Background: Chinese materia medica processing is a distinguished and unique pharmaceutical technique in Traditional Chinese Medicine (TCM) used for reducing side effects, and increasing or even changing therapeutic efficacy of the raw herbs.Changes in the essential components induced by an optimized processing procedure are primarily responsible for the increased efficacy of medicinal plants.The kidney-yang invigorating effect of rice wine-steamed Cistancha deserticola (C. deserticola) was stronger than raw C. deserticola (CD). Methods: A comparison analysis was carried out using the UPLC-Q-TOF-MS^E with the UNIFI informatics platform to determine the influence of processing. In vitro studies were performed for the characterization of constituents as well as metabolites in vivo. The chemical components were determined in CD and its processed products. The multivariate statistical analyses were conducted to evaluate variations between them while OPLS-DA was used for pairwise comparison. Results: The results of this study revealed considerable variations in phenylethanoid glycosides (PhGs) and iridoids after processing. A total of 97 compounds were detected in the extracts of CD and its processed product. PhGs having 4'-O-caffeoyl group in the 8-O-β-d-glucopyranosyl part, like acteoside, cistanoside C, campneoside II, osmanthuside decreased after being processed, while PhGs with 6'-O-caffeoyl group in the 8-O-β-d-glucopyranosyl part, such as isoacetoside, isocistanoside C, isocampneoside I, isomartynoside increased, especially in the CD-NP group. The intensity of echinacoside and cistanoside B whose structure possess 6'-O-β-d-glucopyranosyl moiety also increased. In in vivo study, 10 prototype components and 44 metabolites were detected in rat plasma, feces, and urine. The obtained results revealed that processing leads to the considerable variation in the chemical constituents of CD and affected the disposition of the compounds in vivo, and phase II metabolic processes are the key cascades of each compound and most of the metabolites are associated with echinacoside or acteoside. Conclusions: This is the first global comparison research of raw and processed CD. These findings add to our understanding of the impact of CD processing and give important data for future efficacy investigations.

The desert‐dwelling Cistanche herb was first recorded in the “Shen Nong Herbal Classic” and is listed as the top‐grade herbal medicine in this publication. The Chinese Pharmacopoeia records that pieces of Cistanche deserticola (CD) and rice wine‐steamed Cistanche deserticola (WCD) can be used in the clinic as the main types of decoctions. After being steamed with rice wine, the antiaging and tonifying kidney‐yang effects are enhanced. In this study, we detected the chemical content of CD and WCD and the pharmacological mechanism of invigorating kidney‐yang deficiency in model rats. Aim. The purpose of this study was to examine the effects of CD and WCD on the neuroendocrine‐immune function of kidney‐yang deficiency in glucocorticoid‐overdosed model rats. Materials and methods. Sprague Dawley (SD) rats were selected. The rats were subcutaneously injected with corticosterone water suspension for the glucocorticoid‐overdosed model rats. The positive control rats were gavaged with Jinkuishenqi pills and high‐, medium‐, and low‐dose CD/WCD suspensions (1.646 g/(kg day), 5.48 g/(kg day), 2.74 g/(kg day), and 1.37 g/(kg day), respectively); the blank control (BC) and model control (MC) groups were given the same volume of distilled water as those in the drug group for 40 consecutive days at a dose of 1 mL/100 g. After the last administration, the blood was collected from the abdominal aorta, and serum levels of T, CRH, ACTH, CORT, cortisol, IL‐10, IL‐6, IL‐2, TNF‐α, and IFN‐γ were measured. Organ indexes of the thymus gland and the spleen were calculated. The expression of Bax, Bcl‐2, caspase‐3, Fas, and FasL in the adrenal gland was measured by immunohistochemistry. The pathological changes in the thymus gland and the adrenal gland were observed by HE staining (×200). T lymphocyte subsets in peripheral blood were detected by flow cytometry, and the expression of CaM mRNA in the hypothalamus and hypophysis tissues was also measured by RT‐PCR. Results. Compared with the MC group, the CD and WCD groups exhibited increases in activity, the organ index of the thymus and the spleen, the serum levels of T, CRH, ACTH, CORT, cortisol, IL‐2, and IL‐10, the ratio of CD4+/CD8+, and the expression of Bcl‐2, caspase‐3, Fas, FasL, and CaM in the hypophysis tissue. The CD and WCD groups also exhibited reductions in the IL‐6, TNF‐α, and IFN‐γ levels in serum and the expression of CaM mRNA in the hypothalamus. Conclusions. Each dose of CD and WCD could counteract the dysregulated sex hormone and immune factors in glucocorticoid‐overdosed model rats, enhancing and restoring the effect of the hypothalamic nerve cells and improving immune function.

Introduction: For centuries, Cistanche deserticola Y. C. Ma has been considered to have the effect of “tonifying the kidney and strengthening the yang,” and is used for the prevention and treatment of diseases such as impotence, female infertility, lumbago and senile constipation. Polysaccharides and small molecules of acteoside are the main chemical compounds co-existing in Cistanche deserticola Y. C. Ma with health benefits, but the interaction of these two compounds in vivo is not yet known.Methods: Here, we investigated the effects of unprocessed Cistanche polysaccharides and wine-processed Cistanche polysaccharides on the metabolism of acteoside in vivo through gut microbiota. Male Sprague–Dawley rats were randomly divided into Control group, unprocessed Cistanche polysaccharide group (UCP group) and wine-processed Cistanche polysaccharide group (WCP group). After 21 days of intervention with unprocessed Cistanche polysaccharides and wine-processed Cistanche polysaccharides, rats were given 100 mg/kg of acteoside on day 22. Acteoside and its associated metabolites pharmacokinetically studied were analysed using UPLC-QqQ-MS, and the composition of short-chain fatty acids (SCFAs) in excrement was measured using the technique of GC–MS. The microbiological composition of the intestines was discovered using 16S rRNA gene amplicon sequencing.Results: The results showed that the Cistanche polysaccharides used in this experiment, including unprocessed Cistanche polysaccharides and wine-processed Cistanche polysaccharides, could regulate the diversity of gut microbiota and increase the number of beneficial bacteria, especially wine-processed Cistanche polysaccharides were able to promote the growth of Ligilactobacillus and Duncaniella genus, and improve the production of SCFAs and the absorption of acteoside.Discussion: By exploring the synergistic effects of large molecules Cistanche polysaccharides and small molecule acteoside, this paper provides a new explanation for the scientific use of plant-derived polysaccharides to improve the bioavailability of oral drugs.

Introduction: Kidney Yang Deficiency (KYD) is a metabolic disorder associated with kidney damage. Its slow progression means that causative factors and effective therapeutic agents remain unclear. Extensive evidence links KYD to gut microbiome metabolic diseases and the Hypothalamic-Pituitary-Thyroid (HPT) axis. Cistanche deserticola (CD) is a commonly used traditional Chinese medicine for treating KYD. However, the precise interactions between gut microbiota and KYD, as well as the mechanisms of raw and processed CD total glycosides (CDG) in modulating KYD, require further investigation. This study aims to evaluate the effects and mechanisms of CDG in a KYD rat model using 16S rRNA gene sequencing and fecal metabolomics.Methods: CDG was extracted from both raw and processed CD and analyzed via HPLC. Propylthiouracil-induced KYD rats were used to assess pharmacological effects, including serum levels of T₃, T₄, TSH, TRH, FFA, LPL, and NO; organ indices of the spleen, kidney, and thymus; blood cAMP/cGMP levels; and liver levels of glycogen, SDH, Ca²⁺-ATPase, and Na⁺-K⁺-ATPase. Immunohistochemistry was also performed.Results: Fecal non-targeted metabolomics identified 98 metabolites associated with KYD, while 16S rRNA sequencing revealed 13 key intestinal microbiotas linked to KYD. CDG therapy effectively alleviated KYD symptoms by modulating the gut microbiota, improving metabolic and microbial imbalances in KYD. RG/WG significantly improves KYD rats mainly through the relationship between the intestinal microbiota and arachidonic acid metabolism. The key bacterial genera lleibacterium and Streptococcus observed in the changes of intestinal flora and fecal metabolite content were significantly negatively correlated with phosphatidylcholine and phosphatidylethanolamine.Discussion: This integrative approach of gut microbiome and fecal metabolomics not only provides a scientific basis for CDG’s preventive effects on KYD via the HPT axis but also elucidates the potential mechanisms underlying CDG’s action against KYD.

Polysaccharides from wine-processed Cistanche deserticola Y.C. Ma (WCPs) are gaining increasing interest because of their diverse bioactivities, yet the extraction method critically affects polysaccharide yield, structure, and function. This study was designed to systematically compare seven extraction strategies and establish a clear method–structure–activity relationship for obtaining high-quality WCPs. Seven extraction protocols, hot water (HWE), enzymatic-assisted (EAE), ultrasound-assisted (UAE), ultrasound–microwave synergistic (UMSE), microwave-assisted (MAE), ultrasound–enzyme combined (UESE), and natural deep-eutectic-solvent (NADES-E) extraction, were evaluated in parallel. UV, FT-IR, Congo-red assays, monosaccharide composition, molecular-weight distribution, and SEM were employed to characterize the functional groups and surface ultrastructures of the obtained WCPs. Five deproteinization techniques were also screened for maximal protein removal with minimal polysaccharide loss. Antioxidant (DPPH, ABTS⁺, OH scavenging) and lipid-lowering (sodium cholate, glycodeoxycholate, taurodeoxycholate binding) activities were measured in a concentration-dependent manner. Optimal extraction conditions were determined to maximize polysaccharide yield and activity. HWE achieved the highest polysaccharide yield (28.43 ± 0.61%), significantly higher than other extraction methods (p < 0.05). Structural and structure–activity relationship analyses demonstrated that the increased bioactivity was driven by specific structural alterations resulting from the extraction process. These alterations notably included increases in the total phenolics and uronic acids, as well as changes in monosaccharide composition, particularly increased levels of mannose and galacturonic acid. The extracted polysaccharides therefore showed markedly greater efficacy in both antioxidant and lipid-lowering activity assays. Among the protein removal methods evaluated, the Sevage method was determined to be the most suitable, as it effectively removed proteins while best preserving polysaccharide bioactivity. Quantitative associations among the WCP extraction protocol, structural composition, and bioactivity were established. HWE is recommended as a simple, high-yield route to produce highly active WCPs for functional-food and pharmaceutical development. These findings provide valuable insights into the structure–activity relationships of WCPs and their potential in functional food development. Graphical Abstract

Acute kidney injury (AKI) is a sudden loss of renal function with a high mortality rate and inflammation is thought to be the underlying cause. The phenylpropanoid components acteoside (ACT) and isoacteoside (ISO), which were isolated from Cistanche deserticola Y.C.Ma, have been reported to have preventive effects against kidney disorders. This study aimed to investigate the anti-inflammatory properties and protective mechanisms of ACT and ISO. In this investigation, kidney function was assessed using a semi-automatic biochemical analyzer, histopathology was examined using Hematoxylin-Eosin staining and immunohistochemistry, and the concentration of inflammatory cytokines was assessed using an enzyme-linked immunosorbent assay (ELISA) test. In addition, using Western blot and q-PCR, the expression of proteins and genes connected to the NF-κB signaling pathway in mice with lipopolysaccharide (LPS)-induced AKI was found. The findings showed that under AKI intervention in LPS group, ACT group and ISO group, the expression of Rela (Rela gene is responsible for the expression of NFκB p65 protein) and Tlr4 mRNA was considerably elevated (P<0.01), which led to a significant improvement in the expression of MyD88, TLR4, Iκ-Bɑ and NF-κB p65 protein (P<0.001). The levels of Alb, Crea and BUN (P<0.001) increased along with the release of downstream inflammatory factors such as IL-1β, IL-6, Cys-C, SOD1 and TNF-α (P<0.001). More importantly, the study showed that ISO had a more favorable impact on LPS-induced AKI mice than ACT. In conclusion, by inhibiting NF-κB signaling pathway, ACT and ISO could relieve renal failure and inflammation in AKI, offering a fresh possibility for the therapeutic management of the condition.