Objective To study the liver protection of crocetin against paraquat (PQ) poisoning induced acute liver injury in rats. Methods Fifty-four male Wistar rats were randomly divided into control group, exposure group and treatment group, and the rats in each group were subdivided into the 0.5th, 2nd, and 6th day after exposure subgroups (n = 6). The model of acute liver failure induced by PQ poisoning was reproduced by intraperitoneal injection of 20 mg/kg of 20% PQ, and the rats in control group was injected with the same amount of normal saline. The rats in treatment group were given with intraperitoneal injection of 50 mg/kg crocetin after 0.5 day, once a day until they were sacrificed; the other two groups were injected with the same amount of normal saline. The rats in all groups were sacrificed at the corresponding time points, and blood was collected from inferior vena cava and hepatic tissue was harvested. Hematoxylin and eosin (HE) staining was used to observe the pathological changes in liver tissue on the 6th day under light microscope. Enzyme linked immunosorbent assay (ELISA) was used to detect the serum tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) levels. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the mRNA expressions of inducible nitric oxide synthase (iNOS) and nuclear factor-κB (NF-κB). The activities of apoptosis related factors, including caspase-8, -9, -12, in hepatic tissue were determined on the 6th day with chromogenic substrate method. Results In the liver tissue of exposed group, extensive infiltration of the inflammatory cells and the diffuse fragments necrosis were visible, and the regeneration of the liver cells was not obvious, and severity of the injury in a time dependent way. In the treatment group, the structure of hepatic artery was visible, and the infiltration of necrosis, congestion and inflammatory cells were not obvious. On the 0.5th, 2nd, and 6th day, serum levels of IL-6 and TNF-α, the mRNA expressions of iNOS and NF-κB in liver tissue, and the caspase-8, -9, -12 activities on the 6th day in the exposure group and treatment group were significantly higher than those in the control group. And the parameters in treatment group were significantly lower than those of the exposure group [IL-6 (ng/L): 188.37±64.21 vs. 376.61±82.42 on the 0.5th day, 287.18±58.69 vs. 432.77±96.28 on the 2nd day, 234.24±10.17 vs. 375.41±37.59 on the 6th day; TNF-α (ng/L): 472.36±76.43 vs. 688.33±102.19 on the 0.5th day, 189.32±87.54 vs. 296.21±89.77 on the 2nd day, 99.28±16.13 vs. 168.41±66.78 on the 6th day; iNOS mRNA (gray value): 2.998±0.801 vs. 3.453±0.026 on the 0.5th day, 3.126±0.306 vs. 5.259±0.153 on the 2nd day, 0.841±0.135 vs. 1.225±0.057 on the 6th day; NF-κB mRNA (gray value): 1.569±0.818 vs. 2.361±0.063 on the 0.5th day, 2.345±0.489 vs. 4.668±0.368 on the 2nd day, 2.348±0.316 vs. 3.972±0.449 on the 6th day; caspase-8 (pmol/mg): 126.77±9.97 vs. 199.18±66.48 on the 6th day; caspase-9 (pmol/mg): 213.12±69.06 vs. 321.62±89.39 on the 6th day; caspase-12 (pmol/mg): 183.46±70.52 vs. 219.68±53.93 on the 6th day, all P < 0.05]. Conclusion Crocetin has protective effect on liver in rats with PQ poisoning, which role is related with reducing the blood level of inflammatory factors, inhibiting the hepatic caspase-8, -9, -12 activities and gene expressions of iNOS and NF-κB.

BACKGROUND:Mesenchymal stem cells can regulate the tumor microenvironment by secreting extracellular vesicles containing cytokines,growth factors and exosomes for the precise regulation of biological behavior of tumor cells. OBJECTIVE:To investigate the effects of human umbilical cord-derived mesenchymal stem cell conditioned medium and their released exosomes on the biological properties of hepatocellular carcinoma cells. METHODS:Human umbilical cord mesenchymal stem cell supernatant was collected,centrifuged and filtered at high speed to obtain human umbilical cord mesenchymal stem cell conditioned medium.Human umbilical cord mesenchymal stem cell supernatant was collected and human umbilical cord mesenchymal stem cell exosomes were extracted by ultra-high speed gradient centrifugation.Human umbilical cord mesenchymal stem cell exosomes were labeled with PKH26 and co-cultured with hepatocellular carcinoma cell MHCC97-H.The uptake of exosomes by MHCC97-H cells was observed by fluorescence microscopy.The effects of human umbilical cord mesenchymal stem cell conditioned medium and human umbilical cord mesenchymal stem cell exosomes on biological functions of hepatocellular carcinoma cells were assessed by the CCK-8 proliferation assay,Transwell migration and invasion assay,and the apoptosis assay. RESULTS AND CONCLUSION:(1)Human umbilical cord mesenchymal stem cell exosomes could be uptaken by MHCC97-H cells and was mainly distributed in the cytoplasm.(2)After treatment with human umbilical cord mesenchymal stem cell conditioned medium,MHCC97-H cells showed a significant increase in proliferation,migration,and invasion(P<0.001,P<0.05,P<0.01),and a significant decrease in apoptosis(P<0.001),while after treatment with human umbilical cord mesenchymal stem cell exosomes,MHCC97-H cells showed a decrease in proliferation(P<0.001)and migration,invasion,and apoptosis were significantly enhanced(P<0.001).(3)The results indicated that human umbilical cord mesenchymal stem cell conditioned medium had the ability to promote the proliferation,migration,invasion,and inhibit apoptosis of MHCC97-H cells,while human umbilical cord mesenchymal stem cell exosomes had the properties of promoting the migration,invasion and apoptosis of MHCC97-H cells,inhibiting the proliferation.

Objective To explore the effects of exosomes loaded with ginsenoside Rh2 on the biological functions of hepatocellular carcinoma cells. Methods Both Huh7 and PLC/PRF/5 cell were equally divided into control group, exosome group (Exos group), drug group (G-Rh2 group), and exosomes-loaded-with-ginsenoside Rh2 group (Exos@G-Rh2 group). The effects of each group on the viability, clonogenic ability, migration ability, invasion ability, and apoptotic level of hepatocellular carcinoma cells were detected through CCK-8 assay, colony formation assay, cell scratch assay, Transwell assay, and flow cytometry. Results Compared with the control group, the Exos@G-Rh2 group and G-Rh2 group showed significantly decreased cell viability, clonogenic ability, and migration and invasion capabilities, along with a markedly increased cell apoptosis rate (P<0.05). These changes were more pronounced in the Exos@G-Rh2 group than in the G-Rh2 group (P<0.05). Conclusion Exos@G-Rh2 can effectively inhibit the viability and clonogenic, migration, and invasion abilities of liver cancer cells and induce cell apoptosis. This effect is stronger than that of free G-Rh2 at the same concentration.