Andrographolide impairs alpha-naphthylisothiocyanate-induced cholestatic liver injury in vivo.

To investigate if andrographolide impairs cholestatic liver injury. All rats were randomly divided into six groups-(1) control (n = 6), (2) control + 200 mg/kg andrographolide (n = 6), (3) alpha-naphthylisothiocyanate (ANIT)-control (n = 6), (4) ANIT + 50 mg/kg andrographolide (n = 6), (5) ANIT + 100 mg/kg andrographolide (n = 6), and (6) ANIT + 200 mg/kg andrographolide (n = 6). We gavaged 50 mg/kg ANIT to mimic cholestatic liver injury in rats. Seven days after treatment, all the rats were killed. Serum biochemistry and hepatic histopathological assays were performed to evaluate liver injury. We observed that 200 mg/kg andrographolide significantly decreased the level of alanine transaminase, aspartate aminotransferase, alkaline phosphatase, γ-glutamyltranspeptidase, total bilirubin, and total bile acid in the blood. It also markedly decreased hepatic interleukin-6 and tumor necrosis factor α. Furthermore, 200 mg/kg andrographolide significantly decreased malondialdehyde but increased superoxide dismutase, glutathione, and erythrocyte glutathione peroxidase. Moreover, 200 mg/kg andrographolide effectively increased the accumulation of sirtuin 1 and nuclear erythroid 2-related factor-2. It also attenuated the level of nuclear factor kappa-light-chain-enhancer of activated B and cyclooxygenase-2. These data suggest that andrographolide may impair cholestatic liver injury via anti-inflammatory and anti-oxidative stress.

Anti-inflammatory activity of chitosan nanoparticles carrying NF-κB/p65 antisense oligonucleotide in RAW264.7 macropghage stimulated by lipopolysaccharide.

The purpose of this present study is to prepare NF-κB/p65 antisense oligonucleotide loaded chitosan nanoparticles (NPs) and evaluate their physicochemical characterization and antisense effects in RAW264.7 macrophages. Condensed nanoparticles with mean particle size of 128±16nm, average Zeta potential of 19.6±6.3mV and high entrapment efficiency (EE) of 98.6±0.11% were formed between NF-κB/p65 antisense gene (NAG) and chitosan by complex coacervation method. Trypan blue staining and MTT tests showed that NAG chitosan NPs had no toxic effect on RAW264.7 macrophages when the dose was no more than 20µg/mL. Confocal microscopy images showed that NAG chitosan NPs were capable to deliver NAG into cytoplasm of RAW264.7 macrophages and finally into nucleus. Real-time PCR tests verified that NAG chitosan NPs could significantly decrease the mRNA expression level of NF-κB/p65 and inflammatory cytokines including TNF-ɑ, IL-1 and IL-6. Accordingly, western blot study showed that NAG NPs uptaken in the cells could efficiently reversed the expression of NF-κB/p65 protein induced by LPS. At last, downstream release level of inflammatory factors including TNF-ɑ, IL-1 and IL-6 in LPS stimulated RAW264.7 macrophages was significantly decreased after treated by NAG chitosan NPs. It could be concluded that chitosan NPs were excellent delivery vectors to ferry the NAG into the cytoplasm and nucleus of macrophages. The NAG chitosan NPs might be a novel therapeutic apparatus for the treatment of LPS induced sepsis by inhibiting NF-κB-related pro-inflammatory cytokines secretion.

[Structural and functional changes in islet beta cells in severely scalded rats].

OBJECTIVE: To observe the structural and functional changes in islet beta cells in severely scalded rats, and to explore its relationship with dysfunction of glycometabolism. METHODS: Seventy-two Wistar rats were divided into scald (S) group and sham injury (SI) group according to the random number table, with 36 rats in each group. Rats in group S were inflicted with 50%TBSA full-thickness scald by a 12-s immersion of back and a 6-s immersion of abdomen in 94 °C hot water. Rats in group SI were sham injured through immersion of back and abdomen in 37 °C warm water. At post injury hour (PIH) 6 and on post injury day (PID) 3 and 7, plasma glucose level was measured for intraperitoneal glucose tolerance test (IPGTT) in 12 rats of each group, and the area under the curve (AUC) of plasma glucose level was calculated. After the IPGTT, pancreatic tissue was harvested and subjected to a double immunostaining for insulin and cell nuclei to determine the pancreatic insulin-positive area ratio, and the area and number of beta cells in the islets (referred to as "the three indicators in the islets"). Data were processed with the analysis of repeated measures and factorial designed analysis of variance, and LSD test was applied for paired comparison. RESULTS: (1) At PIH 6 and on PID 3, the overall plasma glucose levels of rats in group S before and after injection of glucose and at each time point were obviously higher than those of rats in group SI (with F values of main effects respectively 79.372 and 32.962, P values all below 0.001; with P values of paired comparison below 0.05 or 0.01). On PID 7, the overall plasma glucose levels in the two groups before and after injection of glucose and at each time point were close (with P values all above 0.05). (2) The overall AUC of plasma glucose levels of rats in group S was higher than that of rats in group SI (main effects: F = 337.87, P < 0.01). Compared with those of rats in group SI [(1019 ± 32), (1003 ± 72) mmol·min·L(-1)], the AUCs of plasma glucose levels of rats in group S were higher at PIH 6 and on PID 3 [(1501 ± 163), (1132 ± 67) mmol·min·L(-1), P values all below 0.001]. The AUCs of plasma glucose levels were close between two groups on PID 7 (P > 0.05). The AUCs of plasma glucose levels on PID 3 and 7 were both lower than that at PIH 6 in rats of group S (with P values all below 0.001). (3) The three indicators in the islets in rats of group S were all lower than those of rats in group SI (with F values of main effects respectively 135.17, 24.75 and 39.35, P values all below 0.01). There were no significant differences in the three indicators in the islets at PIH 6 between two groups (with P values all above 0.05). The three indicators in the islets of rats in group S on PID 3 and 7 [0.47 ± 0.05, 0.51 ± 0.07; (0.032 ± 0.008), (0.037 ± 0.008) mm(2); (303 ± 64), (341 ± 58) cells] were significantly lower than those of rats in group SI [0.63 ± 0.05, 0.64 ± 0.06; (0.043 ± 0.011), (0.044 ± 0.012) mm(2); (398 ± 112), (387 ± 90) cells; P < 0.05 or P < 0.01] and that at PIH 6 within group S (P < 0.05 or P < 0.01). CONCLUSIONS: The number of beta cells is reduced, and the insulin secretion function of beta cells is decreased in the scalded rats, and they may constitute the cause of dysfunction of glycometabolism, mainly manifested as hyperglycemia.