AIMTo study the protective action of ulinastatin against lipopolysaccharide (LPS)-induced acute lung injury in mice and the mechanism of its action.

METHODSMice were intraperitoneally injected with ulinastatin (50 and 100 ku x kg(-1)) or saline at a period of 12 h, separately, 30 min after the last injection of ulinastatin, except normal control, all mice of other groups were injected a dose of LPS 15 mg x kg(-1) via tail vein. The levels of TNFalpha in serum and lung were measured by ELISA. The expression of TNFalpha mRNA and iNOS mRNA in lung was assayed by RT-PCR. The expression of c-Fos and c-Jun protein in lung was measured by Western blotting method. And the NO2- / NO3- level in serum and MDA in lung were measured with kits.

RESULTSThe levels of NO2- / NO3- and TNFalpha in serum, MDA and TNFa in lung all increased after iv injection of LPS. The expressions of TNFa mRNA, iNOS mRNA, c-Fos and c-Jun in lung of LPS-injected mice were enhanced. Pretreatment with ulinastatin 100 ku x kg(-1) decreased the levels of NO2- / NO3- in serum and lung, reduced the index of lung, and inhibited the expressions of iNOS mRNA and c-Jun in lung induced by LPS in mice, while ulinastatin showed no effect on TNFa level in serum and lung.

CONCLUSIONUlinastatin protected mice from acute lung injury induced by lipopolysaccharides via inhibiting the activation of c-Jun and iNOS mRNA expression.

Animals ; Blotting, Western ; Glycoproteins ; administration & dosage ; pharmacology ; Injections, Intraperitoneal ; Lipopolysaccharides ; Lung ; drug effects ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred ICR ; Nitric Oxide Synthase Type II ; biosynthesis ; genetics ; Protective Agents ; administration & dosage ; pharmacology ; Proto-Oncogene Proteins c-fos ; biosynthesis ; Proto-Oncogene Proteins c-jun ; biosynthesis ; RNA, Messenger ; biosynthesis ; genetics ; Respiratory Distress Syndrome, Adult ; chemically induced ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Necrosis Factor-alpha ; biosynthesis ; blood ; genetics

AIMTo study the mechanisms of anti-diabetic nephropathy of rhein on cultured human mesangial cells (HMCs).

METHODSTo mimic the hyperglycemic (HG) environment of diabetic nephropathy, 30 mmol x L(-1) glucose were added to 10% FBS RPMI 1640. The HMCs were treated with rhein for 8, 24, 48 or 72 h, at these time, the bioactivity, total activity of transforming growth factor-beta1 (TGFbeta1), activity of p38MAPK (p38 mitogen-activated protein kinases, by using immunoprecipitate and Western blot), MMP-2 (matrix metalloproteinase-2), and MMP-9 (matrix metalloproteinase-9, by using gelatinase zymography) and the proliferation of HMCs in high glucose media were measured. Meanwhile the levels of secretion of FN in cultured HMCs were measured.

RESULTSThe results showed that rhein markedly inhibit the proliferation of HMCs, significantly reduce the bioactivity of TGFbeta1 and FN secretion in HMCs, and decrease the increased activity of p38MAPK, but showed no action on the activities of MMP-2 and MMP-9.

CONCLUSIONRhein reduced the secretion of FN and inhibited the proliferation of HMCs may through inhibiting the bioactivities of TGFbeta1 and p38MAPK.

Animals ; Anthraquinones ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Culture Media ; Epithelial Cells ; cytology ; metabolism ; Fibronectins ; secretion ; Glomerular Mesangium ; cytology ; metabolism ; Glucose ; antagonists & inhibitors ; pharmacology ; Humans ; Lung ; cytology ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Mink ; Transforming Growth Factor beta ; metabolism ; Transforming Growth Factor beta1 ; p38 Mitogen-Activated Protein Kinases ; metabolism

Aged ; Beijing ; China ; Emergency Service, Hospital ; Hospitalization ; Humans ; Temperature

To reveal the rationality of compatibility of Salviae Miltiorrhizae Radix et Rhizoma(SMRR) and Puerariae Lobatae Radix(PLR) from the perspective of pharmacokinetics, this study established a UPLC-MS/MS method for quantitative determination of PLR flavonoids(3'-hydroxy puerarin, puerarin, puerarin 6″-O-xyloside, 3'-methoxy puerarin, puerarin apioside) and salvianolic acids and tanshinones(salvianolic acid B, cryptotanshinone, and tanshinone Ⅱ_A) in plasma of rats. Rats were given SMRR extract, PLR extract, and SMRR-PLR extract by gavage and then plasma was collected at different time. UPLC separation was performed under the following conditions: Eclipse C_(18) column(2.1 mm×50 mm, 1.8 μm), 0.1% formic acid in water(A)-0.1% formic acid in acetonitrile(B) as mobile phase for gradient elution. Conditions for MS are as below: multiple reaction monitoring(MRM), ESI~(+/-). Comprehensive validation of the UPLC-MS/MS method(specifically, from the aspects of calibration curve, precision, accuracy, repeatability, stability, matrix effect, extract recovery) was performed and the result demonstrated that it complied with quantitative analysis requirements for biological samples. Compared with SMRR extract alone or PLR extract alone, SMRR-PLR extract significantly increased the AUC and C_(max) of PLR flavonoids and tanshinones in rat plasma, suggesting that the combination of SMRR and PLR promoted the absorption of the above components. The underlying mechanism needs to be further studied.
Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal/pharmacokinetics* ; Plant Roots/chemistry* ; Pueraria/chemistry* ; Rats ; Rhizome/chemistry* ; Salvia miltiorrhiza/chemistry* ; Tandem Mass Spectrometry