Scutellarin (SC), mainly extracted from the Chinese herb Erigeron breviscapus (vant.), has been reported to possess various pharmacological activities; however, its effects on Alzheimer's disease (AD) have not been systemically reported. The protective effects of SC on AD were investigated using an L‑glutamic acid (L‑Glu)‑damaged HT22 cell apoptosis model and an aluminum chloride plus D‑galactose‑induced AD mouse model. In L‑Glu‑damaged HT22 cells, SC significantly increased cell viability, inhibited lactate dehydrogenase release, reduced caspase‑3 activity and suppressed apoptosis, which were determined via an MTT assay, an in vitro Toxicology Assay kit, a Caspase‑3 activity assay kit, and propidium iodide and Annexin V staining. Furthermore, SC suppressed the accumulation of intracellular reactive oxygen species (ROS), restored the dissipation of mitochondrial membrane potential, enhanced the expression of antiapoptotic proteins and reduced the expression of pro‑apoptotic proteins, as determined by immunofluorescence assays and western blotting. In AD mice, SC enhanced vertical and horizontal movements in an autonomic activity test, and reduced the escape latency time in the water maze test. SC reduced the deposition of amyloid β1‑42 (Aβ1‑42) and the expression of phosphorylated‑Tau in the hippocampus as determined by immunohistochemistry analysis, but enhanced the serum levels of Aβ1‑42 of AD mice as determined by ELISA. ELISA analyses also revealed that SC enhanced the levels of acetylcholine, and superoxide dismutase in serum and brain lysate, whereas reduced the levels of ROS in brain lysate of AD mice. The present study confirmed that the protective effects of SC in AD in vitro and in vivo are associated with its antioxidant and antiapoptotic properties.

It has been established that dental pulp stem cells (DPSCs) serve an important role in the restoration and regeneration of dental tissues. DPSCs are present in blood vessels and also exist in the vessel microenvironment in vivo and have a close association with endothelial cells (ECs). The present study aimed to evaluate the influence of ECs and their secretory product endothelin‑1 (ET‑1) on the differentiation of DPSCs. In the present study, cells were divided into four groups: i) a DPSC‑only control group; ii) a DPSC with ET‑1 administration group; iii) a DPSC and human umbilical vein endothelial cell (HUVEC) direct co‑culture group; and iv) a DPSC and HUVEC indirect co‑culture group using a Transwell system. Reverse transcription‑quantitative polymerase chain reaction was used to detect the expression of the odontoblastic differentiation‑associated genes, including dentin sialoprotein (DSP) and dentin matrix acidic phosphoprotein 1 (DMP‑1) at days 4, 7, 14 and 21. Alizarin Red S staining, immunofluorescence and western blot analyses were also conducted to assess the differentiation of the DPSCs in each group. The highest expression levels of odontoblastic differentiation‑associated genes were observed on day 7 and in the two co‑culture groups were increased compared with the DPSC‑only and DPSC + ET‑1 culture groups at all four time points. However, expression levels in the DPSC + ET‑1 group were not downregulated as notably as in the co‑culture groups on days 14 and 21. The Transwell group exhibited the greatest ability for odontoblastic differentiation compared with the other groups according to staining with Alizarin Red S, immunofluorescence and western blot analysis results. According to the results of the present study, the culture solution with HUVECs affected the differentiation of DPSCs. In addition, ET‑1 may promote the odontoblastic differentiation of DPSCs.

Fasudil, which is primarily prescribed to treat cerebral vasospasm, may also inhibit systemic inflammation and prevent sepsis‑induced acute lung injury (ALI) in rats, although the mechanisms remain elusive. The purpose of the present study was to investigate the role of the rhodopsin (Rho)/Rho‑associated protein kinase (ROCK) signaling pathway in the protective effects of fasudil on ALI in septic rats. A total of 60 Wistar rats were pretreated with fasudil (30 mg/kg) through intraperitoneal injections 1 h prior to cecal ligation and puncture. Administration of fasudil led to reductions in polymorphonuclear neutrophil counts, and the protein concentrations of tumor necrosis factor‑α, interleukin (IL)‑1β and IL‑6 in the bronchoalveolar lavage fluid of rats with sepsis‑induced ALI. The results demonstrated that fasudil decreased sepsis‑induced bacteremia. In addition, fasudil effectively reduced the Evans blue content, wet/dry lung weight ratio, lung injury score, and expression levels of malondialdehyde and myeloperoxidase. However, the superoxide dismutase activity in the lung tissue of the rats was increased. Activated caspase‑3 activity in lung tissue was reduced to 29% by fasudil. Furthermore, the expression of Rho and ROCK1 was significantly downregulated, and the phosphorylation of myosin phosphatase‑targeting subunit 1 in lung tissues was markedly decreased, whereas the protein expression levels of zonula occludens 1 were increased in fasudil‑treated rats (P<0.05). In the in vitro experiments, vascular endothelial growth factor, intracellular adhesion molecule 1 and vascular cell adhesion molecule 1 secreted from human pulmonary microvascular endothelial cells treated with lipopolysaccharide (LPS) were attenuated by fasudil. Fasudil also reduced the fluorescence intensity of filamentous actin induced by LPS. Taken together, the results of the present study demonstrated that fasudil was able to improve endothelial permeability and inhibit inflammation, oxidative stress and cellular apoptosis in order to alleviate ALI in septic rats through inhibition of the Rho/ROCK signaling pathway.

This study is to reveal the characteristics of autophagy and the effect of neuroserpin (NSP) treatment on autophagy during the process of functional recovery following spinal cord injury (SCI). After the clip compress rat model of SCI had been made, autophagy‑associated proteins, including LC3‑II, beclin‑1 and p62, were evaluated at 2, 4, 24, 72 h, and 168 h in the experimental group, and the sham group as control. Transmission electron microscopy (TEM) was further used for autophagy detection at 4 and 72 h. All the male rats were randomly divided into three groups: Sham, vehicle and NSP group. NSP or an equal volume of saline vehicle was administered via intrathecal injection immediately after SCI. Each group was further divided into subgroups for the following experiments: i)Western blot (LC3‑II and p62); ii) Immunofluorescent double staining (LC3/MAP‑2/DAPI); iii) Nissl staining and Basso Beattie Bresnahan (BBB score) for NSP neuroprotection evaluation. Our results revealed both LC3‑II and p62 expression trended upward at 24, 72 and 168 h after SCI. The LC3‑II peaked at 72 h, while p62 peaked at 24 h. Beclin‑1 dropped significantly at 72 and 168 h. TEM results showed that autophagosomes largely accumulated at 72 h after SCI when compared with the sham group. Western blot analysis showed that LC3‑II and p62 were markedly decreased with NSP treatment at 72 h after injury compared with that of the vehicle‑group. Immunofluorescent double labeling indicated that accumulation of autophagosomes was reduced in the NSP group. Further, post‑SCI treatment with NSP improved the BBB scale and increased the number of anterior horn motor neurons. Together, this study demonstrates that autophagic flux is impaired, meanwhile NSP restores autophagic flux and promotes functional recovery after SCI in rats.