AIM:This study aimed to investigate the differences in hepatic lipid metabolites in ICR mice in-duced by a high-fat diet and treated with Shaqi concentrated pills(SQ).METHODS:Thirty 8-week-old SPF-grade ICR mice were randomly assigned to five groups:the normal(CON,n=6)group,the high-fat diet model(HFD,n=6)group,the low-dose SQ administration(SQL,n=6)group,the high-dose SQ administration(SQH,n=6)group,and the liver-protecting tablets positive control(PLT,n=6)group.The HFD group was fed a diet consisting of 60%fat for 8 weeks to es-tablish a metabolic-dysfunction-associated fatty liver disease model.Upon successful model establishment,the SQL group received a daily gavage of 395 mg/kg for 4 weeks,while the SQH group received 790 mg·kg-1·d-1.The PLT group was ad-ministered liver-protecting tablets at a dosage of 0.655 g/kg via gavage.Body weight and food intake were monitored week-ly.Liver indices,including Lee's index,triglyceride(TG),total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C),alanine aminotransferase(ALT),and aspartate transaminase(AST)levels,were measured in each group.Hematoxylin-eosin(HE)staining and oil red O staining were performed to assess the extent of pathological damage in liver tissues.Western blot analysis was conducted to evaluate the protein expres-sion levels of choline/ethanolamine phosphotransferase-1(CEPT1),adipose triglyceride lipase(ATGL),and diacylglycerol acyltransferase(DGAT)in the liver.A non-targeted lipidomic analysis using LC-MS was employed to detect changes in hepatic lipid content,and multivariate statistical analyses(principal component analysis and orthogonal partial least squares discriminant analysis)were utilized to compare lipid metabolic profiles among the groups and identify differential lipid metabolites.RESULTS:Compared to the CON group,mice in the HFD group exhibited significantly increased body weight,blood glucose levels,serum TG,TC,LDL-C,ALT,and AST levels,accompanied by a marked decrease in HDL-C levels.HE and oil red O staining results revealed significant lipid droplet accumulation in the liver tissues of HFD mice.In contrast,mice in the SQL and SQH groups showed significant reductions in body weight,blood glucose,serum TG,TC,LDL-C,ALT,and AST levels,along with increased HDL-C levels and less lipid accumulation in liver tissues compared to the HFD group.Staining of liver sections confirmed that SQ treatment mitigated the abnormal accumulation of lipid droplets.Lipidomic analysis indicated that SQ treatment normalized 25 aberrantly expressed lipid metabolites to lev-els comparable to the CON group and identified nine representative differential lipid metabolites.Western blot results dem-onstrated that SQ treatment reduced the protein expression levels of ATGL and DGAT while increasing the expression of CEPT1.CONCLUSION:Treatment with SQ can alleviate metabolic dysfunction-associated fatty liver disease(MAFLD)by modulating triglyceride metabolism,phosphatidylcholine metabolism,and dimethylphosphatidylethanolamine lipid me-tabolism,thereby altering the hepatic lipid profile in MAFLD mice.

BACKGROUND:For knee osteoarthritis,synovium inflammation and abnormal proliferation and activation of fibroblast-like synoviocytes caused by insuffcient apoptosis are important driving factors for disease progression.Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signaling pathway plays a key role in regulating inflammatory response and apoptosis of synovial cells.As a traditional Mongolian medicine component,Agiophyllum Oligo Saccharides has anti-inflammatory and immunomodulatory functions,but its specific molecular mechanism has not been fully revealed.OBJECTIVE:To explore the effects and mechanism of Agiophyllum Oligo Saccharides on lipopolysaccharide-induced inflammation and apoptosis of mouse synovial cells.METHODS:The mouse synovial cells were randomly divided into a control group,a model group,an Agiophyllum Oligo Saccharides group and a Bay 11-7082 group.Osteoarthritic synoviocyte model was prepared using 1.0 μg/mL lipopolysaccharide intervention for 36 hour.The Agiophyllum Oligo Saccharides group was treated with 1.0 μg/mL lipopolysaccharide and 32 μg/mL Agiophyllum Oligo Saccharides for 36 hours;and the Bay 11-7082 group was firstly incubated for 4 hours with 1 μmol/L NF-κB pathway inhibitor Bay 11-7082,and then 1.0 μg/mL lipopolysaccharide and 32 μg/mL Agiophyllum Oligo Saccharides for 36 hours.Western blot was used to detect TLR4,MyD88,NF-κBp65,p-NF-κBp65,and IκBα protein expression.The level of tumor necrosis factor-α and interleukin-1β in the supernatant were analyzed by ELISA.To detect the expressions of Bax and Bcl-2,western blot and immunofluorescence analysis were performed.The intracellular reactive oxygen species level was detected by DCFH-DA.RESULTS AND CONCLUSION:(1)Compared with the control group,the levels of tumor necrosis factor-α,interleukin-1β and reactive oxygen species,the protein expressions of TLR4,MyD88,Bcl-2,and p-NF-κBp65/NF-κBp65 increased(P<0.05),whereas the protein expressions of IκBα and Bax decreased(P<0.05)in the model group.(2)Compared with the model group,the levels of tumor necrosis factor-α,interleukin-1β and reactive oxygen species,the protein expressions of TLR4,MyD88,Bcl-2,and p-NF-κBp65/NF-κBp65 decreased(P<0.05),while the expressions of IκBα and BAX increased(P<0.05)in the Agiophyllum Oligo Saccharides group.(3)The levels of tumor necrosis factor-α,interleukin-1β and reactive oxygen species and the protein expression of Bcl-2 in the Bay 11-7082 group were higher than those in the Agiophyllum Oligo Saccharides group(P<0.05),but still lower than those in the model group(P<0.05),whereas the expression of Bax protein was lower than that in the Agiophyllum Oligo Saccharides group(P<0.05),but still higher than that in the model group(P<0.05).(4)Compared with the model group and the Agiophyllum Oligo Saccharides group,the protein expressions of TLR4,MyD88 and p-NF-κBp65/NF-κBp65 decreased(P<0.05),whereas the protein expressions of IκBα increased(P<0.05)in the Bay 11-7082 group.To conclude,Agiophyllum Oligo Saccharides could inhibit inflammation of mouse synovial cells and promote apoptosis of synovial cells with inflammation,and the mechanisms may be correlated with regulating the TLR4/NF-κB signaling pathway.

AIM:This study aimed to investigate the differences in hepatic lipid metabolites in ICR mice in-duced by a high-fat diet and treated with Shaqi concentrated pills(SQ).METHODS:Thirty 8-week-old SPF-grade ICR mice were randomly assigned to five groups:the normal(CON,n=6)group,the high-fat diet model(HFD,n=6)group,the low-dose SQ administration(SQL,n=6)group,the high-dose SQ administration(SQH,n=6)group,and the liver-protecting tablets positive control(PLT,n=6)group.The HFD group was fed a diet consisting of 60%fat for 8 weeks to es-tablish a metabolic-dysfunction-associated fatty liver disease model.Upon successful model establishment,the SQL group received a daily gavage of 395 mg/kg for 4 weeks,while the SQH group received 790 mg·kg-1·d-1.The PLT group was ad-ministered liver-protecting tablets at a dosage of 0.655 g/kg via gavage.Body weight and food intake were monitored week-ly.Liver indices,including Lee's index,triglyceride(TG),total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C),alanine aminotransferase(ALT),and aspartate transaminase(AST)levels,were measured in each group.Hematoxylin-eosin(HE)staining and oil red O staining were performed to assess the extent of pathological damage in liver tissues.Western blot analysis was conducted to evaluate the protein expres-sion levels of choline/ethanolamine phosphotransferase-1(CEPT1),adipose triglyceride lipase(ATGL),and diacylglycerol acyltransferase(DGAT)in the liver.A non-targeted lipidomic analysis using LC-MS was employed to detect changes in hepatic lipid content,and multivariate statistical analyses(principal component analysis and orthogonal partial least squares discriminant analysis)were utilized to compare lipid metabolic profiles among the groups and identify differential lipid metabolites.RESULTS:Compared to the CON group,mice in the HFD group exhibited significantly increased body weight,blood glucose levels,serum TG,TC,LDL-C,ALT,and AST levels,accompanied by a marked decrease in HDL-C levels.HE and oil red O staining results revealed significant lipid droplet accumulation in the liver tissues of HFD mice.In contrast,mice in the SQL and SQH groups showed significant reductions in body weight,blood glucose,serum TG,TC,LDL-C,ALT,and AST levels,along with increased HDL-C levels and less lipid accumulation in liver tissues compared to the HFD group.Staining of liver sections confirmed that SQ treatment mitigated the abnormal accumulation of lipid droplets.Lipidomic analysis indicated that SQ treatment normalized 25 aberrantly expressed lipid metabolites to lev-els comparable to the CON group and identified nine representative differential lipid metabolites.Western blot results dem-onstrated that SQ treatment reduced the protein expression levels of ATGL and DGAT while increasing the expression of CEPT1.CONCLUSION:Treatment with SQ can alleviate metabolic dysfunction-associated fatty liver disease(MAFLD)by modulating triglyceride metabolism,phosphatidylcholine metabolism,and dimethylphosphatidylethanolamine lipid me-tabolism,thereby altering the hepatic lipid profile in MAFLD mice.

BACKGROUND:For knee osteoarthritis,synovium inflammation and abnormal proliferation and activation of fibroblast-like synoviocytes caused by insuffcient apoptosis are important driving factors for disease progression.Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signaling pathway plays a key role in regulating inflammatory response and apoptosis of synovial cells.As a traditional Mongolian medicine component,Agiophyllum Oligo Saccharides has anti-inflammatory and immunomodulatory functions,but its specific molecular mechanism has not been fully revealed.OBJECTIVE:To explore the effects and mechanism of Agiophyllum Oligo Saccharides on lipopolysaccharide-induced inflammation and apoptosis of mouse synovial cells.METHODS:The mouse synovial cells were randomly divided into a control group,a model group,an Agiophyllum Oligo Saccharides group and a Bay 11-7082 group.Osteoarthritic synoviocyte model was prepared using 1.0 μg/mL lipopolysaccharide intervention for 36 hour.The Agiophyllum Oligo Saccharides group was treated with 1.0 μg/mL lipopolysaccharide and 32 μg/mL Agiophyllum Oligo Saccharides for 36 hours;and the Bay 11-7082 group was firstly incubated for 4 hours with 1 μmol/L NF-κB pathway inhibitor Bay 11-7082,and then 1.0 μg/mL lipopolysaccharide and 32 μg/mL Agiophyllum Oligo Saccharides for 36 hours.Western blot was used to detect TLR4,MyD88,NF-κBp65,p-NF-κBp65,and IκBα protein expression.The level of tumor necrosis factor-α and interleukin-1β in the supernatant were analyzed by ELISA.To detect the expressions of Bax and Bcl-2,western blot and immunofluorescence analysis were performed.The intracellular reactive oxygen species level was detected by DCFH-DA.RESULTS AND CONCLUSION:(1)Compared with the control group,the levels of tumor necrosis factor-α,interleukin-1β and reactive oxygen species,the protein expressions of TLR4,MyD88,Bcl-2,and p-NF-κBp65/NF-κBp65 increased(P<0.05),whereas the protein expressions of IκBα and Bax decreased(P<0.05)in the model group.(2)Compared with the model group,the levels of tumor necrosis factor-α,interleukin-1β and reactive oxygen species,the protein expressions of TLR4,MyD88,Bcl-2,and p-NF-κBp65/NF-κBp65 decreased(P<0.05),while the expressions of IκBα and BAX increased(P<0.05)in the Agiophyllum Oligo Saccharides group.(3)The levels of tumor necrosis factor-α,interleukin-1β and reactive oxygen species and the protein expression of Bcl-2 in the Bay 11-7082 group were higher than those in the Agiophyllum Oligo Saccharides group(P<0.05),but still lower than those in the model group(P<0.05),whereas the expression of Bax protein was lower than that in the Agiophyllum Oligo Saccharides group(P<0.05),but still higher than that in the model group(P<0.05).(4)Compared with the model group and the Agiophyllum Oligo Saccharides group,the protein expressions of TLR4,MyD88 and p-NF-κBp65/NF-κBp65 decreased(P<0.05),whereas the protein expressions of IκBα increased(P<0.05)in the Bay 11-7082 group.To conclude,Agiophyllum Oligo Saccharides could inhibit inflammation of mouse synovial cells and promote apoptosis of synovial cells with inflammation,and the mechanisms may be correlated with regulating the TLR4/NF-κB signaling pathway.

Type 2 diabetes mellitus (T2DM) and nonalcoholic fatty liver disease (NAFLD) are the most problematic metabolic diseases in the world. NAFLD encompasses a spectrum of severity, ranging from simple steatosis to non-alcoholic steatohepatitis (NASH) and fibrosis, increasing the risk of cirrhosis and hepatocellular carcinoma. Importantly, NAFLD is closely linked to obesity and tightly interrelated with insulin resistance and T2DM. T2DM and NAFLD (T2DM-NAFLD) are called as the Xike Rixijing Disease and Tonglaga Indigestion Disease respectively, in Mongolian medicine. Xike Rixijing Disease maybe develop into Tonglaga Indigestion Disease. Forturnately many Mongolian medicines show efficient treatment of T2DM-NAFLD, such as Agriophyllum squarrosum, Haliyasu (dried powder of camel placenta), Digeda-4 (herbs of Lomatogonium carinthiacum, rhizomata of Coptis chinensis, ripe fruits of Gardenia jasminoides, herbs of Dianthus superbus), Guangmingyan Siwei Decoction Powder (Halite, ripe fruits of Terminalia chebula, rhizomata of Zingiber officinale, fruit clusters of Piper longum), Tonglaga-5 (ripe fruits of Punica granatum, barks of Cinnamomum cassia, ripe fruits of Amomum kravanh, fruit clusters of Piper longum, flowers of Carthamus tinctorius), Tegexidegeqi (rhizomata of Inula helenium, ripe fruits of Gardenia jasminoides, rhizomata of Platycodon grandiflorum, rhizomata of Coptis chinensis, heartwood of Caesalpinia sappan), Ligan Shiliu Bawei San (ripe fruits of Punica granatum, barks of Cinnamomum cassia, ripe fruits of Amomum kravanh, fruit clusters of Piper longum, flowers of Carthamus tinctorius, ripe fruits of Amomum tsao-ko, rhizomata of Zingiber officinale), etc. Principles of Mongolian medicine in treating diseases: by balancing “three essences or roots” and “seven elements”, strengthening liver and kidney function, transporting nutrients to enhance physical strength and disease resistance, and combined with drugs for comprehensive conditioning treatment. However, their molecular mechanisms remain unclear. In this review, we prospect that Mongolian medicines might be a promising treatment for T2DM-NAFLD by activating P2X7R/NLRP3/NF-κB inflammatory pathway via lipid-sensitive nuclear receptors (i.e., FXR and LXR).

Aim To observe the effect of agiophyllum oligo saccharides ( AOS) on reducing blood sugar, im-proving insulin resistance on diadetic Goto-Kakizaki ( GK) rats, and to explore the possible mechanism. Methods The type 2 diabetes GK rats were divided into six groups: model control group ( MC ) , Glenn benzene urea group ( GLB ) , high agriophyllum squar-rosum coarse oligosaccharides ( AOS-H ) , medium ( AOS-M ) , low dose group ( AOS-L ) , homologous Wistar rats as normal control ( NC ) . All animals were administered with AOS by oral gavage, for 8 weeks. The fasting blood glucose ( FBG) , random blood sugar ( RBG) , glucose tolerance ( OGTT) were tested before and after administration. No fasting sugar load status before and after dosing changes in blood glucose and serum insulin level in rats were measured in the previ-ous 8 weeks. At the end of administration, the fasting serum glucose ( FPG) , insulin ( FINS) , OGTT and in-sulin resistance index ( HOMA IR) in fasting rats were analyzed. Lastly, the pathological changing of pancreas was observed by HE staining. Results The blood glucose of fasting GK rats was not influenced after using AOS. However, the random blood glucose significantly reduced, the glucose tolerance was improved and AUC was obviously reduced (P < 0. 01) after using AOS. The best effect was on AOS-M group, which was similar with Glenn benzene urea. Through our research, we found AOS could promote release of insulin. This best effect was on AOS-M and AOS-L groups, and the time and quantity of release were better than Glenn benzene urea. Finally, AOS inhibited the pathological changes of islet tissue on GK rats, increased the quantity of pancreas and islet cells. Compared with model group, the changing of islet structure was significantly reduced in AOS group. Conclusion AOS could obviously improve insulin resistance and lower blood sugar, and the mechanism of this effect may be related with rapidly promoting insulin release, increasing the islet cell proliferation,and improving the function of islet.

OBJECTIVETo determine the immune repertoires of peripheral CD4+T cell receptor (TCR) Vb CDR3 in primary biliary cirrhosis (PBC) and analyze TCR diversity and preferred usage at sequence-level resolution.

METHODSARM-PCR and high-throughput sequencing were used to obtain millions of TCR Vb CDR3 sequences from peripheral CD4+T cells isolated from 7 patients with PBC and healthy volunteers. All sequencing data were analyzed, together with corresponding clinical information, by bioinformatic software. The Mann-Whitney U test was used for statistical analysis.

RESULTSThe PBC patients showed a lower level of diversity among the peripheral CD4+TCR Vb CDR3 than the healthy volunteers, and patients with higher level progression of the disease showed a greater lack of diversity. In addition, 4 specific preferred-usage amino acid sequences were discovered for the PBC patients: ASSFTGGPVEQY, ASSLISSGNNEQF, ATSRDTLAGGPGDTQY, and SASLEGNTEAF; these sequences were also found in higher frequencies in patients with later stages of PBC.

CONCLUSIONSDecreased TCR Vb CDR3 diversities and specific preferred usage of TCR CDR3 sequences in peripheral CD4+T lymphocytes in PBC suggest that clonal expansion of a large number of CD4+T cells may be an important factor for PBC progression. These data provide a better understanding about the general characteristics of CD4+T cells in PBC patients and related to pathogenesis of the disease, and may provide useful insights into potential targets for immunotherapy.

Amino Acid Sequence ; CD4-Positive T-Lymphocytes ; High-Throughput Nucleotide Sequencing ; Humans ; Liver Cirrhosis, Biliary ; Polymerase Chain Reaction ; Receptors, Antigen, T-Cell