Effects of Curcumin-mediated photodynamic therapy on autophagy and epithelial-mesenchymal transition of lung cancer cells.

BACKGROUND: This study aimed to investigated whether Curcumin-mediated PDT can suppress EMT in lung cancer cells, and explore the roles of autophagy in the process of regulating EMT. METHODS: Lung cancer cell viability was assessed by CCK-8 assay. The expression of epithelial marker and mesenchymal markers, the conversion of LC3-I to LC3-II and the levels of p62 and beclin1 in A549 and SPCA1 cells were measured by Western blotting assay. The Wound healing and Transwell assays were used to detect the migration and invasion abilities of the A549 and SPCA1 cells. Autophagosome formation was detected via observing the colocalization of Lamp-2 with LC3 in A549 cells, and the autophagy ultrastructure was observed by TEM. RESULTS: Curcumin-PDT inhibited EMT, migration and invasion and induced autophagy in lung cancer cells. Curcumin-PDT induced autophagy was involved in the process of PDT inhibiting EMT, but it presented a promoting effect of EMT in lung cancer cells. Curcumin-PDT combined with CQ further inhibited EMT, invasion and migration of lung cancer cells. CONCLUSIONS: The role of PDT-induced autophagy in the regulation of EMT was determined to be a promoting effect in lung cancer. Therefore, Curcumin-mediated PDT combined with autophagy inhibitor further suppressed EMT of lung cancer cells, and may represent a potential strategy against invasion and migration of lung cancer.

Low-intensity pulsed ultrasound inhibits fibroblast-like synoviocyte proliferation and reduces synovial fibrosis by regulating Wnt/ß-catenin signaling.

OBJECTIVE: Synovial fibrosis is a characteristic symptom of osteoarthritis (OA), which is closely associated with joint pain and stiffness. Previous studies have reported that low-intensity pulsed ultrasound (LIPUS) can alleviate cartilage degradation in OA. However, the functions and mechanisms of LIPUS in OA synovial fibrosis are still unknown. METHODS: The destabilization of the medial meniscus (DMM) mouse model of OA was established in C57 male mice and fibroblast-like synoviocytes (FLS) were isolated from synovial tissue of OA patients. The knee joint diameter, Masson's trichrome (MT) and Hematoxylin-eosin (HE) staining were used to evaluate synovial fibrosis and hyperplasia. The Immunohistochemistry (IHC) staining was performed to detected the expression of synovial fibrosis makers and the activation of Wnt/ß-catenin signaling in vivo. FLS were treated with TGF-ß1 to serve as an in vitro model of synovial fibrosis, Wnt3a was used to activate the Wnt/ß-catenin signaling in cells. Cell proliferation was detected by using EdU assay, cell viability was performed by CCK8 assay. The protein levels of α-SMA, CTGF, Col Ⅰ, ß-catenin, active ß-catenin, c-Myc and cyclin D1 were examined by western blot and immunofluorescence staining. RESULTS: Two weeks after the LIPUS treatment, the synovial fibrosis, synovial hyperplasia and synoviocyte proliferation in the DMM model were significantly decreased. In vitro, LIPUS directly inhibited the TGF-ß1-induced fibrotic response and proliferation of FLS. Meanwhile, LIPUS suppressed Wnt/ß-catenin signaling in the synovium of DMM mice and cultured FLS. More importantly, we found that the synovial fibrosis makers, Wnt/ß-catenin pathway downstream proteins and FLS proliferation were significantly decreased in Wnt3a-stimulated FLS following LIPUS treatment. CONCLUSIONS: Our results present a novel role of LIPUS in OA-related synovial fibrosis, which is associated with its ability to repress Wnt/ß-catenin signaling in FLS. THE TRANSLATIONAL POTENTIAL OF THIS ARTICLE: This study provides new insight into the clinical application of LIPUS as a therapeutic option to manage synovial fibrosis in OA.

Curcumin-mediated Photodynamic Therapy Inhibits the Phenotypic Transformation, Migration, and Foaming of Oxidized Low-density Lipoprotein-treated Vascular Smooth Muscle Cells by Promoting Autophagy.

ABSTRACT: Vascular smooth muscle cells (VSMCs) are becoming a hot spot and target of atherosclerosis research. This study aimed to observe the specific effects of curcumin (CUR)-mediated photodynamic therapy (CUR-PDT) on oxidized low-density lipoprotein (ox-LDL)-treated VSMCs and confirm whether these effects are mediated by autophagy. In this study, the mouse aortic smooth muscle cell line and A7r5 cell lines were used for parallel experiments. VSMC viability was evaluated by Cell Counting Kit-8 assay. VSMCs were treated with ox-LDL to establish a model of atherosclerosis in vitro. The autophagy level and the expression of proteins related to phenotypic transformation were detected by western blotting. The migration ability of the cells was detected by using transwell assay. The presence of intracellular lipid droplets was detected by Oil Red O staining. The results showed that VSMCs transformed from the contraction phenotype to the synthetic phenotype when stimulated by ox-LDL, during which autophagy was inhibited. However, CUR-PDT treatment significantly promoted the level of autophagy and inhibited the process of phenotypic transformation induced by ox-LDL. In addition, ox-LDL significantly promoted VSMC migration and increased the number of lipid droplets, whereas CUR-PDT treatment significantly reduced the ox-LDL-induced increase in the migration ability of, and lipid droplet numbers in, VSMCs. When the VSMCs were pretreated with the autophagy inhibitor 3-methyladenine for 24 hours, the effects of CUR-PDT were reversed. Therefore, our study indicated that CUR-PDT can inhibit the phenotypic transformation, migration, and foaming of ox-LDL-treated VSMCs by inducing autophagy.

Excessive application of pig manure increases the risk of P loss in calcic cinnamon soil in China.

Soil phosphorus (P) is a critical factor affecting crop yields and water environmental quality. To investigate the degree of loss risk and forms of soil P in calcic cinnamon soil, the P fraction activities in soils were analysed using chemical methods, combined with an in situ field experiment. Seven treatments were set in this study, including control (unfertilized), no P fertilizer (No-P), mineral P fertilizer (Min-P), low (L-Man) and high (H-Man) quantities of pig manure, Min-P+L-Man, and Min-P+H-Man. The results showed that manure fertilizer could not only significantly increase maize yield but could also enhance the accumulation of soil P in organic and inorganic forms. After 23years of repeated fertilization, the soil Olsen-P contents respectively showed 64.7-, 43.7- and 31.9-fold increases in the Min-P+H-Man, Min-P+L-Man and H-Man treatments, while the soil Olsen-P in Min-P treatment only increased 23.7-fold. The soil Olsen-P thresholds ranged from 22.59 to 32.48mgkg-1 in calcic cinnamon soil to maintain a higher maize yield as well as a lower risk of P loss. Therefore, long-term excessive manure application could obviously raise the content of soil Olsen-P and increase the risk of P loss in calcic cinnamon soil.

[Temporal and spatial variations of soil NO(3-)-N in Orychophragmus violaceus/spring maize rotation system in North China].

The February orchid (Orychophragmus violaceus)-spring maize rotation system is established to resolve the problems caused by the expansion of fallow fields in North China. Based on a site-specific experiment, temporal and spatial variations of soil NO(3-)-N were investigated during the period from February orchid incorporation to maize harvest. The results showed that the nitrate content in soil profiles not only showed a temporal characteristic, i. e., increasing at the beginning of the maize season and decreasing then after, but also showed a spatial characteristic, i. e., the gradual occurrence of the peak of nitrate content from shallower to deeper layer with the growth season of maize. Meanwhile, incorporation of February orchid could affect temporal and spatial variations of soil NO(3-)-N. February orchid planting reduced the soil NO(3-)-N accumulation in the profile of 0180 cm. After incorporation of February orchid, similar characteristics were observed at the seedling and bell stages of maize, i. e., the soil NO(3-)-N mainly stayed in the profile of 0-20 cm, and NO(3-)-N concentrations in the treatments with February orchid were higher in 0-100 cm layer and lower in 100-180 cm layer than those of the treatments without February orchid. After tasseling stage, opposite phenomena were found, and the soil NO(3-)-N content was all relative low. Overall, incorporation of February orchid could increase the storage capacity of soil NO(3-)-N in the profile of 0-180 cm.