanisms of action of TCM treatment and scientific evidence-based verification. Modern medical technologies can be used to explore the precise efficacy of TCM in regulating the endocrine system, immune network, and neurotransmitters, thereby providing a clearer theoretical basis and more practical diagnostic and treatment approaches for TCM intervention in menopausal syndrome. Summary of the Invention

[0006] (a) Technical problems to be solved

[0007] To address the shortcomings of existing technologies, this invention provides a traditional Chinese medicine compound for menopausal syndrome.

[0008] (II) Technical Solution

[0009] To achieve the above objectives, the present invention provides the following technical solution:

[0010] On the one hand, this application provides a traditional Chinese medicine compound for menopausal syndrome, which is made from the following raw materials in parts by weight: 10-36 parts of Codonopsis pilosula, 10-36 parts of Gynostemma pentaphyllum, 8-30 parts of Longan pulp, 5-25 parts of Codonopsis pilosula, 5-30 parts of Paeonia lactiflora, 10-40 parts of Smilax glabra, 6-24 parts of Trichosanthes kirilowii, 2-25 parts of Anemarrhena asphodeloides, 0.5-6 parts of Syzygium aromaticum, and 1-18 parts of Citrus reticulata.

[0011] Specifically, the traditional Chinese medicine compound is made from the following herbal raw materials in the indicated weight proportions: 15-30 parts of Codonopsis pilosula, 15-30 parts of Gynostemma pentaphyllum, 12-28 parts of Longan pulp, 10-20 parts of Codonopsis pilosula, 10-15 parts of Paeonia lactiflora, 15-30 parts of Smilax glabra, 10-15 parts of Trichosanthes kirilowii, 6-18 parts of Anemarrhena asphodeloides, 1-3 parts of Syzygium aromaticum, and 2-10 parts of Citrus reticulata.

[0012] Specifically, the traditional Chinese medicine compound is made from the following herbal raw materials in the indicated weight proportions: 25 parts of Codonopsis pilosula, 25 parts of Gynostemma pentaphyllum, 25 parts of longan pulp, 16 parts of Codonopsis pilosula, 16 parts of Paeonia lactiflora, 12 parts of Smilax glabra, 12 parts of Trichosanthes kirilowii, 12 parts of Anemarrhena asphodeloides, 1.5 parts of Syzygium aromaticum, and 6 parts of Citrus reticulata.

[0013] On the other hand, the present invention provides the application of the traditional Chinese medicine compound in the preparation of a drug for menopausal syndrome. The traditional Chinese medicine compound is made from the above-mentioned weight parts of Chinese herbal medicine as raw materials, supplemented with pharmaceutically acceptable excipients to prepare an oral dosage form. The oral dosage form is any one of decoction, granules, capsules, or concentrated pills.

[0014] The pharmacological effects of various Chinese herbal raw materials in the compound Chinese medicine formula of this invention are as follows:

[0015] Taibai ginseng: The root of *Pedicularis davidii* Franch., *Pedicularis decora* Franch., and *Pedicularis dunniana* Bonati, all belonging to the Scrophulariaceae family. It has a sweet and slightly bitter taste and is warm in nature. It enters the kidney and spleen meridians. It has the effects of nourishing yin and tonifying the kidney, invigorating qi and strengthening the spleen. It is used for deficiency of both spleen and kidney, steaming bone fever, joint pain, and loss of appetite.

[0016] Chickweed (also known as Thousand Needle Thread Herb): This plant belongs to the genus Chickweed in the family Caryophyllaceae. It enters the liver, spleen, and kidney meridians. It has the effects of invigorating qi and nourishing blood, strengthening the spleen and benefiting the kidneys; it is used for symptoms such as qi and blood deficiency, lethargy, dizziness, palpitations, soreness of the waist and knees, and irregular menstruation; it is an essential medicine for treating women's weakness.

[0017] Longan pulp: The aril of *Dimocarpus longan* Lour., a plant belonging to the Sapindaceae family. It has a sweet taste and warm nature; it enters the heart and spleen meridians. It has the effects of nourishing the heart and spleen, replenishing blood, and calming the mind. It is mainly used to treat excessive thinking, overwork damaging the heart and spleen, palpitations, insomnia, and forgetfulness.

[0018] Orchid Ginseng: The root or whole herb of *Wahlenbergia marginata* (Thunb.) A. DC., a plant in the Campanulaceae family. It has a sweet and slightly bitter taste, is neutral in nature, and enters the spleen and lung meridians. It has the effects of invigorating qi and strengthening the spleen, relieving cough and eliminating phlegm, and stopping bleeding. It is mainly used to treat weakness and fatigue, spontaneous sweating, night sweats, infantile malnutrition, leukorrhea in women, colds, coughs, epistaxis, malaria, and scrofula.

[0019] White peony root: The root of Paeonia lactiflora Pall., a plant belonging to the genus Paeonia in the family Ranunculaceae. It tastes bitter and sour, and is slightly cold in nature; it enters the liver and spleen meridians. It has the effects of nourishing blood and regulating menstruation, astringing yin and stopping sweating, softening the liver and relieving pain, and calming liver yang. It is used for blood deficiency and chlorosis, irregular menstruation, spontaneous sweating, night sweats, hypochondriac pain, abdominal pain, limb spasms, headache, and dizziness.

[0020] White Smilax: The tuber of *Smilax glabra*, *Smilax china*, and *Smilax scabra*, all belonging to the Liliaceae family. Sweet, bland, and neutral in nature. It enters the Liver and Stomach meridians. Its functions and indications are clearing heat and dampness, detoxifying and reducing swelling. It is used for dysuria, leukorrhea, and carbuncles and boils.

[0021] Mountain root: The dried tuberous root of *Merremia hungaiensis* (Lingelsh. et Borza) RC Fang, a plant belonging to the Convolvulaceae family. It is sweet and neutral in nature, entering the spleen and stomach meridians. It has the effects of clearing heat, removing dampness, relieving cough, and strengthening the spleen. It is mainly used to treat jaundice, chronic hepatitis, cough due to lung heat, hematochezia, leukorrhea, insufficient lactation, infantile malnutrition, and burns.

[0022] Anemarrhena asphodeloides Bge., a plant belonging to the genus Anemarrhena in the family Liliaceae, is a dried rhizome. It has a bitter and sweet taste, and is cold in nature; it enters the lung, stomach, and kidney meridians. It has the effects of clearing heat and purging fire, nourishing yin and moistening dryness. It is mainly used to treat exogenous febrile diseases, high fever and thirst, dry cough due to lung heat, steaming bone fever, internal heat and thirst, and constipation due to intestinal dryness.

[0023] Clove: The dried flower buds of *Eugenia caryophyllata* Thunb., a plant in the Myrtaceae family. It has a pungent taste and warm properties; it enters the spleen, stomach, lung, and kidney meridians. It has the effects of warming the middle jiao (spleen and stomach), relieving nausea and vomiting, tonifying the kidneys and strengthening yang. It is mainly used to treat spleen and stomach deficiency-cold, hiccups and vomiting, poor appetite and diarrhea, cold pain in the heart and abdomen, and kidney deficiency-induced impotence.

[0024] Mountain orange: The fruit of *Fortunella hindsii* (champ.) Swingle, a plant in the Rutaceae family. It is pungent, bitter, and warm in nature. It has the effect of regulating qi and relieving abdominal distension, and is mainly used to treat stomach pain, food stagnation and abdominal distension, hernia, cough due to wind-cold, and cold asthma.

[0025] (III) Beneficial Effects

[0026] Menopausal syndrome is clinically characterized by hot flashes, night sweats, mood swings, and insomnia. Traditional Chinese medicine believes its root cause lies in the gradual decline of kidney qi and the depletion of yin and yang within the kidneys. Menopausal patients often experience deficiency of essence and blood, depletion of yin and blood due to overwork, insufficient blood to nourish the heart, and restlessness leading to difficulty falling asleep, hence symptoms such as insomnia and palpitations. Yin deficiency and blood deficiency also cause disharmony between the heart and kidneys, resulting in excessive heart fire, which manifests as hot flashes and night sweats. This formula uses *Panax notoginseng*, *Hedyotis diffusa*, and longan pulp as the principal herbs. *Panax notoginseng* nourishes yin and tonifies the kidneys, invigorates qi and strengthens the spleen; *Hedyotis diffusa* invigorates qi and blood, strengthens the spleen and kidneys; longan pulp tonifies the heart and spleen, invigorates qi and blood, and calms the mind, directly addressing the pathogenesis and effectively relieving symptoms such as restlessness and insomnia. The three herbs combined are specifically designed to nourish and replenish, ensuring sufficient essence and blood, vigorous *Ren* and *Chong* meridians, abundant qi and blood, and nourishment of the internal organs. *Panax notoginseng* and white peony root are used as assistant herbs. Orchid ginseng invigorates qi and strengthens the spleen, replenishes deficiencies, and also has the effect of stopping sweating. It assists the principal herb in supporting the body's vital energy and directly addresses night sweats. White peony nourishes blood, softens the liver, astringes yin, and harmonizes the nutritive qi, blood, and yin essence, containing them internally and preventing their dissipation. White scutellaria, wild trichosanthes, anemarrhena, and clove are used as adjuvant herbs. Each of these four herbs performs its specific function, preventing the tonifying herbs from causing stagnation. White scutellaria and wild trichosanthes, when combined, effectively clear heat and promote diuresis, preventing the tonifying herbs from causing stagnation and dampness, and guiding damp-heat pathogens out through urination. Clove, being pungent and warm, acts as a counteracting agent, warming the kidneys and assisting yang. Within a large group of sweet and cold yin-nourishing herbs, it seeks yin within yang, maintaining the yin-yang balance in the kidneys, ensuring a source for the generation of yin essence, while also protecting the yang qi of the spleen and stomach. Anemarrhena asphodeloides clears heat and purges fire, nourishes yin and moistens dryness. When combined with Codonopsis pilosula, it prevents excessive heat and dryness from damaging body fluids. When combined with Codonopsis pilosula and Paeonia lactiflora, its heat-clearing and fire-purging properties alleviate tidal fever, calm the mind and relieve irritability. Combined with the tonifying and astringent effects of Codonopsis pilosula and the yin-nourishing effects of Paeonia lactiflora, it prevents the leakage of body fluids as sweat. Citrus aurantium is used as the primary herb to promote qi circulation, soothe the liver and regulate qi, and smooth the flow of qi. When combined with Paeonia lactiflora, one disperses and the other astringes, it alleviates liver qi stagnation caused by emotional anxiety. The combined effects of these herbs nourish the liver and kidneys, calm the mind and soothe the spirit, regulate the liver and strengthen the spleen, thereby calming the mind, nourishing vital energy, balancing yin and yang, harmonizing qi and blood, clearing the meridians, and coordinating the functions of the internal organs.

[0027] A systematic validation of this traditional Chinese medicine compound was conducted through multidimensional pharmacodynamic studies. The results showed that the compound can inhibit ovarian granulosa cell apoptosis, improve ovarian function, and delay the decline of ovarian function during menopause. Simultaneously, it significantly increases serum estradiol (E2) levels, decreases follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels, and increases the levels of neurotransmitters serotonin (5-HT) and β-endorphin (β-EP), exhibiting a benign regulatory effect on menopausal endocrine disorders and neurotransmitter imbalances. Furthermore, the compound can effectively improve sleep quality in animals and alleviate depressive and anxiety-like behaviors. In summary, this compound demonstrates significant efficacy in multiple aspects, providing not only modern pharmacological experimental evidence for the treatment of menopausal syndrome but also a new reference direction for optimizing clinical medication strategies. Attached Figure Description

[0028] Figure 1 The following data are presented in an aging mouse model: ovarian granulosa cell apoptosis (A), number of growing follicles (B), number of mature follicles (C), number of corpora lutea (D), and ovarian index (E). * P<0.05, ** P<0.01.

[0029] Figure 2 The levels of neurotransmitters 5-HT and β-EP (AB), sleep status (C), and sex hormones E2, FSH, and LH (DF) in an ovariectomized mouse model were compared. * P<0.05, ** P<0.01.

[0030] Figure 3 These are the results of the open field experiment in an ovariectomized mouse model. * P<0.05, ** P<0.01. Detailed Implementation

[0031] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0032] Example 1

[0033] A traditional Chinese medicine compound for menopausal syndrome is made from the following ingredients by weight: 10 parts of Codonopsis pilosula, 10g of Gynostemma pentaphyllum, 8g of longan pulp, 5g of Codonopsis pilosula, 5g of Paeonia lactiflora, 10g of Smilax glabra, 6g of Trichosanthes kirilowii, 2g of Anemarrhena asphodeloides, 0.5g of Syzygium aromaticum, and 1g of Citrus reticulata.

[0034] Example 2

[0035] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine compound is made from the following raw materials in the indicated weights: 36g of Codonopsis pilosula, 36g of Gynostemma pentaphyllum, 30g of Longan pulp, 25g of Codonopsis pilosula, 30g of Paeonia lactiflora, 40g of Smilax glabra, 24g of Trichosanthes kirilowii, 25g of Anemarrhena asphodeloides, 6g of Syzygium aromaticum, and 18g of Citrus reticulata.

[0036] Example 3

[0037] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine compound is made from the following raw materials in the indicated weights: 15g of Codonopsis pilosula, 15g of Gynostemma pentaphyllum, 12g of Longan pulp, 10g of Codonopsis pilosula, 10g of Paeonia lactiflora, 15g of Smilax glabra, 10g of Trichosanthes kirilowii, 6g of Anemarrhena asphodeloides, 1g of Syzygium aromaticum, and 2g of Citrus reticulata.

[0038] Example 4

[0039] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine compound is made from the following raw materials in the indicated weights: 30g of Codonopsis pilosula, 30g of Gynostemma pentaphyllum, 28g of Longan pulp, 20g of Codonopsis pilosula, 15g of Paeonia lactiflora, 30g of Smilax glabra, 15g of Trichosanthes kirilowii, 18g of Anemarrhena asphodeloides, 3g of Syzygium aromaticum, and 10g of Citrus reticulata.

[0040] Example 5

[0041] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine compound is made from the following raw materials in the indicated weights: 25g of Codonopsis pilosula, 25g of Gynostemma pentaphyllum, 25g of Longan pulp, 16g of Codonopsis pilosula, 16g of Paeonia lactiflora, 12g of Smilax glabra, 12g of Trichosanthes kirilowii, 12g of Anemarrhena asphodeloides, 1.5g of Syzygium aromaticum, and 6g of Citrus reticulata.

[0042] Example 6

[0043] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine compound is made from the following raw materials in the indicated weights: 28g of Codonopsis pilosula, 28g of Gynostemma pentaphyllum, 26g of Longan pulp, 18g of Codonopsis pilosula, 20g of Paeonia lactiflora, 35g of Smilax glabra, 18g of Trichosanthes kirilowii, 16g of Anemarrhena asphodeloides, 5g of Syzygium aromaticum, and 12g of Citrus reticulata.

[0044] Example 7

[0045] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine compound is made from the following raw materials in the indicated weights: 34g of Codonopsis pilosula, 32g of Gynostemma pentaphyllum, 10g of longan pulp, 23g of Codonopsis pilosula, 25g of Paeonia lactiflora, 38g of Smilax glabra, 22g of Trichosanthes kirilowii, 21g of Anemarrhena asphodeloides, 2g of Syzygium aromaticum, and 16g of Citrus reticulata.

[0046] Example 8

[0047] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine compound is made from the following raw materials in the indicated weights: 12g of Codonopsis pilosula, 12g of Gynostemma pentaphyllum, 18g of Longan pulp, 8g of Codonopsis pilosula, 28g of Paeonia lactiflora, 20g of Smilax glabra, 8g of Trichosanthes kirilowii, 10g of Anemarrhena asphodeloides, 4g of Syzygium aromaticum, and 8g of Citrus reticulata.

[0048] Example 9

[0049] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine compound is made from the following raw materials in the indicated weights: 20g of Codonopsis pilosula, 20g of Gynostemma pentaphyllum, 20g of Longan pulp, 12g of Codonopsis pilosula, 18g of Paeonia lactiflora, 25g of Smilax glabra, 14g of Trichosanthes kirilowii, 16g of Anemarrhena asphodeloides, 2g of Syzygium aromaticum, and 11g of Citrus reticulata.

[0050] Experiment 1

[0051] 1.1 Laboratory Animals

[0052] Female KM mice aged 12-13 months were selected as the aged mouse group. Mice that showed no estrous cycle were screened by vaginal exfoliated cells and then enrolled in subsequent experiments.

[0053] Meanwhile, 8-10 week old KM mice were selected as the young control group. Mice exhibiting a regular 4-5 day estrous cycle were enrolled for subsequent experiments after screening by vaginal exfoliative cells.

[0054] 1.2 Experimental Drugs

[0055] Traditional Chinese medicine compound (ZYFF): 25g of Codonopsis pilosula, 25g of Gynostemma pentaphyllum, 25g of Longan pulp, 16g of Codonopsis pilosula, 16g of Paeonia lactiflora, 12g of Smilax glabra, 12g of Trichosanthes kirilowii, 12g of Anemarrhena asphodeloides, 1.5g of Syzygium aromaticum, and 6g of Citrus reticulata.

[0056] Comparative Example 1 (ZY-1): Codonopsis pilosula 25g, Rehmannia glutinosa 25g, Longan aril 25g, Codonopsis pilosula 16g, Paeonia lactiflora 16g, Smilax glabra 12g, Trichosanthes kirilowii 12g, Anemarrhena asphodeloides 12g, Syzygium aromaticum 1.5g, Citrus reticulata 6g.

[0057] Comparative Example 2 (ZY-2): 25g of *Hedyotis diffusa*, 16g of *Codonopsis pilosula*, 16g of *Paeonia lactiflora*, 12g of *Smilax glabra*, 12g of *Trichosanthes kirilowii*, 12g of *Anemarrhena asphodeloides*, 1.5g of *Citrus aurantium*, and 6g of *Citrus reticulata*.

[0058] Take the medicinal materials in the above prescription proportions, crush and mix them, add water to decoct and extract, filter and concentrate to a concentrated solution, so that the crude drug content is 5g / mL, and then package it for later experimental testing.

[0059] Kunbao Pills (KBW): Purchased from Beijing Tongrentang Co., Ltd.

[0060] Estradiol Valerate Tablets (EV): Purchased from Bayer Healthcare Co., Ltd., Guangzhou Branch

[0061] 1.3 Grouping and Dosing

[0062] Aged mice were randomly divided into 6 groups: a model group (menopausal group), a traditional Chinese medicine compound group, a control group 1, a control group 2, a Kunbao pill group, and an estradiol valerate tablet group, with 5 mice in each group. A young control group was also included. The traditional Chinese medicine compound group, control group 1, control group 2, the Kunbao pill group, and the estradiol valerate tablet group were administered the medication by gavage at a dose of 23 g / kg once daily; the model group and the young control group were given an equal volume of physiological saline. After 4 weeks of continuous administration, the animals were sacrificed and samples were collected for subsequent analysis of various indicators.

[0063] 1.4 Detection Indicators

[0064] 1.4.1 Detection of ovarian granulosa cell apoptosis

[0065] Ovarian tissue was collected, fixed, dehydrated in a gradient, embedded in paraffin, and then serially sectioned. Staining was performed according to the TUNEL kit instructions. After staining, the tissue was observed under a microscope, and the positive cells were semi-quantitatively analyzed using ImageJ software.

[0066] 1.4.2 Ovarian function status

[0067] Bilateral ovarian tissue was harvested, weighed using an analytical balance to calculate organ indices, and then fixed with tissue fixative. Subsequent procedures included paraffin embedding and HE staining. Microscopic examination was performed to observe the number of follicles at various stages and the number of corpora lutea in the ovaries.

[0068] 1.5 Statistical Analysis

[0069] All experimental data were analyzed using GraphPad Prism 8.0 statistical software, and one-way ANOVA was used for comparisons between groups. A p-value < 0.05 was considered statistically significant, and a p-value < 0.01 was considered extremely statistically significant.

[0070] 1.6 Results

[0071] 1.6.1 Effects of each group on ovarian granulosa cell apoptosis

[0072] Compared with the young control group, the number of apoptotic granulosa cells in the ovaries of aged mice was significantly increased (P<0.01); after intervention with traditional Chinese medicine compound, the number of apoptotic granulosa cells in the ovaries of mice was significantly reduced (P<0.01), suggesting that this traditional Chinese medicine compound can inhibit excessive apoptosis of granulosa cells in the ovaries of aged mice, delay the aging process of the ovaries, and thus improve ovarian function.

[0073] 1.6.2 Effects of each group on ovarian function

[0074] Compared with the young control group, the ovarian function of aged mice was significantly reduced in terms of growth, number of mature follicles, number of corpora lutea, and ovarian index (P<0.01), indicating that ovarian function in aged mice without estrous cycles declines and ovarian atrophy occurs, similar to the symptoms of increased follicular atresia and reduced follicular number during menopause. After intervention in each treatment group, this traditional Chinese medicine compound significantly increased the number of growing follicles, number of mature follicles, number of corpora lutea, and ovarian index (P<0.01), suggesting that this traditional Chinese medicine compound can increase the number of follicles at all stages in the ovaries of aged mice, inhibit excessive follicular depletion during menopause, increase follicular reserve, and restore ovarian function.

[0075] Experiment 2

[0076] 1.1 Laboratory Animals

[0077] The rat castration model is one of the most commonly used models in menopausal syndrome research. This study aims to observe the efficacy of this traditional Chinese medicine compound based on this model. Fifty-six SPF-grade male SD rats, weighing 180-220g, were selected and acclimatized for one week before the experiment.

[0078] 1.2 Experimental Drugs

[0079] Traditional Chinese medicine compound (ZYFF): 25g of Codonopsis pilosula, 25g of Gynostemma pentaphyllum, 25g of Longan pulp, 16g of Codonopsis pilosula, 16g of Paeonia lactiflora, 12g of Smilax glabra, 12g of Trichosanthes kirilowii, 12g of Anemarrhena asphodeloides, 1.5g of Syzygium aromaticum, and 6g of Citrus reticulata.

[0080] Comparative Example 1 (ZY-1): Codonopsis pilosula 25g, Rehmannia glutinosa 25g, Longan aril 25g, Codonopsis pilosula 16g, Paeonia lactiflora 16g, Smilax glabra 12g, Trichosanthes kirilowii 12g, Anemarrhena asphodeloides 12g, Syzygium aromaticum 1.5g, Citrus reticulata 6g.

[0081] Comparative Example 2 (ZY-2): 25g of *Hedyotis diffusa*, 16g of *Codonopsis pilosula*, 16g of *Paeonia lactiflora*, 12g of *Smilax glabra*, 12g of *Trichosanthes kirilowii*, 12g of *Anemarrhena asphodeloides*, 1.5g of *Citrus aurantium*, and 6g of *Citrus reticulata*.

[0082] The above-mentioned medicinal materials were weighed according to the prescription ratio, crushed and mixed to make a concentrated decoction containing 5g / ml of raw medicinal materials, which was used for subsequent experiments.

[0083] Kunbao Pills (KBW): Purchased from Beijing Tongrentang Co., Ltd.

[0084] Estradiol Valerate Tablets (EV): Purchased from Bayer Healthcare Co., Ltd., Guangzhou Branch

[0085] 1.3 Modeling Methods and Grouping

[0086] Fifty-six female SD rats were randomly divided into seven groups: sham-operated group, model group, traditional Chinese medicine compound group, comparative example 1 group, comparative example 2 group, Kunbao pill group, and estradiol valerate tablet group, with eight rats in each group. All rats were anesthetized and fixed in a prone position. The skin was shaved at the costovertebral angle on the back, and routine disinfection was performed. A 1 cm incision was made along the costovertebral angle to expose the fat pad, which was gently pulled out. The pink or yellowish-red ovary was located within the fat pad, the fallopian tube was ligated, and the ovary was completely removed. The fat and uterine horn were then returned to the abdominal cavity. The contralateral ovary was removed using the same method. In the sham-operated group, only the back muscles were incised; the ovary was not removed. Postoperatively, penicillin sodium was injected intramuscularly for three consecutive days to prevent infection. Vaginal smears were examined for five consecutive days postoperatively, with estrous cycle irregularities used as the criterion for successful model establishment.

[0087] After successful model establishment, each treatment group (traditional Chinese medicine compound group, comparative example 1 group, comparative example 2 group, Kunbao pill group, and estradiol valerate tablet group) was administered the corresponding drug by gavage at a dose of 16 g / kg once daily for 30 consecutive days; the sham surgery group and the model group were given an equal volume of physiological saline. After the drug administration was completed, subsequent indicator tests were performed.

[0088] 1.4 Detection Indicators

[0089] 1.4.1 Serum levels of related hormones and neurotransmitters in each group of rats

[0090] Serum estradiol (E2), follicle-stimulating hormone (FSH), luteinizing hormone (LH), serotonin (5-HT), and β-endorphin (β-EP) levels were detected by ELISA.

[0091] 1.4.2 Central Open Field Experiment

[0092] Thirty minutes after the last dose, Xiao Hu was placed in an open field box. After acclimatization, the number of activities within 5 minutes was measured and the activity route was recorded.

[0093] 1.4.3 Effects of sodium pentobarbital on sleep disorders

[0094] Thirty minutes after the last administration, mice in each group were injected intraperitoneally with 30 mg / kg sodium pentobarbital. The number of animals whose righting reflex disappeared for more than 1 minute after 15 minutes was observed and recorded.

[0095] 1.5 Statistical Analysis

[0096] All experimental data were analyzed using GraphPad Prism 8.0 statistical software, and one-way ANOVA was used for comparisons between groups. A p-value < 0.05 was considered statistically significant, and a p-value < 0.01 was considered extremely statistically significant.

[0097] 2 Results

[0098] 2.1 Serum levels of related hormones and neurotransmitters in each group of rats

[0099] Hormonal imbalances primarily stem from ovarian dysfunction. During menopause, declining ovarian function and depleted follicular reserves lead to a sharp decrease in E2 and inhibin secretion, weakening the negative feedback inhibition on the pituitary gland and resulting in compensatory increases in FSH and LH. Therefore, decreased serum E2 levels and increased FSH and LH levels are key endocrine markers reflecting ovarian dysfunction and hypothalamic-pituitary-ovarian axis imbalance. Experimental results showed that compared to the sham-operated group, the model group rats exhibited significantly lower serum E2 levels (P<0.01) and significantly higher LH and FSH levels (P<0.01), indicating a successful establishment of the menopausal model. After intervention with various drugs, E2 levels in all groups of rats recovered to varying degrees, while LH and FSH levels significantly decreased. The traditional Chinese medicine compound showed the best effect, suggesting that it can effectively regulate the endocrine disorder in menopausal rats, delay ovarian aging, and improve ovarian function, thereby achieving the goal of preventing and treating menopausal syndrome.

[0100] The decline in estrogen levels accompanying ovarian dysfunction can further disrupt the neurotransmitter system, leading to typical menopausal symptoms such as hot flashes, insomnia, irritability, and anxiety. Therefore, this study measured neurotransmitter-related indicators. Results showed that compared to the sham-operated group, the serum levels of 5-hydroxytryptamine (5-HT) and β-endorphin (β-EP) in the model group rats were significantly reduced. Compared to the model group, all treatment groups increased the levels of these neurotransmitters, with the traditional Chinese medicine compound group showing the most significant improvement. These results suggest that this traditional Chinese medicine compound can regulate disordered neurotransmitter levels to a relatively stable state, thereby stabilizing hypothalamic function and effectively preventing and alleviating menopausal symptoms such as hot flashes, depression, and anxiety.

[0101] 2.2 Behavioral results of spontaneous activity ability of rats in each group

[0102] Analysis of activity trajectories and dwell time in the central region during the open field test showed that, compared with the sham-operated group, rats in the model group were more inclined to move in the peripheral region, with significantly fewer entries into the central region and shorter dwell times, suggesting that the model group rats exhibited menopausal-related depressive or anxiety-like behaviors. All treatment groups improved the aforementioned behavioral abnormalities to some extent, but the traditional Chinese medicine compound group showed the most significant improvement in increasing the number of entries into the central region and prolonging the dwell time. Compared with the model group, the difference in dwell time in the central region in the traditional Chinese medicine compound group was statistically significant (P<0.01), and superior to other treatment groups.

[0103] 2.3 Effects of sodium pentobarbital on sleep disorders

[0104] In a sleep experiment induced by a subthreshold dose of sodium pentobarbital, mice in the sham-operated group exhibited normal sleep responses, while mice in the model group failed to enter a sleep state at all, suggesting sleep dysfunction. After drug intervention, the sleep onset rate of mice in both the traditional Chinese medicine compound group and the estradiol valerate group was significantly improved, with the traditional Chinese medicine compound group showing the best effect and demonstrating a superior sleep-promoting effect.

[0105] 3. Discussion

[0106] The oophorectomy model, lacking estrogen-secreting organs, leads to a significant decrease in serum sex hormone levels and brain serotonin (5-HT) content, with changes closely matching laboratory indicators of human menopausal syndrome. However, this model cannot directly observe the degenerative changes in ovarian tissue itself, and ovarian aging is the root cause of a series of physiological and pathological changes during menopause. While the natural aging model retains some residual ovarian function, it is prone to co-occurring with other age-related diseases. To overcome the limitations of a single model, this study simultaneously used both models for pharmacodynamic evaluation to ensure the comprehensiveness and reliability of experimental data.

[0107] Based on the pathogenesis and core clinical symptoms of menopausal syndrome, this study conducted a multidimensional pharmacodynamic investigation of a traditional Chinese medicine compound. Using Kunbao pills and estradiol valerate tablets as positive controls, the effects of the compound on sex hormone levels, neurotransmitter content, insomnia symptoms, and emotional abnormalities in menopausal rats were systematically observed. Results showed that the compound significantly reduced serum follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels in model rats and effectively reversed the low estradiol (E2) levels. In the evaluation of insomnia symptoms, the sleep onset rate was significantly improved in the compound group after administration of a subthreshold dose of sodium pentobarbital, suggesting a certain sleep-aiding effect. Open field test results showed that after intervention with the compound, the number of times rats entered the central region and the duration of their stay were significantly increased, and depressive or anxiety-like behaviors were significantly improved.

[0108] In summary, this study comprehensively evaluated the pharmacodynamics of a traditional Chinese medicine compound for treating menopausal syndrome from multiple perspectives, including hormone levels, neurotransmitter content, behavioral manifestations, and ovarian function. The results showed that the compound was significantly effective in multiple aspects, providing not only modern pharmacological evidence for the treatment of menopausal syndrome in women but also new reference directions for optimizing clinical medication strategies.

[0109] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A traditional Chinese medicine compound for menopausal syndrome, characterized in that, The traditional Chinese medicine compound is made from the following herbal raw materials in the indicated weight proportions: 10-36 parts of Codonopsis pilosula, 10-36 parts of Gynostemma pentaphyllum, 8-30 parts of Longan pulp, 5-25 parts of Codonopsis pilosula, 5-30 parts of Paeonia lactiflora, 10-40 parts of Smilax glabra, 6-24 parts of Trichosanthes kirilowii, 2-25 parts of Anemarrhena asphodeloides, 0.5-6 parts of Syzygium aromaticum, and 1-18 parts of Citrus reticulata.

2. The traditional Chinese medicine compound according to claim 1, characterized in that, The traditional Chinese medicine compound is made from the following herbal raw materials in the indicated weight proportions: 15-30 parts of Codonopsis pilosula, 15-30 parts of Gynostemma pentaphyllum, 12-28 parts of Longan pulp, 10-20 parts of Codonopsis pilosula, 10-15 parts of Paeonia lactiflora, 15-30 parts of Smilax glabra, 10-15 parts of Trichosanthes kirilowii, 6-18 parts of Anemarrhena asphodeloides, 1-3 parts of Syzygium aromaticum, and 2-10 parts of Citrus reticulata.

3. The traditional Chinese medicine compound according to claim 1, characterized in that, The traditional Chinese medicine compound is made from the following herbal raw materials in the indicated weight proportions: 25 parts of Codonopsis pilosula, 25 parts of Gynostemma pentaphyllum, 25 parts of longan pulp, 16 parts of Codonopsis pilosula, 16 parts of Paeonia lactiflora, 12 parts of Smilax glabra, 12 parts of Trichosanthes kirilowii, 12 parts of Anemarrhena asphodeloides, 1.5 parts of Syzygium aromaticum, and 6 parts of Citrus reticulata.

4. The application of the traditional Chinese medicine compound according to any one of claims 1 to 3 in the preparation of a drug for menopausal syndrome, characterized in that, The medicine is made from the above-mentioned proportions of Chinese herbal medicines by weight, supplemented with pharmaceutically acceptable excipients to form an oral dosage form, which may be any one of decoction, granules, capsules, or concentrated pills.

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