A traditional Chinese medicine composition for treating endometriosis, a preparation method and application thereof

Abstract

The application discloses a traditional Chinese medicine formula for treating endometriosis, which comprises the following components in the weight fractions: cassia twig 14-16 parts, five spirit fat 8-12 parts, raw safflower 4-8 parts, xiaosha 4-8 parts, blood 4-8 parts, leech 2-5 parts, white peony root 8-12 parts, ophiopogon 8-12 parts, uncaria 12-14 parts, amber 4-8 parts and licorice 4-8 parts. The application is innovatively combined with the combination of heart calming and liver regulating on the basis of the original Pingchongjiangni formula, that is, warming heart yang to calm the rushing qi, calming heart qi to lower the rushing qi, and calming liver qi to follow the rushing qi, so that the purpose of calming the rushing qi is achieved. The traditional Chinese medicine formula can not only effectively relieve the dysmenorrhea of the patients with endometriosis, but also improve the psychological state such as depression and anxiety, and still has certain curative effect after stopping the medicine for three months.

Description

A traditional Chinese medicine formula for treating endometriosis, its preparation method and application Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, specifically to a traditional Chinese medicine formula for treating endometriosis, its preparation method, and its application. Background Technology

[0002] Endometriosis (EMs) is a condition in which endometrial tissue (glands and stroma) appears, grows, infiltrates, and repeatedly bleeds outside the uterus, lining the uterine cavity, leading to pain, nodules, or masses, and even infertility. Endometriosis affects up to 10% of women of reproductive age worldwide. Endometriosis lesions are widespread, diverse in morphology, highly invasive, and recurrent, and are hormone-dependent. Clinical cases related to endometriosis in Western medicine can be traced back to the 17th and 18th centuries. Zhao Ruihua et al. summarized a review of research on traditional Chinese medicine (TCM) treatment of endometriosis (Zhao Ruihua, A Review of Research on Traditional Chinese Medicine Treatment of Endometriosis, Journal of Beijing University of Traditional Chinese Medicine, September 2023, Vol. 46, No. 9), summarizing the current research status of TCM in the nomenclature, etiology, pathogenesis, symptoms, syndromes, and prescriptions of endometriosis. Currently, the pathogenesis of endometriosis is complex, making it difficult to reach a unified understanding.

[0003] Based on the pathogenesis of endometriosis (EMT) characterized by upward reversal of Qi and internal obstruction of blood stasis, and combined with the general treatment principles of "suppressing the reversal" and "suppressing the high" in the *Neijing* (Inner Canon of Medicine), Professor Liang Ruining proposed a treatment method of "suppressing upward reversal, resolving stasis and unblocking the collaterals," which has been well applied in clinical practice. The basic formula for suppressing upward reversal is: Cinnamon Twig 15g, Ophiopogon Root 10g, Leech 3g, Agarwood 6g, Dragon's Blood 3g, Five-Spiritus Dung 10g, and Typha Pollen 6g. Using this decoction in combination with oral mifepristone tablets to treat EMT can improve clinical symptoms, reduce recurrence rate, and increase pregnancy rate (Liu Yuling et al., Treatment of 30 Cases of Endometriosis Based on Traditional Chinese Medicine Suppressing Upward Reversal, Jiangxi Journal of Traditional Chinese Medicine, June 2021, Vol. 52, No. 462). Liu Yuling et al. further applied the Pingchong Jiangni acupoint patch combined with thermosensitive moxibustion to patients with endometriosis, which effectively improved patients' sex hormones and serum indicators while reducing adverse reactions (Liu Yuling, Application of Pingchong Jiangni Acupoint Patch Combined with Thermosensitive Moxibustion in Patients with Endometriosis, Journal of Rare Diseases, December 2023, Vol. 30, No. 12). Chen Danni further explored the effect and preliminary mechanism of Pingchong Jiangni Formula in intervening in pain in EMS model rats (Chen Danni, Jiangxi University of Traditional Chinese Medicine, Master's Thesis, 2023). The Pingchong Jiangni Formula used was: Cinnamon Twig 15g, Blood Dragon 6g, Five-spice Powder 10g, Raw Typha Pollen 6g, Dalbergia Incense 6g, Leech 3g, Ophiopogon japonicus 10g, White Peony Root 10g, Licorice Root 6g. This Pingchong Jiangni Formula can alleviate the pain caused by EMS to a certain extent. Its pain relief method may be through inhibiting the expression of NGF, TRPV1, and PKC. Summary of the Invention

[0004] Purpose of the invention: The technical problem to be solved by the present invention is to provide an improved formula for relieving menstrual pain in patients with endometriosis, which can not only effectively relieve dysmenorrhea in patients with endometriosis, but also improve their psychological state such as depression and anxiety, effectively reduce the size of lesions and the level of inflammatory factors, and is not prone to recurrence after discontinuation of medication.

[0005] To address the aforementioned technical problems, this invention discloses a traditional Chinese medicine formula for treating endometriosis. The formula comprises the following components in parts by weight: 14-16 parts of cinnamon twig, 8-12 parts of five-spice powder, 4-8 parts of raw cattail pollen, 4-8 parts of sandalwood, 4-8 parts of dragon's blood, 2-5 parts of leech, 8-12 parts of white peony root, 8-12 parts of ophiopogon root, 12-14 parts of uncaria, 4-8 parts of amber, and 4-8 parts of licorice root.

[0006] Preferably, the traditional Chinese medicine formula includes the following components in parts by weight: 15 parts cinnamon twig, 10 parts five-spice powder, 6 parts raw cattail pollen, 6 parts sandalwood, 6 parts dragon's blood, 3 parts leech, 10 parts white peony root, 10 parts ophiopogon root, 10 parts uncaria, 6 parts amber, and 6 parts licorice root.

[0007] The present application further provides the use of the above traditional Chinese medicine formula in the preparation of a medicament or preparation for treating endometriosis.

[0008] Specifically, the traditional Chinese medicine formula can relieve pain, reduce the volume of lesions and the levels of inflammatory factors caused by endometriosis.

[0009] Among them, the dosage form of the preparation is any one of liquid preparations, granules, patches, ointments, pills, boluses, powders, and tablets.

[0010] In one embodiment, the dosage form of the preparation is a liquid preparation.

[0011] Specifically, the preparation method of the liquid preparation includes the following steps: Soak the formula amounts of cinnamon twig, weeping forsythia, pollen typhae, leech, white peony root, dwarf lilyturf tuber, amber, and licorice root in water and then decoct. For the first decoction, bring to a boil over high heat and then turn to low heat and simmer, filter out the medicinal juice; for the second decoction, bring to a boil over high heat and then turn to low heat and simmer, add the formula amounts of dalbergia odorifera and uncaria rhynchophylla, filter out the medicinal juice, mix the medicinal juices of the first and second decoctions, filter, concentrate, and add dragon's blood after grinding into powder.

[0012] Preferably, the amount of water added for the first decoction is 8 - 10 times the total mass of the raw materials, the amount of water added for the second decoction is 4 - 6 times the total mass of the raw materials, the first decoction is simmered over low heat for 20 - 30 minutes, the second decoction is simmered over low heat for 10 - 20 minutes, and after adding the formula amounts of dalbergia odorifera and uncaria rhynchophylla, simmer for 4 - 6 minutes.

[0013] In a preferred embodiment, the amount of water added for the first decoction is 10 times the total mass of the raw materials, the amount of water added for the second decoction is 5 times the total mass of the raw materials, the first decoction is simmered over low heat for 20 minutes, the second decoction is simmered over low heat for 15 minutes, and after adding the formula amounts of dalbergia odorifera and uncaria rhynchophylla, simmer for 5 minutes.

[0014] Preferably, the medicinal materials are soaked for 20 - 40 minutes before the first decoction.

[0015] The traditional Chinese medicine formula of the present application takes calming the adverse qi and resolving stasis and dredging collaterals as the core. Cinnamon twig is used heavily to calm the adverse qi as the monarch drug; weeping forsythia and pollen typhae resolve stasis and dredge collaterals as the ministerial drugs; dalbergia odorifera regulates qi and relieves pain and resolves stasis, dragon's blood and leech dispel stasis and dredge collaterals. These three drugs together assist the monarch and ministerial drugs to strengthen the effect of calming the adverse qi and dredging collaterals. White peony root nourishes blood and harmonizes nutrient qi, dwarf lilyturf tuber nourishes yin and moistens dryness. The two drugs used together not only prevent the blood - activating drugs from damaging yin and blood but also restrict the warm - dryness of cinnamon twig. Uncaria rhynchophylla soothes the liver and clears heat, amber tranquilizes the mind and calms the nerves. The two drugs used together not only calm the adverse qi but also clear the heart and calm the mind. These four drugs are all assistant drugs, and licorice root harmonizes all the drugs as the envoy drug. All the drugs are used together, combining cold and heat, combining rigidity and softness, warm but not dry, tonifying but not stagnating, dispelling stasis without damaging yin and blood, and jointly achieving the effects of calming the adverse qi, resolving stasis and dredging collaterals.

[0016] This application proposes an innovative theory of menstrual physiology, which states that menstruation is formed by the orderly infusion of Qi and blood from the Chong meridian into the uterus. The orderly infusion of Qi and blood from the Chong meridian into the uterus is coordinated by multiple organs. Kidney Qi absorbs Qi and blood from the Chong meridian and flows downward, heart Qi descends and helps the Qi and blood from the Chong meridian to flow downward, and liver Qi regulates the orderly infusion of Qi and blood from the Chong meridian.

[0017] Endometriosis is characterized by dysmenorrhea as its main symptom. We are the first to propose that its core pathogenesis is kidney deficiency leading to the upward flow of Qi and blood in the Chong meridian, possibly due to liver Qi stagnation or direct exposure to cold pathogens, which obstruct the flow of Qi and blood from the Chong meridian to the uterus, further exacerbating the disordered flow of Qi and blood in the meridians. Since the Qi and blood in the Chong meridian are particularly abundant during menstruation, lower abdominal pain is severe. This is the so-called "Chong meridian disease, rebellious Qi and abdominal urgency." Therefore, we propose that the main treatment principle for dysmenorrhea in endometriosis is to balance the Chong meridian.

[0018] This application innovatively combines the original Pingchong Jiangni formula with a combination of calming the mind and regulating the liver. Specifically, it warms the heart yang to calm the Chong qi, calms the heart qi to lower the Chong qi, and regulates the liver qi to smooth the Chong qi, achieving a harmonious balance to pacify the Chong qi. The new Pingchong Jiangni formula is the original formula with the addition of Uncaria rhynchophylla and Amber. Uncaria rhynchophylla enters the liver meridian, clearing liver heat and calming liver yang, thus promoting the smooth flow of liver qi and aiding in the smooth circulation of qi and blood in the Chong meridian. Amber enters the heart and liver meridians, calming the mind and relieving anxiety, allowing the heart qi to descend and suppress the rebellious Chong qi. By optimizing the dosage and using a new combination of Cinnamomum cassia, Uncaria rhynchophylla, and Amber, it not only warms the heart yang to calm the Chong qi, calms the heart qi to lower the Chong qi, and regulates the liver qi to smooth the Chong qi, but also further facilitates the smoothing of the rebellious Chong qi. Furthermore, it has the effect of calming the mind and regulating the will. Clinical studies have shown that the new formula not only effectively relieves dysmenorrhea in patients with endometriosis but also improves their psychological state, such as depression and anxiety, and retains some efficacy even three months after discontinuation of medication. Attached Figure Description

[0019] Figure 1 shows the comparison of PWT values ​​in rats before surgery, before drug administration, and 3 weeks after drug administration.

[0020] Figure 2 shows the comparison of ectopic lesion volume in rats with endometriosis after 3 weeks of drug administration.

[0021] Figure 3 shows the concentrations of TNF-α and IL-β in the serum of rats (x ± s).

[0022] Figure 4 shows the comparison of PWT (g) values ​​in rats before surgery, before drug administration, and 3 weeks after drug administration.

[0023] Figure 5 shows the comparison of ectopic lesion volume in rats with endometriosis after 3 weeks of drug administration.

[0024] Figure 6 shows the concentrations of TNF-α and IL-β in rat serum.

[0025] Figure 7 shows the comparison of PWT (g) values ​​in rats before surgery, before drug administration, and 3 weeks after drug administration.

[0026] Figure 8 shows the comparison of ectopic lesion volume in rats with endometriosis after 3 weeks of drug administration.

[0027] Figure 9 shows the concentrations of TNF-α and IL-β in rat serum.

[0028] Figure 10 shows the VAS scores of patients before and after treatment and after a three-month follow-up period.

[0029] Figure 11 shows the number of patients with different VAS scores;

[0030] Figure 12 shows the BDI scores of the patients before and after treatment and after a three-month follow-up period.

[0031] Figure 13 shows the number of patients with different BDI scores;

[0032] Figure 14 shows the BAI scores of patients before and after treatment and after a three-month follow-up period.

[0033] Figure 15 shows the number of patients with different BAI scores.

[0034] Figure 16 shows the POMS scores of the patients before and after treatment and after a three-month follow-up period;

[0035] In the above figures, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, and ns are considered statistically significant. Detailed Implementation

[0036] The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments, and the advantages of the present invention in the above and / or other aspects will become clearer.

[0037] Example 1: Preparation of the traditional Chinese medicine formula.

[0038] Formula: 15g cinnamon twig, 6g dragon's blood, 10g five-spice powder, 6g raw cattail pollen, 6g sandalwood, 3g leech, 10g ophiopogon japonicus, 10g white peony root, 6g licorice root, 10g uncaria rhizome, 6g amber.

[0039] Preparation method: Except for dragon's blood, dalbergia odorifera, and uncaria rhynchophylla, the above traditional Chinese medicines are soaked in water for 30 minutes, then added with 10 times the amount of water and soaked for 0.5 hours. After soaking, the water and medicinal materials are poured into a decocting pot. For the first decoction, bring to a boil over high heat, then turn to low heat and simmer for about 20 minutes, and filter out the medicinal juice. For the second decoction, add 5 times the amount of water, bring to a boil over high heat, then turn to low heat and simmer for 15 minutes, add the later-added traditional Chinese medicines dalbergia odorifera and uncaria rhynchophylla, and finally simmer for 5 minutes, filter out the medicinal juice, mix the medicinal juices of the first and second decoctions, filter, and finally simmer over low heat and concentrate to obtain the juice, add the powdered dragon's blood, and obtain the medicinal liquid.

[0040] Preparation of the traditional Chinese medicine formula in Example 2.

[0041] Formula: Cinnamon twig 14g, dragon's blood 5g, weeping forsythia 12g, pollen typhae 4g, dalbergia odorifera 8g, leech 2g, dwarf lilyturf tuber 8g, white peony root 12g, licorice root 4g, uncaria rhynchophylla 12g, amber 4g.

[0042] The preparation method is the same as that in Example 1.

[0043] Preparation of the traditional Chinese medicine formula in Example 3.

[0044] Formula: Cinnamon twig 16g, dragon's blood 8g, weeping forsythia 8g, pollen typhae 8g, dalbergia odorifera 4g, leech 5g, dwarf lilyturf tuber 12g, white peony root 8g, licorice root 8g, uncaria rhynchophylla 14g, amber 8g.

[0045] The preparation method is the same as that in Example 1.

[0046] Effect of the traditional Chinese medicine formula in Example 4 on mechanical pain threshold (PWT), lesion volume, and levels of inflammatory factors (TNF-α, IL-β) in rats.

[0047] (1) Experimental animals

[0048] Forty-eight female Sprague Dawley rats, 7 weeks old, weighing 180 - 200 g, were purchased from the animal facility of Jiangxi University of Traditional Chinese Medicine (Nanchang, China) [license number: SCXK (Gan) 2023 - 0001]. The feeding environment for these experimental animals was at a temperature of (23 ± 2) °C, relative humidity of 50% - 60%, a 12-hour day-night rhythm, and they had free access to food and water.

[0049] (2) Preparation of experimental drugs

[0050] Original formula drugs: Cinnamon twig 15g, dragon's blood 6g, weeping forsythia 10g, pollen typhae 6g, dalbergia odorifera 6g, leech 3g, dwarf lilyturf tuber 10g, white peony root 10g, licorice root 6g, prepared according to the preparation method in Example 1.

[0051] New formula drugs: The medicinal liquid obtained from the preparation in Example 1.

[0052] β-Estradiol (Beijing Solarbio Technology Co., Ltd., batch number: 107H025) should be used by mixing β-estradiol powder into soybean oil at a dosage of 1 mg / kg.

[0053] (3) Animal model establishment, grouping and administration

[0054] After one week of acclimatization feeding, eight rats were randomly selected as the sham-operated group. In the sham-operated group, only routine laparotomy and closure were performed. The remaining rats underwent autologous endometrial transplantation to establish a rat model of endometriosis. Postoperatively, estradiol (1 mg / kg) was administered subcutaneously twice weekly for three weeks to promote the growth of ectopic lesions. Three weeks postoperatively, the successfully modeled rats were randomly divided into a model group, a low-dose traditional Chinese medicine (TCM) group, a high-dose TCM group, a medium-dose TCM group, and the original formula group. The sham-operated group and the model group were administered physiological saline by gavage daily at a concentration of 10 ml / kg of body weight. The original formula group was administered the original formula at a concentration of 7.56 g / kg of raw herbs by gavage. The low, medium, and high-dose new formula groups were administered the new formula at concentrations of 2.31 g / kg, 4.62 g / kg, and 9.24 g / kg, respectively, for a total of three weeks.

[0055] The concentration setting is based on the weight of the raw medicinal materials. For example, taking the first prescription as an example, the weight of the raw medicinal materials is 88g. The standard weight of an adult female is 60kg. The conversion factor between adults and rats is 6.3. Assuming that one rat weighs 300g, then: the clinical dose converted to rat dose is: (88g ÷ 60kg) × 6.3 = 9.24g of raw medicinal materials / kg (this is the amount of raw medicinal materials required by one rat per day). Based on this calculation, the second prescription is administered by gavage at a concentration of 10.29g / kg, and the third prescription is administered by gavage at a concentration of 9.87g / kg.

[0056] The therapeutic effect of the original Pingchong Jiangni formula on rats with endometriosis has been confirmed. (Refer to Chen, D., Ren, Y., Jin, J., Liu, S., Zhan, X., Li, X., Liang, R., & Ding, Z. (2024). Pingchong Jiangni recipe through nerve growth factor / transient receptor potential vanilloid 1 signaling pathway to relieve pain in endometriosis model rats. Journal of ethnopharmacology, 318(Pt A), 116940. https: / / doi.org / 10.1016 / j.jep.2023.116940.) Experimental results showed that the original Pingchong Jiangni formula (i.e., the original drug) dosage (7.56 g / kg) had the best therapeutic effect. Therefore, from an economic and effectiveness perspective, the relevant experiments on the original formula of Pingchongfang at different concentrations do not need to be re-verified in this experiment. Only the groups using the same dosage as the model group and the groups using the new formula of Pingchongfang at different concentrations are compared, and the experimental results have a certain degree of reliability.

[0057] (4) Behavioral testing

[0058] One day before surgery, one day before drug administration, and three weeks after drug administration, rats in each group were placed in a wire test box for half an hour to adapt. The fiber probe of the measuring instrument was then inserted along the edge of the wire mesh into the rat's front paw, and force was applied evenly. When the rat exhibited behaviors such as leg retraction or escape, the pressure value displayed on the measuring instrument screen at this time was recorded (accuracy of 0.1g). This pressure value is the rat's mechanical pain threshold (PWT).

[0059] (5) Specimen collection

[0060] After administration, the animals were fasted for 12 hours. Then, the rats were injected intraperitoneally with 10% chloral hydrate solution (3 ml / 100 g). After anesthesia, blood was collected from the heart. The whole blood was centrifuged at 3000 rpm for 10 min. The upper serum layer was collected and the levels of inflammatory factors (TNF-α, IL-1β) in the serum were detected by ELISA. The lesions of the rats were measured to calculate the volume. The formula for calculating the lesion volume is V=(π / 6)*m*n*h (m is the maximum transverse diameter, n is the longitudinal diameter perpendicular to the transverse diameter, and h is the thickness).

[0061] (6) Statistical analysis

[0062] All data are expressed as mean ± standard deviation. Statistical analysis and plotting were performed on all data using GraphPad Prism software version 10.0 (GraphPad software Inc., La Jolla, USA). Normality and homogeneity of variance were tested. If normality and homogeneity of variance were found, one-way ANOVA and LSD intergroup comparisons were used. If variances were not homogeneous, Dunne's t-test was used. A p-value < 0.05 was considered statistically significant.

[0063] (7) Experimental Results

[0064] 1) Effects of the new formula and the original formula on the mechanical pain threshold in rats with endometriosis

[0065] The results are shown in Table 1 and Figure 1. Before surgery, there was no statistically significant difference in PWT among the groups (P>0.05). Three weeks after surgery (before drug administration), compared with the sham surgery group, PWT in all model groups decreased significantly (P<0.05), and there was no statistically significant difference in PWT between groups (P>0.05). Three weeks after drug administration, compared with the model group, PWT of both the original and new drug formulas was significantly increased (P<0.05). Compared with the original formula, the high-dose group of the new formula showed a more significant improvement in PWT values ​​in the endometriosis model rats, with a statistically significant difference (P<0.05). This indicates that both the new and original formulas effectively relieved pain in rats with endometriosis, and the new formula, based on the original formula, showed better efficacy in increasing PWT values ​​in the endometriosis model rats. The therapeutic effect of the new Pingchong formula is dose-dependent. The high-dose traditional Chinese medicine formula had the best therapeutic effect and was superior to the original Pingchong formula (7.56g / kg). The low and medium doses of the new Pingchong formula were not as effective as the original Pingchong formula, possibly because the dosage and concentration of the traditional Chinese medicine were too low to exert a significant therapeutic effect.

[0066] Table 1 Comparison of PWT values ​​in rats before surgery, before drug administration, and 3 weeks after drug administration.

[0067]

[0068] Note: Compared with the sham surgery group, #p<0.01; compared with the model group, **p<0.01,**P<0.01; compared with the original prescription group, △p<0.05; compared with the high-dose new prescription group, ◇◇p<0.01.

[0069] 2) Effects of the new formula and the original formula on the lesion volume of rats with endometriosis

[0070] The volume of ectopic lesions in rats in each group was measured after three weeks of drug administration. The results are shown in Table 2 and Figure 2. Compared with the model group, the original formula and the new formula significantly reduced the volume of lesions in rats, and the difference was statistically significant (p<0.05). Compared with the original formula, the high-dose group of the new formula had a more significant effect on the volume of lesions, and the difference was statistically significant (p<0.05). This indicates that the new formula and the original formula can inhibit the growth of ectopic lesions in rats with endometriosis. The new formula has a better therapeutic effect on the size of lesions than the original formula.

[0071] Table 2 Comparison of ectopic lesion volume in rats with endometriosis after 3 weeks of drug administration.

[0072]

[0073] Note: Compared with the model group, **p<0.01; compared with the original prescription group, △p<0.05; compared with the high-dose new prescription group, ◇◇p<0.01.

[0074] 3) Effects of the new formula and the original formula on the levels of inflammatory factors in rats with endometriosis

[0075] ELISA results showed that after three weeks of administration, the serum inflammatory factor levels in the model group were significantly higher than those in the sham-operated group (P<0.05); compared with the model group, the levels of inflammatory factors in rats treated with both the original and new formulas were significantly lower (P<0.05). Compared with the original formula, the high-dose group of the new formula had a more significant effect on inflammation in rats with endometriosis, and the difference was statistically significant (p<0.05), indicating that both the original and new formulas can inhibit the inflammatory level in rats with endometriosis, and the new formula has a better effect on reducing inflammation than the original formula, as shown in Table 3 and Figure 3.

[0076] Table 3. Concentrations of TNF-α and IL-β in rat serum

[0077]

[0078]

[0079] Note: Compared with the sham surgery group, #p<0.01; compared with the model group, **p<0.01; compared with the original prescription group, △p<0.05; compared with the high-dose new prescription group, ◇◇p<0.01.

[0080] Example 5: Effect of different dosage ratios of Uncaria rhynchophylla and amber on pain in rats with endometriosis.

[0081] (I) Experimental drug formulation:

[0082] (1) Prescription 1: Cinnamon twig 15g, blood dragon 6g, five-spice powder 10g, raw cattail pollen 6g, sandalwood 6g, leech 3g, ophiopogon japonicus 10g, white peony root 10g, licorice root 6g, uncaria rhizome 10g, amber 6g;

[0083] (2) Formula 2: Cinnamon twig 15g, blood dragon 6g, five-spice powder 10g, raw cattail pollen 6g, sandalwood 6g, leech 3g, ophiopogon japonicus 10g, white peony root 10g, licorice root 6g, uncaria rhizome 20g, amber 6g;

[0084] (3) Prescription 3: Cinnamon twig 15g, blood dragon 6g, five-spice powder 10g, raw cattail pollen 6g, sandalwood 6g, leech 3g, ophiopogon japonicus 10g, white peony root 10g, licorice root 6g, uncaria rhizome 10g, amber 12g.

[0085] The preparation method is the same as in Example 1.

[0086] (II) Animal model establishment, grouping, and drug administration

[0087] After one week of acclimatization, rats were used to establish a rat model of endometriosis via autologous endometrial transplantation. Post-surgery, estradiol (1 mg / kg) was administered subcutaneously twice weekly for three weeks to promote the growth of ectopic lesions. Three weeks post-surgery, successfully modeled rats were randomly divided into three groups: Group 1 (medication group 1), Group 2 (medication group 2), and Group 3 (medication group 3). Group 1 was administered via gavage at a concentration of 9.24 g / kg, Group 2 at 10.29 g / kg, and Group 3 at 9.87 g / kg, for a total of three weeks.

[0088] (III) Experimental Results

[0089] 1) Effects of different dosages of Uncaria rhynchophylla and amber on the mechanical pain threshold in rats with endometriosis

[0090] As shown in Table 4 and Figure 4, there were no statistically significant differences in PWT among the groups of rats before surgery and before drug administration (P>0.05). After 3 weeks of drug administration, the PWT of rats in each group increased significantly compared with that before drug administration, and the difference was statistically significant (p<0.05). Compared with groups 2 and 3, group 1 had a more significant effect on the PWT value of rats with endometriosis. This indicates that the combination of 10g of Uncaria rhynchophylla and 6g of amber has the best effect on relieving pain in rats with endometriosis.

[0091] Table 4 Comparison of PWT (g) values ​​in rats before surgery, before drug administration, and 3 weeks after drug administration.

[0092]

[0093] Note: Compared with Group 1 (prescription), **p<0.001.

[0094] 2) Effects of different dosages of Uncaria rhynchophylla and amber on the lesion volume in rats with endometriosis.

[0095] The volume of ectopic lesions in each group of rats was measured after three weeks of drug administration. The results are shown in Table 5 and Figure 5. Compared with the lesions in groups 2 and 3, the lesion volume in group 1 was the smallest, and the difference was statistically significant (p<0.05). This indicates that the combination of 10g of Uncaria rhynchophylla and 6g of amber has the best therapeutic effect on inhibiting the growth of ectopic lesions in rats with endometriosis.

[0096] Table 5 Comparison of ectopic lesion volume in rats with endometriosis after 3 weeks of drug administration.

[0097]

[0098] Note: Compared with group 1 (prescription), **p<0.01

[0099] 3) Effects of different dosages of Uncaria rhynchophylla and amber on the levels of inflammatory factors in rats with endometriosis.

[0100] As shown in Table 6 and Figure 6, the ELISA results showed that after three weeks of administration, the serum inflammatory factor levels in rats in Formula 1 were lower than those in Formula 2 and Formula 3, and the difference was statistically significant (p<0.05). This indicates that the combination of 10g of Uncaria rhynchophylla and 6g of amber has the best therapeutic effect on inhibiting the inflammatory level in rats with endometriosis.

[0101] Table 6. Concentrations of TNF-α and IL-β in rat serum

[0102]

[0103]

[0104] Note: Compared with the first group of prescriptions, **p<0.01.

[0105] Animal experiments have verified that both the original and new formulas can effectively increase the mechanical pain threshold, reduce the volume of ectopic lesions, and decrease the expression level of inflammatory factors in rats with endometriosis, thus effectively relieving pain caused by endometriosis. In terms of pulse-wheat tachycardia (PWT), lesion volume, and inflammation level, the new formula is more effective than the original formula; the combination of 10g of Uncaria rhynchophylla and 6g of amber in the new formula showed the best therapeutic effect on pain in rats with endometriosis.

[0106] Example 6: Effects of different Chinese herbal formulas on pain and other symptoms in rats with endometriosis.

[0107] (I) Experimental drug formulation:

[0108] (1) Prescription 1: Cinnamon twig 15g, Dragon's blood 6g, Five-spice powder 10g, Raw cattail pollen 6g, Dalbergia odorifera 6g, Leech 3g, Ophiopogon japonicus 10g, White peony root 10g, Licorice root 6g, Uncaria rhynchophylla 10g, Amber 6g

[0109] (2) Formula 4: Cinnamon twig 15g, Dragon's blood 6g, Five-spice powder 10g, Raw cattail pollen 6g, Dalbergia odorifera 6g, Leech 3g, Ophiopogon japonicus 10g, White peony root 10g, Licorice root 6g, Uncaria rhynchophylla 5g, Amber 3g

[0110] (3) Prescription 5: Cinnamon twig 15g, Dragon's blood 3g, Five-spice powder 5g, Raw cattail pollen 3g, Dalbergia odorifera 3g, Leech 1.5g, Ophiopogon japonicus 5g, White peony root 5g, Licorice root 3g, Uncaria rhynchophylla 10g, Amber 6g

[0111] The preparation method is the same as in Example 1.

[0112] (II) Animal model establishment, grouping, and drug administration

[0113] After one week of acclimatization, a rat model of endometriosis was established using autologous endometrial transplantation. Post-surgery, rats were subcutaneously injected with estradiol 1 mg / kg twice weekly for three weeks to promote the growth of ectopic lesions. Three weeks later, the successfully modeled rats were randomly divided into three groups: Group 1, Group 4, and Group 5. Group 1 received 9.24 g / kg of estradiol daily by gavage, Group 4 received 8.4 g / kg daily by gavage, and Group 5 received 6.25 g / kg daily by gavage for a total of three weeks.

[0114] The effects of different dosage ratios of amber, uncaria, and other drugs on the mechanical pain threshold, lesion volume, and in vivo validation factor levels in rats with endometriosis were investigated using behavioral testing, specimen collection, and statistical analysis methods as described in Example 4.

[0115] (III) Experimental Results

[0116] (1) The effect of different dosage ratios of amber, uncaria, and other drugs on the mechanical pain threshold in rats with endometriosis model, with the principal drug cinnamon twig remaining unchanged.

[0117] As shown in Table 7 and Figure 7, there were no statistically significant differences in PWT among the groups of rats before surgery and before drug administration (p>0.05); after 3 weeks of drug administration, the PWT of rats in each group increased significantly compared with that before drug administration, and the difference was statistically significant (p<0.05); compared with group 4, group 5 was more effective in increasing the PWT value of rats with endometriosis; compared with groups 4 and 5, group 1 had a more significant effect on the PWT value of rats with endometriosis. This indicates that the drug dosage ratio in group 1 has the best effect on relieving pain in rats with endometriosis.

[0118] Table 7 Comparison of PWT (g) values ​​in rats before surgery, before drug administration, and 3 weeks after drug administration.

[0119]

[0120] Note: Compared with Group 1 of the prescription, ****p<0.001.

[0121] (2) The effect of different dosage ratios of amber, uncaria, and other drugs on the lesion volume in rats with endometriosis model, with the principal drug cinnamon twig remaining unchanged.

[0122] As shown in Table 8 and Figure 8, after three weeks of drug administration, the volume of ectopic lesions in each group of rats was measured. It was found that the lesion volume in group 5 was significantly smaller than that in group 4. Compared with groups 4 and 5, the lesion volume in group 1 was the most significantly reduced. All of these results showed statistical differences (p<0.05), indicating that the drug dosage ratio in group 1 had the best effect on reducing the lesion volume in rats with endometriosis.

[0123] Table 8 Comparison of ectopic lesion volume in rats with endometriosis after 3 weeks of drug administration.

[0124]

[0125]

[0126] Note: Compared with group 1 of the prescription, ****p<0.0001

[0127] (3) The effect of different dosage ratios of amber, uncaria, and other drugs on the levels of inflammatory factors in rats with endometriosis model, with the principal drug cinnamon twig remaining unchanged.

[0128] As shown in Table 9 and Figure 9, the ELISA results indicated that after three weeks of administration, compared with group 4, the serum concentrations of inflammatory factors TNF-α and IL-β in rats of group 5 were decreased; compared with groups 4 and 5, the serum expression levels of inflammatory factors TNF-α and IL-β in rats of group 1 were significantly reduced, and the differences were statistically significant (p<0.05). This suggests that the drug dosage ratio in group 1 had the best effect on inhibiting the expression levels of inflammatory factors in rats with endometriosis.

[0129] Table 9. Concentrations of TNF-α and IL-β in rat serum

[0130]

[0131] Note: Compared with Group 1 (prescription), *p<0.05, ***p<0.001, ****p<0.0001.

[0132] Conclusion: Animal experimental results showed that compared with group 5, group 4 improved the PWT value, lesion volume, and inflammation level in endometriosis model rats; compared with groups 4 and 5, group 1 showed the most significant improvement in PWT value, lesion volume, and inflammation level. With the principal herb cinnamon twig unchanged, although different dosage ratios of amber and uncaria with other drugs in groups 1, 4, and 5 effectively increased the mechanical pain threshold, reduced the volume of ectopic lesions, decreased the expression level of inflammatory factors, and effectively alleviated pain caused by endometriosis in rats, the dosage ratio of amber and uncaria with other drugs in group 1 showed the most significant therapeutic effect on pain in rats with endometriosis.

[0133] Example 7: The effects of traditional Chinese medicine formulation on dysmenorrhea, depression, and anxiety in patients with endometriosis.

[0134] (I) Inclusion Criteria

[0135] (1) Diagnostic criteria

[0136] The diagnosis was made in accordance with the "Guidelines for the Diagnosis and Treatment of Endometriosis" (Chinese Medical Association Obstetrics and Gynecology Branch Endometriosis Collaborative Group. Guidelines for the Diagnosis and Treatment of Endometriosis [J]. Chinese Journal of Obstetrics and Gynecology, 2015, 50(3): 161-169. DOI:10.3760 / cma.j.issn.0529-567x.2015.03.001).

[0137] Clinical symptoms and signs: pelvic pain, infertility, pelvic mass, etc.

[0138] Imaging examinations: Color Doppler ultrasound is mainly valuable for the diagnosis of ovarian endometriotic cysts. The typical ultrasound image of ovarian endometriotic cysts is a dense echogenic area within an anechoic region. Transvaginal or rectal ultrasound, CT, and MRI examinations are of certain significance for the diagnosis and assessment of deep lesions infiltrating the rectum or rectovaginal septum.

[0139] Laparoscopy: Currently, the standard method for diagnosing endometriosis is laparoscopic observation of the lesion morphology. During the procedure, careful observation of the pelvic cavity, especially the uterosacral ligaments and ovarian fossa, is crucial. A definitive diagnosis requires pathological examination; histopathological results are fundamental evidence for endometriosis diagnosis (although some clinical cases have definitively diagnosed without histopathological evidence). Pathological diagnostic criteria: Endometrial glands and stroma are visible in the lesions, accompanied by inflammatory reactions and fibrosis.

[0140] Serum CA125 level testing: CA125 level testing is not very useful for the diagnosis of early endometriosis. Elevated CA125 levels are more common in severe endometriosis, patients with significant pelvic inflammation, and those with ruptured endometriotic cysts or adenomyosis.

[0141] For suspected endometriosis of the bladder or intestine, preoperative cystoscopy or colonoscopy with biopsy should be performed to rule out lesions of the organ itself, especially malignant tumors. The probability of diagnosing endometriosis by biopsy is 10%–15%.

[0142] (2) Traditional Chinese Medicine Diagnosis Standards

[0143] Referencing the diagnostic criteria for cold-induced blood stasis in Traditional Chinese Medicine Gynecology: Main symptoms: lower abdominal distending pain, stabbing pain, or cold pain, with a fixed location; lower back distending pain, often worsening before or during menstruation; secondary symptoms: heavy or light menstrual flow, prolonged or irregular menstrual periods, dark red menstrual blood with clots; chest or breast distending pain; low mood, irritability, chest tightness; aversion to cold and cold limbs; profuse white or yellow leukorrhea; tongue and pulse: dark red tongue with white coating, possibly with petechiae or ecchymosis; deep and wiry or wiry and hesitant pulse.

[0144] (3) Inclusion criteria

[0145] Age 22-45 years; meeting both Western medicine diagnostic criteria and traditional Chinese medicine syndrome differentiation criteria; none of them had received treatment or had received treatment but discontinued medication for ≥3 months.

[0146] (4) Exclusion criteria

[0147] Those under 22 years of age or over 45 years of age; those with pelvic inflammatory disease, reproductive organ tumors, or other local or systemic malignant tumors; those who have been ruled out for suspected pregnancy; those with serious primary diseases of the cardiovascular, cerebrovascular, liver, kidney, and hematopoietic systems, or mental illness; those with endometriosis without dysmenorrhea; those with endometriosis ovarian endometriotic cysts larger than 5cm; and those who do not accept traditional Chinese medicine treatment.

[0148] (5) General Information

[0149] All cases were patients with endometriosis-related dysmenorrhea who visited the gynecology outpatient clinic of Jiangxi Provincial Hospital of Traditional Chinese Medicine and the Second Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine between December 2022 and December 2023. A before-and-after self-control method was used to collect 32 cases consistent with endometriosis-related dysmenorrhea.

[0150] (II) Treatment Methods

[0151] All enrolled patients received oral Chinese herbal medicine treatment. The basic formula consisted of: 15g cinnamon twig, 6g dragon's blood, 10g five-spice powder, 6g raw cattail pollen, 6g sandalwood, 3g leech, 10g ophiopogon japonicus, 10g white peony root, 6g licorice root, 10g uncaria rhynchophylla, and 6g amber.

[0152] Begin taking this medicine orally one week before menstruation and continue until the end of the menstrual period, one dose per day. Decoct in water to 400mL, and take twice daily, morning and evening, half an hour after meals, warm. One course of treatment consists of three consecutive menstrual cycles. Discontinue other related medications during treatment and reduce or avoid raw, cold, and spicy foods.

[0153] (III) Observation Indicators

[0154] Dysmenorrhea severity assessment: Visual analog scale (VAS) scoring method was used. Scores of 1-3 indicate mild pain; 4-6 indicate moderate pain; 7-10 indicate severe pain. Psychological state assessment: (1) Beck Self-Rating Depression Scale (BDI) scoring criteria were used. Scores of 0-9 indicate the normal range; 10-18 indicate mild depression; 19-29 indicate moderate depression; 30-63 indicate severe depression. The higher the score, the more severe the depression. (2) Beck Self-Rating Anxiety Scale (BAI) scoring criteria were used. Scores of 0-14 indicate the normal range; 15-25 indicate mild anxiety; 26-35 indicate moderate anxiety; 36 and above indicate severe anxiety. The higher the score, the more severe the anxiety. (3) The POMS scale is used to measure a person's emotional state, including dimensions such as tension, anger, fatigue, depression, energy, panic, and self-related emotions. The higher the score, the worse the psychological and emotional state. The dysmenorrhea and psychological state were evaluated before treatment, three months after medication, and three months after stopping medication, during the first 1-3 days of menstruation.

[0155] (iv) Statistical Methods

[0156] SPSS 22.0 statistical software was used for statistical analysis of the data. Quantitative data (following a normal distribution) were analyzed using... Description: One-way ANOVA was used for comparisons between groups, and P < 0.05 was considered statistically significant.

[0157] (V) Results

[0158] (1) Comparison of VAS, BDI, BAI and POMS scores of patients before and after treatment and after three months of follow-up Table 1 Comparison of VAS, BDI, BAI and POMS scores of patients before and after treatment and after three months of follow-up

[0159]

[0160] Note: n=32, compared with the pre-treatment group * P < 0.05 ** P < 0.01, *** P < 0.001, **** P < 0.0001; compared with the post-treatment group, # P < 0.05.

[0161] (2) VAS scores of patients before and after treatment and after three months of follow-up

[0162] As shown in Figure 10, compared with the pre-treatment group, the VAS scores after treatment and at the three-month follow-up were significantly lower (P < 0.0001), while compared with the post-treatment group, the VAS scores at the three-month follow-up group were higher (P < 0.05). This indicates that the Pingchong Jiangni Decoction plus Uncaria rhynchophylla and Amber can significantly improve the dysmenorrhea condition of patients during treatment and after recovery, but the efficacy decreases after discontinuation of the medication.

[0163] As shown in Figure 11, before treatment, the patients mainly experienced severe dysmenorrhea, while after treatment, the majority experienced mild dysmenorrhea. Compared with the post-treatment group, three months after stopping medication, the number of patients with mild dysmenorrhea decreased, while the number of patients with moderate dysmenorrhea relatively increased.

[0164] (3) Patient's BDI scores before and after treatment and after three months of follow-up

[0165] As shown in Figure 12, compared with the pre-treatment group, the BDI scores of patients decreased after treatment, and the difference was statistically significant (P < 0.001). Compared with the post-treatment group, the BDI scores of patients increased three months after drug withdrawal. This indicates that the Pingchong Jiangni formula with the addition of Uncaria rhynchophylla and amber can significantly improve the patients' depressive condition, and this formula has an antidepressant effect.

[0166] Among the collected cases of endometriosis, a small number of patients had psychological problems such as depression. As shown in Figure 13, compared with the pre-treatment group, the number of patients with low and moderate depression decreased after three months of medication; compared with the post-treatment group, the number of patients with low depression increased after three months of follow-up.

[0167] (4) Patient's BAI score before and after treatment and after three months of follow-up

[0168] As shown in Figure 14, compared with the pre-treatment group, the BAI scores decreased after treatment and at the three-month follow-up (P < 0.05). Three months after discontinuation of medication, the patients' BAI scores increased. This indicates that the Pingchong Jiangni formula combined with Uncaria rhynchophylla and amber can significantly improve the anxiety state of patients with endometriosis.

[0169] As shown in Figure 15, most of the collected endometriosis cases presented with anxiety. Compared to before treatment, after three months of medication, the number of patients with severe, moderate, and mild anxiety decreased, while the number of normal patients increased. However, after three months of discontinuing medication, the number of patients with low and moderate anxiety increased slightly.

[0170] (5) POMS scores of patients before and after treatment and after three months of follow-up

[0171] As shown in Figure 16, compared with before treatment, the POMS scores decreased after treatment and at the three-month follow-up, with statistically significant differences (P < 0.05). This indicates that the new Pingchong Jiangni formula can significantly improve patients' negative psychological emotions such as tension and fatigue. However, three months after stopping the medication, the patients' POMS scores increased, but the difference was not statistically significant.

[0172] Analysis of dysmenorrhea (VAS) and psychological status (BDI, BAI, and POMS) scores before treatment, after treatment, and after a three-month follow-up showed that the new Pingchong Jiangni formula can not only significantly reduce dysmenorrhea symptoms in patients with endometriosis, but also alleviate negative psychological states such as depression and anxiety in patients with endometriosis, and still has a certain therapeutic effect three months after stopping the medication.

[0173] This invention provides a traditional Chinese medicine formula for treating endometriosis. Many methods and approaches exist for implementing this technical solution; the above description is merely a preferred embodiment of the invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of this invention, and these improvements and modifications should also be considered within the scope of protection of this invention. All components not explicitly stated in this embodiment can be implemented using existing technologies.

Claims

1. A traditional Chinese medicine formula for treating endometriosis, characterized in that, The traditional Chinese medicine formula is made from the following components in parts by weight: 15 parts cinnamon twig, 10 parts five-spice powder, 6 parts raw cattail pollen, 6 parts sandalwood, 6 parts dragon's blood, 3 parts leech, 10 parts white peony root, 10 parts ophiopogon root, 10 parts uncaria, 6 parts amber, and 6 parts licorice root.

2. The use of the traditional Chinese medicine formula according to claim 1 in the preparation of drugs or preparations for treating endometriosis.

3. The application according to claim 2, characterized in that, The traditional Chinese medicine formula relieves pain caused by endometriosis, reduces lesion volume and inflammatory factor levels.

4. The application according to claim 2, characterized in that, in, The formulation type is any one of liquid, granule, patch, ointment, pill, powder, or tablet.

5. The application according to claim 2, characterized in that, The dosage form of the preparation is a liquid.

6. The application according to claim 5, characterized in that, The preparation method of the liquid preparation includes the following steps: soaking the prescribed amounts of cinnamon twig, five-spice powder, raw cattail pollen, leech, white peony root, ophiopogon root, amber, and licorice root in water and then decocting them. For the first decoction, bring it to a boil over high heat and then simmer over low heat, filtering out the decoction. For the second decoction, bring it to a boil over high heat and then simmer over low heat, adding the prescribed amounts of sandalwood and uncaria, filtering out the decoction, mixing the decoctions from the first and second decoctions, filtering, concentrating, and adding powdered dragon's blood.

7. The application according to claim 6, characterized in that, When decocting for the first time, add water at a rate of 8 to 10 times the total weight of the raw materials. When decocting for the second time, add water at a rate of 4 to 6 times the total weight of the raw materials. Simmer the first decoction over low heat for 20 to 30 minutes and the second decoction over low heat for 10 to 20 minutes. After adding the prescribed amounts of Dalbergia odorifera and Uncaria rhynchophylla, decoct for another 4 to 6 minutes.

8. The application according to claim 7, characterized in that, For the first decoction, add water at a rate of 10 times the total weight of the raw materials. For the second decoction, add water at a rate of 5 times the total weight of the raw materials. Simmer the first decoction over low heat for 20 minutes and the second decoction over low heat for 15 minutes. After adding the prescribed amounts of Dalbergia odorifera and Uncaria rhynchophylla, simmer for another 5 minutes.

9. The application according to claim 6, characterized in that, After soaking the herbs for 20-40 minutes, decoct them for the first time.

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