A method, device, equipment and medium for evaluating quality of jingugutongxiao pills based on anti-inflammatory effect

Abstract

The application discloses a method, device, equipment and medium for evaluating the quality of Jingu Tongxiao pills based on anti-inflammatory efficacy. The application claims a method for evaluating the quality of Jingu Tongxiao pills based on anti-inflammatory efficacy. The effect component index of Jingu Tongxiao pills is calculated according to a formula. The effect component index is positively correlated with the anti-inflammatory efficacy of Jingu Tongxiao pills. The higher the effect component index, the stronger the anti-inflammatory efficacy of Jingu Tongxiao pills and the better the quality of Jingu Tongxiao pills. In the method, the effect component indexes of the Jingu Tongxiao pills to be evaluated and the standard Jingu Tongxiao pills meeting the quality standard can be determined at the same time. If the effect component index of the Jingu Tongxiao pills to be evaluated is not lower than the effect component index of the standard Jingu Tongxiao pills, the quality of the Jingu Tongxiao pills to be evaluated is evaluated as qualified. In the method, the correlation coefficient of the effect component index and the anti-inflammatory efficacy of Jingu Tongxiao pills is not lower than 0.90, which indicates that the two have a high correlation, and provides strong data support for the reliability and accuracy of the method.

Description

Technical Field

[0001] The present invention belongs to the detection field, relates to quality detection and evaluation of traditional Chinese medicine compound prescriptions, and specifically relates to a method, device, equipment and medium for evaluating the quality of Jingu Tongxiao Pills based on anti-inflammatory pharmacodynamic evaluation. Background Art

[0002] Jingu Tongxiao Pills are a new traditional Chinese medicine developed based on the empirical prescription of the national intangible cultural heritage "Pingle Guo's Bone-setting Method", which is a national protected traditional Chinese medicine variety and has been included in the 2025 edition of Chinese Pharmacopoeia. The complete prescription is composed of 11 traditional Chinese medicinal materials including Salviae Miltiorrhizae Radix et Rhizoma, Spatholobi Caulis, Cyperi Rhizoma, Linderae Radix, Cinnamomi Ramulus, Clematidis Radix et Rhizoma, Gentianae Macrophyllae Radix, Paeoniae Radix Alba, Rehmanniae Radix, Cyathulae Radix, and Glycyrrhizae Radix et Rhizoma [References 1-2]. Its prescription has rigorous compatibility with distinct primary and secondary roles: Salviae Miltiorrhizae Radix et Rhizoma and Spatholobi Caulis act as monarch drugs, which can promote blood circulation to remove blood stasis, relax tendons and relieve pain; Cyperi Rhizoma and Linderae Radix act as minister drugs, which are good at promoting qi circulation to relieve pain, warming channels and dispelling cold; Cinnamomi Ramulus, Clematidis Radix et Rhizoma, and Gentianae Macrophyllae Radix can warm and unblock the channels and collaterals, expel wind and dampness, and subside swelling to relieve pain; Paeoniae Radix Alba and Rehmanniae Radix can nourish blood and astringe yin, tonify deficiency and relieve pain; Cyathulae Radix can promote blood circulation to remove blood stasis and dredge the joints, and the above three groups of drugs together act as adjuvant drugs; Glycyrrhizae Radix et Rhizoma acts as the conductant drug, which has the effects of moderating acute pain and harmonizing all the drugs. The combined use of all drugs exerts the core effects of promoting blood circulation and qi circulation, warming and unblocking the channels and collaterals, and subsiding swelling to relieve pain [References 3-4]. Clinical studies have shown that this drug is mainly used to treat knee joint pain, swelling, limited mobility and other conditions caused by blood stasis and cold coagulation as well as hyperosteogeny, with definite curative effect, few adverse reactions and high medication safety [Reference 4]. At the same time, Jingu Tongxiao Pills, when used alone or in combination with other drugs for intervention, have significant clinical improvement effects on a variety of bone and joint diseases such as scapulohumeral periarthritis, chondromalacia patellae, rheumatoid arthritis, lumbar disc herniation, and knee osteonecrosis [References 5-8].

[0003] At present, studies on the quality control and evaluation of Jingu Tongxiao Pills mainly focus on multi-index quantification and fingerprint analysis [References 9-10]. The content determination of multiple components and multiple indexes is the mainstream mode of traditional Chinese medicine quality control at present, but this traditional quality control method has obvious limitations: it only determines the content of each index component separately, ignores the difference in the contribution of different active components to the overall efficacy, the evaluation system is fragmented, it is difficult to truly reflect the intrinsic quality of medicinal materials, and its correlation with clinical efficacy is weak. In response to the above problems, the team of Xiao Xiaohe first proposed the effect-constituent index (ECI), a new comprehensive quantitative evaluation method for traditional Chinese medicine quality, in 2014 [Reference 11]. Centered on clinical utility, ECI integrates quantitative analysis of chemical components and biological potency testing, relies on the "dose-effect" relationship equation of active components of traditional Chinese medicine, assigns differentiated weight coefficients according to the efficacy contribution degree of each active component, and obtains a single comprehensive quantitative index through weighted calculation [References 12-13]. This evaluation mode effectively makes up for the shortcomings of the traditional traditional Chinese medicine quality control system, breaks the limitation of independent evaluation of multiple indexes, realizes the accurate correlation between the content of traditional Chinese medicine components and the overall efficacy, can scientifically and comprehensively evaluate the intrinsic quality of traditional Chinese medicine, can provide an objective basis for the evaluation of traditional Chinese medicine authenticity, the distinction of commodity specifications and the clinical safe and effective medication, and significantly improves the scientificity and systematicness of traditional Chinese medicine quality evaluation [References 11,14-15].

[0004] In order to overcome the deficiencies of the prior art in the quality control of Jingu Tongxiao Pills and further improve the quality control level of Jingu Tongxiao Pills, the present invention is specially proposed.

[0005] The above reference information corresponds to the following:

[0006] [1] Wu Ailing. Study on the anti-inflammatory effect and mechanism of Jingu Tongxiao Pills. World Journal of Integrated Traditional Chinese and Western Medicine. 2014. 9(10): 1046-1048.

[0007] [2] Chinese Pharmacopoeia Commission. Pharmacopoeia of the People's Republic of China. Volume I [M]. Beijing: China Medical Science Press, 2025: 1770-1771.

[0008] [3] Liang Feng. Clinical Observation on the Treatment of Knee Osteoarthritis with "Hubin Songjin" Tuina Manipulation Combined with Jingu Tongxiao Pills [J]. Acta Chinese Medicine, 2018, 33(03): 503-506.

[0009] [4] Tan Xuyi, Liu Liyun, Gao Shutu, et al. Effects of Jingu Tongxiao Pills on WOMAC Scores, PGE2 and MMP-3 in Patients with Knee Osteoarthritis [J]. Chinese Journal of Traditional Medical Traumatology & Orthopedics, 2014, 22(02): 18-20.

[0010] [5] Hu Pei. Treatment of 30 cases of periarthritis of the shoulder with Jin Gu Tong Xiao Wan combined with Qi Zhu Zhan Jin San [J]. Rheumatology and Arthritis, 2014, 3(03): 21-23.

[0011] [6] Zhang Yunbin. Observation on the efficacy of internal administration of Jin Gu Tong Xiao Wan combined with external application of Er Cao Er Pi Tang and Western medicine in the treatment of patellar chondromalacia [J]. Shaanxi Journal of Traditional Chinese Medicine, 2012, 33(04): 436-437.

[0012] [7] Chen Baohong. Treatment of 50 cases of rheumatoid arthritis with Jin Gu Tong Xiao Wan [J]. Journal of Traditional Chinese Medicine Research, 2007, (07): 31-32.

[0013] [8] Hao Jun, Gao Wenxiang, Zou Chunyu, et al. Treatment of 21 cases of knee osteonecrosis with integrated traditional Chinese and Western medicine [J]. Journal of Traditional Chinese Medicine, 2014, 20(03): 97-99.

[0014] [9] Sun Shi, Wang Yifang, Zhao Xinjie, et al. Establishment of HPLC fingerprint of Jin Gu Tong Xiao Wan and determination of the content of 7 components therein [J]. China Pharmacy, 2021, 32(10): 1235-1240.

[0015]

[10] Chen Yinzi, Hu Jing, Li Ning, et al. Comprehensive quality evaluation of Jin Gu Tong Xiao Wan based on multi-component quantitative combined chemometrics and entropy weight TOPSIS method [J]. Chinese Medicinal Herbs, 2026, (03): 685-691.

[0016]

[11] Xiong Yin, Xiao Xiaohe, Yan Dan, et al. Comprehensive quantitative integration strategy for quality control of traditional Chinese medicine: index of effective components of traditional Chinese medicine [J]. Chinese Herbal Drugs, 2014, 45(01): 1-7.

[0017]

[12] Liu Zhenjie, Tan Xiaoqing, Qiu Haixin, et al. Research on the compatibility of Danshen based on the effect component index [J]. Journal of Traditional Chinese Medicine, 2022, 28(05): 84-87.

[0018]

[13] Zhang Meng, Feng Liang, Jia Xiaobin. Current status and prospect of the development of traditional Chinese medicine quality evaluation technology based on bioactivity and effect benchmarks [J]. World Journal of Traditional Chinese Medicine, 2020, 15(15): 2234-2239.

[0019]

[14] De-Min Li, Rui Yuan,Xue Bai, et al .Effect-constituent index-based quality assessment of Anemarrhenae Rhizoma before and after processing[J]. J Asian Nat Prod Res, 2025: 1-22.

[0020]

[15] Xiaomin Cui, Hui Ren, Wenjing Lu, et al. Construction of anti-oxidant active effect-constituent index for Herba Phlojodicarpi[J]. Central South Pharmacy, 2025, 23(07): 1839-1843. Contents of the Invention

[0021] A first objective of the present invention is to provide a method for evaluating the quality of Jingu Tongxiao pills based on anti-inflammatory pharmacodynamics, a second objective is to provide a device for evaluating the quality of Jingu Tongxiao pills based on the method, a third objective is to provide an electronic device capable of implementing the method, and a fourth objective is to provide a computer-readable storage medium storing a computer program for implementing the method.

[0022] The above objectives of the present invention are achieved through the following technical solutions:

[0023] The present invention provides a method for evaluating the quality of Jingu Tongxiao pills based on anti-inflammatory pharmacodynamics, wherein the effect-constituent index of the Jingu Tongxiao pills is calculated according to Formula (I), the effect-constituent index is positively correlated with the anti-inflammatory pharmacodynamics of the Jingu Tongxiao pills, and a higher effect-constituent index indicates stronger anti-inflammatory pharmacodynamics and better quality of the Jingu Tongxiao pills;

[0024]

[0025] In Formula (I):

[0026] ECI represents the effect-constituent index of Jingu Tongxiao pills;

[0027] i represents the pharmacodynamic constituents paeoniflorin, albiflorin, gentiopicroside, loganic acid, salvianolic acid B, and 1,2,3,4,6-O-pentagalloylglucose;

[0028] n=6, representing the number of pharmacodynamic constituents;

[0029] Xi represents the content of each pharmacodynamic constituent in Jingu Tongxiao pills;

[0030] Wi represents the weight coefficient of each pharmacodynamic constituent, which is calculated according to Formula (II):

[0031]

[0032] In formula (II), the definitions of i and n are the same as in formula (I), and IC30 represents the concentration required for each pharmacodynamic component to inhibit 30% of the expression of inflammatory factors in an inflammation model.

[0033] In this method, the effective component indexes of the Jin Gutongxiao Pill to be evaluated and the standard Jin Gutongxiao Pill that meets the quality standard can be measured simultaneously. If the effective component index of the Jin Gutongxiao Pill to be evaluated is not lower than that of the standard Jin Gutongxiao Pill, the quality of the Jin Gutongxiao Pill to be evaluated is judged as qualified, otherwise it is judged as unqualified.

[0034] In a specific embodiment, the effective component index is positively correlated with the anti-inflammatory efficacy of Jin Gutongxiao Pill, and the correlation coefficient is not less than 0.90. The positive correlation means that the two variables change in the same direction, when one increases, the other also tends to increase, the correlation coefficient r is greater than 0 and less than or equal to 1, and the closer r is to 1, the stronger the positive correlation.

[0035] In a specific embodiment, the effective component index is positively correlated with the anti-inflammatory efficacy of Jin Gutongxiao Pill, and the correlation coefficient is not less than 0.95. The closer the correlation coefficient r is to 1, the stronger the positive correlation.

[0036] In a specific embodiment, the inflammatory factor is selected from one or more of NO, IL-6 and TNF-α. According to the literature [Wu Ailing. Study on the Anti-inflammatory Effect and Mechanism of Jin Gutongxiao Pill. World Journal of Integrated Traditional Chinese and Western Medicine. 2014], the anti-inflammatory effect of Jin Gutongxiao Pill is related to reducing the levels of NO, IL-6 and TNF-α.

[0037] In a specific embodiment, the inflammation model is selected from one or more of in vitro cell inflammation models and in vivo animal inflammation models.

[0038] In a specific embodiment, the inflammation model is an in vitro inflammation model established by lipopolysaccharide-induced RAW264.7 cells, which is the most classic and commonly used in vitro cell inflammation model.

[0039] The present invention provides a Jin Gutongxiao Pill quality evaluation device based on any of the above methods, comprising:

[0040] A data acquisition module, configured to acquire the content Xi of the pharmacodynamic components paeoniflorin, albiflorin, gentiopicroside, loganic acid, salvianolic acid B and 1,2,3,4,6-O-pentagalloylglucose in the Jin Gutongxiao Pill to be evaluated, as well as the IC30 value of the concentration required for each pharmacodynamic component to inhibit 30% of the expression of inflammatory factors in an inflammation model; the inflammation model is selected from one or more of in vitro cell inflammation models and in vivo animal inflammation models, and the inflammatory factor is selected from one or more of NO, IL-6 and TNF-α.

[0041] a processing module, configured to input the IC30 value of each active pharmaceutical ingredient into formula (II) to obtain the weight coefficient Wi of each active pharmaceutical ingredient, then input the weight coefficient Wi of each active pharmaceutical ingredient and the content Xi of each active pharmaceutical ingredient in the Jin Gutongxiao pill to be evaluated into formula (I) to obtain the effective component index of the Jin Gutongxiao pill to be evaluated, and finally compare the effective component index of the Jin Gutongxiao pill to be evaluated with the effective component index of the Jin Gutongxiao pill with standard quality; the formula (I) and the formula (II) are as described above;

[0042] an output module, configured to output whether the quality of the Jin Gutongxiao pill to be evaluated is qualified, wherein the output is qualified if the effective component index thereof is not lower than the effective component index of the Jin Gutongxiao pill with standard quality, otherwise the output is unqualified.

[0043] In a specific embodiment, the effective component index of the Jin Gutongxiao pill with standard quality is obtained by using Jin Gutongxiao pills that meet the quality standards.

[0044] The present invention provides an electronic device, comprising:

[0045] a processor;

[0046] a memory, configured to store a computer program;

[0047] when the computer program is executed by the processor, any one of the above methods is implemented.

[0048] The present invention provides a computer-readable storage medium, on which a computer program is stored, and when the computer program is executed by a processor, any one of the above methods is implemented.

[0049] Beneficial effects:

[0050] 1. Jin Gutongxiao pill has the effects of activating blood and promoting qi, warming meridians and unblocking collaterals, detumescence and relieving pain, and is used for symptoms such as knee joint pain, swelling and limited activity caused by blood stasis and cold coagulation and hyperosteogeny of knee joint. Those skilled in the art know that anti-inflammation is effective and critical for the treatment of symptoms such as knee joint pain, swelling and limited activity. Wu Ailing discussed the anti-inflammatory effect and mechanism of Jin Gutongxiao pill in "Study on the Anti-inflammatory Effect and Mechanism of Jin Gutongxiao Pill" [Wu Ailing. Study on the Anti-inflammatory Effect and Mechanism of Jin Gutongxiao Pill. World Journal of Integrated Traditional Chinese and Western Medicine. 2014]. The method for evaluating the quality of Jin Gutongxiao pill provided by the present invention is exactly based on the anti-inflammatory efficacy. It is particularly rare that the effective component index in this method is positively correlated with the anti-inflammatory efficacy of Jin Gutongxiao pill, and the correlation coefficient is above 0.95. Those skilled in the art know that positive correlation means that two variables change in the same direction, when one increases, the other also tends to increase, the correlation coefficient is greater than 0 and less than or equal to 1, the closer to 1, the stronger the positive correlation, and the more accurate and reliable it is to evaluate one variable through the other variable.

[0051] 2. When screening active components via spectrum-effect relationship: According to the VIP value (VIP>1), Peak 10, Peak 9, Peak 11, Peak 6, Peak 4, and Peak 14 make significant contributions to the inhibition of NO production. However, limited by the prior art, the No. 11 chromatographic peak and No. 14 chromatographic peak have not been identified; according to the VIP value (VIP>1), Peak 10, Peak 9, Peak 11, Peak 6, Peak 4, Peak 14, and Peak 5 make significant contributions to reducing the IL-6 level, and the No. 11 chromatographic peak and No. 14 chromatographic peak are also unidentified; according to the VIP value (VIP>1), Peak 10, Peak 6, Peak 9, Peak 4, Peak 3, and Peak 11 make significant contributions to reducing the TNF-α level, and the No. 11 chromatographic peak is also unidentified. Theoretically, since the compounds corresponding to the unidentified chromatographic peaks make significant contributions to the inhibition of NO, IL-6 and TNF-α, discarding these compounds with strong pharmacodynamic contribution (while adding compounds with weak pharmacodynamic contribution where VIP≤1) will reduce the representativeness of the effective component index for the anti-inflammatory pharmacodynamics of Jingu Tongxiao Pills. However, in practice, the correlation coefficients between the effective component indices based on the final active components, namely paeoniflorin, albiflorin, gentiopicroside, loganic acid, salvianolic acid B and 1,2,3,4,6-O-pentagalloylglucose, and the anti-inflammatory pharmacodynamic indicators of Jingu Tongxiao Pills all reach above 0.95, and such an effect is unexpected to those skilled in the art.

[0052] 3. In Liu Yuan's doctoral dissertation from Hunan University of Chinese Medicine [Liu Yuan. Network Pharmacology Analysis and Mechanism Study of Jingu Tongxiao Pills in the Treatment of Knee Osteoarthritis. Doctoral Dissertation. 2021], network pharmacology was adopted to screen out the top 5 compounds in terms of pharmacodynamic importance of Jingu Tongxiao Pills, which are quercetin, luteolin, kaempferol, naringin and β-sitosterol, and most of these compounds are anti-inflammatory active compounds well known in the art. During the formation of the technical solution of the present invention, when none of the aforementioned compounds was found in the active components screened out through the spectrum-effect relationship, the inventor once doubted the necessity of continuing the research; when further determining the IC50 value of the proposed active components, most of the compounds showed weak anti-inflammatory activity, and the inhibition rate at the maximum non-toxic concentration could not reach 50%, which directly led to the suspension and shelving of this technical solution. The differences between previous studies and literature reports all point to one problem: the proposed active components are most likely not the most representative anti-inflammatory active components of Jingu Tongxiao Pills. Without representativeness, it will most probably lead to a low correlation between the effective component index and the anti-inflammatory pharmacodynamics of Jingu Tongxiao Pills. Later, IC was abandoned 50 and IC was used instead 30 for quantitative evaluation of anti-inflammatory activity, the fact that the correlation coefficient between the effective component index and the anti-inflammatory pharmacodynamics of Jingu Tongxiao Pills is still as high as 0.95 or more far exceeds the inventor's expectation.

[0053] 4. Chen Yinzi et al. provided a method for comprehensively evaluating the quality of Jingu Tongxiao Pills based on the quantitative determination of multi-index components combined with chemometrics and entropy weight technique for order preference by similarity to ideal solution (EW-TOPSIS) method [Chen Yinzi et al. Comprehensive quality evaluation of Jingu Tongxiao Pills based on multi-component quantification combined with chemometrics and entropy weight TOPSIS method. Chinese Herbal Medicines. 2026]. Using salvianolic acid B as the internal reference substance, the quantitative analysis of multi-components by single-marker method was adopted to simultaneously determine the contents of gallic acid, loganic acid, gentiopicroside, albiflorin, paeoniflorin, liquiritin, 1,2,3,4,6-O-pentagalloylglucose, salvianolic acid B and glycyrrhizic acid in Jingu Tongxiao Pills. Cluster analysis and partial least squares discriminant analysis were used to screen the differential components affecting the quality of Jingu Tongxiao Pills, and the entropy weight technique for order preference by similarity to ideal solution (EW-TOPSIS) method was used for comprehensive evaluation. The results of partial least squares discriminant analysis showed that the VIP values of albiflorin, paeoniflorin, salvianolic acid B and loganic acid were greater than 1. Compared with this prior art: firstly, the method of the present invention has only 6 pharmacodynamic components, which is more streamlined; secondly, the quality evaluation method of this prior art is not based on anti-inflammatory pharmacodynamics, and cannot be used to evaluate the quality of Jingu Tongxiao Pills based on anti-inflammatory pharmacodynamics. For example, albiflorin, paeoniflorin, salvianolic acid B and loganic acid are considered important because these four components may be the main variables causing quality differences between different batches of Jingu Tongxiao Pills, which have nothing to do with pharmacodynamics, and provide no technical enlightenment to the technical solution of the present invention. Brief Description of the Drawings

[0054] Figure 1 is the chromatogram of 20 batches of Jingu Tongxiao Pills in Example 1 and the marking of 16 common peaks;

[0055] Figure 2 shows the effects of gradient concentrations of Jingu Tongxiao Pills in Example 1 on the survival rate and NO secretion of LPS-induced RAW264.7 cells;

[0056] Figure 3 is the VIP diagram of the spectrum-effect relationship of the anti-inflammatory activity of Jingu Tongxiao Pills in Example 1;

[0057] Figure 4 is the chromatogram of the mixed reference substances in Example 1;

[0058] Figure 5 is the IC of paeoniflorin, albiflorin, gentiopicroside, loganic acid, salvianolic acid B and 1,2,3,4,6-O-pentagalloylglucose in Example 1 based on inhibiting the expression of inflammatory factors 30 values;

[0059] Figure 6 is the correlation analysis between the effect component index in Example 1 and the IC of NO, IL-6 and TNF-α 30 . Detailed Description of the Embodiments

[0060] The substantive content of the present invention will be described in detail below with reference to specific embodiments. However, those skilled in the art should know that the scope of protection of the present invention should not be limited to these specific embodiments.

[0061] Example 1: A method for evaluating the quality of Jin Gu Tong Xiao Wan (Muscle and Bone Pain Relief Pills) based on its anti-inflammatory efficacy.

[0062] 1. Instruments and Materials

[0063] 1.1 Cells

[0064] RAW264.7 cell line, batch number JF4RFYWU3S, was purchased from Wuhan Pronosei Life Science Technology Co., Ltd.

[0065] 1.2 Instruments

[0066] Agilent 1260 series high performance liquid chromatograph, equipped with VWD detector and Agilent Chem Station workstation, Agilent Technologies, Inc., USA; KQ-400DE CNC ultrasonic cleaner, Kunshan Ultrasonic Instrument Co., Ltd.; SQP 1 / 100,000 electronic balance and SQP 1 / 10,000 electronic balance, Sartorius Scientific Instruments (Beijing) Co., Ltd.

[0067] 1.3 Medicinal Materials and Reagents

[0068] 1.3.1 Medicinal Materials

[0069] Jin Gu Tong Xiao Wan (products numbered 20231005, 20231100, 20231101, 20231103, 20231104, 20231107, 20231110, 20231111, 20231112, 20231113, 20231115, 20231201, 20231202, 20231203, 20231205, 20231208, 20231210, 20240901, 20240902, 20240115), product numbers: S1-S20; all specifications are 6 g per bag, purchased from Henan Luozheng Pharmaceutical Co., Ltd.

[0070] 1.3.2 Reference Standard

[0071] Gallic acid (batch number: PS012605), with an HPLC mass fraction of ≥98%, was purchased from Chengdu Pusi Biotechnology Co., Ltd.; loganic acid (batch number: CHB231024), gentiopicrin (batch number: CHB230822), paeoniflorin (batch number: CHB230923), paeoniflorin (batch number: CHB230901), glycyrrhizin (batch number: CHB231106), 1,2,3,4,6-O-pentagalloylglucose (batch number: CHB230915), and salvianolic acid B (batch number: CHB231102), with an HPLC mass fraction of ≥98%, were all purchased from Chengdu Kloma Biotechnology Co., Ltd.; cryptotanshinone (batch number: 24081091), with an HPLC mass fraction of ≥98%, was purchased from Chengdu Pufeide Biotechnology Co., Ltd.

[0072] 1.3.3 Reagents

[0073] DMEM medium (batch number: C11965500BT), Thermo Fisher Scientific, USA; phosphate-buffered saline (PBS) (batch number: 130114001), Beijing Lanjie Shunyi Technology Co., Ltd.; RAW 264.7 cell culture medium, Wuhan Pronosai Life Science Technology Co., Ltd.; NO reagent kit (batch number: S0021S), Shanghai Beyotime Biotechnology Co., Ltd.; IL-6 ELISA kit (batch number: FN240725), TNF-α ELISA kit (batch number: FN231115), Wuhan Feien Biotechnology Co., Ltd.; acetonitrile was chromatographic grade, and other reagents were analytical grade; water was Wahaha purified water.

[0074] 2. Methods and Results

[0075] 2.1 Establishment of fingerprint map

[0076] 2.1.1 Preparation of the test solution

[0077] Weigh 0.4 g of the sample powder and place it in a 100 mL Erlenmeyer flask. Add 30 mL of 80% methanol. Sonicate at room temperature (frequency 45 kHz, power 600 W) for 45 min. After cooling to room temperature, replenish the lost mass with 80% methanol. Filter the supernatant through a 0.45 μm microporous membrane to obtain the final product.

[0078] 2.1.2 Chromatographic conditions

[0079] Agilent 5TC-C18 column (250 mm × 4.6 mm, 5 μm); mobile phase: acetonitrile-0.1% phosphoric acid aqueous solution, gradient elution (0–15 min, 5%–10% acetonitrile; 15–35 min, 10%–13% acetonitrile; 35–75 min, 13%–32% acetonitrile; 75–90 min, 32%–54% acetonitrile; 90–130 min, 54%–70% acetonitrile); flow rate: 1.0 mL / min; detection wavelength: 230 nm; column temperature: 30 °C; injection volume: 10 μL.

[0080] 2.1.3 Methodological Examination

[0081] 2.1.3.1 Precision Test

[0082] Accurately weigh 0.4 g of the same batch of Jin Gu Tong Xiao Wan (S1) powder and prepare the test solution according to the method in section "2.1.1". Inject the sample six times repeatedly under the chromatographic conditions in section "2.1.2", using peak 10 (tanshinone B) as the reference peak. Calculate the relative retention time and relative peak area of ​​each characteristic peak. Examine the RSD (n=6) of the relative retention time and the RSD (n=6) of the relative peak area for each characteristic peak. The results show that the RSD values ​​of the relative retention time of the common peaks are all <0.08%, and the RSD values ​​of the relative peak areas are all <2.19%, indicating good instrument precision.

[0083] 2.1.3.2 Repeatability Test

[0084] Accurately weigh 0.4 g of the same batch of Jin Gu Tong Xiao Wan (S1) powder and prepare six parallel test solutions using the method described in section "2.1.1". Inject each solution according to the chromatographic conditions described in section "2.1.2", using peak 10 (tanshinone B) as the reference peak. Calculate the RSD (n=6) of the relative retention time and the RSD (n=6) of the relative peak area for each common peak. The results show that the RSD values ​​of the relative retention time for each common peak are all <0.35%, and the RSD values ​​of the relative peak area are all <2.19%, indicating good repeatability of the method.

[0085] 2.1.3.3 Stability Test

[0086] Take the same batch of Jingu Tongxiao Pill (S1) powder, accurately weigh 0.4 g, prepare the test solution according to the method under "2.1.1", perform injection detection at 2, 4, 6, 8, 10, 12, 24 h respectively under the chromatographic conditions under "2.1.2" and record the chromatograms. Using the 10th peak (Salvianolic Acid B) as the reference peak, calculate the relative retention time and relative peak area of each common peak, and investigate the RSD (n=6) of the relative retention time and the RSD (n=6) of the relative peak area of each common peak respectively, so as to investigate the stability of the test solution. The results show that the RSD values of the relative retention time of the common peaks are all less than 0.28%, and the RSD values of the relative peak area are all less than 2.54%, indicating that the test solution is stable within 24 h.

[0087] 2.1.4 Establishment of fingerprint and similarity evaluation

[0088] Accurately pipette 10 µL of each of the 20 batches of Jingu Tongxiao Pill test solutions, and perform injection determination under the chromatographic conditions under "2.1.2". Import the relevant data into the "Similarity Evaluation System Software for Traditional Chinese Medicine Fingerprints", adopt the median method for multi-point correction, and then automatically match to generate the control fingerprint (R) of Jingu Tongxiao Pill. Using the 10th peak as the reference, 16 common peaks are identified, and similarity evaluation is performed on 30 batches of samples. The results show that the similarity of the fingerprints of the 20 batches of Jingu Tongxiao Pill samples are all greater than 0.95, as shown in Figure 1 .

[0089] 2.1.5 Anti-inflammatory effect on LPS-induced RAW264.7 cells

[0090] 2.1.5.1 Preparation of test sample

[0091] Weigh 0.6 g of each of the 20 batches of Jingu Tongxiao Pill powder, and weigh accurately. Place in a stoppered conical flask, accurately add 10 mL of pure water, stopper tightly, and weigh the mass; perform ultrasonic treatment (power 250 W, frequency 40 kHz) for 0.5 h, and weigh the mass again; make up the lost mass with water, and take the subsequent filtrate. Use complete medium to prepare the medicated medium with the 20 batches of Jingu Tongxiao Pill at a concentration of 60 mg·mL -1 .

[0092] 2.1.5.2 Effect on inflammatory factors in LPS-induced RAW264.7 cells

[0093] Take RAW264.7 cells in the logarithmic growth phase, and adjust the cell density to 1.5×10 5 ~2.0×10 5cells / mL, and seeded into a 96-well plate. The cells were allowed to adhere by standing, the blank group was added with 200 μL of complete medium; the model group was added with LPS at a final concentration of 1 μg / mL; the administration groups were respectively added with complete medium containing 0, 2.34, 4.69, 9.38, 18.75, 37.5, 75.0, 150.0 μg / mL of Jingu Tongxiao Pills. Except for the blank group, each group was added with LPS at a final concentration of 1 μg / mL. After conventional culture for 24 h, the medium was aspirated, 100 μL of CCK8 solution was added to each well, the absorbance (A) value was measured at 450 nm, and the cell viability was calculated. Cell viability = (A 给药 -A 空白 ) / (A 对照 -A 空白 ). The Griess method [References 16-17] was used to determine the NO concentration in each group, as shown by Figure 2 that when the final mass concentration of Jingu Tongxiao Pills was in the range of 0~150 μg / mL, the cell viability was all above 90%. The detection of NO by the Griess method found that when the final mass concentration was 150 μg / mL and 75 μg / mL, the NO concentration decreased significantly compared with the model group (P<0.05, 0.001). The results of the cell viability test and the Griess NO detection test showed that the optimal administration concentration of Jingu Tongxiao Pills was determined to be 75 μg / mL for subsequent tests. The ELISA method [References 18-19] was used to detect the concentrations of IL-6 and TNF-α in each group. The results are shown in Table 1 and the continuation of Table 1 (where the inhibition rate is the inhibition rate at the dose of 75 μg / mL Jingu Tongxiao Pills).

[0094] References 16-19 are as follows:

[0095]

[16] Gabriella Iula, Panagiota M Kalligosfyri, Ada Raucci, et al. Evaluating in-vitro cellular toxicity through the analysis of nitrite ions variation at printed electrochemical strips[J]. Talanta, 2026, 299: 129073.

[0096]

[17] Ying Feiyu, He Shiyan, Li Li, et al. UPLC-Q-TOF-MS / MS combined with network pharmacology, molecular docking technology and experimental verification to predict the quality markers (Q-Marker) of the anti-inflammatory effect of Yao medicine Sannaimu[J]. Drug Evaluation Research, 2025, 48(1): 121-135.

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[19] Peiyu Wang, Zhen Jia, Jie Gao, et al. Effects of Yinhuang Lotion on IL-18, TNF-α and HIF-1α / VEGF pathway in rats with diabetic ulcers[J]. Acta Chinese Medicine and Pharmacology, 2026, 54(2): 15-21.

[0099] Table 1 Fingerprint peak areas and anti-inflammatory activities of Jingu Tongxiao Pills

[0100] batch number Peak 1 Peak 2 Peak 3 Peak 4 Peak 5 Peak 6 Peak 7 Peak 8 Peak 9 Peak 10 Peak 11 S1 126.7 46.5 339.2 792.1 55.5 804.8 114.8 82.5 414.6 1243.3 94.4 S2 130.8 37.7 309.4 727.9 40.1 690.2 105.8 75.9 94.9 675.6 32.1 S3 130.8 37.7 309.4 727.9 40.1 690.2 105.8 75.9 94.9 675.6 32.1 S4 130.8 37.7 309.4 727.9 40.1 690.2 105.8 75.9 94.9 675.6 32.1 S5 130.8 37.7 309.4 727.9 40.1 690.2 105.8 75.9 94.9 675.6 32.1 S6 130.8 37.7 309.4 727.9 40.1 690.2 105.8 75.9 94.9 675.6 32.1 S7 130.8 37.7 309.4 727.9 40.1 690.2 105.8 75.9 94.9 675.6 32.1 S8 128.2 38.5 301.4 730.4 45.3 688.0 105.1 70.0 381.2 1138.4 92.8 S9 127.9 38.2 305.1 734.9 46.3 738.1 115.7 69.7 244.7 770.0 75.1 S10 130.8 37.7 309.4 727.9 40.1 690.2 105.8 75.9 94.9 675.6 32.1 S11 130.8 37.7 309.4 727.9 40.1 690.2 105.8 75.9 94.9 675.6 32.1 S12 130.8 37.7 309.4 727.9 40.1 690.2 105.8 75.9 94.9 675.6 32.1 S13 130.8 37.7 309.4 727.9 40.1 690.2 105.8 75.9 94.9 675.6 32.1 S14 130.8 37.7 309.4 727.9 40.1 690.2 105.8 75.9 94.9 675.6 32.1 S15 130.8 37.7 309.4 727.9 40.1 690.2 105.8 75.9 94.9 675.6 32.1 S16 130.8 37.7 309.4 727.9 40.1 690.2 105.8 75.9 94.9 675.6 32.1 S17 130.8 37.7 309.4 727.9 40.1 690.2 105.8 75.9 94.9 675.6 32.1 S18 126.7 46.5 339.2 792.1 55.5 804.8 114.8 51.0 414.6 1243.3 102.7 S19 128.2 38.5 301.4 730.4 45.3 688.0 105.1 70.0 381.2 1138.4 92.8 S20 128.2 38.5 301.4 730.4 42.7 688.0 105.1 70.0 381.2 1138.4 92.8

[0101] Table 1 (continued) Fingerprint peak areas and anti-inflammatory activities of Jingu Tongxiao Pills

[0102] batch number Peak 12 Peak 13 Peak 14 Peak 15 Peak 16 NO inhibition rate IL-6 inhibition rate TNF-α inhibition rate S1 24.8 166.0 106.9 69.3 201.6 34.17% 45.18% 11.82% S2 31.1 152.8 83.0 59.4 182.9 33.23% 56.04% 8.51% S3 31.1 152.8 83.0 59.4 182.9 40.44% 52.54% 11.80% S4 31.1 152.8 83.0 59.4 182.9 50.31% 53.23% 13.98% S5 31.1 152.8 83.0 59.4 182.9 48.59% 43.78% 14.57% S6 31.1 152.8 83.0 59.4 182.9 37.77% 73.90% 9.23% S7 31.1 152.8 83.0 59.4 182.9 32.45% 42.73% 8.76% S8 34.9 148.4 95.2 62.8 178.5 50.31% 42.37% 10.39% S9 34.7 150.7 92.2 65.2 179.0 32.45% 71.10% 11.06% S10 31.1 152.8 83.0 59.4 182.9 43.26% 51.48% 12.57% S11 31.1 152.8 83.0 59.4 182.9 40.44% 49.73% 9.60% S12 31.1 152.8 83.0 59.4 182.9 41.38% 51.13% 15.53% S13 31.1 152.8 83.0 59.4 182.9 30.56% 59.18% 14.52% S14 31.1 152.8 83.0 59.4 182.9 36.83% 73.20% 12.69% S15 31.1 152.8 83.0 59.4 182.9 38.71% 49.37% 7.17% S16 31.1 152.8 83.0 59.4 182.9 37.77% 75.29% 5.59% S17 31.1 152.8 83.0 59.4 182.9 35.11% 91.07% 11.01% S18 24.8 166.0 106.9 69.3 201.6 42.32% 65.49% 6.58% S19 34.9 148.4 95.2 62.8 178.5 41.38% 64.43% 11.68% S20 34.9 148.4 83.0 62.8 178.5 40.44% 65.84% 16.23%

[0103] 2.1.5.3 Screening of pharmacodynamic components via spectrum-effect relationship

[0104] The orthogonal partial least squares-discriminant analysis (OPLS-DA) method was adopted to investigate the spectrum-effect relationship between the fingerprint peak area (X) of Jingu Tongxiao Pills and the anti-inflammatory activity (Y). The peak areas and anti-inflammatory activities of Jingu Tongxiao Pills are shown in Table 1 and Table 1 (continued). The VIP values of 16 variables in the OPLS-DA model were extracted to study the influence of common peaks on the difference in anti-inflammatory activity, and the results are shown in Figure 3 . As shown Figure 3 in A, according to the VIP values (VIP>1), Peak 10, Peak 9, Peak 11, Peak 6, Peak 4, and Peak 14 have significant contributions to the inhibition of NO production. Except that the chromatographic Peak 11 and chromatographic Peak 14 are not identified, the other chromatographic peaks are identified as salvianolic acid B, 1,2,3,4,6-O-pentagalloylglucose, paeoniflorin and gentiopicroside, respectively. As shown Figure 3As shown in B, according to the VIP value (VIP>1), Peak 10, Peak 9, Peak 11, Peak 6, Peak 4 and Peak 14 make significant contributions to reducing the IL-6 level. Among them, except that the chromatographic peaks No. 11 and No. 14 are not identified, the other chromatographic peaks are identified as salvianolic acid B, 1,2,3,4,6-O-pentagalloylglucose, paeoniflorin, gentiopicroside and albiflorin respectively. As shown Figure 3 As shown in C, according to the VIP value (VIP>1), Peak 10, Peak 6, Peak 9, Peak 4, Peak 3 and Peak 11 make significant contributions to reducing the TNF-α level. Among them, except that the chromatographic peak No. 11 is not identified, the other chromatographic peaks are identified as salvianolic acid B, paeoniflorin, 1,2,3,4,6-O-pentagalloylglucose, gentiopicroside and loganic acid respectively. Combining the OPLS-DA results, the pharmacodynamic components are finally determined to be the components of Peak 3, Peak 4, Peak 5, Peak 6, Peak 9 and Peak 10. After comparison with the reference substances (the chromatogram of the reference substances is shown in Figure 4 , where: 1-gallic acid; 3-loganic acid; 4-gentiopicroside; 5-albiflorin; 6-paeoniflorin; 8-liquiritin; 9-1,2,3,4,6-O-pentagalloylglucose; 10-salvianolic acid B; 13-cryptotanshinone), the above 6 peak components are designated as loganic acid, gentiopicroside, albiflorin, paeoniflorin, 1,2,3,4,6-O-pentagalloylglucose, and salvianolic acid B respectively.

[0105] 2.2 Determination of the content of effective components in Jingu Tongxiao Pills

[0106] 2.2.1 Preparation of solutions

[0107] Appropriate amounts of loganic acid, gentiopicroside, albiflorin, paeoniflorin, 1,2,3,4,6-O-pentagalloylglucose and salvianolic acid B are weighed precisely, dissolved in acetonitrile-0.1% phosphoric acid aqueous solution (5:95) after precise weighing, to prepare a mixed reference substance solution containing loganic acid (0.189 mg / mL), gentiopicroside (0.243 mg / mL), albiflorin (0.201 mg / mL), paeoniflorin (0.235 mg / mL), 1,2,3,4,6-O-pentagalloylglucose (0.211 mg / mL), and salvianolic acid B (0.394 mg / mL). The above reference substance solution and the sample solution of Jingu Tongxiao Pills are taken for injection and determination according to the chromatographic conditions under the item of "2.1.2", and the chromatograms are recorded, see Figure 1 and Figure 4 . The baseline of the chromatogram is stable, the retention time of the 6 components in the test solution is consistent with that of the reference substances, and the separation effect from adjacent chromatographic peaks is good.

[0108] 2.2.2 Methodological validation

[0109] 2.2.2.1 Investigation of linear relationship

[0110] Precisely pipette the mixed reference substance stock solution under the item "2.2.1" and dilute it to different concentrations, then perform the measurement under the chromatographic conditions under the item "2.1.2". Perform linear regression with mass concentration as the abscissa (X) and peak area as the ordinate (Y). The obtained regression equation results are shown in Table 2, and it can be seen that each component has a good linear relationship within its respective range.

[0111] Table 2 Linear equations of 6 compounds

[0112] compound Linear range (mg / mL) Regression equation R Strychnos nuciferic acid 0.00295~0.189 Y = 15273X – 9.2793 0.9999 Gentianoside 0.00379~0.243 Y = 25825X – 17.672 0.9999 Paeoniflorin 0.00314~0.201 Y = 11209X – 19.864 0.9993 paeoniflorin 0.00367~0.235 Y = 27628X – 43.675 0.9998 1, 2, 3, 4, 6-O-pentagalloylglucose 0.00329~0.211 Y = 14279X – 12.947 0.9999 Tanshinone B 0.00612~0.394 Y = 15309X – 28.565 0.9999

[0113] 2.2.2.2 Investigation of precision, stability and repeatability

[0114] Take the test sample solution (S1), and investigate the precision and repeatability of the method according to the method under the item "2.1.2". The results show that the RSD values of the peak areas of the 6 components are all less than 1.00%. In addition, take the test sample solution of the same batch, and measure the peak areas of the 6 components according to the method under the item "2.1.2" after 0, 3, 6, 12, 16, and 24 h respectively. The results show that the RSD values of the peak areas of the 6 components are all less than 2.10%, indicating that the method has good precision, stability and repeatability.

[0115] 2.2.2.3 Sample recovery test

[0116] Separately and precisely weigh 0.2 g of Jingu Tongxiao Wan samples (such as S1) with known contents of the components to be tested, for a total of 9 portions, and precisely add an appropriate amount of each reference substance solution respectively, so that the added amount of the reference substance is about 80%, 100% and 120% of the original content in the test sample. Then prepare the spiked test sample solution according to the method under the item "2.1.1", inject the sample for detection according to "2.1.2", and calculate the sample recovery rate of each component to be tested. The results show that the average sample recovery rate of each active component ranges from 95.70% to 104.58%, and the RSD value ranges from 2.20% to 4.80%, which all comply with relevant regulations, indicating that the method has good accuracy.

[0117] 2.2.3 Content determination of components

[0118] Accurately weigh the test solutions from 20 batches of samples, and determine the contents of loganic acid, gentiopicroside, paeoniflorin, paeoniflorin, 1,2,3,4,6-O-pentagalloylglucose, and salvianolic acid B in the 20 batches of samples according to the method under section "2.1.2". The results showed that the contents of loganic acid, gentiopicroside, paeoniflorin, paeoniflorin, 1,2,3,4,6-O-pentagalloylglucose, and salvianolic acid B in each batch of Jin Gu Tong Xiao Wan ranged from 3.325 to 4.000, 1.839 to 2.288, 2.536 to 3.700, 4.229 to 4.822, 0.302 to 0.430, and 3.396 to 3.884 mg·g, respectively. -1 See Table 3.

[0119] Table 3. Results of content determination of six components in Jin Gu Tong Xiao Wan (mg·g) -1 (n=3)

[0120] batch Strychnos nuciferic acid Gentianoside Paeoniflorin paeoniflorin 1,2,3,4,6-O-pentagalloylglucose Tanshinone B Total content S1 3.755 1.905 2.611 4.411 0.423 3.466 16.571 S2 3.644 1.839 2.759 4.501 0.362 3.449 16.554 S3 3.700 2.171 2.549 4.401 0.421 3.518 16.76 S4 3.833 2.250 2.647 4.519 0.358 3.553 17.16 S5 3.782 2.220 2.925 4.516 0.392 3.752 17.587 S6 3.897 2.288 2.536 4.615 0.373 3.884 17.593 S7 3.530 2.169 2.709 4.620 0.303 3.544 16.875 S8 3.394 2.136 2.538 4.304 0.302 3.421 16.095 S9 3.325 2.166 2.636 4.229 0.351 3.396 16.103 S10 3.545 2.259 2.859 4.437 0.369 3.423 16.892 S11 4.000 2.348 2.795 4.724 0.335 3.586 17.788 S12 3.622 2.126 2.857 4.565 0.354 3.539 17.063 S13 3.890 2.283 2.918 4.517 0.430 3.504 17.542 S14 3.757 2.205 2.806 4.528 0.372 3.763 17.431 S15 3.785 2.222 2.728 4.603 0.390 3.476 17.204 S16 3.744 2.198 2.727 4.676 0.354 3.414 17.113 S17 3.954 2.321 2.927 4.815 0.420 3.569 18.006 S18 3.745 2.198 2.691 4.822 0.349 3.522 17.327 S19 3.687 2.164 3.700 4.718 0.317 3.732 18.318 S20 3.614 2.121 3.600 4.556 0.398 3.612 17.901

[0121] 2.3 Anti-inflammatory effects of pharmacological components on LPS-induced RAW264.7 cells

[0122] 2.3.1 Preparation of the test solution

[0123] Take appropriate amounts of paeoniflorin, paeoniflorin lactone, gentiopicrin, loganic acid, salvianolic acid B, and 1,2,3,4,6-O-pentagalloylglucose reference standards, and prepare stock solutions of each at a concentration of 2 mM using pure water. Then, serially dilute these solutions with DMEM medium (containing sodium pyruvate) to concentrations of 100 µM, 50 µM, 25 µM, 12.5 µM, 6.25 µM, and 3.125 µM, respectively, for later use.

[0124] 2.3.2 Effects of pharmacological components on LPS-induced inflammatory factors in RAW264.7 cells

[0125] RAW 264.7 cells were cultured using standard methods, and cells in the logarithmic growth phase were seeded into 96-well plates. After static incubation to allow cell adhesion, 0.2 mL of complete culture medium was added to the control group; LPS was added to the model group at a final concentration of 1 μg / mL; and the drug-treated groups were treated with different concentrations of the six active ingredients. Except for the control group, all groups received LPS at a final concentration of 1 μg / mL. After 24 h of standard culture, the culture medium was aspirated, and the NO concentration in each group was determined using the Griess method [References 16-17, ibid.]. The TNF-α and IL-6 concentrations in each group were detected using ELISA [References 18-19, ibid.]. Because most compounds had weak anti-inflammatory activity and the maximum non-toxic concentration inhibition rate did not reach 50%, IC50 was used.30 The anti-inflammatory activity is quantitatively evaluated for the calculation of the effect component index. That is, the concentrations of paeoniflorin, albiflorin, gentiopicroside, loganic acid, salvianolic acid B, and 1,2,3,4,6-O-pentagalloylglucose at an inhibition rate of 30% are respectively calculated. This concentration is defined as the IC based on the pharmacological effect of inhibiting the expression of inflammatory factors NO, TNF-α, and IL-6 30 value. The results are shown in Figure 5 (Note: PGG-1, 2, 3, 4, 6-O-pentagalloylglucose).

[0126] 2.4 Construction of anti-inflammatory activity ECI for Jingu Tongxiao Pills

[0127] 2.4.1 Determination of anti-inflammatory weight coefficients of pharmacodynamic components

[0128] A custom weight coefficient method is adopted. Based on the IC of paeoniflorin, albiflorin, gentiopicroside, loganic acid, salvianolic acid B, and 1,2,3,4,6-O-pentagalloylglucose under the levels of inhibiting NO, TNF-α, and IL-6 30 values, the weight coefficients are respectively calculated according to the following formula. The results show that for the inflammatory factor NO, the order of the anti-inflammatory contribution rates of the 6 pharmacodynamic components is paeoniflorin = 1,2,3,4,6-O-pentagalloylglucose > gentiopicroside > loganic acid > salvianolic acid B > albiflorin; the order of contribution rates for TNF-α is paeoniflorin = 1,2,3,4,6-O-pentagalloylglucose > salvianolic acid B = albiflorin = gentiopicroside = loganic acid; the order of contribution rates for IL-6 is albiflorin = gentiopicroside > paeoniflorin = 1,2,3,4,6-O-pentagalloylglucose > loganic acid = salvianolic acid B. The results are shown in Table 4.

[0129]

[0130] Where W i is the weight coefficient of the pharmacodynamic component, n is the number of pharmacodynamic components, and i is the name of the pharmacodynamic component. The weight coefficients of paeoniflorin, albiflorin, gentiopicroside, loganic acid, salvianolic acid B, and 1,2,3,4,6-O-pentagalloylglucose based on inhibiting the expression of inflammatory factors NO, IL-6, and TNF-α calculated by the formula are shown in Table 4.

[0131] Table 4 Anti-inflammatory weight coefficients of pharmacodynamic components

[0132] Medicinal ingredients NO weighting coefficient IL-6 weighting coefficient TNF-α weighting coefficient paeoniflorin 0.37 0.15 0.41 Paeoniflorin 0.05 0.21 0.03 Gentianoside 0.08 0.21 0.03 Strychnos nuciferic acid 0.07 0.14 0.03 Tanshinone B 0.06 0.14 0.08 1, 2, 3, 4, 6-O-pentagalloylglucose 0.37 0.15 0.41

[0133] 2.4.2 Construction of anti-inflammatory effect component index

[0134] Using the comprehensive index evaluation method, the calculation formula for the anti-inflammatory effect component index is established as follows: adopting the custom weight coefficient method [Reference 19, supra], based on the IC of 6 pharmacodynamic components 30 values, according to the following formula:

[0135]

[0136] It can be written in detail as ECI (NO) = 0.37X 芍药苷 + 0.05X 芍药内酯苷 + 0.08X 龙胆苦苷 + 0.07X 马钱苷酸 + 0.06X 丹酚酸B + 0.37X 1, 2, 3, 4, 6-O-五没食子酰基葡萄糖 , ECI (IL-6) = 0.15X 芍药苷 + 0.21X 芍药内酯苷 + 0.21X 龙胆苦苷 + 0.14X 马钱苷酸 + 0.14X 丹酚酸B + 0.15X 1, 2, 3, 4, 6-O-五没食子酰基葡萄糖 , ECI (TNF-α) = 0.41X 芍药苷 + 0.03X 芍药内酯苷 + 0.03X 龙胆苦苷 + 0.03X 马钱苷酸 + 0.08X 丹酚酸B + 0.41X 1, 2, 3, 4, 6-O-五没食子酰基葡萄糖 , the ECI of 20 batches of Jingu Tongxiao Pills was calculated, as shown in Table 5. In the formula, ECI refers to the effect component index of Jingu Tongxiao Pills; Xi refers to the content of each pharmacodynamic component; Wi refers to the weight coefficient of each pharmacodynamic component; n refers to the number of pharmacodynamic components.

[0137] Table 5 Effect Component Index (ECI) of 20 Batches of Jingu Tongxiao Pills Medicinal Materials

[0138] serial number ECI (NO) ECI (IL-6) ECI (TNF-α) serial number ECI (NO) ECI (IL-6) ECI (TNF-α) S1 2.542 2.507 2.684 S11 2.695 2.635 2.901 S2 2.546 2.517 2.688 S12 2.599 2.558 2.787 S3 2.555 2.511 2.725 S13 2.641 2.581 2.869 S4 2.598 2.546 2.794 S14 2.618 2.498 2.840 S5 2.630 2.580 2.871 S15 2.635 2.518 2.805 S6 2.661 2.617 2.851 S16 2.640 2.527 2.791 S7 2.590 2.554 2.753 S17 2.760 2.637 2.941 S8 2.445 2.404 2.627 S18 2.697 2.590 2.820 S9 2.436 2.393 2.636 S19 2.703 2.575 3.025 S10 2.555 2.504 2.771 S20 2.652 2.528 2.956

[0139] When the content of each component in Jingu Tongxiao Pills is known, the anti-inflammatory effect component index value of Jingu Tongxiao Pills can be calculated according to the established effect component index formula, and the quality of the medicinal materials can be evaluated according to the magnitude of this value. In this formula, a larger weight coefficient of a pharmacodynamic component indicates that the component has a greater contribution rate to the efficacy of the medicinal material. It can be seen from the above established effect component index formula that paeoniflorin and 1,2,3,4,6-O-pentagalloylglucose have the largest weight coefficients, as high as 0.41, indicating that paeoniflorin and 1,2,3,4,6-O-pentagalloylglucose have a relatively large contribution rate to the anti-inflammatory effect of the medicinal materials. However, the indicator components in the current existing quality standard for Jingu Tongxiao Pills are cryptotanshinone reference substance, tanshinone I reference substance and tanshinone ⅡA, which shows that the current quality standard has the deficiency of being unable to reflect the pharmacodynamic effect.

[0140] 2.5 Verification of the anti-inflammatory activity ECI of Jingu Tongxiao Pills

[0141] To verify the reliability of the effective component index, GraphPad Prism software was used to conduct a correlation analysis between the ECI values of 20 batches of Jingu Tongxiao Pills and their effect values of inhibiting the expression of inflammatory factors NO, IL-6 and TNF-α in LPS-induced RAW264.7 cells. The results showed that the correlation coefficients of the two were 0.970 (linear equation: Y=4.400X-4.400), 0.951 (linear equation: Y=87.08X-193.5), and 0.960 (linear equation: Y=1587X+8.545), respectively. It can be seen that the ECI value is significantly positively correlated with the anti-inflammatory efficacy, indicating that the larger the effective component index, the lower the expression of inflammatory factors, that is, the stronger the efficacy and the better the quality of the finished drug. It can be seen that the quality of the finished drug can be controlled according to the effective component index. The correlation analysis is shown in Figure 6 .

[0142] In summary, this embodiment provides a method for evaluating the quality of Jingu Tongxiao Pills based on anti-inflammatory efficacy. The effective component index of Jingu Tongxiao Pills is calculated according to formula (I). The effective component index is positively correlated with the anti-inflammatory efficacy of Jingu Tongxiao Pills, and a higher effective component index indicates stronger anti-inflammatory efficacy and better quality of Jingu Tongxiao Pills;

[0143]

[0144] In formula (I):

[0145] ECI represents the effective component index of Jingu Tongxiao Pills;

[0146] i represents the pharmacodynamic components paeoniflorin, albiflorin, gentiopicroside, loganic acid, salvianolic acid B, and 1,2,3,4,6-O-pentagalloylglucose;

[0147] n=6, representing the number of pharmacodynamic components;

[0148] Xi represents the content of each pharmacodynamic component in Jingu Tongxiao Pills;

[0149] Wi represents the weight coefficient of each pharmacodynamic component, which is calculated according to formula (II):

[0150]

[0151] In formula (II), the definitions of i and n are the same as those in formula (I), and IC30 represents the concentration required for each pharmacodynamic component to inhibit 30% of the expression of inflammatory factors in the inflammation model.

[0152] In this method, the effective component indices of the Jin Gutongxiao pill to be evaluated and the standard Jin Gutongxiao pill that meets the quality standard can be measured simultaneously. If the effective component index of the Jin Gutongxiao pill to be evaluated is not lower than that of the standard Jin Gutongxiao pill, the quality of the Jin Gutongxiao pill to be evaluated is judged as qualified; otherwise, it is judged as unqualified.

[0153] Example 2: A device for evaluating the quality of Jin Gutongxiao pill based on the method of Example 1

[0154] The device for evaluating the quality of Jin Gutongxiao pill based on the method of Example 1 provided in this embodiment includes:

[0155] A data acquisition module, configured to acquire the content Xi of the active components paeoniflorin, albiflorin, gentiopicroside, loganic acid, salvianolic acid B and 1,2,3,4,6-O-pentagalloylglucose in the Jin Gutongxiao pill to be evaluated, as well as the IC30 value, which is the concentration required for each active component to inhibit 30% of the expression of inflammatory factors in an inflammation model; the inflammation model is selected from one or more of in vitro cell inflammation models and in vivo animal inflammation models, and the inflammatory factor is selected from one or more of NO, IL-6 and TNF-α.

[0156] A processing module, configured to input the IC30 value of each active component into formula (II) to obtain the weight coefficient Wi of each active component, then input the weight coefficient Wi of each active component and the content Xi of each active component in the Jin Gutongxiao pill to be evaluated into formula (I) to obtain the effective component index of the Jin Gutongxiao pill to be evaluated, and finally compare the effective component index of the Jin Gutongxiao pill to be evaluated with the effective component index of the standard quality Jin Gutongxiao pill in magnitude; formula (I) and formula (II) are as described previously; the effective component index of the standard quality Jin Gutongxiao pill is obtained using a Jin Gutongxiao pill that meets the quality standard;

[0157] An output module, configured to output whether the quality of the Jin Gutongxiao pill to be evaluated is qualified, wherein if its effective component index is not lower than that of the standard quality Jin Gutongxiao pill, the output is qualified, otherwise the output is unqualified.

[0158] Example 3: An electronic device

[0159] This embodiment provides an electronic device, including:

[0160] A processor;

[0161] A memory, configured to store a computer program;

[0162] When the computer program is executed by the processor, the method for evaluating the quality of Jin Gutongxiao pill based on anti-inflammatory efficacy in Example 1 is implemented.

[0163] Example 4: A computer-readable storage medium

[0164] This embodiment provides a computer-readable storage medium storing a computer program thereon. When the computer program is executed by a processor, it implements the method for evaluating the quality of Jin Gu Tong Xiao Wan based on the anti-inflammatory efficacy in Embodiment 1.

[0165] The purpose of the above embodiments is to specifically illustrate the substantive content of the present invention, but those skilled in the art should know that the scope of protection of the present invention should not be limited to the specific embodiments.

Claims

1. A method for evaluating the quality of Jingu Tongxiao Pills based on anti-inflammatory pharmacodynamic effect, characterized in that: The effective component index of Jingu Tongxiao pills is calculated according to formula (I), and the effective component index is positively correlated with the anti-inflammatory efficacy of Jingu Tongxiao pills, where a higher effective component index indicates stronger anti-inflammatory efficacy and better quality of Jingu Tongxiao pills; In formula (I): ECI represents the effective component index of Jingu Tongxiao pills; i represents the pharmacodynamic components: paeoniflorin, albiflorin, gentiopicroside, loganic acid, salvianolic acid B, and 1,2,3,4,6-O-pentagalloylglucose; n=6, representing the number of pharmacodynamic components; Xi represents the content of each pharmacodynamic component in Jingu Tongxiao pills; Wi represents the weight coefficient of each pharmacodynamic component, which is calculated according to formula (II): In formula (II), the definitions of i and n are the same as those in formula (I), and IC30 represents the concentration required for each pharmacodynamic component to inhibit 30% of the expression of inflammatory factors in the inflammation model.

2. The method according to claim 1, characterized in that: The effective component index is positively correlated with the anti-inflammatory efficacy of Jingu Tongxiao pills, with a correlation coefficient of no less than 0.

90.

3. The method according to claim 2, characterized in that: The effective component index is positively correlated with the anti-inflammatory efficacy of Jingu Tongxiao pills, with a correlation coefficient of no less than 0.

95.

4. The method according to claim 1, characterized in that: The inflammatory factor is selected from one or more of NO, IL-6 and TNF-α.

5. The method according to claim 1, characterized in that: The inflammation model is selected from one or more of in vitro cell inflammation models and in vivo animal inflammation models.

6. The method according to claim 5, characterized in that: The inflammation model is an in vitro inflammation model established by lipopolysaccharide-induced RAW264.7 cells.

7. A quality evaluation device for muscle and bone pain relief pills based on the method of any one of claims 1 to 6, characterized in that, Comprising: A data acquisition module, configured to acquire the content Xi of the pharmacodynamic components paeoniflorin, albiflorin, gentiopicroside, loganic acid, salvianolic acid B and 1,2,3,4,6-O-pentagalloylglucose in the Jingu Tongxiao pills to be evaluated, and the IC30 value of the concentration required for each pharmacodynamic component to inhibit 30% of the expression of inflammatory factors in the inflammation model; A processing module, configured to input the IC30 value of each pharmacodynamic component into formula (II) to obtain the weight coefficient Wi of each pharmacodynamic component, then input the weight coefficient Wi of each pharmacodynamic component and the content Xi of each pharmacodynamic component in the Jingu Tongxiao pills to be evaluated into formula (I) to obtain the effective component index of the Jingu Tongxiao pills to be evaluated, and finally compare the effective component index of the Jingu Tongxiao pills to be evaluated with the effective component index of standard quality Jingu Tongxiao pills; An output module, configured to output whether the quality of the Jingu Tongxiao pills to be evaluated is qualified, wherein it is output as qualified if the effective component index thereof is not lower than the effective component index of the standard quality Jingu Tongxiao pills, otherwise it is output as unqualified.

8. The quality evaluation device for Jin Gu Tong Xiao Wan according to claim 7, characterized in that: The effective component index of the standard quality Jingu Tongxiao pills is obtained by using Jingu Tongxiao pills that meet the quality standards.

9. An electronic device, characterized in that, Comprising: A processor; A memory, configured to store a computer program; When the computer program is executed by the processor, the method according to any one of claims 1 to 6 is implemented.

10. A computer-readable storage medium, characterized in that: A computer program is stored thereon, which when executed by a processor, implements the method according to any one of claims 1 to 6.

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