Treatment of Bone and Joint Diseases with Gancao Fuzitang： A Review / 中国实验方剂学杂志

Gancao Fuzitang originates from the Treatise on Febrile Diseases and Miscellaneous Diseases （《伤寒杂病论》） and is mainly used to treat pain in the bones and joints and symptoms such as no flexion or extension. It has the effect of tonifying the spleen and kidney and removing dampness and turbidity， so it is widely used in the clinical treatment of various bone and joint diseases. This article reviewed the clinical research and mechanism of Gancao Fuzitang in the treatment of bone and joint diseases. The research has found that this prescription has good efficacy in treating bone and joint diseases such as rheumatoid arthritis， rheumatoid arthritis， ankylosing spondylitis， gout， and intervertebral disc herniation. Its mechanism of action may be related to regulating the level of inflammatory factors， antioxidation， and the protein expression of inflammatory and apoptotic cell-related pathways， improving bone and joint diseases， and alleviating related symptoms. This study can provide a reference for further deepening the research on the prevention and treatment of bone and joint diseases with Gancao Fuzitang.

Separation, characterization and anti-psoriasis effect of self-assembled nanoparticles from Shaoyao Gancao Decoction / 中国中药杂志

This study aims to separate and characterize self-assembled nanoparticles(SAN) from Shaoyao Gancao Decoction(SGD) and determine the content of active compounds. Further, we aimed to observe the therapeutic effect of SGD-SAN on imiquimod-induced psoriasis in mice. The separation of SGD was performed by dialysis, and the separation process was optimized by single factor experiment. The SGD-SAN isolated under the optimal process was characterized, and the content of gallic acid, albiflorin, paeoniflorin, liquiritin, isoliquiritin apioside, isoliquiritin, and glycyrrhizic acid in each part of SGD was determined by HPLC. In the animal experiment, mice were assigned into a normal group, a model group, a methotrexate group(0.001 g·kg~(-1)), and SGD, SGD sediment, SGD dialysate, and SGD-SAN groups of different doses(1, 2, and 4 g·kg~(-1)) respectively. The psoriasis grade of mice was evaluated based on the pathological changes of skin lesions, the content of inflammatory cytokines, organ index and other indicators. The results showed that SAN obtained by centrifugation at 13 000 r·min~(-1) for 30 min was stable after dialysis for 4 times, which were uniform spherical nanoparticles with the particle size of(164.43±1.34) nm, the polydispersity index of(0.28±0.05), and the Zeta potential of(-12.35±0.80) mV. The active compound content accounted for more than 70% of SGD. Compared with the model group, SAN and SGD decreased the skin lesion score, spleen index, and inflammatory cytokine levels(P<0.05 or P<0.01) and alleviated the skin thickening and infiltration of inflammatory cells. However, the sediment group and the dialysate group had no obvious effect. SGD showed a good therapeutic effect on imiquimod-induced psoriasis in mice, and SAN demonstrated the effect equivalent to SGD in a dose-dependent manner. Therefore, we conclude that the SAN formed during decocting is the main active form of SGD, which can lower the levels of inflammatory cytokines, promote the normal differentiation of keratinocytes, and reduce the infiltration of inflammatory cells in the treatment of psoriasis lesions in mice.

Based on the macromolecules in traditional Chinese medicine exploring the material basis and mechanism of the analgesic effect of Shaoyao Gancao Decoction / 药学学报

In order to study the analgesic effect of Shaoyao Gancao Decoction, this paper discussed material basis and mechanism from the perspective of macromolecules in traditional Chinese medicine. Inspired by the phenomenon of turbidity after boiling Chinese medicine, this experiment took Shaoyao Gancao Decoction as the research object to study the formation process of precipitation during boiling. The results showed that aggregates with a certain shape were formed in the solvent during the boiling process, and the precipitate was obtained by standing and centrifuging. Analysis found that the precipitation was mainly composed of small molecules such as paeoniflorin, albiflorin, liquiritin, glycyrrhizic acid, isoliquiritin and gallic acid, and macromolecules such as protein and polysaccharide. The composition of precipitate was consistent with that of Shaoyao Gancao Decoction, but the analgesic effect of Shaoyao Gancao Decoction after removing the precipitate was significantly reduced. Based on these results, we isolated small molecular compounds, polysaccharides and protein from Shaoyao Gancao Decoction and their contents are 60.4, 700.7 and 207.2 mg·g-1 respectively. We get the ratio, polysaccharide: small molecule = 11.6∶1, protein: small molecule = 3.4∶1, the precipitate is prepared in the state of boiling. The characterization results showed that the particle size of the precipitate will change significantly after co-heating, and the content determination results showed that the content of the six small molecular compounds which was free in solvent was significantly reduced after the formation of the precipitate. The acetic acid writhing experiment proved that the precipitate has a good analgesic effect, and effectively reduced the levels of inflammatory factors prostaglandin E2 and nitric oxide, and increased the level of anti-inflammatory factor interleukin-10. These results proved that the precipitate in Shaoyao Gancao Decoction is an important material basis for analgesic effect, and macromolecules such as protein and polysaccharide are the main components of the precipitate. The study of macromolecules in the precipitate of Shaoyao Gancao Decoction not only provides new ideas and methods for elucidating the pharmacodynamic material basis of Shaoyao Gancao Decoction, but also provides a reference for analyzing the scientificity of traditional decoction.

Effects of Shaoyao gancao decoction on intestinal flora in rats with acute lung injury / 中国药房

OBJECTIVE To study the improvement effects of Shaoyao gancao decoction （SGD） on acute lung injury （ALI） in rats and its effects on the intestinal flora. METHODS Sixty SD rats were randomly divided into normal group （CON group， normal saline）， model group （MOD group， normal saline）， positive control group （DEX group， 5 mg/kg dexamethasone）， SGD low-dose， medium-dose and high-dose groups （SGD-L， SGD-M， SGD-H groups， 5.8， 11.6， 23.2 g/kg， calculated by crude drug）， with 10 rats in each group. Each group was given relevant medicine 10 mL/kg intragastrically， for 7 consecutive days. Thirty minutes after the last administration， CON group was given constant volume of normal saline via airway infusion， and other groups were given lipopolysaccharide （5 mg/kg） via airway infusion to induce ALI model. After 12 hours of modeling， the lung tissue wet/dry weight ratio was calculated， and the contents of interleukin 1β （IL-1β）， IL-6 and tumor necrosis factor α（TNF-α） in rat bronchial alveolar lavage fluid （BALF） were all detected； the pathological changes of lung tissue were observed after hematoxylin-eosin staining. The intestinal flora of rat feces was analyzed by 16S rRNA sequencing technology， and the correlation of differential bacteria genera with inflammatory factors was also analyzed. RESULTS Compared with MOD group， the infiltration of inflammatory cells in the lung tissue of rats in each SGD dose group was decreased， and the thickening of alveolar septum and pulmonary edema improved； lung tissue wet/dry weight ratio， the levels of IL-1β， IL-6 and TNF-α in BALF significantly decreased （P＜0.05 or P＜0.01）. SGD （low dose） could improve the intestinal flora disorder in ALI rats， restore the diversity and richness of intestinal flora， regulate the structure of flora， reduce the abundance of Lactobacillus， Streptococcus and Escherichia-Shigella， and increase the abundance of Firmicutes， Lachnospira， Ruminococcus， Clostridia，Dubosiella and Akkermansia. Through correlation analysis， it was found that the relative abundance of Lactobacillus， Streptococcus and Escherichia-Shigella was positively related to the levels of inflammatory factor IL-1β， IL-6 and TNF-α （P＜0.05 or P＜0.01）. The relative abundance of Lachnospira， Dubosiella， Firmicutes was significantly negatively correlated with the levels of inflammatory factors mentioned above （P＜0.05 or P＜0.01）. CONCLUSIONS SGD may improve ALI by reducing lung tissue injury and inflammatory response and regulating flora structure in rats.

Therapeutic mechanism of Guizhi Gancao Decoction for heart failure: a network pharmacology-based analysis / 南方医科大学学报

OBJECTIVE@#To predict the targets and pathways in the therapeutic mechanism of Guizhi Gancao Decoction (GZGCD) against heart failure (HF) based on network pharmacology.@*METHODS@#The chemical components of GZGCD were analyzed using the databases including TCMSP, TCMID and TCM@Taiwan, and the potential targets of GZGCD were predicted using the SwissTargetPrediction database. The targets of HF were obtained using the databases including DisGeNET, Drugbank and TTD. The intersection targets of GZGCD and HF were identified using VENNY. Uniport database was used to convert the information, and the components-targets-disease network was constructed using Cytoscape software. The Bisogene plug-in, Merge plug-in, and CytoNCA plug-in in Cytoscape software were used for protein-protein interaction (PPI) analysis to acquire the core targets. Metascape database was used for GO and KEGG analysis. The results of network pharmacology analysis were verified with Western blot analysis. Three factors (PKCα, ERK1/2 and BCL2) were screened according to the degree value of network pharmacology results and the degree of correlation with heart failure process. The pentobarbtal sodium was dissolvein H9C2 cells treated with serum-free high glucose medium to simulate the ischemic anoxic environment of heart failure. The total proteins of myocardial cells were extracted. The protein contents of PKCα, ERK1/2 and BCL2 were determined.@*RESULTS@#We identified a total of 190 intersection targets between GZGCD and HF using Venny database, involving mainly the circulatory system process, cellular response to nitrogen compounds, cation homeostasis, and regulation of the MAPK cascade. These potential targets were also involved in 38 pathways, including the regulatory pathways in cancer, calcium signal pathway, cGMP-PKG signal pathway, and cAMP signal pathway. Western blot analysis showed that in an in vitro H9C2 cell model of HF, treatment with GZGCD downregulated PKCα and ERK1/2 expressions and upregulated BCL2 expression.@*CONCLUSION@#The therapeutic mechanism of GZGCD for HF involves multiple targets including PRKCA, PRKCB, MAPK1, MAPK3, and MAPK8 and multiple pathways including the regulatory pathway in cancer and the calcium signaling pathway.

Effects of self-assembled nanoparticles from Shaoyao gancao decoction on the in vitro release and intestinal absorption of the main components of Glycyrrhiza uralensis / 中国药房

OBJECTIVE To study the effects of self-assembled nanoparticles from Shaoy ao gancao decoction （SGD-SAN）on the in vitro release and intestinal absorption of the main components of Glycyrrhiza uralensis . METHODS Gancao single decoction （GSD），Shaoyao single decoction （SSD），mixed suspension of Shaoyao and Gancao single decoction （MSSGD）and SGD （i.e. Shaoyao-Gancao decoction ）were prepared ，and SAN was characterized. HPLC method was adopted to determine the contents of 7 main components （liquiritin apioside ， liquiritin， isoliquiritin apioside ， isoliquiritin， liquiritigenin， glycyrrhizic acid ， isoliquiritigenin）in G. uralensis . The dialysis bag method was used to investigate the effects of the formation of SGD-SAN on in vitro release of 7 main components in G. uralensis with pH 1.2 HCl solution and pH 6.8 phosphate buffered solution （PBS）as release media. Single-pass intestinal perfusion study was performed to investigate the effects of the formation of SGD-SAN on the intestinal absorption of 7 main components from G. uralensis . RESULTS SAN with particle size of 200-300 nm and polydispersity index of 0.3-0.5 was found in GSD ，MSSGD and SGD. GSD-SAN and MSSGD-SAN were in rod shape while SGD-SAN was irregularly spherical under transmission electron microscope. The results of in vitro release study showed that the formation of SGD-SAN could significantly increase in vitro release of liquiritigenin ，isoliquiritigenin and glycyrrhizic acid ，and had no effect on other components of G. uralensis in pH 1.2 HCl solution. The formation of SGD-SAN also had no effect on the release of each component from G. uralensis in pH 6.8 PBS. The results of intestinal perfusion experiments showed that the formation of SGD-SAN could significantly promote the absorption of each component from G. uralensis in the ileum. CONCLUSIONS- The formation of SGD-SAN significantly improves the in vitro release of poorly soluble components from G. uralensis and promotes the intestinal absorption of main components from G. uralensis ，which is the physical structure basis for the compatibility and synergy of Paeonia lactiflora and G. uralensis .

Spectrum-effect relationship of anti-gastric ulcer effect of Shaoyao gancao decoction / 中国药房

OBJECTIVE To study the sp ectrum-effect relationship of anti-gastric ulcer effect of Shaoyao gancao decoction. METHODS Eleven batches of Shaoyao gancao decoction were prepared ；gastric ulcer model was established by anhydrous ethanol modeling method. Using Weikangling as positive control ，the effects of Shaoyao gancao decoction on the contents of defensive factors [nitric oxide （NO），epidermal growth factor （EGF），superoxide dismutase （SOD）] and attack factor [malondialdehyde （MDA）] in gastric tissue of model rats were investigated. HPLC fingerprints of 11 batches of Shaoyao gancao decoction were established and similarity evaluation was performed with Similarity Evaluation System of Traditional Chinese Medicine Chromatographic Fingerprint （2004A edition ）； common peaks were identified by comparing with mixed control. The spectrum-effect relationship of Shaoyao gancao decoction against gastric ulcer was analyzed based on the grey correlation analysis. RESULTS Eleven batches of Shaoyao ganyao decoction could significantly decrease the content of MDA in gastric tissue ，while increased the contents of NO ，EGF and SOD in gastric ulcer model rats （P＜0.01），and had a certain inhibitory effect on the gastric ulcer. There were 23 common peaks in chromatograms of 11 batches of samples ，and the similarity with the control fingerprint was not less than 0.9. By comparing with mixed control ，7 common peaks were identified ，namely gallic acid （peak 5），albiflorin（peak 9），paeoniflorin（peak 10），liquiritin（peak 12），isoliquiritin apioside （peak 14），isoliquiritoside（peak 15）， glycyrrhizic acid （peak 22）. The average correlation degree of 7 identified common peaks and pharmacodynamic indexes were greater than 0.6，of which peak 22（glycyrrhizic acid ），peak 10（paeoniflorin）and peak 12（liquiritin）had the largest correlation ， and their values were 0.807，0.772 and 0.770 respectively. RESULTS The anti-gastric ulcer effect of Shaoyao gancao decoction is the result of the synergistic effect of multiple components ，among which glycyrrhizic acid ，paeoniflorin and liquiritin may be the main pharmacodynamic components.

Quantitative identification of TCM Q-markers based on analytical hierarchy and the entropy weight comprehensive method by taking Shaoyao Gancao decoction as an example / 药学学报

The quality markers (Q-markers) of traditional Chinese medicine (TCM) have become a topic of interest in TCM research in recent years. Nonetheless, there is still no consensus on how to scientifically characterize TCM Q-markers. Our study establishes an identification method for TCM Q-markers based on the analytical hierarchy process (AHP) and the entropy weight comprehensive method. By constructing an evaluation system encompassing the target layer, the factor layer and the control layer, AHP can be used to analyze the weight of three core TCM quality attributes, including effectiveness, testability and specificity. Following that, the entropy weight method is employed to analyze the specific indicators for each attribute based on the literature and experimental data. Finally, the comprehensive weight of each index is obtained by combining the two weights, and the comprehensive weight and the specific value of each component is multiplied and summed to obtain the integrated score ranking, and thereby identify the TCM Q-markers. Taking Shaoyao Gancao decoction as an example, the analysis revealed that the top 8 components are as follows: paeoniflorin > quercetin > albiflorin > glycyrrhizic acid > naringenin > liquiritin > oxypaeoniflorin > benzoylpaeoniflorin, and can be identified as Q-markers of Shaoyao Gancao decoction. This study not only provides support for the establishment of quality standards and process quality control of TCM formulae, but also provides innovative ideas and methods for quantitative evaluation and accurate identification of TCM Q-markers.

Mechanisms of Gancao Xiexin Decoction on protecting intestinal epithelial cell barrier under stress state by regulating PERK-elF2α-CHOP signaling pathway / 中国药理学通报

Aim To observe the protective effect of Gancao Xiexin Decoction (GXD) on intestinal epithelial cells (IECs) under endoplasmic reticulum stress from the perspective of PERK-elF2α-CH0P signaling pathway. Methods Caco-2 cells were cultured and divided into normal control group (NC), model control group(MC), low-dose GXD group(GXD-L), mediumdose GXD group (GXD-M) and high-dose GXD group (GXD-H). Models of the stress epithelial cells were induced by tunicamycin(Tm), and GXD groups were treated with GXD contained serum at the same time. The cell survival rate was assessed by CCK-8 method, the cell apoptotic rate and cell cycle distribution were determined by flow cytometry, and the cell barrier permeability was determined by TEER and FITC-dextran method; the expression levels of core proteins of PERK-elF2α-CH0P signaling pathway were detected by Western blot. Results Compared with MC group, GXD intervention could improve cell survival rate(P < 0. 05), reduce their apoptotic rate(P <0. 01), relieve cell cycle arrest(P < 0. 01), improve cell barrier permeability by increasing cell TEER value (P < 0. 01) and decreasing FITC-dextran concentration (P < 0.05), and the levels of p-PERK, p-elF2α, ATF4 and CHOP in GXD-M and GXD-H groups all visibly descended (P < 0. 01). Conclusions GXD can reduce the excessive apoptosis of IECs and protect intestinal epithelial cell barrier homeostasis by inhibiting the signal transduction of PERK-elF2α-CH0P apoptotic pathway.

Effect of self-assembled nanoparticles from Shaoyao Gancao Decoction on release and absorption of main components of Baishao / 中国中药杂志

To study the effect of self-assembled nanoparticles from Shaoyao Gancao Decoction(SGD-SAN) on the encapsulation, in vitro release and intestinal absorption of the main components of Baishao. Particle size analysis and morphological observation were used to verify the formation of SGD-SAN in the decoction. The entrapment efficiency(EE) of SGD-SAN on the main components of Baishao was determined by ultrafiltration centrifugation. The dialysis bag method was used to study the in vitro release of the main components of Baishao with pH 6.8 phosphate buffer solution as the release media. Single-pass intestinal perfusion study was performed to investigate the effect of SGD-SAN on the absorption of the main components of Baishao. The results showed that there were nanoparticles in the SGD, and the particle sizes and PDI of SGD-SAN were about 200 nm and 0.38, respectively. SGD-SAN was irregularly spherical under transmission electron microscope(TEM). The EEs of albiflorin, paeoniflorin and benzoylpaeoniflorin in SGD-SAN were 33.78%±1.03%,33.61%±0.90%,88.53%±0.58%, respectively. The release characteristics of albiflorin, paeoniflorin and benzoylpaeoniflorin from SGD-SAN showed a slow-release effect on pH 6.8 phosphate buffer solution media. SGD-SAN could significantly enhance the absorption of albiflorin, paeoniflorin and benzoylpaeoniflorin in the ileum. The results of this study indicated that SAN could be formed during the mixed decoction of Baishao and Gancao, and SGD-SAN could encapsulate the components of Baishao, with a certain slow-release effect, and the formation of SAN facilitated the absorption of drugs in the ileum.

Effect of Gancao Fuzi Decoction on osteoarthritis and proteomics of articular cartilage in rats / 中国中药杂志

The rat osteoarthritis model was replicated by injection of sodium iodoacetate into the knee joint cavity, and the effects of Gancao Fuzi Decoction on rat osteoarthritis and the proteome of articular cartilage were investigated. Sixty SD rats weighing 230-250 g were randomly divided into normal group, model group, glucosamine sulfate group, and Gancao Fuzi Decoction high, medium and low dose groups. Osteoarthritis model was induced by intra-articular injection of sodium iodoacetate(3 mg on each leg) in all groups except the normal group. After modeling, each administration group was given intragastric administration for 1 month. During the administration period, joint pain test and joint width measurement were performed every week to observe the autonomous behavior of rats. Enzyme linked immunosorbent assay(ELISA) method was used to detect the contents of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), matrix metalloproteinase-3(MMP-3), and matrix metalloproteinase tissue inhibitor(TIMP-1) in rat joint lavage fluid. Hematoxylin-eosin(HE) staining was used to observe bone and joint morphology. Nano-LC-LTQ-Orbitrap system was used to detect arti-cular cartilage proteins. The results showed that, compared with the model group, Gancao Fuzi Decoction could significantly improve joint pain and joint swelling in osteoarthritis rats, significantly reduce the contents of TNF-α, IL-1β and MMP-3 in the joint cavity la-vage fluid, increase the content of TIMP-1, and relieve inflammatory diseases such as enlarged joint space, rough cartilage edge, different thickness of cartilage layer, and disordered arrangement of chondrocytes. After comparing the proteins between the groups, 273 differential proteins were screened out. KEGG analysis found that the above differential proteins involved 43 signaling pathways such as systemic lupus erythematosus, among which 11 signaling pathways were related to osteoarthritis. The above results indicated that Gancao Fuzi Decoction had a preventive effect on osteoarthritis, and its mechanism of action may be accomplished by regulating the protein expression of osteoarthritis-related signal pathways.

Mechanism of Shaoyao Gancao Decoction in treatment of rheumatoid arthritis based on network pharmacology / 中草药

Objective: To study the mechanism of Shaoyao Gancao Decoction in the treatment of rheumatoid arthritis (RA) based on network pharmacology. Methods: The active components of Shaoyao Gancao Decoction were obtained from traditional Chinese medicine systems pharmacology (TCMSP) database, and human target proteins corresponding to active components were searched in Swiss Target Prediction and TCMSP database. The targets of RA were collected through Therapeutic Target Database and Drugbank database. The key targets of Shaoyao Gancao Decoction to treat RA were screened by building the Venn diagram. And the key protein interaction (PPI) network model was constructed by STRING database. The gene ontology (GO) and pathway enrichment analysis were performed by DAVID database. All of the correlative results were visualized by Cytoscape 3.7.2, and the network feature analysis was made by Network Analyzer. Results: A total of 102 compounds were obtained from Shaoyao Gancao Decoction, with 310 corresponding targets, and 68 common targets of Shaoyao Gancao Decoction-RA were obtained. Herb-compound-target-pathway network showed that kaempferol, quercetin, formononetin, naringenin, isorhamnetin were the key compounds of Shaoyao Gancao Decoction in the treatment of RA, and PTGS2, NOS2, MAPK14, PPARG, IL-6, TNF, and IL-1β were the key targets. GO entries included 28 biological process entries, two cellular component entries, and 13 molecular function entries. There were 40 pathways involving TNF signaling pathway, osteoclast differentiation, T cell receptor signaling pathway, MAPK signaling pathway, steroid hormone biosynthesis and toll-like receptor signaling pathway. Conclusion: The results of this study verify the multi-component, multi-target and multi-pathway regulation characteristics of Shaoyao Gancao Decoction, preliminarily predict material basis and mechanism of Shaoyao Gancao Decoction in the treatment of RA, which provides reference for further research.

Potential synergic mechanism of Wutou-Gancao herb-pair by inhibiting efflux transporter P-glycoprotein / 药物分析学报

Wutou-Gancao herb-pair is extensively used to attenuate the toxicity and enhance the efficacy of aconite. In this study, potential synergic mechanism of the herb pair was investigated by utilizing multiple ap-proaches. In silico and in vitro Caco-2 cell models were applied to study the potential binding mode of bioactive ingredients existing in liquorice with P-glycoprotein (P-gp), as well as the inhibition effects on P-gp. Additionally, anti-inflammatory activity of aconitine (AC) combined with active ingredients of liquorice, as well as pharmacokinetic patterns of AC after co-administration was investigated. Anti-inflammatory effect of AC (1 mg/kg) in rats was enhanced in combination with bioactive ingredients of liquorice (10 mg/kg). In the meanwhile, the exposure of AC in vivo was altered, in terms of Cmax and AUC. For instance, the Cmax and AUC were increased to 1.9 and 1.3 folds, respectively, when used in combination with liquiritigenin. The in silico study revealed the potential binding mode with outward facing conformation of P-gp. The resulting data obtained from transport of rhodamine-123 (Rh-123) across Caco-2 cell monolayer further indicated that the function of P-gp was inhibited by chemicals in liquorice. The synergic effect was therefore proposed to be attributed to inhibition of P-gp by liquorice since AC has been demonstrated to be the substrate of P-gp. The resuls revealed that potential synergic mechanism of Wutou-Gancao herb-pair by inhibiting function of key efflux transporter P-gp to enhance the exposure of AC in systematic circulation, and further the anti-inflammatory effect, which helps clarify the compatibility rationale of these two herbs.

Mechanism of cAMP-PKA Signaling Pathway Mediated by Shaoyao Gancao Decoction () on Regulation of Aquaporin 5 and Muscarinic Receptor 3 Levels in Sjögren's Syndrome / 中国结合医学杂志

OBJECTIVE@#To investigate the mechanism of cAMP-PKA signaling pathway mediated by Chinese medicine formula Shaoyao Gancao Decoction (, SGD) on the regulation of aquaporin 5 (AQP5) and muscarinic receptor 3 (M3R) levels in Sjögren's syndrome (SS).@*METHODS@#Of the 30 mice, 5 were randomly selected as control, and others were used for creating SS model. After successful modeling, mice were randomly divided into 5 groups (n=5 per group) and intragastrically administered with saline (8 mL/kg), pilocarpine (1.4 mg/kg), or low, medium and high doses SGD (0.14, 0.21, 0.35 g/kg Radix paeoniae with 0.01 g/kg Radix glycyrrhizae, respectively) for 6 weeks. Human labial gland acinar cells were treated with pilocarpine or varying doses of SGD with saline as the placebo. Hematoxylin and eosin staining was used to observe the histopathological changes of the submandibular glands of mice. The serum levels of anti-SS antigen A (SS-A), anti-SS antigen B (SS-B), M3R, and α-fodrin in submandibular glands of mice were measured by enzyme-linked immunosorbent assay. Immunofluorescence staining was used to observe the spatial localization of AQP5 and M3R in acinar cells. Reverse transcriptase polymerase chain reaction and Western blot were used to detect the expressions of PKA, cAMP, Epac1, AQP5, M3R, nuclear factor kappa-B (NF-κB), and tumor necrosis factor (TNF)-α in submandibular gland tissues and cells of each group.@*RESULTS@#Compared to normal mice, body weight, 5-min salivary secretion, 30-min secretion of tears and breakup time of tear film of model mice decreased at 1-6 weeks after immunization (all P<0.05), whereas water intake increased (all P<0.05). In the model group, glands of the submandibular glands showed atrophy, accompanied by acini of different sizes, decreased numbers and loose arrangement, with catheter dilatation and different degrees of lymphocyte infiltration. Conditions of mice in SGD groups were improved. The positive expression of AQP5 and M3R were higher in the acinar cells treated with all doses SGD compared to the normal group; serum levels of SS-A, SS-B, and α-fodrin were lower, and that of M3R was higher in all doses SGD treated animals than the model or pilocarpine treated ones (all P<0.05). Compared to the model and pilocarpine groups, the mRNA and protein levels of NF-κB and TNF-α were lower in mice or cells treated with medium or high-dose SGD (all P<0.05), while those of PKA, Epac1, AQP5 and M3R were higher (all P<0.05).@*CONCLUSION@#SGD can improve symptoms of SS by regulating the cAMP-PKA signaling pathway and increasing AQP5 and M3R levels.

Effect of Modified Shaoyao Gancao Tang on MPTP-induced Parkinson's Disease Model Mice / 中国实验方剂学杂志

Objective:To study the effect of modified Shaoyao Gancao Tang on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced Parkinson's disease model mice. Method:The sixty C57/BL6 mice were randomly divided into normal group, model group, Madopar group (50 mg·kg-1) and low, medium and high doses modified Shaoyao Gancao Tang (1,2,4 g·kg-1). The modeling method was to intraperitoneally inject C57/BL6 mice with MPTP(40 mg·kg-1) once a day for 7 days. Except normal group and model group were given normal saline daily,drug-administered group was intragastrical administered once a day, and the third day after the drug was administered according to the above method (except normal group). The behavior of each group of mice (climbing test, swimming test, and tail suspension test) was examined up to 15 days after administration. Subsequently, the levels of Cystatin-C (Cys-C) and neuron-specific enolase (NSE) in the serum of each group were detected by enzyme-linked immunosorbent assay (ELISA). The levels of superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) in striatum of each group were detected by spectrophotometry. The protein expression of α-synuclein (α-syn), tyrosine hydroxylase (TH),B lymphoma-2 gene (Bcl-2) and Bcl-2 related X protein (Bax) in the striatum of each group was detected by Western blot. Result:Compared with normal group, the swimming experiment scores and suspension experiments scores in the behavioral experiments of model group mice were significantly lower (PPPPPPPα-syn and Bax expression in striatum were significantly increased (PPPPPPPPα-syn and Bax expression in striatum were significantly decreased (PPPPPPPPα-syn expression and Bax expression in the striatum were significantly decreased (PConclusion:The neuroprotective effect of modified Shaoyao Gancao Tang on MPTP-induced Parkinson's disease model mice may be achieved by inhibiting oxidative damage and apoptosis.

Effect of Gancao Fuzitang on Synovial Fibroblast-like Cell Proliferation in Adjuvant Arthritis Mice Model / 中国实验方剂学杂志

Objective:To investigate the effect of Gancao Fuzitang on the proliferation of fibroblast-like synoviocytes (FLS) in synovial tissue of mice with adjuvant-induced arthritis (AA). Method:The 48 male Balb/c mice were randomly divided into 4 groups:normal control group, model group(arthritis induced through injection with adjuvant), Gancao Fuzitang group and triptolide group. The normal control group and the model control group were orally given physiological saline. After the successful modeling, the mice of the other two groups were treated with Gancao Fuzitang or triptolide orally for 18 days. Then the animals were euthanized, the paw edema of the animals was measured, and arthritic ankles were observed by histological examination. Tumor necrosis factor (TNF)-α was examined by immunoassays. The proliferation of synovial cells of foot joints was detected by immunohistochemical expressions of Vimentin. The expressions of Cyclin D1, proliferating cell nuclear antigen (PCNA), p53 and p21 were detected by Western blot. Result:Compared with the normal control group, the mice of synovial hyperplasia and bone erosion in model group were obvious. The joint swelling degree, TNF-α level and proliferation of FLS were increased (P1,PCNA were up-regulated(PPα, cell cycles (Cyclin D1 and PCNA), and increased protein expressions of p53 and p21 (PConclusion:Gancao Fuzitang has a therapeutic effect on AA mice, and the mechanism might be associated with its anti-inflammatory effect and the effects in regulating Cyclin D1, PCNA, p53 and p21 expressions and inhibiting FLS proliferation.

Clinical Efficacy of Modified Guizhi Gancao Longgu Muli Tang in Treating Menopausal Insomnia and Its Effect on Sleep Quality and Neurotransmitter / 中国实验方剂学杂志

Objective: To explore the clinical efficiency of modified Guizhi Gancao Longgu Muli Tang in treating menopausal insomnia.Method: A total of 134 female cases with menopausal insomnia in our hospital from January 2016 to June 2018 were selected as research objectives and randomly divided into observation group (67 cases) and control group (67 cases).Oral zopiclone was provided to control group,and modified Guizhi Gancao Longgu Muli Tang combined with oral zopiclone was provided to observation group.All cases were treated for 4 weeks.The two groups'clinical effects,sleep quality[pittsburgh sleep quality index (PSQI) score],neurotransmitter[glutamate (Glu),γ-aminobutyric acid (GABA),5-hydroxytryptamine (5-HT),dopamine (DA)]levels before and after treatment,as well as adverse reactions were compared.Result: Observation group's overall effective rate was 92.5%(62/67),which was significantly higher than 80.6%(54/67) of control group (PPPPPPConclusion: Modified Guizhi Gancao Longgu Muli Tang can improve sleep quality,alleviate symptoms of insomnia,and correct neurotransmitter metabolic disorder for patients with menopausal insomnia,with an exact curative effect and a higher tolerance of patients.

Effect of Shoutaiwan and Shaoyao Gancao Tang on Recurrent Spontaneous Abortion / 中国实验方剂学杂志

Objective: To observe the effect of Shoutaiwan and Shaoyao Gancao Tang on recurrent spontaneous abortion due to spleen and kidney Qi deficiency. Method: Totally 80 eligible patients were randomly divided into the control group and the treatment group, with 90 cases in each group. The control group was treated with lymphocyte immune; the Shoutaiwan and Shaoyao Gancao Tang combined with lymphocyte immune was applied in the treatment group. Coagulation function[the part plasminogen activator inhibitor-1 (PAI-1), fibrinogen (FIB), type plasminogen activator (t-PA), prothrombin time (PT), D-dimer (D-D) and tissue factor (TF), fibrin peptide A (FPA), fibrinogen (Fig), Plasminase-antifibrinolytic complex (PAP), glycoprotein platelet particle-140 (GMP-140), thrombin-antithrombin complex (TAT)], serum closed antibody (CD3+,CD4+,CD8+,CD4+/CD8+), inflammatory factor interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), Interleukin-1β (IL-1β), interleukin-10 (IL-10)]were scored after the treatment. The rates of birth and adverse reactions in each group were compared. The traditional Chinese medicine(TCM) syndrome differentiation of abortion spleen and kidney Qi deficiency was scored. Result: 5 cases were lost during the study period. The term delivery rate was 92.0%in the treatment group, which was higher than 73.6%in control group (PP+, CD4+, CD4+/CD8+), inflammatory factors (IFN-γ,TNF-α,IL-1β) and TCM syndrome scores were significantly lower in the treatment group than that in control group (P+), inflammatory factors IL-10 were significantly increased compared with the control group (PPConclusion: Shoutaiwan and Shaoyao Gancao Tang have a remarkable effect on recurrent spontaneous abortion case by spleen and kidney Yang deficiency.

Effect of Gancao Xiexintang Combined with Vitamin B12 on T Lymphocyte Subset, Ulcer Area, Oral Flora in Children with Recurrent Oral Ulcer / 中国实验方剂学杂志

Objective:To investigate the clinical efficacy of Gancao Xiexintang combined with vitamin B12 on recurrent oral ulcer (ROU) in children. Method:Totally 116 children with ROU admitted in Jiangxi Provincial Children's Hospital from June 2016 to June 2018 were divided into observation group and control group on the basis of random number table,with 58 cases in each group. The control group was orally treated with vitamin B12, while the observation group was orally treated with Gancao Xiexintang in addition to the therapy of the control group. All of the children were treated for 14 days. Clinical efficacy, changes of T lymphocyte subset (CD3+, CD4+, CD8+, CD4+/CD8+), ulcer area, content of oral flora (veillonella, streptococcus) before and after treatment, and side effect were compared between the two groups. Result:The overall effective rate of the observation group was 96.6%(56/58), which was much higher than 84.5%(49/58) of the control group (P+, CD4+, CD4+/CD8+in peripheral blood, but lower CD8+compared with those before treatment (P+, CD4+, CD8+, CD4+/CD8+) indexes in observation group was improved more significantly (PPPPPConclusion:In treating ROU in children, the combination of Gancao Xiexintang and vitamin B12 can significantly correct imbalance of T lymphocyte subset, promote the recovery of oral ulcer, and positively regulate oral micro-ecological environment, with an exact curative effect and high patient tolerance.

Pharmacodynamics and mechanism of Shaoyao Gancao Intragastric Floating Tablets based on unification of medicines and excipients / 中草药

Objective: To study the pharmacodynamic effect of “unification of medicines and excipients” and the mechanism of action about Shaoyao Gancao Intragastric Floating Tablets (SGIFT). Methods: Rabbits were randomly divided into control group and model group, Shenmei Yangwei Granule group (1.0 g/kg), Shaoyao Gancao Decoction extract group (1.0 g/kg), low-dose SGIFT group (1.0 g/kg), high-dose SGIFT group (1.5 g/kg), chitosan blank excipients group (1.0 g/kg). The acute gastric ulcer model was established in the blank excipient group without chitosan (1.0 g/kg). Six rats in each group, fasting for 24 h after 13 d of continuous administration, and gastric administration of anhydrous ethanol (2.5 mL/single). The blood and gastric tissue were taken out after 1.5 h to observe the pathological damage in each group. The expression of GAS, MDA, PG, epidermal growth factor (EGF) in serum and EGF, PGE2, TFF1 in gastric tissue of each group were detected. Results: Compared with the model group, the other groups can improve the gastric histopathology of rabbits with acute gastric ulcer effectively. The content of EGF in serum and EGF, PGE2, and TFF1 in gastric tissue of rabbits was increased in varying degrees, and the content of GAS, MDA, and PG in serum was all decreased in different degrees. Conclusion: SGIFT had certain therapeutic effects on rabbits with anhydrous ethanol gastric ulcer. The mechanism might be related to protecting gastric mucosa, increasing the content of protective factors, and reducing the content of attack factors.