KHẢO SÁT THÀNH PHẦN CÁC HOẠT CHẤT NHÓM IRIDOID TRONG DỊCH CHIẾT TỪ RỄ CỦ ĐỊA HOÀNG (Rehmannia glutinosa Libosch.) BẰNG UHPLC VÀ LC-MSD-Trap-SL

Dung môi đóng vai trò rất quan trọng trong việc trích li các hợp chất tự nhiên bởi dung môi ảnh hưởng lớn đến thành phần của dịch chiết. Nghiên cứu này nhằm so sánh sự khác nhau về thành phần các hoạt chất nhóm iridoid trong các dịch chiết nước và ethanol của rễ củ địa hoàng (Rehmannia glutinosa Libosch.) bằng phương pháp sắc kí lỏng hiệu năng cao (UHPLC) và sắc kí lỏng ghép khối phổ (LC-MSD-Trap-SL). Theo các số liệu của LC-MSD-Trap-SL, Jioglutoside B, Leonuride và Jioglutin E là những iridoid phân biệt sự khác nhau giữa hai dịch chiết. Trong khi đó, phân tích UHPLC cũng chứng minh rằng hàm lượng catalpol, 8-epiloganic axit và glutinoside trong dịch chiết nước cao hơn nhiều so với của dịch chiết ethanol (lớn hơn lần lượt là 126; 4,46 và 11,14 lần). Vì vậy, nghiên cứu này đề xuất sử dụng nước để chiết các hoạt chất iriodid có hoạt tính sinh học cao từ rễ củ địa hoàng, trong đó có catalpol, một iridoic glycoside nổi trội trong rễ củ địa hoàng.

Ethnopharmacological Survey of Traditional Chinese Medicine Pharmacy Prescriptions for Dysmenorrhea

Chinese herbal medicines have long been used for the treatment of dysmenorrhea. The treatment experiences of traditional Chinese medicine (TCM) pharmacies passed down through generations have contributed to a wealth of prescriptions for dysmenorrhea that have achieved significant therapeutic effects in countless Taiwanese women. Therefore, surveying and analyzing these prescriptions may enable us to elucidate the core medication combinations used in TCM prescriptions for dysmenorrhea. In the present study, a field investigation was conducted on various TCM pharmacies in Taiwan. A total of 96 TCM pharmacies were sampled, and 99 prescriptions for dysmenorrhea containing 77 different medicinal materials were collected. Compositae (8%) was the most common botanical source of the medicinal materials, and the predominant TCM property and flavor of the materials were warm (45%) and sweet (73%), respectively. The blood-activating and stasis-dispelling effect (23%) and the qi-tonifying effect (23%) were the most prevalent traditional effects, and the modern pharmacological effects most commonly found in the materials were anti-inflammatory (73%), antitumor (59%), and analgesic (12%) effects. Network analysis of the 77 medicinal materials used in the prescriptions, which was performed using the Traditional Chinese Medicine Inheritance Support System, yielded seven core medicinal materials and the corresponding network diagram. The seven core medicinal materials ranked in order of relative frequency of citation (RFC) were Angelica sinensis (Oliv.) Diels (Dang Gui), Ligusticum chuanxiong Hort (Chuan Qiong), Rehmannia glutinosa Libosch (Di Huang), Paeonia lactiflora Pall (Bai Shao), Hedysarum polybotrys Hand.-Mazz (Hong Qi), Lycium chinense Mill (Gou Qi Zi), and Cinnamomum cassia (L.). J. Presl (Gui Zhi). A total of 58 combinations, each consisting of two to five of the seven medicinal materials and 107 association rules among the materials, were identified. This study provides a record of valuable knowledge on TCM pharmacy prescriptions for dysmenorrhea. The rich medicinal knowledge of TCM pharmacies in Taiwan is worthy of further exploration, and the results of this study can serve as a basis for future pharmacological research and the development of naturally derived medications for dysmenorrhea.

Optimization of Spray Drying Without Drying Carriers from Rehmannia glutinosa L. Root and Coix lachryma- jobi L. Trunk

Spray drying is a well-known method for preparing dried extracts from herbs. Rehmannia glutinosa root and Coix lachryma-jobi trunk are widely researched especially for diabetes treatment. The optimization of extraction conditions, namely the alcohol concentration, treated herbs and solvent ratio, extraction temperature, and time. The extracts are dried by Labplant machine with two specifications (drying inlet air temperature and flow- rate). Rehmannia glutinosa root is extracted by refluxing method for 60 minutes with 40% alcohol, the ratio of treated herbs and solvent was 1:3, at 800C by three times extract and its dried extract is prepared at a drying inlet air temperature of 90oC, 8 cycles per minute. Coix lachryma-jobi trunk is also extracted by the same method for 60 minutes with 80% alcohol, the ratio of treated herbs and solvent was 1:6, extraction temperature and time are respective 80oC and two times, spray drying conditions were 80oC (drying inlet air temperature) and 4 cycles per minute (flow- rate). The obtained dried extracts of Rehmannia glutinosa root and Coix lachryma-jobi trunk have average moisture contents of 4.52% and 3.84%, respectively. Therefore, this study is a novel approach to significantly decreasing the amount of carriers to reduce research's funds and saving dried extracts' price.

Transcriptome-Wide Identification of WRKY Transcription Factor and Functional Characterization of RgWRKY37 Involved in Acteoside Biosynthesis in Rehmannia glutinosa

WRKYs play important roles in plant metabolism, but their regulation mechanism in Rehmannia glutinosa remains elusive. In this study, 37 putative WRKY transcription factors (TFs) with complete WRKY domain from R. glutinosa transcriptome sequence data were identified. Based on their conserved domains and zinc finger motif, the R. glutinosa WRKY TFs were divided into five groups. Structural feature analysis shows that the 37 RgWRKY proteins contain WRKYGQK/GKK domains and a C2H2/C2HC-type zinc finger structure. To identify the function of RgWRKY members involved in acteoside biosynthesis, transcriptional profiles of 37 RgWRKYs in hairy roots under salicylic acid (SA), methyl jasmonate (MeJA), and hydrogen peroxide (H2O2) treatments were systematically established using RNA-seq analysis. Based on the correlationship between the expression levels of RgWRKY genes and acteoside content, RgWRKY7, RgWRKY23, RgWRKY34, RgWRKY35, and RgWRKY37 were suggested to be involved in acteoside biosynthesis in R. glutinosa, and RgWRKY37 was selected for gene functional research. Overexpression of RgWRKY37 increased the content of acteoside and total phenylethanoid glycosides (PhGs) in hairy roots and enhanced the transcript abundance of seven enzyme genes involved in the acteoside biosynthesis pathway. These results strongly suggest the involvement of the WRKY transcription factor in the regulation of acteoside biosynthesis.

Study of Terpenoid Synthesis and Prenyltransferase in Roots of Rehmannia glutinosa Based on iTRAQ Quantitative Proteomics

Rehmannia glutinosa has important medicinal value; terpenoid is one of the main active components in R. glutinosa. In this study, iTRAQ technique was used to analyze the relative abundance of proteins in roots of R. glutinosa, and 6,752 reliable proteins were quantified. GO enrichment results indicated that most proteins were involved in metabolic process or cellular process, 57.63% proteins had catalytic activity, and 65.80% proteins were enriched in membrane-bounded organelle. In roots of R. glutinosa, there were 38 KEGG enrichments with significance, more DEPs were found in some pathways, especially the proteasome pathway and TCA cycle with 15.0% DEPs between elongation stage and expansion stage of roots. Furthermore, five KEGG pathways of terpenoid synthesis were found. Most prenyltransferases belong to FPP/GGPP synthase family, involved in terpenoid backbone biosynthesis, and all interacted with biotin carboxylase CAC2. Compared with that at the elongation stage, many prenyltransferases exhibited higher expression at the expansion stage or maturation stage of roots. In addition, eight FPP/GGPP synthase encoding genes were cloned from R. glutinosa, namely FPPS, FPPS1, GGPS, GGPS3, GGPS4, GGPS5, GPPS and GPPS2, introns were also found in FPPS, FPPS1, GGPS5 and GGPS2, and FPP/GPP synthases were more conservative in organisms, especially in viridiplantae, in which the co-occurrence of GPPS or GPPS2 was significantly higher in plants. Further analysis found that FPP/GGPP synthases of R. glutinosa were divided into three kinds, GGPS, GPPS and FPPS, and their gene expression was significantly diverse in different varieties, growth periods, or tissues of R. glutinosa. Compared with that of GGPS, the expression of GPPS and FPPS was much higher in R. glutinosa, especially at the expansion stage and maturation stage. Thus, the synthesis of terpenoids in roots of R. glutinosa is intricately regulated and needs to be further studied.

The Role of Root Tissue Membrane Proteins in Replanting Stress in Rehmannia glutinosa

The perennial herbaceous plant, Rehmannia glutinosa Libosch, is one of traditional Chinese medicines with a long history of cultivation and medicinal use. However, in production of R. glutinosa, replanting disease severely affected its yield and medicinal quality. Replanting disease is the special stress including biotic and abiotic factors. The membrane proteins system plays the important role in process of plants responding to stress factors. In this study, the differentially expressed root tissue membrane proteins between first planted and replanted R. glutinosa were identified through the isobaric tag for relative and absolute quantitation (iTRAQ). As a result, the membrane protein extraction kit could highly effectively extract the membrane proteins from R. glutinosa root tissue. A total of 698 differential membrane proteins between first planted and replanted R. glutinosa were obtained. Functional analysis revealed that the differential membrane proteins were involved in various metabolic pathways, including transport and breakdown, signal transduction, membrane trafficking and environmental response. Two important molecular events that occurred in cellular membrane of replanted R. glutinosa including the imbalance of ROS (Reactive Oxygen Species) metabolism and immune response were identified in this study. When replanted R. glutinosa plants faced the complex environment factors in rhizosphere, the proteins located in cellular membrane were often first activated to response to stress stimulus, resulted in the upregulated expression of a large number of LRR-RLKs (Leucine-rich repeat receptor-like kinases) receptor proteins. Meanwhile, the Ca2+ signal proteins and related receptor proteins transmitted and responded to the replanting stress, which induced severe oxidative stress response in the cell membrane of R. glutinosa, membrane peroxidation, intracellular signal disorder, and eventually produce replanting disease. Our findings provided the theoretical and data foundation for elucidating the key mechanisms associated with replanting stress. © 2021 Friends Science Publishers