Sarmentosin Induces Autophagy-dependent Apoptosis via Activation of Nrf2 in Hepatocellular Carcinoma.

Background and Aims: Hepatocellular carcinoma (HCC) is a common and deadly cancer. Accumulating evidence supports modulation of autophagy as a novel approach for determining cancer cell fate. The aim of this study to evaluate the effectiveness of sarmentosin, a natural compound, on HCC in vitro and in vivo and elucidated the underlying mechanisms. Methods: Cell functions and signaling pathways were analyzed in HepG2 cells using western blotting, real-time PCR, siRNA, transmission electron microscopy and flow cytometry. BALB/c nude mice were injected with HepG2 cells to produce a xenograft tumour nude mouse model for in vivo assessments and their tumors, hearts, lungs and kidneys were isolated. Results: We found that autophagy was induced by sarmentosin in a concentration- and time-dependent manner in human HCC HepG2 cells by western blot assays and scanning electron microscopy. Sarmentosin-induced autophagy was abolished by the autophagy inhibitors 3-methyladenine, chloroquine, and bafilomycin A1. Sarmentosin activated Nrf2 in HepG2 cells, as shown by increased nuclear translocation and upregulated expression of Nrf2 target genes. Phosphorylation of mTOR was also inhibited by sarmentosin. Sarmentosin stimulated caspase-dependent apoptosis in HepG2 cells, which was impaired by silencing Nrf2 or chloroquine or knocking down ATG7. Finally, sarmentosin effectively repressed HCC growth in xenograft nude mice and activated autophagy and apoptosis in HCC tissues. Conclusions: This study showed sarmentosin stimulated autophagic and caspase-dependent apoptosis in HCC, which required activation of Nrf2 and inhibition of mTOR. Our research supports Nrf2 as a therapeutic target for HCC and sarmentosin as a promising candidate for HCC chemotherapy.

Simultaneous Evaluation of Dissolution and Absorption Study of Compound Danshen Tablets and Capsules Based on Cellular Electrical Sensing Model.

This study explores the dissolution mechanism and absorption process of compound Danshen tablets (CDTs) and compound Danshen capsules (CDCs) in vitro. Taking the cell index as the evaluation index of dissolution and absorption of multi-component solid preparations of CDTs and CDCs, it breaks through the idea of traditional research. We used real-time cell-based assay (RTCA) to provide a new idea and method for the consistency evaluation of traditional Chinese medicine (TCM) compound preparations. The drug dissolution and absorption simulation system (DDASS) was established to obtain the dissolution and absorption samples of compound Danshen solid preparations at different time points. The cell index (CI) of the sample to H9C2 cells was detected by RTCA technology, and the dissolution and absorption percentage were calculated based on this index to obtain the dissolution and absorption kinetics model. Meanwhile, one batch of tablets and one batch of capsules (batch numbers ZKC1816 and 202101001) were selected to conduct the overall animal pharmacodynamic experiment to verify the feasibility of drug effect evaluation with cell index as an indicator. The best fitting model of dissolution curves of each batch of CDTs and CDCs is the Weibull model. There was a good correlation (r > 0.86) between the dissolution-absorption-pharmacodynamic curve. Based on RTCA technology, we have established the comprehensive evaluation method for cell biology of compound Danshen solid preparations in line with the overall concept of TCM and a synchronous evaluation system of dissolution and absorption in vitro of new TCM compound solid preparations.

[Mechanism of Qingwei Powder in treatment of periodontitis based on UPLC-Q-TOF-MS, GC-MS, network pharmacology and molecular docking].

The present study explored the mechanism of Qingwei Powder(QP) in the treatment of periodontitis based on chromatography-mass spectrometry and network pharmacology-molecular docking techniques. UPLC-Q-TOF-MS and GC-MS were used to identify the chemical constituents of QP. The active components and targets were screened out through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), and their targets were predicted using SwissTargetPrediction. Targets related to periodontitis were obtained from GeneCards, OMIM, and DisGeNET. Venn diagram was constructed using Venny 2.1 to obtain the intersection targets. Cytoscape 3.7.2 was used to construct the "chemical component-target-disease" network. The targets were analyzed for Gene Ontology(GO) function and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment by clusterProfiler R, and the "chemical component-target-pathway" network was constructed. The binding activity of the active components to the target proteins was verified by molecular docking. A total of 189 chemical components were obtained by UPLC-Q-TOF-MS and GC-MS, including 39 active components with 180 potential targets related to periodontitis. Target enrichment analysis of the active components yielded 92 KEGG pathways. Twenty KEGG pathways, 34 active components, and 99 targets were involved in the "chemical component-target-pathway" network. Molecular docking verified a good binding ability of the key targets to the key compounds. This study preliminarily indicates that QP is effective in treating periodontitis through multiple components, multiple targets, and multiple pathways, which reflects the complex system of Chinese medicine. This study provides the theoretical foundation for the subsequent research on the material basis and key quality attributes of QP.

[Quality value transmitting of substance benchmarks in Danggui Buxue Decoction].

To clarify the key quality attributes of substance benchmarks in Danggui Buxue Decoction(DBD), this study prepared 21 batches of DBD substance benchmarks, and established two methods for detecting their fingerprints, followed by the identification of peak attribution and similarity range as well as the determination of extract and transfer rate ranges and contents of index components ferulic acid, calycosin-7-O-ß-D-glucoside, and astragaloside Ⅳ. The mass fractions and transfer rates of DBD substance benchmarks from different batches were calculated as follows: ferulic acid(index component in Angelicae Sinensis Radix): 0.037%-0.084% and 31.41%-98.88%; astragaloside Ⅳ(index component in Astragali Radix): 0.021%-0.059% and 32.18%-118.57%; calycosin-7-O-ß-D-glucoside: 0.002%-0.023% and 11.51%-45.65%, with the extract rate being 18.4%-36.1%. The similarity of fingerprints among 21 batches of DBD substance benchmarks was all higher than 0.9. The quality control method for DBD substance benchmarks was preliminarily established based on the HPLC fingerprint analysis and index component determination, which has provided a basis for the subsequent development of DBD and the quality control of novel related preparations.

Simultaneous determination of eight flavonoids in Sedum sarmentosum Bunge from different areas by UHPLC with triple quadrupole MS/MS.

Sedum sarmentosum Bunge (SSB) is a traditional Chinese herbal medicine containing multiple components that has been extensively used clinically to treat chronic viral hepatitis and some inflammatory diseases. Total flavonoids are major pharmacologically active components of SSB. To gain a deeper understanding of SSB resources, we analyzed eight chemical constituents in 33 batches of SSB from 11 regions in China. An accurate, precise and sensitive ultra-high-performance liquid chromatography coupled with triple quadrupole electrospray tandem mass spectrometry method was developed for the determination of eight flavonoids in SSB. Under the optimized chromatographic conditions, good separation for the eight target components was obtained on an Agilent Zobax SB C18 (50 × 2.1 mm, 5 µm) column within 4 min. The established methods were validated with good linearity (r ≥ 0.9988), precision (RSD ≤ 2.68%), stability (1.43-3.28%) and repeatability (1.14-2.89%). Moreover, the average recoveries were 95.91-100.68%, and the RSDs were 1.50-3.80%. In addition, the analytical conditions of UPLC-ESI-MS/MS provided better sensitivity with a shorter analysis time when compared with the HPLC-DAD method. Hierarchical clustering analysis and principal component analysis were performed to estimate and classify these samples based on the contents of the eight chemical constituents. This study provided the theoretical basis and scientific evidence for the development and utilization of SSB resources.

Celastrol inhibits colorectal cancer through TGF-ß1/Smad signaling.

BACKGROUND: There are few clinical challenges associated with the treatment of colorectal cancer (CRC). Studies have shown that TGF-ß plays a crucial role in CRC. Importantly, celastrol, a major components of the root extract of the traditional Chinese herb Tripterygium wilfordii Hook F, has been shown to inhibit the growth, adhesion, and metastasis of human CRC cells through the inhibition of TGF-ß1/Smad signaling. MATERIALS AND METHODS: Real-time PCR and Western blot tests were proceeded to present TGF-ß1, TGF-ß receptor type I (TGFßRI), TGF-ß receptor type II (TGFßRII), Smad2/3, p-Smad2/3, Smad4, and glyceraldehyde-3-phosphate dehydrogenase expression in human colon cancer cell samples. RESULTS: Our results indicated that celastrol can reduce the expression levels of TGF-ß1, TGFßRI, and TGFßRII in HCT116 and SW620 cells. Furthermore, celastrol could also prevent the increase in Smad4 and p-Smad2/3 in HCT116 and SW620 cells. CONCLUSION: Celastrol could inhibit tumor growth through TGF-ß1/Smad signaling and might be a promising therapeutic component against CRC.

Hyperoside alleviated N-acetyl-para-amino-phenol-induced acute hepatic injury via Nrf2 activation.

N-acetyl-para-amino-phenol (APAP) acute hepatic injury is receiving increasing attention. In the present study, we examined the effects of Hyperoside (Hype) on APAP-induced acute hepatic injury. Oral administration of Hype dose-dependently attenuated the index of hepatic injury, including the production of AST, ALT, and ALP. Increased glutathione (GSH) and decreased ROS production induced by Hype demonstrated its potential antioxidant capacity. In addition, Nrf2 and its downstream genes were markedly activated by Hype. Furthermore, enhanced levels of SOD, GST, and GSH-Px were markedly suppressed by Hype in a dose-dependent manner. At the same time, decreased LPO was also detected in Hype-treated mice. The in vitro study verified a protective effect of Hype on APAP-induced injuries in LO2 cells. Moreover, the regulatory effect was found to be mostly dependent on Nrf2 which decreased LDH and ALT generation and increased cell viability. Nrf2-silenced LOS cells were sensitive to APAP-induced injury, while Hype did not exhibit any further effects on LO2 cells, which demonstrate the critical role of Nrf2 in this process. Taken together, our results demonstrated the ability of Hype to inhibit APAP-induced acute hepatic injury and its potential use in the treatment of Nrf2-associated diseases.

Application of Real-Time Cell Electronic Analysis System in Modern Pharmaceutical Evaluation and Analysis.

Objective: We summarized the progress of the xCELLigence real-time cell analysis (RTCA) technology application in recent years for the sake of enriching and developing the application of RTCA in the field of Chinese medicine. Background: The RTCA system is an established electronic cellular biosensor. This system uses micro-electronic biosensor technology that is confirmed for real-time, label-free, dynamic and non-offensive monitoring of cell viability, migration, growth, spreading, and proliferation. Methods: We summarized the relevant experiments and literature of RTCA technology from the principles, characteristics, applications, especially from the latest application progress. Results and conclusion: RTCA is attracting more and more attention. Now it plays an important role in drug screening, toxicology, Chinese herbal medicine and so on. It has wide application prospects in the area of modern pharmaceutical evaluation and analysis.

Study on the Quality Evaluation of Compound Danshen Preparations Based on the xCELLigence Real-Time Cell-Based Assay and Pharmacodynamic Authentication.

Objective: To perform a preliminary study on the quality evaluation of compound Danshen preparations based on the xCELLigence Real-Time Cell-based Assay (RTCA) system and make a pharmacodynamics verification. Methods: The compound Danshen was discussed as a methodological example, and the bioactivity of the compound Danshen preparations were evaluated by real-time cell electronic analysis technology. Meanwhile, an in vivo experiment on an acute blood stasis rat model was performed in order to verify this novel evaluation through the curative effect of dissipating blood stasis. Results: We determined the cell index (CI) and IC50 of the compound Danshen preparations and produced time/dose-dependent cell response profiles (TCRPs). The quality of the three kinds of compound Danshen preparations was evaluated through the RTCA data. The trend of CI and TCRPs reflected the effect of drugs on the cell (promoting or inhibiting), and it was verified that the results correlated with the biological activity of the drugs using a pharmacodynamics experiment. Conclusion: The RTCA system can be used to evaluate the quality of compound Danshen Preparations, and it can provide a new idea and new method for quantitatively characterizing the biological activity of traditional Chinese medicines (TCMs).

A novel release kinetics evaluation of Chinese compound medicine: Application of the xCELLigence RTCA system to determine the release characteristics of Sedum sarmentosum compound sustained-release pellets.

Purpose: To establish a novel release kinetics evaluation method of Chinese compound medicine (Sedum Sarmentosum compound) with xCELLigence Real-Time Cell-based Assay (RTCA) system. Methods: Cell lines sensitive to Sedum Sarmentosum compound are screened, and cell index-time (CI-T) graphs and cell index release kinetics models are established based on the cell index (CI) monitored. The methodological studies of precision and repeatability were processed by the cell monitors system. The release profiles of the sustained-release Sedum Sarmentosum compound were determined. Consequently, the sustained-release property was characterized by the kinetic parameters based on the cell-index. Results: The accumulative release rate based on cell index of Sedum sarmentosum compound sustained-release preparation was determined and it had a good correlation with time, fitting better with First-order model, Higuchi model and Ritger-Peppas model, and fitting best with Weibull model. It indicated that the release rate is proportional with the diffusion coefficient. Conclusion: The new method of cell-index release kinetics may provide a quantitative description for the release of the multi active agents from Traditional Chinese Medicines. The application of xCELLigence RTCA system for evaluating the release kinetics of Chinese compound medicine is feasible.

PSN-PC: A Novel Antimicrobial and Anti-Biofilm Peptide from the Skin Secretion of Phyllomedusa-camba with Cytotoxicity on Human Lung Cancer Cell.

Peptides derived from amphibian skin secretion are promising drug prototypes for combating widespread infection. In this study, a novel peptide belonging to the phylloseptin family of antimicrobial peptides was isolated from the skin secretion of the Phyllomedusa camba, namely phylloseptin-PC (PSN-PC). The biosynthetic precursor was obtained by molecular cloning and the mature peptide sequence was confirmed through tandem mass spectrometry (MS/MS) fragmentation sequencing in the skin secretion. The synthetic replicate exhibited a broad spectrum antimicrobial activity against Staphylococcus aureus, methicillin-resistant Staphylococcus aureus,Escherichia coli, Pseudomonas aeruginosa, Candida albicans at concentrations of 2, 2, 8, 32 and 2 µM, respectively. It also showed the capability of eliminating S. aureus biofilm with a minimal biofilm eradication concentration of 8 µM. The haemolysis of this peptide was not significant at low concentrations but had a considerable increase at high concentrations. Additionally, this peptide showed an anti-proliferation effect on the non-small cell lung cancer cell line (NCI-H157), with low cytotoxicity on the human microvascular endothelial cell line (HMEC-1). The discovery of the novel peptide may provide useful clues for new drug discoveries.

[Nasal resorption of prim-O-glucosylcimifugin and 5-O-methylvisammioside in rats].

In this experiment, rat nasal mucosa absorption characteristics of prim-O-glucosylcimifugin and 5-O-methylvisammioside were studied to provide a basis for drug delivery of Toutongning nasal spray. The nasal mucosa absorption test in rats was conducted with in situ nasal perfusion method after pH 6 buffer solution was used to prepare high, medium and low concentrations of prim-O-glucosylcimifugin, 5-O-methylvisammioside mixed solution as liquid circulation in nasal cavity. Then the concentrations of the circulating liquid compositions to be measured were determined by HPLC, and the absorption rates of prim-O-glucosylcimifugin and 5-O-methylvisammioside under different pH conditions were also investigated. According to the results, the absorption rate constant was (0.588±0.041)×10⁻³, (0.547±0.023)×10⁻³, (0.592±0.063)×10⁻³ min⁻¹ for prim-O-glucosylcimifugin high, middle and low concentrations, and (0.438±0.041)×10⁻³, (0.407±0.023)×10⁻³, and (0.412±0.063)×10⁻³ min⁻¹ for 5-O-methylvisammioside high, middle and low concentrations. There was no significant difference among high, middle and low concentration groups, and the absorption under pH 6 was better than that under other pH conditions. Therefore, we can get the conclusion that the main active ingredient of Toutongning nasal sprays can be absorbed through the nasal mucosa, and it is feasible to make nasal spray; in addition, pH 6 of nasal spray is scientific and reasonable.

[Screen absorption enhancer for intranasal administration preparations of paeoniflorin based on nasal perfusion method in rats].

This experiment was aimed to screen the absorption enhancer for intranasal administration preparations of paeoniflorin. In this study, HPLC method for determination of paeoniflorin in perfusion liquid was established and the improved model of nasal perfusion in rats was used to screen out the species and amounts of absorption enhancer. In order to avoid the influence of the secretion and absorption of nasal cavity on the volume of perfusion fluid, the residual dose was calculated by using the volume correction method. Linear regression was carried out between the logarithm to the percentage of the residual dose and the corresponding time, and the slope of the regression line was exactly the absorption rate constant. Experimental results showed that hydroxypropyl-ß-cyclodextrin and water-soluble azone can significantly improve the nasal absorption of paeoniflorin. Furthermore, water-soluble azone had the highest absorption rate constant and the best promoting penetration effect on intranasal administration preparations of paeoniflorin. It was also found that when the mass concentration of water-soluble azone in the perfusion liquid increased from 5 g•L⁻¹ to 20 g•L⁻¹, the absorption rate constant was gradually increased and peaked at 20 g•L⁻¹. When the mass concentration was increased to 30 g•L⁻¹, the absorption rate constant was decreased, indicating that the best mass concentration of water-soluble azone was 20 g•L⁻¹.

In-silico ADME Studies for New Drug Discovery: From Chemical Compounds to Chinese Herbal Medicines.

Nowadays, in silico tools are widely used to provide the potential structure of the metabolites formed depending on the site of metabolism. These methods can also highlight the molecular moieties that help to direct the molecule into the cytochrome cavity so that the site of metabolism is in proximity to the catalytic center. In this minireview, we summarized three aspects of the in silico methods in the application of prediction of ADME (absorption, distribution, metabolism and excretion) properties of compounds: structure-based approaches for predicting molecular modeling of drug metabolizing enzymes; in silico metabolite prediction; and pharmacophore models for analysis substrate specificity. Moreover, we also extended the in silico studies in Chinese herbal medicines (CHM) research.

[Research on Biological Assessment for Panax notoginseng Quality on Real-time Cell Electronic Analysis].

OBJECTIVE: To establish the quality control and evaluation methods of Panax notoginseng on promoting blood circulation and removing blood stasis effects, by determining cell index to evaluate the quality of Panax notoginseng from different habitats. METHODS: Using the real-time cell electronic analysis technology (RTCA) to examine the biological activity of specific cell-dependent cell lines on Panax notoginseng extracts. Changing trends and laws of the samples within a certain time were analyzed, and the cell index at the optimum time was determined. RESULTS: In four batches of Panax notoginseng from different habitats, cell index of Panax notoginseng from Chuxiong of Yunnan Province at the optimal time of 38 h was the highest, and the biological activity was the strongest. Cell index of Panax notoginseng from Wenshan of Yunnan Province at the optimal time of 38 h cells was the lowest, and the biological activity was the weakest. CONCLUSION: The method based on the real-time cell electronic analysis technology can initially be used in the detection of biological activity of Panax notoginseng.

[In vitro transdermal delivery of Qingfei Xiaocuo gel based on principal component analysis].

The objective of the present study was to establish a method based on principal component analysis (PCA) for the study of transdermal delivery of Chinese medicinal formulae, and to choose the best penetration enhancers for Qingfei Xiaocuo gel depend on this method. Using improved Franz type diffusion cell and excised rat skin in vitro as transdermal barrier, the receptive solution fingerprint was established by HPLC, harvesting the areas of the common peaks in the fingerprint, then the total factor scores of the concentrations at different times were calculated using PCA and were employed instead of the concentrations to compute the cumulative amounts (Q12) and enhancement ratio (ER), the latter of which were considered as the indexes for optimizing penetration enhancers. Compare to the control group, the ER of the other groups increased significantly and furthermore, 2.5% azone with 2.5% menthol manifested the best effect. PCA represent most information in the receptive solution, the method above could choose the best penetration enhancers, it could be a reference for the study of transdermal delivery of Chinese medicinal formulae.

Preparation and pharmacokinetics in beagle dogs of ganershu sustained-release pellets.

BACKGROUND: The active ingredients of Ganershu compound recipe, which are effective for hepatitis treatment in liver protection and transaminase reduction. However, the active ingredients of Ganershu compound recipe are poor absorption, which conduct it has a low oral bioavailability. OBJECTIVE: We prepared Ganershu sustained-release pellets (GSPs) by fluidized-bed on central composite design-response surface methodology and increase its bioavailability in beagle dogs. MATERIALS AND METHODS: In this study, GSPs were successfully prepared. The Drug-loaded pellets and sustained-release coated were carried out in fluidized-bed machine. GSP was optimized for fitting release, roundness, and the overall desirability by central composite design-response surface methodology. RESULTS: To optimize cumulative release profile, the outermost ethyl cellulose coating layer and the hydroxypropyl methyl cellulose (HPMC) swelling layer were employed, which were respectively given coating levels in terms of weight gain of 22% and 6%, the concentration of HPMC is 4.5% (g/ml). The pharmacokinetics of Ganershu normal pellets (GNPs) and GSP was studied in beagle dogs after oral administration. The naringenin as an index, the area under the curve0-∞ of naringenin in GSP was 1.38 times greater than that of GNP. Meanwhile, Tmax of GSP was prolonged for about 74%. CONCLUSION: This study can clearly indicate that we enhanced the oral bioavailability of Ganershu by preparing the GSP, which had the sustained dissolution and improved the potential of it for clinical application.

Hyaluronic acid-coated bovine serum albumin nanoparticles loaded with brucine as selective nanovectors for intra-articular injection.

OBJECTIVE: To evaluate the potential of hyaluronic acid (HA)-coated bovine serum albumin nanoparticles (BSANPs) as a novel chondrocyte-targeting drug-delivery nanomedicine. METHODS: The HA-BSANPs were characterized by dynamic light scattering, transmission electron microscopy, differential scanning calorimetry, and X-ray diffraction. Fluorescence imaging was used to visualize the distribution of nanoparticles after intra-articular injection. The chondrocyte-targeting efficiency and cellular uptake mechanism of HA-BSANPs were investigated using endocytic inhibitors. RESULTS: HA-BSANPs were successfully prepared with HA coating the surface and amorphous drug in the core. Compared with BSANPs, HA-BSANPs exhibited improved uptake by chondrocytes through a receptor-mediated active uptake mechanism. The endocytosis process of BSANPs and HA-BSANPs involved clathrin-mediated endocytosis, caveolae-mediated endocytosis, and macropinocytosis. No apparent thickening or hyperplasia of the synovium was observed in either BSANPs or HA-BSANPs. The HA-BSANPs could reside in the articular cavity of rats for more than 14 days, which was significantly longer than BSANPs. CONCLUSION: HA-BSANPs are a promising carrier for articular-related diseases due to elongated articular residence and improved chondrocytic accumulation.

[Study on HPLC fingerprint of Ganershu intermediate and determination of three active components].

OBJECTIVE: To establish the fingerprint analysis method of Ganershu intermediate by HPLC. METHODS: An analysis was performed on a sunFire C18 (4. 6 mm x 250 mm, 5 microm) column with acetonitrile-0.1% phosphate aqueous as the mobile phase by gradient elution. The flow rate was 1.0 mL/min, the detection wavelength was 320 nm and detection time was 80 min. The column temperature was 35 degrees C. In the recorded chromatogram of Ganershu intermediate, neohesperidin was used as reference substance, and RSD of the relative retention time and the relative peak areas of all peaks compared with its peak were measured. The similarity of 10 batches of Ganershu intermediate was appraised by the similarity evaluation system. Using the external standard method, the contents of chlorogenic acid, neohesperidin and naringin were determined in 10 batches of intermediate. RESULTS: Tweenty-three peaks were separated on HPLC fingerprint in Ganershu intermediate, degree of similarity of fingerprint for ten batches of Ganershu intermediate were greater than 0.90. Three compounds' contents were almost the same in each batch of intermediate sample. CONCLUSION: The method is stable, accurate, reliable and can be used as a quality control for Ganershu intermediate and sustained-release capsules.

Development of brucine-loaded microsphere/thermally responsive hydrogel combination system for intra-articular administration.

Intra-articular drug delivery system could directly deliver a drug to an affected joint and offer the possibility of reaching high drug concentrations at the site of action with limited systemic toxicity. However, depending on their chemical structure, some active compounds were rapidly cleared from the joint, thus requiring numerous injections, which could cause infection or joint disability. To control the release behavior for prolonged time periods, a novel biologically based drug delivery vehicle was designed for intra-articular using microsphere/thermally responsive hydrogel combination system in this paper. And brucine was the test drug. The system was constructed by dispersing the brucine microspheres which was prepared by using a spray-drying method in a thermally responsive biopolymer hydrogel contained with chitosan-glycerol-borax. The microspheres were spherical as evidenced by the scanning electron microscopy (SEM) photographs. And the entrapment rate was 98.60% w/w with an average size range of 0.9-4.5 µm. Fourier transforms infrared (FT-IR) spectroscopy and X-ray diffraction (XRD) revealed the absence of drug-polymer interaction and amorphous nature of an entrapped drug. From the in vitro drug release study we could see that there was a burst release of microsphere, which was obviously retarded when dispersed in hydrogel. And the studies of biocompatibility with synovium showed that no apparent thickening or hyperplasia of the synovium, a small quality of phlogocyte imbibitions was observed. The results of FX imaging in rats showed that by intra-articular injection the BMH could stay in articular for over 7 days were consistent with our in vitro release. And the results of pharmacodynamics revealed the BMH could benefit OA joint by suppressing the levels of TNF-α and IL-1ß, protect the damaged joint from degradation. The novel microsphere/thermoresponsive hydrogel combination system could be a promising treatment option for OA and RA. In conclusion, the system appears to be generally biocompatible with synovium and could control the drug release for several days; hence it might be suitable for the development of treatment strategies for rheumatic diseases.