The Human Host Defense Peptide LL-37 Overexpressed in Lung Cell Lines by Methanolic Extract of Valeriana officinalis

Abstract The present study investigated the effects of valerian methanolic extract and valerenic acid on the expression of LL-37 gene and protein in A549 and MRC5 line cells. After preparing Valerian seeds, sowing them in March 2020, and harvesting the rhizome in October 2020, the extract was prepared from the valerian rhizome by maceration method. Valerian acid content was determined using high performance liquid chromatography (HPLC). Two cell lines (A549 and MRC-5) were used to study the effects of valerian extract, and the MTT test was used to evaluate cell viability. The expression of LL-37 mRNA and protein was assessed by Real-Time PCR and western blot, respectively. In vivo safety assessments and histopathological analysis were also conducted. Data was analyzed by Graphpad Prism 8 software. Valerian methanolic extract and valerenic acid upregulated the LL-37 mRNA and protein expression in both treated cell lines. Valerenic acid showed a greater effect on upregulating LL-37 expression than valerian methanolic extract. A549 cells were more sensitive to valerian methanolic extract compared to MRC5 cells, and its cell viability was reduced. Furthermore, liver and kidney-related safety assessments showed that valerian methanolic extract had no toxic effects. In general, it was concluded that the methanolic extract of valerian as well as the resulting valerenic acid as the most important component of the extract has the ability to upregulate LL-37expression. Therefore, methanolic extract of valerian and valerenic acid can be considered for improving the immune system.

High throughput single cell separation and identification using a self-priming isometric and Equant screw valve-based (SIES) microfluidic chip.

The emergence of various single cell separation and identification platforms has greatly promoted the development of single cell research. Among these platforms, microfluidic chip-based strategies occupy a significant position in single cell separation and identification. Here, we proposed a self-priming isometric and Equant screw valve-based microfluidic chip (SIES chip) for high throughput single cell isolation and identification. With several special designs, such as a peripheral water tank to balance negative pressure distribution in a marginal area of the chip, a screw valve to preserve the suction power during the step-by-step sample loading, and multistage branching "T" shape channels to separate cells evenly into the chambers, up to 2000 single cells can be well dispersed and analyzed at the same time using this chip. We applied this chip for the isolation and identification of single A549 cells targeting the activated leukocyte cell adhesion molecule (ALCAM) gene. The results showed that only a small proportion (approximately 5.1%) of A549 cells expressed ALCAM, which can potentially provide a reference for A549 cell reclassification. Besides being inexpensive, user-friendly and portable, our chip can be used in some resource-limited settings and may have a great potential in POC (Point-of-Care) applications.