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Objective:To observe clinical effect on chronic atrophic gastritis （CAG） with deficiency of stomach Yin when modified Shaoyao Gancaotang combined with Yiguanjian used on patients， and to study influence to prognostic risk. Method:One hundred and ten cases patients were randomly divided into control group 54 cases and observation group 56 cases. Two groups got treatment of eradication of <italic>Helicobacter pylori </italic>（Hp）. Patients in control group got Yangweishu capsules， 3 grains/time， 2 times/day. Patients in observation groups when Shaoyao Gancaotang combined with Yiguanjian modified， 1 dose/day. The treatment continued for 16 weeks. Before and after treatment， gastroscope was discussed and operative link for gastritis assessment （OLGA） and operative link on intestinal-metaplasia assessment （OLGIM） were recorded， scores of atrophic degree， intestinal metaplasia degree， Hp infection， chronic inflammatory reaction， activity， endoscopic mucosal signs， dyspepsia symptoms， reported outcome patients reported outcomes （PRO） of patients with gastrointestinal diseases were graded. And to test levels of pepsinogenⅠ （PG Ⅰ），PGⅡ， gastrin-17 （G-17）， cytotoxin associated gene A （CagA） and vacuolar cytotoxin （VacA）. Result:The effect of patients treated in observation group was better than the effect of patients received measures in control group （<italic>Z</italic>=2.068， <italic>P</italic><0.05）. Scores of atrophic degree， intestinal metaplasia degree， chronic inflammatory reaction and activity were lower than the scores of patients received measures in control group （<italic>P</italic><0.01）. Levels of OLGA （<italic>Z</italic>=2.037， <italic>P</italic><0.05） and OLGIM （<italic>Z</italic>=2.014， <italic>P</italic><0.05） of patients treated in observation group were better than the levels of patients received measures in control group. And levels of PGⅠ， G-17 and PG Ⅰ/PGⅡ of patients treated in observation group were higher than the dates of patients received measures in control group （<italic>P</italic><0.01）. Positive rates of CagA and VacA were all lower than the dates in control group （<italic>P</italic><0.05）. And gastroscopic mucosal signs and dyspepsia symptoms and scores of PRO were all lower than dates of patients received measures in control group （<italic>P</italic><0.01）. Conclusion:On the basis of eradicating Hp， treatment of Shaoyao Gancaotang with Yiguanjian modified can relieve the clinical symptoms， improve the quality of life of patients， promote mucosal repair， improve the pathological changes， reduce the degree and scope of gastric mucosal atrophy， which is conducive to delay the progression of lesions and reduce the risk of canceration when it used on CAG. It has good clinical efficacy and was worthy of further study
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Objective:Aiming at the residue of Shaoyao Gancaotang， the extraction， qualitative and quantitative study of the small molecule resource components were carried out to clarify the residual small molecule chemical components in the residue and explore the ways of its resource utilization. Method:The ultra-performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry （UPLC-QTOF-MS/MS） was used to qualitatively identify the residual small molecule substances in the dregs of Shaoyao Gancaotang. Agilent C<sub>18</sub> reversed-phase chromatographic column （3.0 mm×100 mm， 2.7 µm） was used at the flow rate of 0.4 mL·min<sup>-1</sup>， the injection volume was 5 µL， and the mobile phase was gradient eluted with 0.05% formic acid aqueous solution （A）-acetonitrile （B） （0-1 min， 14%-17.5%B； 1-3 min， 17.5%-19%B； 3-4 min， 19%-20%B； 4-5 min， 20%B； 5-6 min， 20%-21%B； 6-9 min， 21%B； 9-22 min， 21%-36%B； 22-23 min， 36%B； 23-32 min， 36%-43%B）， electrospray ionization （ESI） was employed with negative ion mode scanning and scanning range of <italic>m</italic>/<italic>z</italic> 50-1 200. A high performance liquid chromatography （HPLC） was established for the quantitative analysis of its main components with Agilent C<sub>18</sub> reversed-phase chromatographic column （4.6 mm×150 mm， 5 µm）， the detection wavelength was set at 235 nm， the flow rate was 0.8 mL·min<sup>-1</sup>， and the injection volume was 5 µL. Mobile phase was 0.05% phosphoric acid （A）-acetonitrile （B） for gradient elution （0-1 min， 14%-19%B； 1-4 min， 19%B； 4-18 min， 19%-50%B）. The content changes of main components in the residue of Shaoyao Gancaotang were compared before and after two different techniques of organic solvent extraction and enzymatic extraction. Result:A total of 16 chemical components in the residue of Shaoyao Gancaotang were qualitatively analyzed， and quantitative analysis found that there were many chemical components in the residue， among which the residues of 6 index components such as paeoniflorin and liquiritin reached more than 70% in the original decoction piece. After enzymolysis by cellulase， liquiritin in the residue could be converted into liquiritigenin. The content of crude polysaccharide in enzymatic extract of the residue was 6 times higher than that in the blank group， and the content was up to 12%. Conclusion:There are still many small molecule resource components in the residue of Shaoyao Gancaotang， which has great development potential. Organic solvents can be used to re-extract the target components in the residue， and liquiritin can be converted into liquiritigenin by biological fermentation technology， and the crude polysaccharide from the residue can be extracted by enzymatic method to develop animal feed. This study can provide reference basis and approach for reusing the residues of Shaoyao Gancaotang preparations and dispensing granules， so as to realize the high-value utilization of Shaoyao Gancaotang.
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Objective:To investigate the spectrum-effect relationship of effect of material components in Shaoyao Gancaotang on the levels of malondialdehyde (MDA), superoxide dismutase (SOD) and adenosine triphosphatase (ATPase) in brain tissue of epileptic mice, and to reveal the material basis of the antiepileptic effect of Shaoyao Gancaotang. Method:HPLC was used to establish the fingerprint of 15 batches of Shaoyao Gancaotang, the mobile phase was consisted of acetonitrile (A)-0.1% phosphoric acid aqueous solution (B) for gradient elution (0-15 min, 19%A; 15-45 min, 19%A-50%A; 45-46 min, 50%A-90%A; 46-48 min, 90%A), the flow rate was 1.0 mL·min-1, and the detection wavelength was 237 nm. The mouse epilepsy model was induced by penicillin, the protective effect of 15 batches of Shaoyao Gancaotang on MDA, SOD and ATPase levels in brain tissue of epileptic mice was investigated. Grey correlation analysis was used to analyze the correlation between the peak areas of characteristic peaks in the fingerprint of 15 batches of Shaoyao Gancaotang and three pharmacodynamic indexes (the activities of MDA, SOD and ATPase), and the mathematical model of spectrum-effect relationship between different material components and pharmacodynamic indexes was established. Result:Shaoyao Gancaotang could increase the contents of ATPase and SOD and reduce the content of MDA in the brain tissue of epileptic mice, and most of the differences were significant (P<0.05, P<0.01). There were 28 common peaks in the fingerprint of 15 batches of Shaoyao Gancaotang, and 15 of them were characteristic peaks. Grey correlation analysis found that the characteristic peaks contributed to the activities of MDA, SOD and ATPase included peak 2 (albiflorin), peak 3 (paeoniflorin), peak 5 (liquiritin), and so on, the specific ranking was peak 3>peak 6>peak 12>peak 8>peak 2>peak 5>peak 9>peak 4>peak 10>peak 7>peak 13>peak 11>peak 15>peak 1>peak 14. Conclusion:Shaoyao Gancaotang can affect the activities of SOD, MDA and ATPase in brain tissue of epileptic mice by multi-component synergy. Simultaneously, introducing the grey correlation analysis into the correlation evaluation of the spectrum-effect relationship between components and efficacy of Chinese herbal compounds, it can objectively reflect the essence of the synergistic action of multiple components in traditional Chinese medicine (TCM), and it is an effective analysis method for screening and predicting the pharmacodynamic components of TCM.
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Shaoyao Gancaotang, first seen in Treatise on Febrile and Miscellaneous Diseases, is composed of Paeoniae Radix Alba and Glycyrrhizae Radix et Rhizoma Praeparata cum Melle in equal proportion.It has the functions of preserving Yin to nourish blood, harmonizing liver and spleen, relieving spasm and pain.This formula is applied in leg and foot spasm and abdominal pain caused by blood deficiency, body fluid consuming and the unmoistened muscles and veins.It has been highly praised and used by medical experts throughout the ages and has extended its application scope.Modern pharmacological studies have shown that Shaoyao Gancaotang has significant effects in antispasmodic, analgesic, antitussive and other areas, and is used to treat spastic diseases, painful diseases, inflammatory diseases and so on.This paper will systematically elaborate the historical evolution, compatibility analysis, pharmacological and pharmacodynamic studies, modern clinical application of Shaoyao Gancaotang, in order to provide theoretical basis and reference for the development of this famous classical formula.
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Objective::To analyze and identify the metabolites of major components of Shaoyao Gancaotang in urine and feces of normal and polycystic ovary syndrome (PCOS) rats, and to explore the effect of PCOS on the metabolism of Shaoyao Gancaotang. Method::Female SD rats were randomly divided into normal group and PCOS group. PCOS rat model was prepared by administration of letrozole solution for 21 days, and the estrous cycle of rats was observed. UHPLC-QTRAP-MS/MS technique and LightSight 2.3 software were used to analyze and identify the metabolites of major components of Shaoyao Gancaotang in urine and feces of normal and PCOS rats. The mobile phase was water-methanol for gradient elution, the flow rate was 0.3 mL·min-1, electrospray ion source was employed under negative ion mode. Result::Phase Ⅰ and phase Ⅱ reactions mainly occurred in drug metabolism. A total of 27 metabolites were detected in urine of normal rats, and 34 metabolites were detected in urine of PCOS rats. A total of 29 metabolites were detected in feces of normal rats, and 27 metabolites were detected in feces of PCOS rats. Conclusion::The metabolites in the urine of PCOS rats are more diverse than those of normal rats, and the disease status of PCOS may affect the in vivo metabolic pathway of active ingredients in Shaoyao Gancaotang.
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Object:To analyze the effect of muscular tension injury in spasticity of cerebral apoplexy(SCA) rats with spasticity and the expression of γ-aminobutyric acid receptor (GABAα), γ-aminobutyric acid transporter (GAT-1) protein and mRNA expression in cerebral cortex modified Shaoyao Gancaotang, electroacupuncture Quchi, Yanglingquan and acupuncture combined with herb therapies alleviate the effect of SCA. Method:Randomly divided into two groups, divided into blank group, sham operation group, model group, modified Shaoyao Gancaotang group, acupuncture combined with herb group, electroacupuncture group, selected qualified SD rats into the group, 9 rats in each group. The modified Zea-Longa thread embolism method+internal capsule injection of NMDA receptor method was used to prepare a rat model. After the behavioral score confirms the success of the model,dispose of each group separately,electroacupuncture group (electroacupuncture on both sides of Yangling Spring and Quchi,1 time/d for 5 days), modified Shaoyao Gancaotang group (with Shaoyao Gancaotang 1 time/d,10 mL·kg-1 for 5 days),acupuncture combined with herb (modified Shaoyao Gancaotang was given, then electroacupuncture treatment was continued for 5 days). After treatment, the muscle tension was detected by behavioral analysis. Real-time fluorescent quantitative PCR（Real-time PCR） and Western blot were used to detect the expression of GABAα and GAT-1 related mRNA and protein in the cortex. Result:Compared with normal group, the muscle tension of the model group increased (P<0.01), the expression of GABAα mRNA and protein decreased (P<0.01), and the expression of GAT-1 mRNA and protein increased (P<0.01), compared with model group, the muscle tone score of the each treatment group decreased (P<0.05), the expression of GABAα mRNA and protein increased (P<0.05，P<0.01), and the expression of GAT-1 mRNA and protein decreased (P<0.05，P<0.01), of which the corresponding expression in the acupuncture combined with herb group was the most significant. Conclusion:Electroacupuncture Quchi, Yanglingquan,modified Shaoyao Gancaotang and acupuncture combined with three therapies can alleviate the muscle tension of limbs spasm of stroke,and the combination of acupuncture and medicine treatment on the cortical γ-aminobutyric acid receptor the highest expression efficiency suggests that it has the best effect on improving the excitatory spasticity of stroke limbs.
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Objective:To investigate the therapeutic effect and possible mechanism of four types of Chinese herbal moisturizers made in laboratory for atopic dermatitis induced by 2，4-dinitrofluorobenzene （DNFB） in mice. Method:According to the body weight， BALA/c mice were randomly divided into normal group， model group， blank cream group （moisturizer A）， Shaoyao Gancaotang group （moisturizer B）， Shaoyao Gancaotang with Portulacae Herba，Ginseng Radix et Rhizoma and Honeysuckle Stem group （moisturizer C）， and Shaoyao Gancaotang with Ginseng Radix et Rhizoma and Honeysuckle Stem group （moisturizer D） ， with the dose of 25 g·kg-1 per day， as well as tacrolimus ointment group of 3 g·kg-1 per day， with 10 to 12 mice in each group. Except the normal group， the mice in the other groups were treated with 0.5% DNFB in the hair removal skin of back， 100 μL each for 7 days. Starting from the 7th day， each group was given the appropriate skin cream for external use intervention， once per day， for 15 consecutive days， except for the normal and the model groups. The animal body mass was measured once a week， and the animal back skin was graded three times a week， and the skin lesion score was recorded. After the mice were killed， the left and right ears were taken， the weight of both ears was punched and the degree of swelling was calculated. The back skin was fixed and stained with hematoxylin-eosin（HE） method， and then pathologic examination was conducted to observe and score the pathological changes of mouse back skin. Blood was obtained after the last dose and enzyme-linked immunosorbent assay （ELISA） was used to determine the immunoglobulin（Ig）E content in serum. Western blot was used to measure the expression of signal transduction and activator of transcription 3 （STAT3）， phosphorylation （p）-STAT3 in the skin tissue. Result:Compared with the normal group， the body mass decreased continuously， a series of inflammatory changes such as erythema， edema， dryness， desquamation and callus exfoliation and so on occurred in the modeling area， and the skin lesion score increased significantly in the model group. Additionally， the cuticle of ear skin was thickened and the degree of ear swelling was obviously increased in the model group. Microscopically， the occurred changes in the model mice included the local necrosis of the epidermis， epidermal thickening， epidermal hyperplasia， and the hyperkeratosis and hypokeratosis in the cuticle， as well as the subcutaneous inflammatory cell infiltration and so on. Furthermore， the content of serum IgE andthe expression of p-STAT3 in skin tissues increased significantly in the model group. Compared with the model group， the body mass of mice in group C and D was significantly increased （P<0.01）， and the skin lesion status score was decreased （P<0.05，P<0.01）.The degree of auricle swelling was significantly reduced in group B， C and D compared with that in the model group （P<0.01）.The degree of skin necrosis and defect and epidermal hyperplasia of mice in moisturizer C group was significantly reduced compared with that in model group （P<0.05，P<0.01）. Serum IgE levels of mice in group C and D were significantly lower than those in the model group （P<0.05，P<0.01）. The expression of p-STAT3 protein in skin tissues of mice in moisturizer C group was significantly lower than that in model group （P<0.05）. Conclusion:The moisturizers B， C and D all have certain therapeutic effect on atopic dermatitis， among which moisturizers C has the most obvious therapeutic effect. The possible mechanism may be that it reduces the level of inflammatory cytokines by inhibiting the increase of serum IgE content and the phosphorylation of STAT3.
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Objective:To establish the HPLC fingerprint test method for the medicinal materials, decoction pieces and substance benchmarks of Shaoyao Gancaotang for investigating the quality transmitting of substance group in preparation process of the medicinal materials-decoction pieces-substance benchmarks, then to evaluate the scientificity and rationality of preparation process of substance benchmarks of Shaoyao Gancaotang by combining with yields of dry extract, transfer rates of effective components and other indexes. Method:Substance benchmarks of Shaoyao Gancaotang was prepared according to the method recorded in ancient medical books, fingerprints of 15 batches of medicinal materials, decoction pieces and substance benchmarks were detected by HPLC, and the contents of effective ingredients were determined. At the same time, the correlation analysis of quality transmitting of substance group during the preparation of substance benchmarks was carried out by combining the yields of dry extract and transfer rates of effective components. Result:The established HPLC fingerprint method has good precision, repeatability and stability, it can be used for the simultaneous determination of fingerprint of medicinal materials, decoction pieces and substance benchmarks. In the fingerprint of substance benchmarks, 16 common peaks were determined by taking liquiritin as the reference peak, of which 6 chromatographic peaks belong to Paeoniae Radix Alba and 11 chromatographic peaks belong to Glycyrrhizae Radix et Rhizoma, 7 major chromatographic peaks were identified. The similarities of fingerprints of 15 batches of medicinal materials, decoction pieces and substance benchmarks of Shaoyao Gancaotang were good by comparing with their respective reference fingerprints(≥ 0.90), the average dry extract rate of 15 batches of substance benchmarks was 24.81%, and no discrete data were found; the average transfer rates of paeoniflorin, liquiritin and glycyrrhizic acid from decoction pieces to substance benchmarks were 79.68%, 63.70% and 51.20%, respectively, and no discrete data were found. Conclusion:In this paper, a scientific and reasonable method for evaluating the process of substance benchmarks of Shaoyao Gancaotang is established by means of the fingerprint method controlled by the whole substance group, the research idea of quality transmitting of substance group in the preparation process, and the evaluation of technical and economic indicators. It can be used as a reference for the evaluation and research of material benchmarks in other famous classical formulas.
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Objective: To discuss the effect of Juantong decoction on phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)signaling pathway proteins in rats with endometriosis (EMS). Method: The EMS Rat model was established by autologous endometrial transplantation. And 48 rats were randomly divided into 6 group, namely the sham operation group, the model group, the low-dose Juantong decoction group, the middle-dose Juantong decoction group, high-dose Juantong decoction group, and the PI3K pathway blocker group (LY294002). Then, the high-dose Juantong decoction group, the middle-dose Juantong decoction group and the low-dose Juantong decoction group were given different doses (42.9,14.3,4.8 g·kg-1). The pathway blocker group (LY294002) was given LY294002 (0.04 g·kg-1) through peritoneal injection weekly. The sham operation group and the model group were given saline irrigation (10 mL·kg-1). The administration lasted for 28 days. At last, the ectopic endometrial tissues in rats were observed by transmission electron microscope, the proteins of PI3K, Akt and mTOR in the tissue were detected by immunohistochemical method, and the p70 ribosomal S6 kinase (p70S6K) mRNA expression of ectopic endometrial tissue was tested by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) method. Result: Compared with the normal group, the protein expressions of PI3K, Akt and mTOR and the mRNA expression of p70S6K of ectopic endometrium in the model group increased significantly (PPConclusion: The proteins of PI3K/Akt/mTOR signaling pathway participate in the occurrence of endometriosis. Juantong decoction can inhibit the activity of epithelial mesenchymal cells and promote apoptosis by reducing the protein expressions of PI3K, Akt and mTOR and the mRNA expression of P70S6K in endometriotic tissues of model rats, thus inhibiting the PI3K/Akt/mTOR signaling pathway in the treatment of endometriosis.
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Objective:To observe the effect of Shaoyao Gancaotang and its disassembled prescriptions on the expressions of transient receptor potential vanilloid 3 (TRPV3) and transient receptor potential vanilloid 4 (TRPV4) in myocardium and hypothalamus of rats in the ultra-high temperature environment, in order to explore the mechanism of Shaoyao Gancaotang and its disassembled prescriptions against the high temperature. Method:Seventy five male SPF-grade SD rats were randomly divided into 5 groups according to their body weight, namely normal control group, high temperature normal control group, Shaoyao Gancaotang group (2.16 g·kg-1), single Paeoniae Radix Alba(PRA) group (1.08 g·kg-1) and single Glycyrrhizae Radix et Rhizoma(GRR) group (1.08 g·kg-1). Except normal control group, the remaining groups were fed in artificial climate boxes for 13 days. The normal control group and the high temperature normal control group were given the same volume of pure water. Western blot and quantitative real-time fluorescence polymerase chain reaction(Real-time PCR) were used to determine expressions of TRPV3, TRPV4 protein and mRNA in myocardium and hypothalamus of each group. Result:Compared with the normothermic control group, the expressions of TRPV3 protein, TRPV4 mRNA, TRPV3 protein and TRPV3 mRNA in the myocardium of the hyperthermic control group were significantly higher than those of the normal control group (PPPConclusion:TRPV3, TRPV4 channels in rat myocardium and hypothalamus were activated in the high temperature environment. The anti-hyperthermic mechanism of Shaoyao Gancaotang and its disassembled prescriptions was related to the regulation of TRPV3, TRPV4 channel protein and mRNA expressions in rat myocardium and hypothalamus.