Effect of Jingfang Granules on carrageenan-induced tail thrombosis in mice based on ERK/p38 MAPK signaling pathway / 中国中药杂志

The present study explored the anti-inflammatory and anti-thrombotic mechanism of Jingfang Granules on tail thrombosis induced by carrageenan in mice. Thirty-two male ICR mice were randomly divided into a control group, a model group, a Jingfang Granules group, and a positive drug(aspirin) group, with eight mice in each group. The thrombosis model was induced by intraperitoneal injection of carrageenan(45 mg·kg~(-1)) combined with low-temperature stimulation, and the mice were treated with drugs for 7 days before modeling. Twenty-four hours after modeling, blood was detected for four blood coagulation indices in each group. The enzyme-linked immunosorbent assay(ELISA) was used to detect the activity of plasma interleukin-6(IL-6), interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), and other inflammatory factors. The tails of mice in each group were cut off to observe tail lesions and measure the length of the thrombus. The protein expression and phosphorylation level of extracellular signal-regulated kinase 1/2(ERK1/2) and p38 mitogen-activated protein kinase(p38 MAPK) in spleen tissues were detected by Western blot. The results showed that dark red thrombus appeared in the tails of mice in each group. The length of the black part accounted for about 40% of the total tail in the model group. Additionally, the model group showed prolonged prothrombin time(PT), increased fibrinogen(FIB) content, and shortened activated partial thromboplastin time(APTT). Compared with the model group, the groups with drug intervention displayed shortened black parts in the tail and improved four blood coagulation indices(P<0.05). As revealed by ELISA, the expression levels of TNF-α, IL-1β, and IL-6 in the mouse plasma were significantly up-regulated in the model group, and those in the groups with drug intervention were reduced as compared with the model group(P<0.05). As demonstrated by Western blot, the protein expression and phosphorylation levels of ERK1/2 and p38 MAPK in the spleen tissues were significantly elevated in the model group, while those in the Jingfang Granules group were down-regulated as compared with the model group with a significant difference. Jingfang Granules can inhibit tail thrombosis of mice caused by carrageenan presumedly by inhibiting the activation of ERK1/2 and p38 MAPK signaling pathways.

Metabonomics study of Shouhui Tongbian Capsules in slow transit constipation based on UPLC-ESI-QE-Orbitrap-MS / 中国中药杂志

The effect of Shouhui Tongbian Capsules(SHTB) on the endogenous metabolites of colon tissue in mice with slow transit constipation was analyzed by metabolomics methods to explore its mechanism in the treatment of constipation. ICR mice were randomly divided into normal group, model group and SHTB group according to the body weight. The mice were given diphenoxylate to establish the slow transit constipation model. Mouse carbon ink pushing rate, first defecation time and the number of defecation particles in 12 h were observed. The mouse colon tissue was separated and the mucous cells were detected by Periodic acid Schiff and Alcian blue(AB-PAS) staining. Ultra-high-performance liquid chromatography electrospray ionization orbitrap tandem mass spectrometry(UPLC-ESI-Orbitrap-MS/MS) technology was used to characterize the differences in tissue metabolism to screen out the potential different metabolites and possible metabolic pathways in colon tissue. The results indicated that SHTB could significantly shorten the first defecation time and the number of defecations, and increase the number of intestinal peristalsis and mucous cells in the colonic mucosa compared to the model mice. Metabolomics results showed that, compared with the normal group, a total of 17 potential biomarkers, including L-kynurenine, N6,N6,N6-trimethyl-L-lysine, L-formylkynurenine, N6-acetyl-L-lysine, L-phenylalanine, phenylacetaldehyde, xanthoxin, thymidine, glycyl-L-leucine, cystathionine,(R)-1-aminopropan-2-ol, deoxycytidine, gamma-glutamyl-gamma-aminobutyraldehyde, D-galactose, L-arginine, L-proline and pyruvate, were found and identified in colon tissue. Treated with SHTB, these metabolic differences tended to return to normal levels. Therefore, it could be made a conclusion that the therapeutic effect of SHTB on chronic transit constipation may be related to regulating phenylalanine metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, arginine and proline metabolism, cysteine and methionine metabolism, tyrosine metabolism, arginine biosynthesis, pyruvate metabolism, glycolysis, pyrimidine metabolism, tricarboxylic acid cycle and galactose metabolism.

Hydrogen rich water protects against myocardial ischemia/reperfusion injury in rats / 中国组织工程研究

BACKGROUND: Increasing evidence has indicated that low-concentration hydrogen or hydrogen rich water or hydrogen saturated saline exerts a protective effect on various diseases, such as myocardial ischemia/reperfusion injury. OBJECTIVE: To explore the protective effect of hydrogen rich water on myocardial ischemia/reperfusion injury. METHODS: Forty-eight Wistar rats were equally randomized into control and hydrogen-rich groups, and then subdivided into ischemic preconditioning, ischemia, and ischemia/reperfusion groups (n=8 rats in each subgroup). The myocardial ischemia/reperfusion model was established in the heart of each rat by the following procedures: reverse perfusion for 10 minutes, room temperature for 20 minutes, and reperfusion for 20 minutes. The control rats was perfused with pre-oxygenated (95% O2plus 5% CO2) 37 ℃ K-R solution and the hydrogen-rich group was perfused with pre-oxygen-equilibrated (95% O2plus 5% CO2) 37 ℃ K-R solution plus hydrogen-rich water (0.6 mmol/L, pH=7.3). Subsequently, the heart was removed, the pathological changes of the myocardial tissues were observe by hematoxylin-eosin staining, the activities of lactic dehydrogenase and creatine kinase in the myocardial tissues were determined, and the levels of tumor necrosis factor-α and interleukin-1β were detected by ELISA. RESULTS AND CONCLUSION: In the control group, the activity of lactic dehydrogenase at the ischemic and ischemia/reperfusion stages was significantly higher than that at the ischemic preconditioning stage (P < 0.05), and the activity of creatine kinase at the ischemia/reperfusion stage was significantly higher than that at the ischemic preconditioning and ischemic stages (P < 0.05). In the hydrogen-rich group, there was no significant difference in the activities of lactic dehydrodenase and creatine kinase at each stage, but the activities of at the ischemia/reperfusion stage was significantly lower than those in the control group (P < 0.05). In the two groups, the order of the levels of tumor necrosis factor-α and interleukin-1β was as follows: the ischemia/reperfusion stage > ischemic stage > ischemic preconditioning stage (P < 0.05). The levels of above factors in the hydrogen-rich group were significantly lower than those in the control group (P < 0.05). Our findings imply that hydrogen rich water has protective effect on myocardial ischemia/reperfusion injury of the rat hearts in vitro,which may be by reducing the expression of tumor necrosis factor-α and interleukin-1β, and further alleviating the inflammatory response.