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To improve the utilization of by-products of the Hyriopsis cumingii, this study investigated the effect of Hyriopsis cumingii polysaccharide (HCP) on the proliferation and migration of normal cells, as well as its reparative effects on ultraviolet radiation-B (UVB)-damaged cells. First, HCP was isolated and purified using DEAE Cellulose-52 column chromatography, Sephadex G-100 column chromatography and nanofiltration. Then, structural analysis was conducted using high-performance liquid chromatography, high-performance gel permeation chromatography, and infrared spectroscopy. Following this, cell proliferation experiments, cell scratch tests, UVB damage experiments, and protein expression analyses were employed to explore the anti-UVB damage effects and corresponding mechanisms of polysaccharide components. The results indicated the three substances obtained by isolation and purification were all homogeneous polysaccharide components. Among them, HCP-2 played a major role in promoting cell proliferation in HCP and facilitated the migration of HaCaT cells. The recovery of UVB-damaged cell vitality was raised by the increasing concentration of HCP-2. Furthermore, the reparative effect was more substantial than the protective effect at the same sample concentration. It was also observed that HCP-2 up-regulated the expression of the anti-apoptotic protein bcl-2 while inhibited the expression of apoptosis-related proteins casp9, p-Akt, and p-p38, indicating that HCP-2 could repair HaCaT cells by regulating apoptosis of cells damaged by UVB. In conclusion, HCP had the ability to inhibit cell apoptosis which provide a theoretical basis for its future application in the development of food and health products. [ABSTRACT FROM AUTHOR]

To improve the utilization of by-products of the Hyriopsis cumingii, this study investigated the effect of Hyriopsis cumingii polysaccharide (HCP) on the proliferation and migration of normal cells, as well as its reparative effects on ultraviolet radiation-B (UVB)-damaged cells. First, HCP was isolated and purified using DEAE Cellulose-52 column chromatography, Sephadex G-100 column chromatography and nanofiltration. Then, structural analysis was conducted using high-performance liquid chromatography, high-performance gel permeation chromatography, and infrared spectroscopy. Following this, cell proliferation experiments, cell scratch tests, UVB damage experiments, and protein expression analyses were employed to explore the anti-UVB damage effects and corresponding mechanisms of polysaccharide components. The results indicated the three substances obtained by isolation and purification were all homogeneous polysaccharide components. Among them, HCP-2 played a major role in promoting cell proliferation in HCP and facilitated the migration of HaCaT cells. The recovery of UVB-damaged cell vitality was raised by the increasing concentration of HCP-2. Furthermore, the reparative effect was more substantial than the protective effect at the same sample concentration. It was also observed that HCP-2 up-regulated the expression of the anti-apoptotic protein bcl-2 while inhibited the expression of apoptosis-related proteins casp9, p-Akt, and p-p38, indicating that HCP-2 could repair HaCaT cells by regulating apoptosis of cells damaged by UVB. In conclusion, HCP had the ability to inhibit cell apoptosis which provide a theoretical basis for its future application in the development of food and health products.

Objective To investigate the effects of expression of microRNA-223（miR-223) on cell proliferation, cell cycle, apoptosis and inflammatory reaction of psoriasis and their mechanism. Methods Human immortalized keratinocytes HaCaT were cultured in vitro. The cells were divided into the control group, model group, model+miR-NC group, model+miR-223 inhibitor group. Except for the control group, cells in the other groups were induced to establish psoriasis cell models by M5 method. Real-time quantitative PCR （RT-qPCR） was used to detect the expression of miR-223 in each group. CCK-8 method and flow cytometry were used to detect cell proliferation activity, cell cycle, and apoptosis in each group. ELISA was used to detect the levels of inflammatory factors such as IL-6, IL-1β and TGF-β1 in supernatant. Western blotting was used to detect the expression of phosphatase and tensin homolog deleted on chromosome ten （PTEN) protein. Results Compared with the control group, the expression level of miR-223 significantly increased （P<0. 05), OD values at 48, 72 and 96 h and the proportion of cells in the cell cycle S phase significantly increased （all P<0. 05), the proportion of cells in the cell cycle G0/G1 and apoptosis rate significantly decreased （all P<0. 05), the levels of IL-6, IL-1β and TGF-β1 significantly increased (all P<0. 05), while the expression of PTEN protein decreased in the model group （P<0. 05）. Compared with the model +miR-NC group, the expression level of miR-223 inhibitor group significantly decreased （P<0. 05)，OD values at 48, 72 and 96 h and the proportion of cells in the S phase decreased （P<0. 05), the proportion of cells in the cell cycle G0/G1 and apoptosis rate significantly increased （P<0. 05), the levels of IL-6, IL-1β and TGF-β1 significantly decreased （P<0. 05), and PTEN protein expression increased in the model +miR-223 inhibitor group （P<0. 05）. There were no significant differences in the above indexes between the model group and the model +miR-NC group （all P>0. 05). Conclusions The expression level of miR-223 increases in psoriasis cells. Down-regulating the level of miR-223 can inhibit the proliferation and inflammatory response of psoriasis cells, and promote their apoptosis and block the cell cycle in G0/G1 phase， which may be related to its regulation of PTEN. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Guangdong Pharmaceutical University is the property of Journal of Guangdong Pharmaceutical University and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

This study aims to investigate the protective effects of the extract of Sophora japonica L. (Sophora extract) and vitamin C (VC) on UV-induced damage in HaCaT cells. HaCaT cells were cultured in vitro and divided into different groups: Control group (without UV irradiation or treatment with Sophora extract, VC, or their combination), UV radiation group, 125 μg/mL Sophora extract group (with UV irradiation and treatment with Sophora extract), 125 μg/mL vitamin C group (with UV irradiation and treatment with VC), and 125 μg/mL combination group (with UV irradiation, Sophora extract and VC at a mass ratio of 1:1). The inhibitory effects of Sophora extract, VC, and their combination on reactive oxygen species (ROS) production and mitochondrial ROS in UV-irradiated HaCaT cells were measured using fluorescence staining and enzyme-linked immunosorbent assay. Immunofluorescence staining and enzyme-linked immunosorbent assay were also employed to evaluate the expression levels of cyclobutane pyrimidine dimers (CPDs) and the inflammatory cytokine IL-6 in UV-irradiated HaCaT cells treated with Sophora extract, VC, or their combination. The results showed that the total ROS inhibition rates of Sophora extract, VC, and the combination in UVA-treated HaCaT cells were 38.23%±9.23%, 27.20%±6.87%, and 64.13%±4.08%, respectively. The combination significantly enhanced the total ROS inhibition rate compared to the individual treatments (P

Copyright of Science & Technology of Food Industry is the property of Science & Technology of Food Industry Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The collagen was extracted by acid, enzymolysis and hot water methods respectively from the body wall of Sipunculus nudus, and their physicochemical properties were analyzed. The three collagen extracts were compared respectively by amino acid composition, UV spectroscopy, Fourier infrared spectroscopy and SEM scanning electron microscopy. The effects of the three collagen zynolytes on the growth viability of HaCaT cells were investigated. The results show that the extraction rate of collagen by hot water method is higher than the other two methods, and all three collagens contain the collagen characteristic amino acids Gly, Pro and Hyp. The UV and IR spectrograms show that all three collagens have obvious absorption peaks near 222 nm, and there are absorption peaks of amide A and amide B and characteristic peaks of amide Ⅰ, Ⅱ and Ⅲ, suggesting that all three collagens belong to type I collagen with a three-stranded helical structure. And the results of scanning electron microscopy show that the three-stranded helical structure of collagen extracted by acid method is more complete and maintains the original structure of collagen fibers better than that of enzymatic method and hot water method. The collagen peptides extracts have the ability to significantly increase the viability of normal and UVB-damaged HaCaT cells in the range of 50‒100 µg/mL, indicating that the enzymatic products of collagen of Sipunculus nudus have good potential applications in skin photoaging repair. [ABSTRACT FROM AUTHOR]

Objective To observe the effects of T follicular helper cells and the main functional cytokines IL-21 on the proliferation and cell cycle of HaCaT cells cultured in vitro in patients with psoriasis. Methods Fifteen patients with psoriasis vulgaris and 10 healthy controls were defined as psoriasis group and healthy control group, respectively. CD4+CXCR5+ Tfh cells from psoriatic patients and healthy controls were separated by flow cytometry and co-cultured with HaCaT cells at 1∶1. IL-21 or IL-21 neutralizing antibodies were added and the HaCaT cells were collected after 72 h of intervention. The cell proliferation was measured by the CCK-8 and the cell cycle and Ki-67 expression of HaCaT cells were detected by flow cytometry. Results IL-21 promoted the proliferation of HaCaT cells （P < 0.05） and increased the proportion of HaCaT cells in S phase and decreased the proportion of cells in G0/G1 phase （P < 0.05）. When Tfh cells were co-cultured with HaCaT cells, the proliferation activity of HaCaT cells in psoriasis group and healthy control group was significantly higher than that in control group （P < 0.000 1）, and the proliferation activity of HaCaT cells in psoriasis group was higher than that in IL-21 neutralization antibody group （P < 0.000 1）. In psoriasis group, IL-21 neutralizing antibody group compared with Tfh-HaCaT co-culture group show that it could block HaCaT cells in G0/G1 phase, reduce the number of cells in S phase, and then inhibit the proliferation of HaCaT cells. Conclusion IL-21 promoted the HaCaT cell proliferation invitro; Tfh cells inpsoriasispatients promoted the proliferation of keratinocytes invitro, and blocking IL-21 inhibitd this promoting effect. In summary, Tfh cells may promote theproliferation of keratinocytes through producing IL-21, thereby promoting the development and progression of psoriasis. [ABSTRACT FROM AUTHOR]

BACKGROUND: The mechanisms underlying the occurrence of pressure injuries are complex, and it is not entirely clear which factors play a central role in the development of pressure injuries and how these factors operate. OBJECTIVE: To investigate the relationship between Piezo-type mechanosensitive ion channel component 1 (Piezo1) and the occurrence of pressure injuries. METHODS: (1) Cellular experiment: Human immortalized keratinocytes (HaCaT) were treated with Yoda1, a Piezo1 agonist, at different concentrations. Cell viability, calcium ion influx, Piezo1, and apoptosis-related protein expression were detected. (2) Animal experiment: Twelve Sprague-Dawley rats were randomly divided into a control group and three experimental groups, with three rats in each group. The control group was not subjected to pressure, while in the three experimental groups, magnets with a thickness of 1, 2, and 3 mm were used to press on both sides of the rats’ back for 1 hour, respectively, to establish the animal models of pressure injuries. After modeling, all traumatic tissues were excised and subjected to hematoxylin-eosin, Masson, immunofluorescence staining and western blot assay. RESULTS AND CONCLUSION: Cellular experiments: The results of live/dead cell staining showed that HaCaT cell apoptosis increased with the increase of Yoda1 concentration (0, 2.5, 5, and 10 μmol/L), and calcium ion influx increased with the increase of Yoda1 concentration (0, 5, and 10 μmol/L), as well as with the prolongation of treatment time. Western blot assay results showed an increase in the expression of BAX, TG2, and PIEZO1 and a decrease in the expression of the expression of Bcl-2 protein in HaCaT cells in 5 and 10 μmol/L Yoda1 groups compared with the control group (0 μmol/L Yoda1). Animal experiments: The results of hematoxylin-eosin and Masson staining showed that the skin structure of the three experimental groups was damaged at the compression site, there was subcutaneous fat liquefaction and necrosis, and collagen was sparse and disorganized, and damage to the skin structure at the compression site was aggravated with the increase of magnet thickness. Immunofluorescence staining and western blot results showed that compared with the control group, the expression of BAX, TG2, Yap1 and PIEZO1 proteins was elevated, and the expression of Bcl-2 proteins was lowered in the three experimental groups. Moreover, the expression of related proteins showed more significant changes with the increase of magnet thickness (pressure). To conclude, skin compression activates PIEZO1, leading to a significant influx of calcium ions. As the pressure increases, this ultimately results in cell apoptosis due to calcium overload. [ABSTRACT FROM AUTHOR]

[Objective] The present paper aimed to investigate the inhibition effect of raspberry pulp oil on ultraviolet B (UVB) induced photoaging of human keratinocytes (HaCaT cells) . [Method] The raspberry pulp oil was extracted by supercritical CO2 and its chemical compositions were determined by gas chromatography-mass spectrometry (GC-MS) .The photoaging model was established by UVB, the cell viability of HaCaT cells were determined by CCK-8 method, and the concentrations of matrix metalloproteinase-3 (MMP-3), Collagen type 1 (COL1A1), Interleukin-1β (IL-1β), Interleukin-6 (IL-6), Tumor necrosis factor α (TNF-α), Thioredoxin (Trx) and Superoxide dismutase (SOD) were detected by Enzyme-linked immunosorbent assays (ELISA) and microplate assay. [Result] The analysis of GC-MS showed that raspberry pulp oil consisted of 53 chemical components, mainly including fatty acids, ester of fatty acid, tocopherols, and phytosterols.Oleic acid, linoleic acid, ethyl Z-9-oleate, vitamin E and sterol were closely associated with inhibition effects of raspberry pulp oil on photoaging of UVB-induced HaCaT cells.Compared with the model group, raspberry pulp oil significantly improved the cell viability of photodamaged HaCaT cells induced by UVB (P＜0.01) and showed in a does-dependent manner.In addition, raspberry pulp oil significantly increased concentrations of COL1A1 and reduced the concentrations of MMP-3 to promote skin elasticity (P＜0.05, P＜0.01), reduced the levels of IL-1β, IL-6 and TNF-α to alleviate the inflammatory response (P＜0.05, P＜0.01), increased the contents of Trx and SOD to improve skin oxidative stability (P＜0.05, P＜0.01) in the UVB-induced photodamaged HaCaT cells. [Conclusion] Raspberry pulp oil exhibits good anti-photoaging activity probably by alleviating inflammatory reactions, easing the degradation of extracellular matrix components and reducing oxidative stress in the photodamaged HaCaT cells. [ABSTRACT FROM AUTHOR]

Copyright of Shipin Kexue/ Food Science is the property of Food Science Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Asian Journals of Ecotoxicology is the property of Gai Kan Bian Wei Hui and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

[Objective] The aim of this paper is to study the effect of Naringenin ( NAR) on human immortalized HaCaT cells and its antioxidant activity. [Method] Cell activity was detected by MIT. H & E staining was used to observe the effect of naringin treatment on the morphology of oxidative damage HaCaT cells. Cell migration ability was detected by scratches. Cell proliferation was detected by cloning. Western blot was used to detect the protein expression. SOD and MDA levels were measured with the oxidation kit. [Result] Naringenin had no obvious inhibitory effect on the activity of HaCaT cells, and the migration of HaCaT cells damaged by H2O2 was accelerated after the intervention of naringienin. After the treatment of naringenin on HaCaT cells damaged by H2O2, the expression level of SOD and CDK protein increased, the activity of SOD increased and the content of MDA decreased. [Conclusion] Naringenin can restore the proliferation and migra-tion of HaCaT cells damaged by H2O2, affect the expression of SOD2 and CDK2 protein, improve SOD activity and reduce the content of MDA. Subsequently, it may be developed into a potential skin protective drug to delay aging. [ABSTRACT FROM AUTHOR]

Objective: To observe the effects of curcumin at different concentrations on the proliferation and apoptosis of HaCaT cells, and to explore its possible mechanism. Methods: HaCaT cells in the logarithmic growth phase were randomly divided into low-dose, medium-dose and high-dose curcumin groups, and control group. Cells in the low-dose, medium-dose and high-dose curcumin groups were added with 10, 20 and 40 μmol/L curcumin, respectively, while the cells in the control group were added with the same amount of medium. After 48 hours of treatment, MTS assay was used to detect the cell proliferation, plate cloning assay was used to detect cell cloning ability, flow cytometry was used to detect the apoptosis and cell cycle ratio, and Western blotting was used to detect the relative expression of vascular endothelial growth factor (VEGF), cyclin D1 (Cyclin D1), and B cell lymphoma/leukemia-2 (Bcl-2) protein. Results The inhibition rate of cell proliferation and apoptosis in the curcumin groups and control group decreased in turn, and the number of clone formation increased in turn, with statistically significant difference between these two groups (all P< 0. 05). The percentage of cells in the G0/G1 phase in high-dose curcumin group was higher, while the percentage of cells in the G2/M phase was lower than those in the medium-dose and low-dose curcumin groups and control group (all P< 0.0 5). The relative expression of VEGF, Cyclin D1 and Bcl-2 protein in the high-dose, medium-dose and low-dose curcumin groups and control group increased in turn, with statistically significant difference between these two groups (all P< 0. 05). Conclusion Curcumin can inhibit the proliferation and promote apoptosis of HaCaT cells in a dose-dependent manner, and the mechanism may be related to the inhibition of the expression of VEGF, Cyclin D1 and Bcl-2 protein. [ABSTRACT FROM AUTHOR]

Objective: To observe the effect of aucubin on the expression of Bax/Bcl-2/Caspase-3 in UVB induced HaCaT cells and further investigate its mechanisms. Methods: HaCaT cells were randomly divided into the normal group,model group, 10-7, 10-6, 10-5 mol· L-1 AU treatment groups, model was established by 64 mJ· cm-2 UVB irradiation. After the modeling, final concentration of 10-7, 10-6, 10-5 mol· L-1 aucubin treatment in modeling HaCaT cells.The content of ROS, rate of apoptosis, activity of Caspase-3, protein expression of Bcl-2, Bax were detected. Results: After UV irradiation, the ROS content, apoptotic rate, Caspase-3 activity, Bcl-2, Bax protein expression increased and Bcl-2/Bax value decreased (P＜0.01). The ROS content, rate of apoptosis, activity of Caspase-3, Bax protein expression were decreased, Bcl-2 protein expression and Bcl-2/Bax value were increased in HaCaT cells induced by aucubin, significant differences were found between 10-6, 10-5 mol ·L-1 of aucubin group and model control group (P＜0.05,0.01). Conclusion: Decrease of ROS, Bax, Caspase-3 and increase of Bcl-2 expression might be involved in the anti-photoaging of aucubin. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effects of tumor necrosis factor-alpha (TNF-α)/extracellular signal-regulated kinase (ERK) pathway on the migration ability of HaCaT cells and full-thickness skin defects in mice. Methods: The experimental research method was adopted. According to the random number table (the same below), HaCaT cells were divided into the normal oxygen group and the hypoxia group cultured under hypoxia (with oxygen volume fraction of 1%, the same below) condition. After 24 hours of culture, the significantly differentially expressed genes between the 2 groups were screened using the microarray confidence analysis software SAM4.01. The significance of the number of each gene in the signaling pathway was analyzed through the Kyoto encyclopedia of genes and genomes to screen the significantly differentially signaling pathways ( n =3). HaCaT cells were cultured for 0 (immediately), 3, 6, 12, and 24 h under hypoxia condition. The secretion level of TNF-α was detected by enzyme-linked immunosorbent assay (ELISA), and the number of samples was 5. HaCaT cells were divided into normal oxygen group, hypoxia alone group, and hypoxia+inhibitor group cultured with FR180204 (an ERK inhibitor) and under hypoxia condition. The cells were cultured for 3, 6, 12, and 24 h. The migration ability of the cells was detected by scratch test ( n =12). The expressions of phosphorylated nuclear factor kappa B (p-NF-κB), phosphorylated p38 (p-p38), phosphorylated ERK1/2 (p-ERK1/2), N-cadherin, and E-cadherin in HaCaT cells were detected by Western blotting under hypoxic condition for 0, 3, 6, 12, and 24 h ( n =3). Sixty-four BALB/c male mice aged 6 to 8 weeks were used to make a full-thickness skin defect wound model on the dorsum of the mice. The mice were divided into the blank control group and the inhibitor group treated with FR180204, with 32 mice in each group being treated accordingly. On post injury day (PID) 0, 3, 6, 9, 12, and 15, the wound conditions of mice were observed and the healing rate was calculated ( n =8). On PID 1, 3, 6, and 15, hematoxylin-eosin staining was used to observe neovascularization, inflammatory cell infiltration, and epidermal regeneration on wound, Masson staining was used to observe collagen deposition on wound, the expressions of p-NF-κB, p-p38, p-ERK12, N-cadherin, and E-cadherin in wound tissue were detected by Western blotting ( n =6), the number of Ki67 positive cells and the absorbance value of vascular endothelial growth factor (VEGF) were detected by immunohistochemistry ( n =5), the protein expressions of interleukin 6 (IL-6), IL-10, IL-1β, and CCL20 in wound tissue were detected by ELISA ( n =6). Data were statistically analyzed with one-way analysis of variance, analysis of variance for repeated measurement, factorial design analysis of variance, Tukey test, least significant difference test, and independent sample t test. Results: After 24 hours of culture, compared with normal oxygen group, 7 667 genes were up-regulated and 7 174 genes were down-regulated in cells in hypoxic group. Among the above differentially expressed genes, the TNF-α signaling pathway had significant change ( P <0.05) with large number of genes. Under hypoxia condition, the expression of TNF-α at 24 h of cell culture was (11.1±2.1) pg/mL, which was significantly higher than (1.9±0.3) pg/mL at 0 h ( P <0.05). Compared with normal oxygen group, the migration ability of cells in hypoxia alone group was significantly enhanced at 6, 12, and 24 h of cell culture (with t values of 2.27, 4.65, and 4.67, respectively, P <0.05). Compared with hypoxia alone group, the migration ability of cells in hypoxia+inhibitor group was significantly decreased at 3, 6, 12, and 24 h of cell culture (with t values of 2.43, 3.06, 4.62, and 8.14, respectively, P <0.05). Under hypoxia condition, the expressions of p-NF-κB, p-ERK1/2, and N-cadherin were increased significantly at 12 and 24 h of cell culture compared with 0 h of culture ( P <0.05), the expression of p-p38 was significantly increased at 3, 6, 12, and 24 h of cell culture ( P <0.05), the expression of E-cadherin was significantly decreased at 6, 12, and 24 h of cell culture ( P <0.05), the expression of p-ERK1/2, p-NF-κB, and E-cadherin was time-dependent. Compared with blank control group, on PID 3, 6, 9, 12, and 15, the wound healing rate of mice in inhibitor group was significantly decreased ( P <0.05); there were more inflammatory cell infiltration around the wound edge of mice in inhibitor group on PID 3, 6, and 15, especially on PID 15, a large number of tissue necrosis and discontinuous new epidermal layer were observed on the wound surface, and collagen synthesis and new blood vessels were reduced; the expression of p-NF-κB in the wound of mice in inhibitor group was significantly decreased on PID 3 and 6 (with t values of 3.26 and 4.26, respectively, P <0.05) but significantly increased on PID 15 ( t =3.25, P <0.05), the expressions of p-p38 and N-cadherin were significantly decreased on PID 1, 3, and 6 (with t values of 4.89, 2.98, 3.98, 9.51, 11.69, and 4.10, respectively, P <0.05), the expression of p-ERK1/2 was significantly decreased on PID 1, 3, 6, and 15 (with t values of 26.69, 3.63, 5.12, and 5.14, respectively, P <0.05), the expression of E-cadherin was significantly decreased on PID 1 ( t =20.67, P <0.05) but significantly increased on PID 6 ( t =2.90, P <0.05); the number of Ki67 positive cells and absorbance value of VEGF of wound in inhibitor group were significantly decreased on PID 3, 6, and 15 (with t values of 4.20, 7.35, 3.34, 4.14, 3.20, and 3.73, respectively, P <0.05); the expression of IL-10 in the wound tissue of the inhibitor group was significantly decreased on PID 6 ( t =2.92, P <0.05), the expression of IL-6 was significantly increased on PID 6 ( t =2.73, P <0.05), the expression of IL-1β was significantly increased on PID 15 ( t =3.46, P <0.05), and CCL20 expression levels were significantly decreased on PID 1 and 6 (with t values of 3.96 and 2.63, respectively, P <0.05) but significantly increased on PID 15 ( t =3.68, P <0.05). Conclusions: The TNF-α/ERK pathway can promote the migration of HaCaT cells, and regulate the healing of full-thickness skin defect wounds in mice by affecting the expression of inflammatory cytokines and chemokines.

Objective: To investigate the effect and mechanism of exosomes derived from human amniotic epithelial cells (hAEC-Exos) on the proliferation and migration of HaCaT in high glucose environment. Methods: The experimental research method was adopted. The amniotic membrane tissue was collected from 10 healthy pregnant women at full term delivery in the Department of Obstetrics and Gynecology of Fujian Medical University Union Hospital from January to June 2019, and the primary human amniotic epithelial cells (hAECs) were isolated. The growth status and morphological changes of the primary hAECs on the 2nd, 4th, and 7th day of culture were observed, and the expressions of the cells surface markers of CD73, CD90, CD29, CD34, and human leukocyte antigen DR (HLA-DR). The 2nd to 4th passages of hAECs were used in the following experiments. The hAEC-Exos were separated by ultracentrifugation method. The HaCaT and hAEC-Exos were co-cultured for 3 h, and the uptake of hAEC-Exos by HaCaT was observed by inverted fluorescence microscopy. The HaCaT were divided into phosphate buffer solution (PBS) group and hAEC-Exos group or dimethyl sulfoxide (DMSO)+PBS group, DMSO+hAEC-Exos group, and LY294002+hAEC-Exos group, which were dealt correspondingly, with 3 wells in each group. Cell counting kit 8 (CCK-8) method was used to detect cell proliferation activity after 0 (immediately), 12, 24, 36, 48, and 60 h of culture. The scratch test was conducted to detect the scratch healing at 0, 24, 48, and 72 h after the scratch, and the scratch healing rate was calculated, respectively. The Transwell experiment was conducted to detect the number of transmembrane cells after 48 h of culture. The Western blotting was used to detect the protein expressions of mammalian target of rapamycin (mTOR), phosphorylated mTOR (p-mTOR), protein kinase B (Akt), and phosphorylated Akt (p-Akt) related to phosphatidylinositol 3-kinase-Akt-mTOR (PI3K-Akt-mTOR) pathway after 24 h of culture. Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, and independent sample t test. Results: Most of the primary hAECs were oval and uniform in size on the 2nd day of culture. The hAECs were arranged in a typical cobblestone-like monolayer on the 4th and 7th day of culture. The primary hAECs highly expressed CD73, CD90, and CD29 of mesenchymal stem cell related surface markers, and were with no or low expressions of CD34 and HLA-DR of hematopoietic stem cell related surface markers. After 3 h of culture, hAEC-Exos were successfully endocytosed by HaCaT into the cytoplasm and gathered around the nucleus. After 12, 24, 36, 48, and 60 h of culture, the proliferation activity of HaCaT in hAEC-Exos group was significantly higher than that in PBS group ( t =3.691, 10.861, 12.121, 10.531, 14.931, P <0.01). At 24, 48, and 72 h after scratch, the scratch healing rates of HaCaT in PBS group were significantly lower than those in hAEC-Exos group (t =3.342, 6.427, 5.485, P <0.05 or P <0.01). After 48 h of culture, the number of transmembrane HaCaT in hAEC-Exos group was significantly more than that in PBS group ( t =5.385, P <0.01). After 24 h of culture, the protein expressions of p-mTOR and p-Akt in HaCaT of hAEC-Exos group were significantly higher than those in PBS group ( t =4.240, 5.586, P <0.01), while the protein expressions of mTOR and Akt in HaCaT of the two groups were similar ( P >0.05). After 24 h of culture, the protein expressions of p-mTOR and p-Akt in HaCaT of DMSO+hAEC-Exos group were significantly higher than those in DMSO+PBS group ( t =6.155, 8.338, P <0.01) and LY294002+hAEC-Exos group ( t =5.030, 3.960, P <0.01), while the protein expressions of mTOR and Akt in HaCaT of the three groups were similar ( P >0.05). The proliferation activity of HaCaT in DMSO+hAEC-Exos group at 12, 24, 36, 48, and 60 h of culture was 0.78±0.05, 1.23±0.07, 1.60±0.09, 1.86±0.09, and 2.03±0.08, which was significantly higher than 0.46±0.04, 0.69±0.07, 0.98±0.08, 1.16±0.08, and 1.26±0.11 in DMSO+PBS group ( t =4.376, 7.398, 8.488, 9.766, 10.730, P <0.01). The proliferation activity of HaCaT in DMSO+hAEC-Exos group at 24, 36, 48, and 60 h was significantly higher than 0.96±0.09, 1.20±0.08, 1.39±0.08, and 1.55±0.10 in LY294002+hAEC-Exos group ( t =3.639, 5.447, 6.605, 6.693, P <0.05 or P <0.01). The scratch healing rates of HaCaT in DMSO+hAEC-Exos group at 24, 48, and 72 h after scratch were significantly higher than those in DMSO+PBS group ( t =4.003, 6.349, 7.714, P <0.01) and LY294002+hAEC-Exos group ( t =3.805, 4.676, 4.067, P <0.05 or P <0.01). After 48 h of culture, the number of transmembrane HaCaT in DMSO+hAEC-Exos group was significantly more than that in DMSO+PBS group and LY294002+hAEC-Exos group, respectively ( t =7.464, 1.232, P <0.01). Conclusions: PI3K-Akt-mTOR pathway can promote the proliferation and migration of HaCaT in high glucose environment by mediating hAEC-Exos.