Effects of Pogostemon cablin on gastrointestinal function of rats with syndrome of damp retention in middle-jiao / 中国中药杂志

To investigate the effects of Pogostemon cablin(patchouli) on gastrointestinal function of rats with the syndrome of damp retention in middle-jiao, and explore its therapeutic mechanism. In this study, gastrointestinal function of rats with the syndrome of damp retention in middle-jiao was evaluated by multiple assays including gastric remnant rate, small intestine propelling rate, gastric juice quantity, pepsin activity and gastrointestinal tissue morphology. ELISA was used to detect gastrointestinal hormones including MTL, GAS, VIP and cytokines including TNF-α and interleukin 10 in rat serum. Real-time fluorescent quantitative PCR technique was used to detect relative mRNA expression of AQP3, AQP4 and AQP8 in gastric and colonic tissues to explore the mechanism of P. cablin in treatment of gastrointestinal functions. The results showed that middle and high dose of P. cablin (3.24, 6.48 g•kg⁻¹) could obviously decrease the gastric remnant rate, promote gastric emptying, increase the small intestine propelling rate(P<0.05), speed up the propulsive movement of gastrointestinal tract, increase the secretion and acidity of gastric juice, increase the activity of pepsin, and improve the injury of gastrointestinal tissue. All the doses of P. cablin could increase the concentration of MTL and GAS in serum, reduce the concentration of VIP, TNF-α and IL-10 in serum, decrease the mRNA expression of AQP3 in gastric and colonic tissues, and increase the expression levels of AQP4 and AQP8 in colonic tissues. The regulatory effects were better in middle and high dose groups. In conclusion, regulation of the levels of gastrointestinal hormones, inflammatory cytokines and aquaporins may be the paths for P. cablin to maintain normal gastrointestinal function of rats with the syndrome of damp retention in middle-jiao. The results of the study laid a foundation for clarifying the treatment mechanism of aromatic damp-resolving drugs for indications including damp retention in middle-jiao and transformation failure of spleen.

Attenuation mechanism of gamboges during processing based on inflammatory toxicity and AQP3, AQP4 protein and mRNA expressions in rat gastric and duodenal tissues / 中国中药杂志

To expand the clinical application of gamboges, it is necessary to study crude gamboges' toxicity after oral administration and attenuation mechanism during processing. In this study, crude gamboges' toxicity was judged by multiple assays, including inflammatory mediums [such as nitric oxide (NO), tumor necrosis factor alpha (TNF-α), and interleukin 6 (IL-6)] released by macrophage RAW264.7, and pathological manifestations of rat stomach and duodenal tissues after oral administration with crude and processed gamboges. The attenuation mechanism during processing was studied by detecting AQP3, AQP4 protein and mRNA expression in rat gastric and duodenal tissues using immunohistochemical assay and real-time fluorescent quantitative PCR technique. According to the results, crude gamboges group showed promotion in release of NO, TNF-α and IL-6 by macrophage RAW264.7 in a dose-dependent manner; Compared with crude gamboges group, processed gamboges group showed reduction in release of NO and IL-6, with increase in TNF-α. Crude gamboges could cause rat diarrhea, white blood cells increase, lymphocytes decrease, hyperemia and edema in rat gastric mucosa, duodenal mucosal necrosis and inflammatory cells infiltration. All of these results proved that gamboges had the inflammatory toxicity in gastric and duodenal tissues after oral administration in a dose-dependent manner, which however reduced after processing. In addition to the inflammatory toxicity, the mRNA and protein expressions of aquaporin 3 (AQP3), aquaporin 4 (AQP4) in gastric and duodenal tissues of high-dose crude gamboges group were increased significantly (P<0.05), while the protein and mRNA expressions of AQP3, AQP4 were weakened in processed gamboges group. The results showed that AQP3, AQP4 protein and mRNA expressions were positively correlated with the inflammatory toxicity. In conclusion, down-regulation of AQP3, AQP4 protein and mRNA expressions may be one of attenuation mechanisms in processing gamboges.

Determination of amino acids in Galli Gigerii endothelium corneum by HPLC with pre-column derivatization / 中国中药杂志

Hydrolytic amino acids were extracted by acid hydrolysis method, then derivatized with phenyl isothiocyanate (PITC). And the samples were analysed by HPLC on an Ultimate Prime C18 (4.6 mm x 250 mm, 5 microm) column with gradient elution of 0.1 mol x L(-1) sodium acetate buffer solution (adjusted to pH 6. 5)-acetonitrile (93:7) (A) and acetonitrile-water (8:2) (B) at a flow rate of 1.0 mL x min(-1). Column temperature was 40 degrees C and the detected wavelength was 254 nm. Amino acids derivative solution remained stable in 36 hours. The response was linear for 16 amino acids with a correlation coefficient r > 0.999 5. The average recoveries were 98.01% -101.8%. The method is reliable with good accuracy and repeatability, which is useful for the determination of amino acids in Galli Gigerii Endothelium Corneum.

Study on tissue distribution of psoraleae fructus decoction in rats / 中国中药杂志

Advanced technologies are used to clarify the meridian tropism theory of traditional Chinese medicine is an important part of theoretical studies of traditional Chinese medicine. In this article, modern pharmacokinetic method was used to investigate tissue distribution characteristics of psoralen and isopsoralen of Psoraleae Fructus decoction in rats, in order to provide research ideas and experimental basis for the meridian tropism theory. In this study, various tissue samples such as heart, liver, spleen, lung, kidney, brain and spermary were collected at different times after oral administration with FP decoction, in order to determine concentration of psoralen and isopsoralen by HPLC. Pharmacokinetic parameters were calculated by DAS 2.0 software. The study results showed that HPLC indexes of psoralen and isopsoralen in various tissues of rats met the determination requirements of biological samples. Both components were distributed in all of the tissues, with AUC(0-t) order of liver > lung approximately kidney > heart > brain approximately spleen > spermary. There was significant difference between liver, kidney, lung and other tissues (P < 0.05). MRT(0-t) of both psoralen and isopsoralen were about 10 h. Therefore, psoralen and isopsoralen showed stronger targeting selection in liver, kidney and lung.