The Protective Role of Xanthophyll Cycle in Resurrection Angiosperm Boea hygrometrica During Dehydration and Rehydration

The protective role of xanthophyll cycle in resurrection angiosperm Boea hygrometrica (Bunge) R.Br. was investigated by analysis of the changes of chlorophyll fluorescence and xanthophyll cycle components in response to dehydration and rehydration in detached leaves under very weak light condition (3 mumol photons.m(-2).s(-1)) and in the dark. With declines in the values of PSII photochemical efficiency (Fv/Fm), PSII actual quantum yield (Phi(PSII)), photochemical quenching (qP) and non-photochemical quenching (NPQ) during dehydration, zeaxanthin significantly increased in control Boea leaves under very weak light condition, while no zeaxanthin accumulation was detected in Boea leaves treated with dithiothreitol (DTT) and Boea leaves in the dark, and after 3 d rehydration, the parameters Fv/Fm, Phi(PSII), qP and NPQ showed full recovery in control Boea leaves under very weak light condition, but the parameters only underwent partial recovery in Boea leaves treated with DTT and Boea leaves in the dark, suggesting that the recovery of photosystem II (PSII) photochemical activities in Boea leaves was obviously affected by treatments with DTT and darkness, therefore, zeaxanthin may play an important protective role in desiccated Boea leaves even under very weak light conditions.

Protection of ghrelin postconditioning on hypoxia/ reoxygenation in gastric epithelial cells

AIM: To investigate the protective effect and mechanisms of ghrelin postconditioning against hypoxia/reoxygenation (H/R)-induced injury in human gastric epithelial cells. METHODS: The model of H/R injury was established in gastric epithelial cell line (GES-1) human gastric epithelial cells. Cells were divided into seven groups: normal control group (N); H/R postconditioning group; DMSO postconditioning group (DM); ghrelin postconditioning group (GH); D-Lys3-GHRP-6 + ghrelin postconditioning group (D + GH); capsazepine + ghrelin postconditioning group (C + GH); and LY294002 + ghrelin postconditioning group (L + GH). 3-(4,5-dimethylthazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to detect GES-1 cell viability. Hoechst 33258 fluorochrome staining and flow cytometry were conducted to determine apoptosis of GES-1cells. Spectrophotometry was performed to determine release of lactate dehydrogenate (LDH). Protein expression of Bcl-2, Bax, Akt, and glycogen synthase kinase (GSK)-3β was determined by western blotting. Expression of vanilloid receptor subtype 1 (VR1), Akt and GSK-3β was observed by immunocytochemistry. RESULTS: Compared with the H/R group, cell viability of the GH group was significantly increased in a dosedependent manner (55.9% ± 10.0% vs 69.6% ± 9.6%, 71.9% ± 17.4%, and 76.3% ± 13.3%). Compared with the H/R group, the percentage of apoptotic cells in the GH group significantly decreased (12.38% ± 1.51% vs 6.88% ± 0.87%). Compared with the GH group, the percentage of apoptotic cells in the D + GH group, C + GH group and L + GH groups significantly increased (11.70% ± 0.88%, 11.93% ± 0.96%, 10.20% ± 1.05% vs 6.88% ± 0.87%). There were no significant differences in the percentage of apoptotic cells between the H/R and DM groups (12.38% ± 1.51% vs 13.00% ± 1.13%). There was a significant decrease in LDH release following ghrelin postconditioning compared with the H/R group (561.58 ± 64.01 U/L vs 1062.45 ± 105.29 U/L). There was a significant increase in LDH release in the D + GH, C + GH and L + GH groups compared with the GH group (816.89 ± 94.87 U/L, 870.95 ± 64.06 U/L, 838.62 ± 118.45 U/L vs 561.58 ± 64.01 U/L). There were no significant differences in LDH release between the H/R and DM groups (1062.45 ± 105.29 U/L vs 1017.65 ± 68.90 U/L). Compared with the H/R group, expression of Bcl-2 and Akt increased in the GH group, whereas expression of Bax and GSK3β decreased. Compared with the GH group, expression of Bcl-2 decreased and Bax increased in the D + GH, C + GH and L + GH groups, and Akt decreased and GSK-3β increased in the L + GH group. The H/R group also upregulated expression of VR1 and GSK-3β and downregulated Akt. The number of VR1-positive and Akt-positive cells in the GH group significantly increased, whereas the number of GSK-3β-positive cells significantly decreased. These effects of ghrelin were reversed by capsazepine and LY294002.CONCLUSION: Ghrelin postconditioning protected against H/R-induced injury in human gastric epithelial cells, which indicated that this protection might be associated with GHS-R, VR1 and the PI3K/Akt signaling pathway.

Study on the effect of four kinds of raw materials in hypertonic dehydration cell model

It aims to investigate the protective effects of sodium hyaluronate,panthenol,Portulaca oleracea L.and Calendula officinalis L.on hyperosmotic dehydration-induced injury of human immortalized keratinocytes(HaCaT).The safety mass concentrations of four raw materials were screened by detecting cell viability,and the secretion of hyaluronic acid(HA)was determined using the ELISA method.The expression of HaCaT barrier function related genes(OVOL1,EREG,TGM1,TGM2,IVL,IRF6,THBS1,CASP14)was detected at the mRNA level to explore the regulatory effect of four raw materials on these genes.The results demonstrate that pretreatment with the four kinds of raw materials could increase the cell viability after hyperosmotic dehydration,promote the secretion of HA,and improve the expression of barrier function related genes after hyperosmotic dehydration,among which panthenol and Calendula officinalis L.are better.The results show that the four raw materials have a certain protective effect on the hyperosmotic dehydration cell model,which provides data support for its application in cosmetics.

Protective effect of astrocyte-conditioned medium on neurons following hypoxia and mechanical injury

Objective： To investigate the protective effect of mouse astrocyte-conditioned medium （ACM） on hypoxic and mechanically injured neurons by a cell model in vitro, and to explore the possible mechanism. Methods： The model of hypoxic neuronal injury was caused by 3% 02 in three-gas incubator. Neurons were cultured with ordinary medium or 20% ACM respectively and randomly divided into hypoxic group （hypoxia for 4, 8, 24 h and marked as H4R0, H8R0, H24R0） and hypoxia reoxygenation group （H4R24, H8R24, H24R24）. Mechanical injury model was developed by scratching neurons cultured in 20% ACM or ordinary medium to different degrees. Neu- rons in both medium were divided into normal control group, mild, moderate and severe injury groups. The 20% ACM was added 24 h before hypoxia/reoxygenation or mechanical injury. The morphology and survival of neurons were observed and counted by trypan blue staining. The concentration of NO, lactic dehydrogenase （LDH） and membrane ATPase activity were detected by corresponding kits. Results： It was showed that 20% ACM can obviously promote the survival rate of hypoxia/reoxygenated neurons and scratched neurons as well. The morphology and num- ber of neurons exposed to hypoxia or scratch injury showed great difference between groups with or without ACM treatment. Compared with control group, the concentration of NO and LDH was much lower in hypoxic/reoxygenated neurons treated with 20% ACM, and the ATPase activity was higher. For the mechanical injury model, neurons with moderate injury also revealed a lower NO and LDH concen- tration than the control group. All the differences were sta- tistically significant （P〈0.05）. Conclusion： ACM can promote the survival and func- tional recovery of neurons following hypoxia or scratching to a certain degree. The mechanism may be associated with reducing the synthesis and release of NO and LDH as well as increasing the activity of membrane ATPase.