Objective: To probe into the mechanism and interventional effects of silybin-phospholipid complex on amiodarone-induced steatosis in mice. Methods: Eight-week-old male C57BL/6 mice were divided into three groups (5 mice in each group): a control group (WT) with normal diet, a model group with amiodarone 150mg/kg/d by oral gavage (AM), and an intervention group on amiodarone 150mg/kg/d combined with silybin-phospholipid complex(AM+SILIPHOS. All mice were fed their assigned diet for one week. Then, one week later, serum alanine aminotransferase, aspartate aminotransferase, triglyceride, total cholesterol and high-density lipoprotein were detected of each group. A liver pathological change was observed by oil red O and H&E staining. Ultrastructural pathological changes of hepatocytes were observed to evaluate the intervention effect by transmission electron microscopy. RT-q PCR was used to detect the expression of peroxisome proliferator-activated receptor alpha and its regulated lipid metabolism genes CPTI, CPTII, Acot1, Acot2, ACOX, Cyp4a10 and Cyp4a14 in liver tissues. Intra-group comparison was done by paired t-test. One-way ANOVA was used for comparison between groups and semi-quantitative data were tested using Mann-Whitney U test. Results: Oil Red O and H&E staining results of liver tissue in the intervention group showed that intrahepatic steatosis was significantly reduced when compared to model group. Transmission electron microscopy showed that the model group had pyknotic nuclei, mitochondrial swelling, structural damage, and lysosomal degradation whereas the intervention group had hepatic nucleus without pyknosis, reduced mitochondrial swelling and slight structural damage than that of model group. RT-q PCR results showed that the expression of peroxisome proliferator-activated receptor alpha, CPTI, CPTII, Acot1, Acot2, ACOX, Cyp4a10 and Cyp4a14 were increased in the model group but the expression of CPTI, Cyp4a14, Acot1 and peroxisome proliferator-activated receptor alpha were decreased in the intervention group (P < 0.05). Conclusion Silybin-phospholipid complex can alleviate amiodarone-induced steatosis, and its mechanism may play a role in protecting mitochondrial function and regulating fatty acid metabolism. Thus, silybin-phospholipid complex has potential intervention effect on amiodarone-induced fatty liver.

OBJECTIVE: To elucidate the action mechanism of Xingnaojing Injection (, XNJI) for sepsis, and to target screen the potential bioactive ingredients. METHODS: An integrated protocol that combines in silico target screen (molecular docking) and database mapping was employed to find the potential inhibitors from XNJI for the sepsis-related targets and to establish the compound-target (C-T) interaction network. The XNJI's bioactive components database was investigated and the sepsis-associated targets were comprehensively constructed; the 3D structure of adenosine receptor A2a and 5-lipoxygenase proteins were established and evaluated with homology modeling method; system network pharmacology for sepsis treatment was studied between the bioactive ingredients and the sepsis targets using computational biology methods to distinguish inhibitors from non inhibitors for the selected sepsis-related targets and C-T network construction. RESULTS: Multiple bioactive compounds in the XNJI were found to interact with multiple sepsis targets. The 32 bioactive ingredients were generated from XNJI in pharmacological system, and 21 potential targets were predicted to the sepsis disease; the biological activities for some potential inhibitors had been experimentally confirmed, highlighting the reliability of in silico target screen. Further integrated C-T network showed that these bioactive components together probably display synergistic action for sepsis treatment. CONCLUSIONS The uncovered mechanism may offer a superior insight for understanding the theory of the Chinese herbal medicine for combating sepsis. Moreover, the potential inhibitors for the sepsis-related targets may provide a good source to find new lead compounds against sepsis disease.
Arachidonate 5-Lipoxygenase ; metabolism ; Computer Simulation ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; therapeutic use ; Humans ; Injections ; Phytochemicals ; therapeutic use ; Receptor, Adenosine A2A ; metabolism ; Reproducibility of Results ; Sepsis ; drug therapy ; metabolism

OBJECTIVE: To study placenta-derived mesenchymal stem cells with HLA-G (Human Leukocyte Antigen, HLA-G) positive expression induce Treg (regulatory T cell, Treg) in vitro. METHODS: placenta-derived mesenchymal stem cells were separated from neonatal placenta; PEGFP - N1 -HLA-G plasmid was transfected in placenta-derived mesenchymal stem cells by liposome transfection.The cells were divided into 3 groups including control group, PEGFP-N1 group and PEGFP-N1-HLA-G group, 5 complex walls in each group. Expression of HLA-G protein was detected by Western Blotting; after identification of cells, healthy human peripheral blood CD4 T lymphocytes were cultured with placenta-derived mesenchymal stem cells with HLA-G positive expression, and the ratio of CD4CD25Foxp3Treg in T lymphocytes was accounted. RESULTS: After transfection of PEGFP-N1-HLA-G, the placenta-derived mesenchymal stem cells can express HLA-G protein significantly, compared with the control group and PEGFP - N1 group (<0.01). After HLA-G positive placenta-derived mesenchymal stem cells and CD4 + T lymphocytes were cultured for 24 h, the ratio of CD4CD25Foxp3Treg in T lymphocytes was (16.41±0.94)%. After HLA - G positive placenta-derived mesenchymal stem cells and CD4 T lymphocytes were cultured for 48 h, the ratio of CD4CD25Foxp3Treg in T lymphocytes was (16.46±0.59)% significantly, compared with the control group and PEGFP - N1 group (<0.01). CONCLUSIONS Placenta-derived mesenchymal stem cells modified by HLA-G gene can effectively induce CD4CD25Foxp3Treg in vitro.
Female ; Forkhead Transcription Factors ; HLA-G Antigens ; Humans ; Mesenchymal Stem Cells ; Placenta ; Pregnancy ; T-Lymphocytes, Regulatory

Aim To investigate the effects of Periplane-ta americana extract Ento-A on the immune function in immunosuppressed mice . Methods Immunosup-pressed mouse model was induced by intraperitoneal injection of cyclophosphamide in KM mice .To evalu-ate the effects of Ento-A on the immune function in im-munosuppressed mice , neutral red method and MTT assay were used respectively to detect the effects of En-to-A on the phagocytosis of peritoneal macrophages and T cell proliferation rate in mice; with sheep red blood cell as immunogen , the effects of Ento-A on the pro-duction of serum hemolysin were evaluated;peripheral blood was tested and immune organ index calculated . Results Compared with model control group , the high, medium and low doses of Ento-A could improve the expression of serum hemolysin in immunosup-pressed mice ( P<0.01 ) , and increase the spleen in-dex(P<0.01) and thymus index (P>0.05), signifi-cantly increased the content of WBC ( P<0.01 ) , PLT ( P<0.01 ) , HGB ( P<0.01 ) , while the contents of RBC was on the rise , with no significant difference ( P>0.05 ) in peripheral blood , significantly enhanced phagocytic function and T lymphocyte proliferative abil-ity in a dose-dependent manner ( P<0.01 ) .Conclu-sion Ento-A can enhance the immune function of im-munosuppressed mice .

Aim To study the effect of the Kangfuxin liquid on 2,4,6-trinitrobenzene sulfonic acid (TNBS) induced ulcerative colitis in rats, and to explore its mechanism. Methods Male SD rats were divided into normal control, model control, SASP groups, and Kangfuxin low,medium and high dose groups. In ad-dition to the normal control group, other groups were induced ulcerative colitis with TNBS solution. Disease activity index (DAI), organ index, colon mucosa damage index (CMDI) and histopathological score (HS),the expression levels of IL-4, IL-17 in serum, and MPO, EGF, TGF-β1 in colonic mucosa were de-termined. Results The DAI score showed that the model was successful. Compared with the normal group,the level of IL-4 and IL-17, EGF and TGF-β1 in the model group were reduced significantly, while the CMDI score,HS score,colon index and MPO were elevated significantly. The DAI, CMDI, HS and MPO were reduced (P<0.05 or P<0.01), and the levels of IL-4, IL-17, EGF and TGF-β1 increased signifi-cantly(P <0.01). Conclusions Kangfuxin liquid can effectively alleviate ulcerative colitis induced by TNBS in rats. The mechanism may be related to the down-regulation of MPO expression and up-regulation of IL-4,IL-17,EGF and TGF-β1 levels.

Our preliminary study showed that the total flavonoids in Isodon amethystoides(TFIA), a local medicinal herb in Suzhou, had a certain therapeutic effect on adjuvant arthritis, and this therapeutic effect may be achieved through the up-regulation of miR-152 expression. In this paper, the molecular mechanism of TFIA on the pathogenesis of adjuvant arthritis(AA) rats was further studied. AA rats were prepared with complete Freund's adjuvant, and then treated with TFIA by intragastric administration. Real-time qPCR was used to detect the effects of TFIA on the negative regulatory loop of miR-152, methylase DNMT1 and methyl-CpG binding protein MeCP2 in fibroblast like synoviocytes(FLS) of AA rats, as well as the effects of TFIA on the classic Wnt signaling pathway and the expression of fibronectin gene in AA rats. Intragastric administration of TFIA significantly inhibited the expression of DNMT1 and reversed the negative regulatory loop composed of miR-152, DNMT1 and MeCP2 in the pathology of AA rats. After transfection of miR-152 inhibitors into the FLS in treatment group, DNMT1 expression was significantly restored. TFIA significantly up-regulated the expression of SFRP4 and inhibited the expression of β-catenin, C-myc and ccnd1, the key genes of canonical Wnt signaling pathway. TFIA also significantly inhibited the expression of fibronectin, an AA gene. The effect of TFIA on the expression of SFRP4, β-catenin, C-myc, ccnd1 and fibronectin was reversed after transfection with miR-152 inhibitors in the treatment group FLS. TFIA may inhibit the DNMT1 expression, up-regulate the SFRP4 expression, inhibit the expression of classical Wnt signaling genes β-catenin, C-myc, and ccnd1 as well as the RA gene fibronectin expression through the up-regulation of miR-152 expression.

Coronavirus ; chemistry ; ultrastructure ; Microscopy, Electron ; Models, Molecular ; Ribonucleoproteins ; chemistry ; ultrastructure

To study the effect of pulchinenoside (PULC) on the Frizzled (FZD) expression of adjuvant arthritis ( AA) rats. AA rats were prepared through the toe injection with complete Freund's adjuvant to culture fibroblast-like synoviocytes (FLS). The effect of the oral administration with PULC on the FZD8 expression was detected by the real time qPCR. The effect of FZD8 knockout on the expressions of IL-1, IL-6, IL-8 were detected by MTT and ELISA. The role of miR-375 in the abnomal expression of FZD8 was detected by the real time qPCR. The results showed signfiicant decrease in the FZD8 expression among AA rats, FLS proliferation ater FZD8 knockout and IL-1, IL-6, IL-8 expressions and notable increase in miR-375 expression after the oral administration with PULC. The up-regulated miR-375 expression can inhibit the FZD8 expression. PULC may inhibit the FZD8 expression by up-regulating the miR-375 expression.
Animals ; Arthritis, Experimental ; drug therapy ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, Cell Surface ; genetics ; metabolism ; Saponins ; administration & dosage

This study aims to analyze and compare the effect of cell wall-broken decoction pieces, conventional decoction pieces and conventional powder of Rhodiolae Crenulatae Radix et Rhizoma on the intestinal flora of normal mice. The conventional bacterial culture and PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) were adopted for the mice after the oral administration for 14 days. According to the bacterial culture results, the 1/8 dose cell wall-broken decoction pieces group showed fewer Enterococcus and Escherichia coli bacillus but more Lactobacillus and Bifidobacterium than the conventional decoction pieces group and the traditional powder group (P <0.05). Meanwhile, on the basis of the PCR-DGGE results, the 1/8 dose cell wall-broken decoction pieces group revealed the highest Shannon-Wiener index (H) and species richness (S) among the seven groups, with extremely significant differences compared with the normal group (P <0.01), significant differences compared with the conventional decoction pieces group and the conventional powder group (P <0.05) and a high intra-group similarity. In conclusion, the long-term intake of 1/8 dose Rhodiolae Crenulatae Radix et Rhizoma cell wall-broken decoction pieces showed a certain effect in regulating intestinal tract by promoting the growth of Lactobacillus and Bifidobacterium. Furthermore, the intestinal flora community will become more stable.
Animals ; Bifidobacterium ; drug effects ; genetics ; growth & development ; Cell Wall ; Denaturing Gradient Gel Electrophoresis ; Intestines ; microbiology ; Lactobacillus ; drug effects ; genetics ; growth & development ; Mice ; Mice, Inbred C57BL ; Polymerase Chain Reaction ; Rhizome ; Rhodiola

The role of flavonoids of Echinps latifolius (FELT) in Wnt signaling was investigated in adjuvant arthritis (AA) rats. The therapeutic effects of FELT on AA rats were detected by rat arthritis score and MTT. The effect of FELT gavage treatment on the Wnt signaling key gene β-catenin, C-myc and cyclin D1 in synovium from AA rats was detected by Real-time qPCR, and the effects of FELT gavage treatment on the upstream negative regulation gene SFRP 1,2,4,5 in synovium from AA rats were detected by Real-time qPCR. The results showed that FELT gavage treatment significantly inhibited arthritis score and MTT values in AA rats, significantly inhibited the expression of the Wnt signaling gene β-catenin, C-myc and cyclin D1, significantly up-regulated the expression of the up- stream negative regulation gene SFRP 1,2,4. FELT has a better therapeutic effect for AA rats.
Animals ; Arthritis, Experimental ; drug therapy ; genetics ; metabolism ; Asteraceae ; chemistry ; Disease Models, Animal ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; Flavonoids ; administration & dosage ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Male ; Membrane Proteins ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Synovial Membrane ; drug effects ; metabolism ; Wnt Signaling Pathway ; drug effects ; beta Catenin ; metabolism