Objective:To discuss the effects of Buzhong Yiqi Decoction medicated serum combined with cisplatin on the expression of Nrf2 and its effects of downstream antioxidant proteins in human pulmonary adenocarcinoma A549/DDP cells.Methods:Totally 60 rats were divided into blank group, low-dosage group, medium-dosage group, and high-dosage group using a random number table method, with 15 rats in each group. The low-, medium-, and high-dosage groups were orally administered with 3.29, 6.57, and 13.13 g/kg Buzhong Yiqi Decoction for gavage, once a day, for 7 days. 1 hour after the last administration, blood was collected from the abdominal aorta to prepare the serum containing Buzhong Yiqi Decoction. A549/DDP cells were divided into 5 groups: blank group (rat serum), model group (rat serum+cisplatin), low-dosage group (low-dosage Buzhong Yiqi Decoction medicated serum+cisplatin), medium-dosage group (medium-dosage Buzhong Yiqi Decoction medicated serum +cisplatin), and high-dosage group (high-dosage Buzhong Yiqi Decoction medicated serum+cisplatin). And 24 h after the Buzhong Yiqi Decoction intervention, CCK-8 method was used to detect the IC 50 of cisplatin in each group. The fluorescence expression intensity of p-Nrf2 in A549/DDP cells of each group was determined by confocal fluorescence localization method. The mRNA relative expression of Nrf2 in A549/DDP cells of each group was detected by RT-qPCR. The relative expressions of Nrf2, p-Nrf2, GPX4, PRX1 and SOD in A549/DDP cells of each group were detected by Western blot. Results:Compared with the model group, the IC 50 of A549/DDP cells to cisplatin decreased in the low-, medium-, and high-dosage groups ( P<0.01); the fluorescence intensity of p-Nrf2 decreased ( P<0.01); the level of Nrf2 mRNA decreased ( P<0.01); the protein expressions of Nrf2, p-Nrf2, SOD, PRX1, and GPX4 were down-regulated ( P<0.01), with the effect being more significant in the medium- and high-dosage groups ( P<0.05). Conclusion:Buzhong Yiqi Decoction medicated serum can improve cisplatin resistance in human pulmonary adenocarcinoma cancer by inhibiting the active expression of Nrf2 in A549/DDP cells and down-regulating the target genes of SOD, PRX1 and GPX4 in vivo.

ObjectiveThe biosynthetic pathways of benzylisoquinoline alkaloids（BIAs） in Nelumbo nucifera are of great theoretical and economic value. In this paper， N. nucifera O-methyltransferase（NnOMT） and N. nucifera N-methyltransferase（NnNMT） gene families were identified and analyzed by bioinformatics in order to facilitate the biosynthetic pathway of BIAs in N. nucifera. MethodBased on the whole genome of N. nucifera， UniPort and National Center for Biotechnology Information（NCBI） databases were used to identify the NnOMT and NnNMT gene families of N. nucifera， and analyze their physicochemical properties and subcellular localization， then TBtools， MEME， MEGA 11.0， FigTree 1.4.4 and other tools were used to analyze the phylogeny， sequence characteristics， gene structure， functional annotation and cis-acting elements of NnOMT and NnNMT genes identified in the previous stage. ResultA total of 61 NnOMT and NnNMT genes were identified in this paper， the number of amino acids encoded by these genes ranged from 168 aa to 580 aa， the isoelectric point ranged from 4.76 to 9.16， and the relative molecular weight ranged from 18 699.52 Da to 64 934.53 Da， most of which showed acidic and mostly hydrophilic proteins. There were 10 conserved motifs， Kyoto Encyclopedia of Genes and Genomes（KEGG） analysis enriched a total of 12 pathways， including metabolism， biosynthesis of phenylpropane and isoquinoline alkaloids， etc. And Visualization of Gene Ontology（GO） enrichment results showed that 61 NnOMT and NnNMT genes were annotated to 32 items， which included 16 molecular functions［such as reduced nicotinamide adenine dinucleotide（NADH） activity and exopeptidase activity］ and 16 biological processes（such as metabolic process of carbon tetrachloride， anaerobic carbon tetrachloride metabolic process and responses to exogenous biological stimuli）. There were a variety of cis-acting elements in the promoter regions of NnOMT and NnNMT genes， mainly promoter and enhancer regions element， light responsive element and methyl jasmonate responsive element. ConclusionIn this study， a comprehensive bioinformatics analysis of 61 NnOMT and NnNMT genes is carried out based on the genome data of N. nucifera， which lays a foundation for research on the gene structure and function of NnOMT and NnNMT gene families， and provides a reference for biosynthetic pathway elucidation of BIAs in N. nucifera.

ObjectiveTo investigate the mechanism of Buzhong Yiqitang in attenuating cisplatin resistance in non-small cell lung cancer by observing the effects of Buzhong Yiqitang on endoplasmic reticulum stress-related molecules in human lung adenocarcinoma cells （A549） and cisplatin-resistant cells in human lung adenocarcinoma cells （A549/DDP） via the nuclear factor E₂-related factor 2（Nrf2）/reactive oxygen species（ROS） pathway. MethodsThe serum containing Buzhong Yiqitang was prepared and A549 cells and A549/DDP cells were cultured. The cells were randomized into groups A （A549 cells+blank serum）， B （A549 cells+20 mg·L^-1 cisplatin+blank serum）， C （A549 cells+20 mg·L^-1 cisplatin+10% Buzhong Yiqitang-containing serum）， D （A549/DDP cells+blank serum）， E （A549/DDP cells+20 mg·L^-1 cisplatin+blank serum）， and F （A549/DDP cells+20 mg·L^-1 cisplatin+10% Buzhong Yiqitang-containing serum）. The cell counting kit-8 （CCK-8） method was used to detect the half maximal inhibitory concentration （IC₅₀） of cisplatin. The protein levels of Nrf2 and p-Nrf2 were determined by Western blotting. The DCFH-DA fluorescent probe was used to measure the content of reactive oxygen species （ROS） in each group. The protein levels of glucose-regulated protein 78 （GRP78）， activated transcription factor 6 （ATF6）， and C/EBP-homologous protein （CHOP） were determined by Western blot. ResultsCompared with group B， group C showed a reduction in IC₅₀ of cisplatin （P<0.05）， which held true in group E compared with group F （P<0.05）. Moreover， the IC₅₀ of cisplatin to A549/DDP cells was higher than that to A549 cells before and after Buzhong Yiqitang intervention （P<0.05）. Compared with group A， group B showed up-regulated protein levels of Nrf2 and p-Nrf2 （P<0.05）. Compared with group B， group C showed down-regulated protein levels of Nrf2 and p-Nrf2 （P<0.05）. Compared with group D， group E showed up-regulated protein levels of Nrf2 and p-Nrf2 （P<0.05）， which， however， were significantly down-regulated in group F （P<0.05）. The ROS content and the protein levels of GRP78， ATF6， and CHOP followed a descending trend of group C > group B > group A in A549 cells and group F > group E > group D in A549/DDP cells （P<0.05）. Moreover， the ROS content and the protein levels of GRP78， ATF6， and CHOP in A549 cells were higher than those in A549/DDP cells before and after Buzhong Yiqitang intervention （P<0.05）. ConclusionBuzhong Yiqitang may regulate endoplasmic reticulum stress via the Nrf2/ROS pathway to attenuate cisplatin resistance in non-small cell lung cancer.