Genkwa Fols, Kansui Radix, and Euphorbiae Pekinensis Radix in Shizao Decoction(SZD) are toxic to intestinal tract. Jujubae Fructus in this prescription can alleviate the toxicity, but the mechanism is still unclear. Therefore, this study aims to explore the mechanism. To be specific, 40 normal Sprague-Dawley(SD) rats were classified into the normal group, high-dose and low-dose SZD groups, and high-dose and low-dose SZD without Jujubae Fructus(SZD-JF) groups. The SZD groups were given(ig) SZD, while SZD-JF groups received the decoction without Jujubae Fructus. The variation of body weight and spleen index were recorded. The patho-logical changes of intestinal tissue were observed based on hematoxylin and eosin(HE) staining. The content of malondialdehyde(MDA) and glutathione(GSH) and activity of superoxide dismutase(SOD) in intestinal tissue were measured to evaluate the intestinal injury. Fresh feces of rats were collected to detect intestinal flora structure by 16S ribosomal RNA gene(16S rDNA) sequencing technology. The content of fecal short chain fatty acids and fecal metabolites was determined by gas chromatography-mass spectrometer(GC-MS) and liquid chromatography-mass spectrometer ultra-fast liquid chromatography-quadrupole-time-of-flight mass spectrometer(UFLC-Q-TOF-MS), separately. Spearman's correlation analysis was employed to analyze the differential bacteria genera and differential metabolites. RESULTS:: showed that high-dose and low-dose SZD-JF groups had high content of MDA in intestinal tissue, low GSH content and SOD activity, short intestinal villi(P<0.05), low diversity and abundance of intestinal flora, variation in the intestinal flora structure, and low content of short chain fatty acids(P<0.05) compared with the normal group. Compared with high-dose and low-dose SZD-JF groups, high-dose and low-dose SZD groups displayed low content of MDA in intestinal tissue, high GSH content and SOD activity, recovery of the length of intestinal villi, increased abundance and diversity of intestinal flora, alleviation of dysbacteria, and recovery of the content of short chain fatty acids(P<0.05). According to the variation of intestinal flora and fecal metabolites after the addition of Jujubae Fructus, 6 differential bacterial genera(Lactobacillus, Butyricimonas, Clostridia＿UCG-014, Prevotella, Escherichia-Shigella, Alistipes),4 differential short chain fatty acids(such as acetic acid, propionic acid, butyric acid, valeric acid) and 18 differential metabolites(such as urolithin A, lithocholic acid, and creatinine) were screened out. Beneficial bacteria such as Lactobacillus were in positive correlation with butyric acid and urolithin A(P<0.05). The pathogenic bacteria such as Escherichia-Shigella were in negative correlation with propionic acid and urolithin A(P<0.05). In summary, SZD-JF caused obvious intestinal injury to normal rats, which could lead to intestinal flora disorder. The addition of Jujubae Fructus can alleviate the disorder and relieve the injury by regulating intestinal flora and the metabolites. This study discusses the effect of Jujubae Fructus in relieving the intestinal injury caused by SZD and the mechanism from the perspective of intestinal flora-host metabolism, which is expected to serve as a reference for clinical application of this prescription.
Rats ; Animals ; Rats, Sprague-Dawley ; Propionates/pharmacology* ; Gastrointestinal Microbiome ; Fatty Acids, Volatile/pharmacology* ; Butyrates/pharmacology*

OBJEVTIVETo synthesize phenoxybutyric acid derivatives as 5α-reductase inhibitors and test their biological activities in vitro.

METHODSEight analogues as nonsteroidal 5α-reductase inhibitors were designed and synthesized by substitution reaction of 6-(4-phenyl-piperazine-1-yl)-3(2H)-pyridazinone with phenoxybutyric acid derivatives.

RESULTS AND CONCLUSIONThe structures of the compounds were characterized by 1H-NMR and MS. Biological evaluation indicated that 7 out of the 8 compounds exhibited moderate 5α-reductase inhibitory activities, especially the compounds A1 and A7 with inhibition rates reaching 12.50% and 19.64% at the concentration of 3.3 × 10⁻⁵ mol/L, respectively.

5-alpha Reductase Inhibitors ; chemical synthesis ; pharmacology ; Butyrates ; chemistry ; pharmacology ; Drug Design

OBJECTIVETo explore the effect and mechanism of sodium butyrate on expression of human clotting factor VIII in vitro.

METHODSMouse NIH/3T3 cell line was transfected with recombinant plasmid vector pRC/RSV-BDD-hFVIII, which enclosed B-domain deleted (760aa approximately 1 639aa) human factor VIII cDNA (BDD-hFVIII cDNA). Then cells were incubated in Dulbecco's modification of Eagle's medium (DMEM) containing sodium butyrate for 24 hours, hFVIII: C and hFVIII: Ag in the cell culture medium were measured by ELISA assay and one-stage method, respectively. In addition, the effect of sodium butyrate on transcription of cDNA encoding the whole hFVIII, heavy and light chain of hFVIII was also investigated by means of run-on assay.

RESULTSAfter stimulation of sodium butyrate, the levels of hFVIII: C and hFVIII: Ag increased 70% than those of control. Run-on assay showed that sodium butyrate enhanced the transcription of cDNA which encoded heavy chain of hFVIII.

CONCLUSIONSodium butyrate can improve the expression of hFVIII through enhancing the transcription of hFVIII heavy chain encoding cDNA. It demonstrated that sodium butyrate had potential utility in inducing the expression of hFVIII in vitro.

3T3 Cells ; Animals ; Butyrates ; pharmacology ; Factor VIII ; genetics ; Gene Expression Regulation ; drug effects ; Humans ; Mice

OBJECTIVETo explore the molecular mechanisms of apoptotic NB4 cells induced by the histone deacetylase inhibitor, sodium butyrate(SB).

METHODSSB was exposed to NB4 cells at a final concentration of 1.0 mmol/L. The untreated and treated cells were analysed with FACS and 2-dimensional electrophoresis (2-DE) at 0, 12, 24, 48 and 72 h. The changed protein spots were identified with MALDI-TOF-MS and ESI-TOF-MS/MS.

RESULTSSB induced apoptosis of NB4 cells. Twenty-one changed proteins involving apoptotic signal transduction, immunological regulation, transcriptional control, cellular metabolism, molecular transport and so on were identified. Thirteen of them had been reported to be related to apoptosis.

CONCLUSIONSB can induce apoptosis and many functional protein changes in tumor cells. These results pave the way to further explore the anti-tumor mechanism of SB.

Apoptosis ; drug effects ; genetics ; Butyrates ; pharmacology ; Cell Line, Tumor ; Flow Cytometry ; Humans ; Proteomics

OBJECTIVETo study the effects of sodium butyrate (SB) on growth, apoptosis and telomerase activity in Hep-2 cells.

METHODSGrowth inhibition effect of SB on Hep-2 cells was assessed by methyl thiazolyl tetrazolium (MTT) assay. Morphological alterations were observed by electronic microscope. Cell apoptosis was confirmed by terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling (TUNEL) method, DNA fragmentation and flow cytometry (FCM). Cell cycle was analyzed by FCM. Telomerase activity was examined by telomeric repeat amplification protocol (TRAP)-silver staining. The expression status of telomerase subunits was analyzed by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSA time-and dose-dependent inhibition was detected in cells treated with SB. Typical morphological changes of apoptotic cells were observed under electronic microscopy. The characteristic DNA fragmentation of apoptotic cells was detected by agarose gel electrophoresis. Apoptosis and the changes of cell cycle were confirmed by TUNEL method and FCM. The apoptosis indexes of the cells before treatment and at 72 h after SB (2.5 mmol/L) treatment were 2.27 +/- 1.18 and 33.50 +/- 2.75 respectively, the apoptosis rates were 2. 86% and 31. 28% respectively, the proportion of the cells at G0/G1 stage were 50.38% and 70.88% respectively, the proportion of the cells at S stage were 27.40% and 8.20% respectively, and the proliferation indexes of the cells were 49.62% and 29.12% respectively. Telomerase activity and expression level of human telomerase reverse transcriptase (hTERT), the key subunit of telomerase, decreased after SB treatment. No significant changes were observed in the expression of human telomerase RNA (hTR) and human telomerase associated protein (hTP1), the other two subunit of telomerase.

CONCLUSIONSB could inhibit growth of Hep-2 cells and induce apoptosis in the cells, and inhibit telomerase activity by decrease expression level of hTERT.

Apoptosis ; drug effects ; Butyrates ; pharmacology ; Cell Cycle ; drug effects ; Hep G2 Cells ; Humans ; Sodium ; pharmacology ; Telomerase ; metabolism

AIMTo design and synthesize new phenyloxyisobutyric acid analogues as antidiabetic compounds.

METHODSEight new target compounds were synthesized by combination of lipophilic moieties and acidic moiety with nucleophilic replacement or Mitsunobu condensation. The eight compounds were confirmed by 1H NMR, 13C NMR, IR and MS.

RESULTSIn vitro insulin-sensitizing activity (3T3-L1 adipocyte) demonstrated, that the cultured glucose concentration of up-clear solution detected with GOD-POD assay were 5.942, 6.339, 6.226 and 6.512 mmol x L(-1), respectively, when rosiglitazone, pioglitazone, compounds A and B were added to the insulin-resistant system.

CONCLUSIONIn vitro insulin-sensitizing activity of target compound A is in between that of rosiglitazone and pioglitazone, and activity of target compound B is slightly less than that of pioglitazone.

3T3-L1 Cells ; Adipocytes ; drug effects ; Animals ; Butyrates ; chemical synthesis ; chemistry ; pharmacology ; Hypoglycemic Agents ; chemical synthesis ; chemistry ; pharmacology ; Insulin ; pharmacology ; Mice ; Molecular Structure ; PPAR gamma ; agonists ; pharmacology

OBJECTIVETo elucidate the effects of sodium butyrate on rat hepatic oval cell differentiation in vitro.

METHODSHepatic oval cells were isolated from rats fed with a choline-deficient diet supplemented with 0.1% (w/w) ethonine for 4 to 6 weeks. The cultured hepatic oval cells were identified by immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR). After hepatic oval cells were treated with sodium butyrate, the morphological changes were studied through Giemsa staining and the albumin expression level was tested by Western blot.

RESULTSImmunohistochemical results showed the isolated cells were positive for both mature hepatocyte marker albumin and bile duct cell marker cytokeratin-19. Furthermore, RT-PCR results showed that the cells expressed stem cell marker c-kit, but not hematopoietic stem cell marker CD34. In short, the isolated cells were rat hepatic oval cells. 0.75 mmol/L sodium butyrate induced obvious phenotype changes of hepatic oval cells, including enlargement of the oval cells, a decrease in nucleus to cytoplasm ratio, and a 50% increase in the number of binucleated cells. Western blot results showed that 0.75 mmol/L sodium butyrate markedly raised the expression of albumin.

CONCLUSIONSodium butyrate, a differentiation promoting agent, can induce rat hepatic oval cells (liver progenitor cells) to differentiate into mature hepatocytes in vitro.

Animals ; Butyrates ; pharmacology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Hepatocytes ; cytology ; Liver ; cytology ; Rats ; Stem Cells ; cytology

The study was purposed to investigate the role of extracellular signal-regulated kinase (ERK) pathway in the differentiation of human MDS cell lines SKM-1 induced by sodium butyrate (NaB), and to elucidate the molecular mechanism of differentiation in SKM-1 cells induced by NaB. The expression levels of total ERK and phosphorylated-ERK were determined by Western blot. The effect of NaB in combination with the ERK inhibitor PD98059 on the proliferation/differentiation of SKM-1 cells was studied, and then the expression levels of the P21 and HDAC protein were detected by Western blot. The results showed that the expression level of phosphorylated ERK was down-regulated by the 1 mmol/L NaB, and the level of total ERK had not changed. NaB or combination of the MEK inhibitor PD98059 with NaB could increase the differentiation of the SKM-1 cells and up-regulated the levels of the P21 and HDAC protein, but the effect of combination of NaB with PD98059 was higher than that of NaB alone. It is concluded that the inhibition of ERK may be involved in sodium butyrate inducing differentiation in SKM-1 cells.
Butyrates ; pharmacology ; Cell Transformation, Neoplastic ; drug effects ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Humans ; Myelodysplastic Syndromes ; enzymology ; pathology ; Tumor Cells, Cultured

OBJECTIVETo explore the effect of sodium butyrate (SB) on the expression of costimulatory molecules in acute leukemia cells and its mechanism.

METHODSThe expression of CD86 and CD80 was examined on the surfaces of NB4, HL-60, Kasumi-1, U937 and Jurkat cells by flow cytometric analysis after treated by SB or not. Allogeneic mixed lymphocyte reaction was used to evaluate the immunomodulatory effects of cells treated by SB. Activated NF-kappaB was measured with an NF-kappaB assay kit.

RESULTSUp-regulation of CD86 and CD80 at various levels was observed on these leukemia cells treated by SB. The ratio of CD86 expressing cell in NB4 cells treated by 0.5 mmol/L SB was 36.8 times higher than that in control. Up-regulation of NF-kappaB was similar to that of CD86. Allogeneic lymphocyte proliferation was strongly stimulated by the SB treated cells.

CONCLUSIONSB can improve the expression of CD86 in acute leukemia cells. NF-kappaB was an important transcription factor involved in the up-regulation of CD86.

B7-1 Antigen ; biosynthesis ; B7-2 Antigen ; biosynthesis ; Butyrates ; pharmacology ; Cell Line, Tumor ; Humans ; Leukemia ; metabolism ; pathology ; NF-kappa B ; metabolism ; Up-Regulation ; drug effects

OBJECTIVETo study the effect of Radix Scutellariae on the growth, metabolism of Porphyromonas endodontalis (P.e), as a preparation for studying the mechanism of Radix Scutellariae in treating pulp and periapical diseases.

METHODSP.e was chosen as the experimental bacteria. Radix Scutellariae was extracted by means of reflux with 80% ethanol. The value of MIC of Radix Scutellariae was measured by minute amount serial dilusion test, and the production of butyrate was measured by high liquid chromatograph(HPLC).

RESULTSRadix Scutellariae could inhibit the growth of P.e, of which the MIC was 100 mg/L. Following the increase in concentration of Radix Scutellariae, the amount of butyrate decreased to (3.527 +/- 0.009) mg/L, (3.048 +/- 0.005) mg/L, (2.490 +/- 0.011) mg/L, (2.209 +/- 0.016) mg/L, respectively (P < 0.05).

CONCLUSIONRadix Scutellariae could inhibit the growth and metabolism of P.e and might be an effective agent in treating pulp and periapical diseases.

Bacteroidaceae Infections ; microbiology ; Butyrates ; analysis ; Chromatography, High Pressure Liquid ; Dental Pulp ; microbiology ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Microbial Sensitivity Tests ; Porphyromonas endodontalis ; metabolism ; Scutellaria ; chemistry