Amelioration of Sensorineural Hearing Loss through Regulation of Trpv1, Cacna1h, and Ngf Gene Expression by a Combination of Cuscutae Semen and Rehmanniae Radix Preparata.

Sensorineural hearing loss (SNHL) is a common condition that results from the loss of function of hair cells, which are responsible for converting sound into electrical signals within the cochlea and auditory nerve. Despite the prevalence of SNHL, a universally effective treatment has yet to be approved. To address this absence, the present study aimed to investigate the potential therapeutic effects of TS, a combination of Cuscutae Semen and Rehmanniae Radix Preparata. To this end, both in vitro and in vivo experiments were performed to evaluate the efficacy of TS with respect to SNHL. The results showed that TS was able to protect against ototoxic neomycin-induced damage in both HEI-OC1 cells and otic hair cells in zebrafish. Furthermore, in images obtained using scanning electron microscopy (SEM), an increase in the number of kinocilia, which was prompted by the TS treatment, was observed in the zebrafish larvae. In a noise-induced hearing loss (NIHL) mouse model, TS improved hearing thresholds as determined by the auditory brainstem response (ABR) test. Additionally, TS was found to regulate several genes related to hearing loss, including Trpv1, Cacna1h, and Ngf, as determined by quantitative real-time polymerase chain reaction (RT-PCR) analysis. In conclusion, the findings of this study suggest that TS holds promise as a potential treatment for sensorineural hearing loss. Further research is necessary to confirm these results and evaluate the safety and efficacy of TS in a clinical setting.

Investigation of anti-osteoporosis mechanisms of Rehmanniae Radix Preparata based on network pharmacology and experimental verification.

BACKGROUND: Rehmanniae Radix Preparata (RRP) can effectively improve the symptoms of osteoporosis, but its molecular mechanism for treating osteoporosis is still unclear. The objective of this study is to investigate the anti-osteoporosis mechanisms of RRP through network pharmacology. METHODS: The overlapping targets of RRP and osteoporosis were screened out using online platforms. A visual network diagram of PPI was constructed and analyzed by Cytoscape 3.7.2 software. Molecular docking was used to evaluate the binding activity of ligands and receptors, and some key genes were verified through pharmacological experiments. RESULTS: According to topological analysis results, AKT1, MAPK1, ESR1, and SRC are critical genes for RRP to treat osteoporosis, and they have high binding activity with stigmasterol and sitosterol. The main signal pathways of RRP in the treatment of osteoporosis, including the estrogen signaling pathway, HIF-1 signal pathway, MAPK signal pathway, PI3K-Akt signal pathway. Results of animal experiments showed that RRP could significantly increase the expression levels of Akt1, MAPK1, ESR1, and SRC1 mRNA in bone tissue to increase bone density. CONCLUSION: This study explained the coordination between multiple components and multiple targets of RRP in the treatment of osteoporosis and provided new ideas for its clinical application and experimental research.

Integrated gut microbiota and fecal metabolomics reveal the renoprotective effect of Rehmanniae Radix Preparata and Corni Fructus on adenine-induced CKD rats.

Rehmanniae Radix Preparata (RR) and Corni Fructus (CF), well-known traditional Chinese medicines (TCMs), were generally used together in the clinical practices to treat chronic kidney disease (CKD) with synergistic effects for thousands of years, but their combination mechanism remains largely unknown so far. Recent evidences have implicated intestinal flora as potential targets for the therapy of CKD. In this study, the CKD rat model was induced by adenine. The levels of proteinuria, serum creatine (SCr), blood urea nitrogen (BUN) and creatinine clearance (Ccr) were used to assess the cooperation effect of RR and CF. Furthermore, high-throughput 16S ribosomal RNA (rRNA) gene sequencing combined with fecal metabonomics based on UPLC-Q-TOF-MS/MS were applied to explore the variations of intestinal flora and their metabolic profiles. 16S rRNA gene sequencing data indicated that CKD rats treated with RR, CF and RC showed the differences in the composition of gut microbiota. The abundance of beneficial bacteria including Ruminococcaceae UCG-014, Ruminococcus 1, Prevotellaceae_NK3B31_group, Lachnospiraceae NK4A136 group and Lachnospiraceae UCG-001 were elevated in various degrees, while the opportunistic pathogen such as Desulfovibrio was markedly decreased after the treatment. Moreover, fecal metabolite profiles revealed 15 different metabolites associated with CKD. These metabolites were mainly involved in the related metabolic pathways such as amino acid metabolism, bile acids metabolism and glycerophospholipid metabolism. The results implied that gut flora and their metabolites might play a vital role in the progress of CKD, which provided a potential target for the development of novel drugs for the therapy of CKD.

The synergic renoprotective effect of Rehmanniae Radix Preparata and Corni Fructus on adenine-induced chronic kidney disease rats based on integrated plasma metabolomics and network pharmacology approach.

AIMS: Rehmanniae Radix Preparata (RR) and Corni Fructus (CF) are commonly used together for the treatment of chronic kidney disease (CKD) in the clinical practices for thousands of years. However, little information on their synergy mechanism is available. MAIN METHODS: In this study, an integrated approach combining ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS)-based metabonomics and network pharmacology was adopted to elucidate the cooperation mechanism of RR and CF on the amelioration of CKD. Furthermore, the targets from network pharmacology and metabolism pathways were jointly analyzed. Finally, the activities of key metabolic enzymes were experimentally validated by ELISA. KEY FINDINGS: Metabolic profiling indicated that the metabolic disturbance in plasma was markedly alleviated after treatment. Nine putative biomarkers mainly involving in phenylalanine, tyrosine and tryptophan biosynthesis and tyrosine metabolism were identified. Moreover, the compound-target-pathway network of RR and CF for CKD treatment was constructed by network pharmacology, which was related to tyrosine metabolism and arginine and proline metabolism. The results were partly consistent with the findings of plasma metabolomics. SIGNIFICANCE: In conclusion, this study solidly supported and enhanced current understanding of the synergy effects of RR and CF on CKD. Meanwhile, it also confirmed the feasibility of combining metabolomics and network pharmacology to identify active components and elucidate the pharmacological effects of traditional Chinese medicines (TCMs).

A new alkaloid from Rehmanniae Radix Preparata / 药学学报

The chemical constituents of Rehmanniae Radix Preparatawere prepared according to the traditional method of "jiu zheng jiu shai" and investigated using multiple chromatographic methods. Six alkaloids were isolated and their structures were elucidated from spectral data and physicochemical properties, as follows: rehmanniae alkaloid A (4-{[(5-O-á-D-galactopyranosyloxy)methyl]-1H-pyrrole-2-carbaldehyde-1-yl}butyric acidmethyl ester) (1), baimantuoluoamide B (2), capparisine C (3), harman-3-carboxylic acid (4), (2S)-1-[2-(furan-2-yl)-2-oxoethyl]-5-oxopyrrolidine-2-carboxylate (5), and 1-[2-(furan-2-yl)-2-oxoethyl]pyrrolidin-2-one (6). Among them, compound 1 is a new alkaloid. Compounds 2-6 were newly isolated from Rehmannia glutinosa Libosch.The effect of compounds 1-6 on NRK-52e cell injury induced by LPS was investigated. The results show that compounds 1-3 exhibit protective effects against LPS-induced damage to NRK-52e cells.

Comparison of Antioxidant Effect of Rehmannia glutinosa Decoction Pieces and Its Polysaccharides on Aging Model Rats before and after Processing Based on the Characteristic Ancient Method / 中国药房

OBJECTIVE：To compare the difference in the antioxid ant effect of fresh Rehmannia glutinosa ，dried R. glutinosa ， R. glutinosa preparata during ancient characteristic processing and its polysaccharides before and after processing on aging model rats，and to provide reference for the processing of R. glutinosa . METHODS ：The sample of R. glutinosa preparata was prepared according to ancient characteristic method. During the processing ，the fresh and dried R. glutinosa samples were retained. Then crude polysaccharide were extracted from fresh R. glutinosa and Rehmanniae radix preparata by water extraction and alcohol precipitation. Totally 96 rats were divided into blank group （water），model group （water），positive control group [vitamine C ，100 mg/（kg·d）]，fresh R. glutinosa group [ 700 mg/（kg·d）]，dried R. glutinosa group [ 135 mg/（kg·d）] ，Rehmanniae radix preparata group [ 135 mg/（kg·d）]，fresh R. glutinosa polysaccharide group [ 1 400 mg/（kg·d），by the weight of fresh R. glutinosa ] and Rehmanniae radix preparata polysaccharide group mg/（kg·d），by the weight of Rehmanniae radix preparata] ， with 12 rats in each group. Except for blank group ，other 制。E-mail：zhouyan1221@163.com groups were given D-galactose [ 125 mg/（kg·d）] on neck and back to induce sub-acute aging model. At the same time ，they were given relevant medicine in tragastrically，once a day ，for consecutive 56 days. After last admin istration，the liver ，brain，kidney，spleen，heart and thymus indexes were determined. The total antioxidant capacity （T-AOC），superoxide dismutase （SOD）activity，catalase（CAT）activity and MDA content in serum ， liver，brain and kidney were determined. RESULTS ：Compared with blank group ，organ indexes of rats in the model group were decreased significantly （P＜0.05 or P＜0.01）；T-AOC，SOD activity and CAT activity in serum ，brain，liver and kidney tissue were decreased significantly （P＜0.01），while MDA content increased significantly （P＜0.01）. Compared with model group ，the organ indexes of brain ，liver and kidney ，SOD activity in serum and kidney of fresh R. glutinosa group were not significantly increased （P＞0.05）；kidney index ，T-AOC in serum and brain ，SOD activity in serum ，liver and kidney tissue were not significantly increased in the dried R. glutinosa group（P＞0.05）；kidney index ，T-AOC in serum and cerebral tissue ，SOD activity in serum were not significantly increased in fresh R. glutinosa group（P＞0.05）；other organ indexes ，T-AOC，SOD activity and CAT activity in serum and tissues were increased significantly in other groups （P＜0.05 or P＜0.01），while MDA content in serum and tissues were decreased significantly in all administration groups （P＜0.05 or P＜0.01）. Compared with fresh R. glutinosa group，T-AOC in serum was decreased significantly in dried R. glutinosa group（P＜0.01），and there was no significant difference in other indexes （P＞0.05）；kidney and spleen indexes of rats in Rehmanniae radix preparata group were increased significantly （P＜0.05），T-AOC in renal tissue ，SOD activity in serum ，cerebral tissue and renal tissue ，CAT activity in cerebral and liver tissue were increased significantly （P＜0.05 or P＜0.01），while MDA in cerebral and liver tissue were significantly decreased （P＜ 0.01）. CAT in cerebral tissue and liver tissue of rats in Rehmanniae radix preparata group were significantly higher than those in positive control group （P＜0.01）. Compared with fresh R. glutinosa polysaccharide group ，spleen and renal indexes of rats in Rehmanniae radix preparata group were increased significantly （P＜0.05 or P＜0.01），T-AOC，SOD activity and CAT activity in serum and cerebral ，liver，renal tissues were increased significantly （P＜0.05 or P＜0.01）. T-AOC and CAT activity of cerebral ， liver and renal tissues in Rehmanniae radix preparata group were all significantly higher than those in positive control group （P＜ 0.05 or P＜0.01）. CONCLUSIONS ：In the aspect of increasing organ index and improving the activity of antioxidant enzymes in serum，cerebral，liver and

Metabolomics Profiling Reveals Rehmanniae Radix Preparata Extract Protects against Glucocorticoid-Induced Osteoporosis Mainly via Intervening Steroid Hormone Biosynthesis.

Rehmanniae Radix Preparata (RR), the dry rhizome of Rehmannia glutinosa Libosch., is a traditional herbal medicine for improving the liver and kidney function. Ample clinical and pharmacological experiments show that RR can prevent post-menopausal osteoporosis and senile osteoporosis. In the present study, in vivo and in vitro experiments, as well as a UHPLC-Q/TOF-MS-based metabolomics study, were used to explore the preventing effect of RR on glucocorticoid-induced osteoporosis (GIOP) and its underlying mechanisms. As a result, RR significantly enhanced bone mineral density (BMD), improved the micro-architecture of trabecular bone, and intervened in biochemical markers of bone metabolism in dexamethasone (DEX)-treated rats. For the in vitro experiment, RR increased the cell proliferation and alkaline phosphatase (ALP) activity, enhanced the extracellular matrix mineralization level, and improved the expression of runt-related transcription factor 2 (RUNX2) and osteopontin (OPN) in DEX-injured osteoblasts. For the metabolomics study, a total of 27 differential metabolites were detected in the DEX group vs. the control group, of which 10 were significantly reversed after RR treatment. These metabolites were majorly involved in steroid hormone biosynthesis, sex steroids regulation, and amino acid metabolism. By metabolic pathway and Western blotting analysis, it was further ascertained that RR protected against DEX-induced bone loss, mainly via interfering steroid hormone biosynthesis, as evidenced by the up-regulation of cytochrome P450 17A1 (CYP17A1) and aromatase (CYP19A1), and the down-regulation of 11ß-hydroxysteroid dehydrogenase (HSD11B1). Collectively, these results indicated that RR had a notable preventing effect on GIOP, and the action mechanism might be related to steroid hormone biosynthesis.

Rehmanniae Radix Preparata suppresses bone loss and increases bone strength through interfering with canonical Wnt/ß-catenin signaling pathway in OVX rats.

Rehmanniae Radix Preparata (RRP) improves bone quality in OVX rats through the regulation of bone homeostasis via increasing osteoblastogenesis and decreasing osteoclastogenesis, suggesting it has a potential for the development of new anti-osteoporotic drugs. INTRODUCTION: Determine the anti-osteoporotic effect of RRP in ovariectomized (OVX) rats and identify the signaling pathway involved in this process. METHODS: OVX rats were treated with RRP aqueous extract for 14 weeks. The serum levels of tartrate-resistant acid phosphatase (TRAP), receptor activator of nuclear factor kappa-Β ligand (RANKL), alkaline phosphatase (ALP), and osteoprotegerin (OPG) were determined by ELISA. Bone histopathological alterations were evaluated by H&E, Alizarin red S, and Safranin O staining. Bone mineral density (BMD) and bone microstructure in rat femurs and lumbar bones were determined by dual-energy X-ray absorptiometry and micro-computed tomography. Femoral bone strength was detected by a three-point bending assay. The expression of Phospho-glycogen synthase kinase 3 beta (p-GSK-3ß), GSK-3ß, Dickkopf-related protein 1 (DKK1), cathepsin K, OPG, RANKL, IGF-1, Runx2, ß-catenin, and p-ß-catenin was determined by western blot and/or immunohistochemical staining. RESULTS: Treatment of OVX rats with RRP aqueous extract rebuilt bone homeostasis demonstrated by increasing the levels of OPG as well as decreasing the levels of TRAP, RANKL, and ALP in serum. Furthermore, RRP treatment preserved BMD and mechanical strength by increasing cortical bone thickness and epiphyseal thickness as well as improving trabecular distribution in the femurs of OVX rats. In addition, RRP downregulated the expression of DKK1, sclerostin, RANKL, cathepsin K, and the ratio of p-ß-catenin to ß-catenin, along with upregulating the expression of IGF-1, ß-catenin, and Runx2 and the ratio of p-GSK-3ß to GSK-3ß in the tibias and femurs of OVX rats. Echinacoside, jionoside A1/A2, acetoside, isoacetoside, jionoside B1, and jionoside B2 were identified in the RRP aqueous extract. CONCLUSION: RRP attenuates bone loss and improves bone quality in OVX rats partly through its regulation of the canonical Wnt/ß-catenin signaling pathway, suggesting that RRP has the potential to provide a new source of anti-osteoporotic drugs.

Research Progress on Rehmanniae Radix Preparata / 世界科学技术-中医药现代化

This paper is based on the literature of the last ten years by summarizing and analyzing the progress in the study of Rehmanniae Radix, focusing on the research progress are reviewed at the dynamic changes of the main Chemical indicators of Rehmanniae Radix Preparata, objective expression of four kinds of processed products (Steamed, steamed with wine, wine stew, ancient law (steamed for nine times and shined for nine times) ) and the objective expression of four kinds of processed products and the change of the pharmacological effects.There are many objective expression methods for processing end points of different processed products of Rehmanniae Radix Preparata, But processing are unclear until now. Innovative research on standardized unification and new research ideas are needed.

Decoction Amount Changes of Verbascoside and Gallic Acid in Single Decoction and Combined Decoction of Rehmanniae Radix Preparata,Corni Fructus and Poria / 中国药房

OBJECTIVE:To investigate the decoction amount changes of verbascoside,gallic acid in single decoction and com-bined decoction of rehmanniae radix preparata,corni fructus and poria,and provide scientific basis for further study of effective substance basis. METHODS:The single decoction,combined decoction with each 2 medicine and combined decoction with the 3 medicines of rehmanniae radix preparata,corni fructus and poria,were respectively prepared. HPLC was adopted to detect and com-pare the decoction amount of active component verbascoside in rehmanniae radix preparata and active component gallic acid in cor-ni fructus in each group. RESULTS:Compared with single decoction of rehmanniae radix preparata,the decoction amount of ver-bascoside was decreased in combined decoction of poria+rehmanniae radix preparata(P<0.01),decoction amount of verbascoside was increased in combined decoction of corni fructus+rehmanniae radix preparata or combined decoction with the 3 medicines(P<0.01). Compared with single decoction of corni fructus,the decoction amount of gallic acid in each combined decoction was de-creased (P<0.01). CONCLUSIONS:The combined decoction of rehmanniae radix preparata,corni fructus and poria has certain promotion and inhibition effects on the decoction of verbascoside in rehmanniae radix preparata and inhibition effect on the decoc-tion of gallic acid in corni fructus. It is speculated verbascoside may be one of the main components in combined decoction playing the role of effectiveness.

Effects of Rehmanniae Radix Preparata and Poria on decoction amounts of three constituents in Corni Fructus / 中成药

AIM To study the effects of Rehmanniae Radix Preparata (A) and Poria (B) on decoction amounts of loganin,morroniside and 5-hydroxymethylfurfural in Corni Fructus (C).METHODS These three medicinal materials were combined one another and divided into seven groups (A,B,C,A + B,A + C,B + C and A + B + C).Then the contents of three constituents were determined by HPLC.RESULTS Compared with the single decoction of Corni Fructus,the decoction amounts of loganin and 5-hydroxymethylfurfural were decreased,and that of morroniside was increased at the time of mixed decoction of Rehmanniae Radix Preparata and Corni Fructus,or Rehmanniae Radix Preparata,Poria and Corni Fructus.This situation was just the contrary at the time of mixed decoction of Poria and Corni Fructus.CONCLUSION The mixed decoction of Corni Fructus,Rehmanniae Radix Preparata and Poria increases the decoction amount of morroniside,which may make mixed decoction liquid show better efficacy.

Simultaneous analysis of furfural metabolites from Rehmanniae radix preparata by HPLC-DAD-ESI-MS.

A method based on HPLC with diode array detector (DAD) and electrospray ionisation mass spectrometry (ESI-MS) was established for the simultaneous determination of 5-HMF and its derivatives, including a new 5-HMF derivative, in Rehmanniae radix preparata. Validation parameters, such as linearity, limit of detection, limit of quantification, recovery, accuracy, and precision, were successfully obtained. In addition, the efficiencies of diverse extraction methods were compared for the development of a standard analytical method. The verified method was successfully applied to the quantitative determination of four representative metabolites in eighteen R. radix preparata samples from Korea and China. Additionally, the increase in the amount of 5-HMF derivatives was monitored during the processing of three dried R. radix samples. The results showed that a newly isolated diglycosylated 5-HMF derivative, 5-(α-D-galactopyranosyl-(1→6)-α-D-galactopyranosyloxymethyl)-2-furancarboxaldehyde, appeared in concentrations comparable to that of 5-HMF, suggesting its potential to serve as a marker compound in R. radix preparata.