1.Processing History and Modern Research of Bile-processed Coptidis Rhizoma: A Review
Zhaowei DONG ; Jing YANG ; Qinwan HUANG ; Jin WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):269-278
Bile-processed Coptidis Rhizoma(B-pCR), first documented in Shengji Zonglu, is a unique processed products of Coptidis Rhizoma(CR) characterized by "mutual enhancement processing" and "enhancing the cold property of cold-natured herbs". Pig bile can enhance the bitter and cold properties of CR, yielding potent effects in purging excess fire from the liver and gallbladder. The processing increases the dissolution of alkaloids such as berberine, coptisine, and palmatine, while introducing bile acids from pig bile, including taurine-type and glycine-type cholic acids. This enhances its pharmacological effects, such as antipyretic activity, regulation of glucose and lipid metabolism disorders, and intestinal absorption. Traditional processing techniques and quality standards for B-pCR are outlined in the Shanghai Traditional Chinese Medicine(TCM) Decoction Pieces Processing Standard and the Gansu TCM Processing Standard. However, incomplete specifications for critical process parameters and quality criteria significantly impact its production and clinical application. A review of research over the past two decades on the processing history, process optimization, quality evaluation, material basis, and changes in pharmacological effects and properties of B-pCR reveals that the pretreatment method and dosage of pig bile, and processing temperature are key factors influencing its quality. Furthermore, current quality standards lack specific indicators. Additionally, the enhancement of the cold property and medicinal efficacy direction of B-pCR is not only associated with changes in alkaloid groups but also depend on the synergistic effects of bile acids. This review can provide insights for improving the quality evaluation system of B-pCR.
2.Flos Buddlejae self-heating steam eye mask combined with sodium hyaluronate eye drops in the treatment of dry eye disease
Zhaodan TAN ; Qian LI ; Yan SHI ; Kangyuan HU ; Jin HUANG
Journal of Pharmaceutical Practice and Service 2026;44(2):96-102
Objective To assess the clinical efficacy of sodium hyaluronate (0.3%) eye drops combined with herbal self-heating steam eye mask in the treatment of dry eye disease. Methods A prospective randomized controlled clinical trial was performed on 60 patients diagnosed with dry eye at the ophthalmic clinic of a Grade A, Class Ⅲ hospital in Shanghai from June 2023 to September 2024. Specifically, patients were randomly divided into control group and study group. Patients in the control group were treated with sodium hyaluronate (0.3%) eye drops for six weeks; while in the study group, patients received the eye drops combined with the herbal self-heating steam eye mask mainly containing powders of Flos Buddlejae. Subsequently, comparisons and analysis were performed before and after treatment between the two groups in the clinical symptom questionnaire score traditional Chinese medicine (TCM syndrome score),the Chinese dry eye questionnaire score and determination of tear film fluorscein breakup time (FBUT), and curative effect. Results The quality control standard of the herbal powder in the self-heating steam eye mask was established through TLC and HPLC, and good heating behavior of the herbal self-heating steam eye mask was ascertained heating temperature (43±5)℃; heating duration (≥20 min), meeting requirements of the product quality control. After treatment for 6 weeks, FBUT was increased, while TCM syndrome score and the Chinese dry eye questionnaire score were both decreased in the study group (P<0.001). Besides, compared with the control group, TCM syndrome score and the Chinese dry eye questionnaire score were much lower, while the FBUT were higher in the study group (P<0.001). Moreover, the overall response rate in the study group (81.7%) was much better than that in the control group (25.9%). Conclusion The combination of sodium hyaluronate (0.3%) eye drops with herbal self-heating steam eye mask could be applied to the clinical treatment of dry eye disease due to its good clinical effects on relieving dry eye symptoms.
3.Mechanisms of Anemarrhenae Rhizoma Water Extract in Ameliorating Neuroinflammation in Alzheimer's Disease Model Rats via SIRT1/HMGB1/NF-κB Signaling Pathway
Fei WU ; Yuexia LI ; Qi HUANG ; Tianshi LI ; Chuanshan JIN ; Kai MA
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):230-240
ObjectiveTo investigate the therapeutic effects of the Anemarrhenae Rhizoma water extract (AR) on Alzheimer's disease (AD) model rats and to explore its potential underlying mechanisms. MethodsMale rats were intraperitoneally injected with D-galactose (100 mg·kg-1) for 42 days, and on day 14, 1 μL of β-amyloid (Aβ25-35, 2 g·L-1) solution was injected into the hippocampus. Rats were randomly divided into a model group, low-dose AR (0.6 g·kg-1), medium-dose AR (1.2 g·kg-1), high-dose AR (2.4 g·kg-1), and a positive control group (donepezil, 5 mg·kg-1). Healthy rats receiving only a hippocampal injection of 1 μL of sterile saline served as the sham-operated group. From day 21, rats in the treatment groups were administered the corresponding drugs by gavage once daily for 21 consecutive days, while the blank control and model groups received an equal volume of saline. Learning and memory abilities were assessed using the Morris water maze. Brain tissue damage was observed by hematoxylin and eosin (HE) staining, and neuronal apoptosis was evaluated by terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining. Levels of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and interleukin-10 (IL-10) in brain tissues were measured by enzyme-linked immunosorbent assay (ELISA). BV2 microglial cells were co-cultured with Aβ25-35 (40 μmol·L-1) for 2 h, and cell viability was determined by the CCK-8 assay to screen the optimal concentration of AR-containing serum (S-AR). Cells were divided into blank control, Aβ25-35, S-AR, EX527 [silent information regulator 1 (SIRT1) inhibitor], and S-AR+EX527 groups. Immunofluorescence staining was used to detect the expression of CD16, CD206, and high-mobility group box 1 (HMGB1). Western blot analysis was performed to measure the protein expression of CD16, inducible nitric oxide synthase (iNOS), CD206, arginase (Arg), and proteins related to the SIRT1/HMGB1/nuclear factor-κB (NF-κB) signaling pathway. ResultsIn vivo experiments showed that, compared with the sham-operated group, the model group exhibited reduced platform crossings and time spent in the target quadrant (P<0.01), prolonged escape latency, increased hippocampal neuronal apoptosis (P<0.01), and obvious hippocampal damage. The expression levels of IL-6, TNF-α, IL-10, CD16, and iNOS in brain tissues were significantly elevated (P<0.01), while CD206 and Arg protein expression showed an increasing trend without statistical significance. Compared with the model group, all AR-treated groups significantly increased platform crossings and target quadrant time (P<0.05, P<0.01), alleviated hippocampal damage, reduced escape latency and neuronal apoptosis, downregulated the expression of TNF-α, IL-6, CD16, and iNOS (P<0.05, P<0.01), and upregulated the expression of IL-10, CD206 and Arg (P<0.05, P<0.01). In vitro experiments demonstrated that, compared with the blank control group, the Aβ25-35 group showed increased fluorescence intensity of CD206, CD16, and HMGB1, as well as elevated protein expression of iNOS and CD16 (P<0.01), while CD206 and Arg protein expression exhibited an increasing trend without statistical significance. After S-AR intervention, CD206 fluorescence intensity and the protein expression of Arg and CD206 were significantly increased (P<0.01), whereas the fluorescence intensity of CD16 and HMGB1 and the protein expression of iNOS and CD16 were significantly decreased (P<0.01). These effects were reversed by EX527 (P<0.05, P<0.01). Furthermore, compared with the blank control group, the Aβ25-35 group showed significantly increased cytoplasmic HMGB1 expression and p-p65/p65 ratio (P<0.01), along with significantly decreased SIRT1 and nuclear HMGB1 expression (P<0.01). In contrast, the S-AR group exhibited opposite trends compared with the Aβ25-35 group, and the regulatory effects of S-AR on these proteins were reversed by EX527 (P<0.01). ConclusionAR exerts neuroprotective effects in AD model rats by regulating microglial polarization and alleviating neuroinflammation, potentially through modulation of the SIRT1/HMGB1/NF-κB signaling pathway.
4.Effects of Modified Buyang Huanwu Tang on Mice with Cerebral Ischemia-reperfusion Injury by Regulating PINK1/Parkin Signaling Pathway-mediated Mitochondrial Autophagy
Li GUO ; Hengwen CHEN ; Cun ZHAN ; Zhenzhen YING ; Zuomin WU ; Shaoju JIN ; Shangmei CAO ; Shengming HUANG ; Jin WANG ; Xiaotao YU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(11):34-43
ObjectiveTo investigate the effects of modified Buyang Huanwu Tang on cerebral ischemia-reperfusion injury (CI/RI) in mice via the PTEN-induced putative kinase 1/E3 ubiquitin ligase (PINK1/Parkin) signaling pathway-mediated mitophagy, and to explore the underlying mechanism by which modified Buyang Huanwu Tang improves CI/RI. MethodsSeventy-two male C57BL/6J mice were randomly divided into six groups (n = 12 per group): Sham-operated group, middle cerebral artery occlusion/reperfusion (MCAO/R) model group, low-, medium-, and high-dose modified Buyang Huanwu Tang groups (8.84, 17.68, 35.36 g·kg-1·d-1), and an aspirin group (13.00 mg·kg-1·d-1). Neurological deficit scores were assessed using the Zea-Longa method. Cerebral infarct volume ratio was measured by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Histopathological changes and neuronal injury in brain tissues were observed using hematoxylin-eosin (HE) staining and Nissl staining. Apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. Mitochondrial ultrastructure in brain tissue was observed by transmission electron microscopy (TEM). Serum levels of superoxide dismutase (SOD) and malondialdehyde (MDA) were determined by enzyme-linked immunosorbent assay (ELISA). The mRNA and protein expression levels of PINK1, Parkin, microtubule-associated protein 1 light chain 3B (LC3B, LC3Ⅱ/Ⅰ), and p62 in brain tissues were detected by real-time quantitative reverse transcription PCR (Real-time PCR) and Western blot, respectively. ResultsCompared with the sham-operated group, the MCAO/R model group showed significantly increased neurological deficit scores and cerebral infarct volume ratios (P<0.01). Severe cortical injury on the infarct side was observed, characterized by decreased neuronal density, cytoplasmic vacuolation, nuclear pyknosis, a marked reduction in Nissl bodies, dissolution of Nissl bodies in the cytoplasm of some pyramidal neurons, and blurred cellular boundaries. The number of TUNEL-positive cells increased significantly (P<0.01). Mitochondria exhibited cristae membrane rupture and matrix vacuolation, with rupture of the outer mitochondrial membrane and formation of autophagosomes, the number of which increased significantly. Serum SOD activity decreased significantly (P<0.01), while MDA content increased significantly (P<0.01). In infarcted brain tissues of model mice, the relative mRNA expression and protein levels of PINK1, Parkin and LC3B were significantly increased (P<0.05, P<0.01), whereas p62 mRNA and protein expression were significantly decreased (P<0.05, P<0.01), showing statistical significance. Compared with the model group, all treatment groups showed significantly decreased neurological deficit scores and cerebral infarct volume ratios (P<0.01). Neuronal density increased significantly, cytoplasmic vacuolation was alleviated, nuclear morphology tended to be more regular and clearer, Nissl body density increased significantly with reduced dissolution and improved contour clarity. The mitochondrial cristae structure was partially restored, with some mitochondria showing autophagosome encapsulation, and the degree of mitochondrial damage was alleviated. Serum SOD activity increased significantly (P<0.01), while MDA content decreased significantly. The mRNA and protein expression levels of PINK1, Parkin, and LC3Ⅱ/Ⅰ were significantly increased (P<0.05, P<0.01), while p62 mRNA and protein expression in the low- and medium-dose modified Buyang Huanwu Tang groups were significantly decreased (P<0.05, P<0.01), showing statistical significance. ConclusionModified Buyang Huanwu Tang can upregulate the protein expression levels of PINK1, Parkin, and LC3Ⅱ/Ⅰ and downregulate p62 protein expression, suggesting that it may improve CI/RI by regulating the expression of proteins related to the PINK1/Parkin signaling pathway. Regulation of the mitophagy pathway may be one of the mechanisms by which modified Buyang Huanwu Tang alleviates CI/RI in mice.
5.Eucommia ulmoides promotes alveolar bone formation in ovariectomized rats
Lin ZHENG ; Wenjun JIN ; Shanshan LUO ; Rui HUANG ; Jie WANG ; Yuting CHENG ; Zheqing AN ; Yue XIONG ; Zipeng GONG ; Jian LIAO
Chinese Journal of Tissue Engineering Research 2025;29(6):1159-1167
BACKGROUND:Eucommia ulmoides has a certain osteogenic effect,which can promote the proliferation and differentiation of osteoblasts.However,it is unclear whether Eucommia ulmoides has effects on alveolar bone formation and Wnt/β-Catenin signaling pathway. OBJECTIVE:To investigate the mechanism by which Eucommia ulmoides promotes alveolar bone formation in ovariectomized rats based on the Wnt/β-Catenin signaling pathway. METHODS:Sixty female Sprague-Dawley rats were selected and randomly divided into five groups:blank control group,sham-operation group,model group,low-dose group Eucommia ulmoides group,and high-dose Eucommia ulmoides group,with twelve rats in each group.Osteoporosis animal models were constructed by bilateral oophorectomy in the model group and the low-dose and high-dose Eucommia ulmoides groups.The sham-operation group underwent the same method to remove adipose tissue of equal mass around the bilateral ovaries.Three months after surgery,the low-and high-dose Eucommia ulmoides groups were given 2.1 g/kg/d and 4.2 g/kg/d Eucommia ulmoides by gavage,respectively.The sham-operation group and model group were given the same amount of physiological saline by gavage.After 12 weeks of drug intervention,the changes in alveolar bone mass of rats in each group were observed through Micro-CT;hematoxylin-eosin staining was used to observe the pathological structural changes of alveolar bone in rats;enzyme linked immunosorbent assay was used to detect the expression levels of alkaline phosphatase and osteocalcin in the serum of rats;western blot was used to detect the expression levels of β-Catenin and Frizzled9 receptor proteins in the alveolar bone of rats;and real-time fluorescence quantitative PCR was used to detect the expression of osteocalcin,Runt-related transcription factor 2(Runx2),alkaline phosphatase,β-catenin,and frizzled9 mRNAs in alveolar bone tissues of rats. RESULTS AND CONCLUSION:Compared with the blank control group,bone volume fraction,trabecular number,trabecular thickness,and bone mineral density were reduced in the model group(P<0.05),and trabecular separation was elevated(P<0.05).Pathological observation showed that the arrangement of trabeculae was disordered and irregular,the trabeculae were thinned or broken,and the marrow cavity was enlarged in the model group,with a significant reduction in bone volume;the level of alkaline phosphatase in the serum was increased(P<0.05),and the level of osteocalcin was decreased(P<0.05);mRNA expression of alkaline phosphatase,osteocalcin,Runx2,β-catenin,and frizzled9 were decreased(P<0.05);protein expression of β-Catenin and Frizzled9 was decreased(P<0.05).Compared with the model group,the low-and high-dose Eucommia ulmoides groups showed an increase in bone volume fraction,trabecular number,trabecular thickness,and bone mineral density(P<0.05)and a decrease in trabecular separation(P<0.05).In the low-and high-dose Eucommia ulmoides groups,bone trabeculae were slightly aligned and thickened,with a significant increase in bone mass.Compared with the model group,the serum level of alkaline phosphatase was reduced(P<0.05)and the serum level of osteocalcin was elevated(P<0.05)in the low-and high-dose Eucommia ulmoides groups.Compared with the model group,the mRNA expression of alkaline phosphatase,osteocalcin,Runx2,β-catenin,and frizzled9 were increased in the low-and high-dose Eucommia ulmoides groups(P<0.05).Compared with the model group,the protein expression of Frizzled9 was increased in the low-dose Eucommia ulmoides group(P<0.05),while the protein expression of β-Catenin and Frizzled9 was increased in the high-dose Eucommia ulmoides group(P<0.05).Compared with the low-dose Eucommia ulmoides group,the high-dose Eucommia ulmoides group had a more significant improvement in the above indexes.To conclude,Eucommia ulmoides can effectively promote the alveolar bone formation,and its mechanism of action might be related to the activation of the Wnt/β-catenin signaling pathway.
6.Role and mechanism of platelet-derived growth factor BB in repair of growth plate injury
Hongcheng PENG ; Guoxuan PENG ; Anyi LEI ; Yuan LIN ; Hong SUN ; Xu NING ; Xianwen SHANG ; Jin DENG ; Mingzhi HUANG
Chinese Journal of Tissue Engineering Research 2025;29(7):1497-1503
BACKGROUND:In the initial stage of growth plate injury inflammation,platelet-derived growth factor BB promotes the repair of growth plate injury by promoting mesenchymal progenitor cell infiltration,chondrogenesis,osteogenic response,and regulating bone remodeling. OBJECTIVE:To elucidate the action mechanism of platelet-derived growth factor BB after growth plate injury. METHODS:PubMed,VIP,WanFang,and CNKI databases were used as the literature sources.The search terms were"growth plate injury,bone bridge,platelet-derived growth factor BB,repair"in English and Chinese.Finally,66 articles were screened for this review. RESULTS AND CONCLUSION:Growth plate injury experienced early inflammation,vascular reconstruction,fibroossification,structural remodeling and other pathological processes,accompanied by the crosstalk of chondrocytes,vascular endothelial cells,stem cells,osteoblasts,osteoclasts and other cells.Platelet-derived growth factor BB,as an important factor in the early inflammatory response of injury,regulates the injury repair process by mediating a variety of cellular inflammatory responses.Targeting the inflammatory stimulation mediated by platelet-derived growth factor BB may delay the bone bridge formation process by improving the functional activities of osteoclasts,osteoblasts,and chondrocytes,so as to achieve the injury repair of growth plate.Platelet-derived growth factor BB plays an important role in angiogenesis and bone repair tissue formation at the injured site of growth plate and intrachondral bone lengthening function of uninjured growth plate.Inhibition of the coupling effect between angiogenesis initiated by platelet-derived growth factor BB and intrachondral bone formation may achieve the repair of growth plate injury.
7.Development of a RP scoring system for predicting perioperative outcomes in robot-assisted partial nephrectomy by optimizing RENAL and MAP scores
Liang ZHENG ; Bohong CHEN ; Haoxiang HUANG ; Cong FENG ; Jin ZENG ; Wei CHEN ; Dapeng WU
Journal of Modern Urology 2025;30(1):53-58
[Objective] To establish a new scoring system to predict the perioperative outcomes (operation time, intraoperative blood loss, and trifecta achievement) in patients undergoing robot-assisted partial nephrectomy (RAPN) by integrating the RENAL and Mayo adhesive probability (MAP) scores. [Methods] Clinical data of 178 patients with renal cell carcinoma who underwent RAPN performed by the same surgeon in our hospital during Jan.2015 and Jan.2022 were retrospectively analyzed.The RENAL and MAP scores of all patients were calculated.Linear regression and logistic regression were used to evaluate the associations between the components of the RENAL and MAP scores (a total of 6 variables) and perioperative outcomes.The factors with significant associations were then included into logistic regression analysis to identify independent predictors for constructing an assessment system for perioperative outcomes, and the receiver operating characteristic (ROC) curve was plotted to calculate the area under the curve (AUC) to predict its efficacy. [Results] Multivariate linear regression analysis showed that tumor size (β=6.14, 95%CI: 1.93—10.34, P=0.004), exophytic rate (β=10.60, 95%CI: 3.44—17.76, P=0.004), and perinephric fat thickness (β=16.48, 95%CI: 8.52—24.45, P<0.001) were significantly associated with operation time.Tumor size (β=10.55 95%CI: 5.60—15.49, P<0.001) was associated with both intraoperative blood loss and trifecta achievement (OR=1.73, 95%CI: 1.26—2.36, P=0.001). Multivariate logistic regression analysis of these 3 factors identified tumor size (OR=9.07, 95% CI: 1.18—69.45, P=0.03) and perinephric fat thickness (OR=2.28, 95%CI: 1.86—6.04, P=0.01) as independent predictors of perioperative outcomes.Based on these findings, the tumor size and perinephric fat thickness (RP) scoring was constructed, which demonstrated better predictive ability than RENAL score or MAP score alone (RP vs.RENAL vs.MAP: 0.766 vs.0.548 vs.0.684). [Conclusion] The RP score includes fewer variables than the RENAL and MAP scores but outperforms them.
8.Effect of neuromuscular electrical stimulation on quadriceps muscle strength and walking for patients after anterior cruciate ligament reconstruction
Zhijiao FAN ; Lanqi JIN ; Zhibin HUANG ; Yige LI ; Sihan YAO ; Yubao MA
Chinese Journal of Rehabilitation Theory and Practice 2025;31(2):242-248
ObjectiveTo investigate the effect of neuromuscular electrical stimulation (NMES) on quadriceps muscle strength and walking for patients after anterior cruciate ligament reconstruction (ACLR). MethodsThirty-four patients after ACLR were selected at Beijing Rehabilitation Hospital of Capital Medical University from July, 2022 to October, 2023, and randomly divided into control group (n = 17) and experimental group (n = 17). Both groups received routine rehabilitation and functional training, and the experimental group received NMES during the functional training, while the control group received sham NMES, for eight weeks. Quadriceps peak torque-to-weight ratio, single-leg support phase and plantar impulses during walking were measured before and after intervention. ResultsTwo cases in the control group and three in the experimental group dropped down. Quadriceps peak torque-to-weight ratio improved in both groups after intervention (|t| > 17.578, P < 0.001), and improved more in the experimental group than in the control group (t = 4.714, P < 0.001); while the affected single-leg support phase and the affected/unaffected single-leg support phase ratio improved in both groups (|t| > 16.882, P < 0.001), and improved more in the experimental group than in the control group (t > 3.234, P < 0.01); and plantar impulses of all zones optimized in both groups (t > 9.221, P < 0.001), and were better in the experimental group than in the control group(|t| > 2.852, P < 0.01). ConclusionNMES may further improve quadriceps muscle strength, plantar pressure distribution during walking and single-leg support in patients after ACLR.
9.Analysis of Differential Compounds of Poria cocos Medicinal Materials by Integrated Qualitative Strategy Based on UPLC-Q-Orbitrap-MS
Jiayuan WANG ; Xiaohan FAN ; Xiaoxiao WEI ; Rong CAO ; Jin WANG ; Lei WANG ; Fengqing XU ; Shunwang HUANG ; Deling WU ; Hongsu ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):148-156
ObjectiveTo establish a rapid analytical method for identifying the differential components in Poria cocos medicinal materials based on ultra performance liquid chromatography-quadrupole-electrostatic field orbital trap high-resolution mass spectrometry(UPLC-Q-Orbitrap-MS), combined with mass defect filtering(MDF) and molecular network integration techniques. MethodsUPLC-Q-Orbitrap-MS was used for MS data acquisition and identification of P. cocos medicinal materials, with the help of MDF for the study of cleavage behavior and structural identification of triterpenoids. According to the similarity of MS/MS fragmentation patterns of each component, global natural product social molecular network(GNPS) was established, and Cytoscape 3.6.1 was used to screen molecular clusters with similar structures and the the structure of main compound classes were identified and confirmed. Multivariate statistical analyses such as principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to screen the differential components of the five P. cocos medicinal materials with the variable importance in the projection(VIP) value>1 and P<0.05 as the criteria. ResultsA total of 66 compounds were identified by database comparison, 8 compounds were newly identified by MDF, 28 compounds were newly identified by GNPS, and a total of 102 chemical compounds were identified, including 43 triterpenoids, 16 saccharides, 26 amino acids and peptides, 3 nucleosides, and 14 other compounds. Triterpenoids were predominant in Poriae Cutis and wild Fushen, amino acids and peptides were the most abundant in Poria and cultivated Fushen, carbohydrates were the most abundant in Poriae Cutis. Type Ⅰ and Ⅱ triterpenoids had higher amounts in Poria and cultivated Fushen, type Ⅲ triterpenoids were more abundant in Poriae Cutis, all four types of triterpenoids were higher in Fushenmu, and type Ⅰ, Ⅱ, and Ⅳ triterpenoids were higher in wild Fushen. A total of 12 common differential chemical constituents were screened, including serine, guanosine, gallic acid, 2-octenal, maltotriose, trametenolic acid, dehydroeburicoic acid, dehydrotrametenolic acid, poricoic acid A, poricoic acid B, poricoic acid E and G, but the relative contents of them varied significantly among different medicinal materials. ConclusionAmong the five P. cocos medicinal materials, the types of constituents are generally similar, but their relative contents differed significantly among these medicinal materials, especially in the distribution of triterpenoids. The integration of UPLC-Q-Orbitrap-MS, MDF and GNPS can provide a reference for the rapid qualitative analysis of other Chinese medicines.
10.Analysis of Differential Compounds of Poria cocos Medicinal Materials by Integrated Qualitative Strategy Based on UPLC-Q-Orbitrap-MS
Jiayuan WANG ; Xiaohan FAN ; Xiaoxiao WEI ; Rong CAO ; Jin WANG ; Lei WANG ; Fengqing XU ; Shunwang HUANG ; Deling WU ; Hongsu ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):148-156
ObjectiveTo establish a rapid analytical method for identifying the differential components in Poria cocos medicinal materials based on ultra performance liquid chromatography-quadrupole-electrostatic field orbital trap high-resolution mass spectrometry(UPLC-Q-Orbitrap-MS), combined with mass defect filtering(MDF) and molecular network integration techniques. MethodsUPLC-Q-Orbitrap-MS was used for MS data acquisition and identification of P. cocos medicinal materials, with the help of MDF for the study of cleavage behavior and structural identification of triterpenoids. According to the similarity of MS/MS fragmentation patterns of each component, global natural product social molecular network(GNPS) was established, and Cytoscape 3.6.1 was used to screen molecular clusters with similar structures and the the structure of main compound classes were identified and confirmed. Multivariate statistical analyses such as principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to screen the differential components of the five P. cocos medicinal materials with the variable importance in the projection(VIP) value>1 and P<0.05 as the criteria. ResultsA total of 66 compounds were identified by database comparison, 8 compounds were newly identified by MDF, 28 compounds were newly identified by GNPS, and a total of 102 chemical compounds were identified, including 43 triterpenoids, 16 saccharides, 26 amino acids and peptides, 3 nucleosides, and 14 other compounds. Triterpenoids were predominant in Poriae Cutis and wild Fushen, amino acids and peptides were the most abundant in Poria and cultivated Fushen, carbohydrates were the most abundant in Poriae Cutis. Type Ⅰ and Ⅱ triterpenoids had higher amounts in Poria and cultivated Fushen, type Ⅲ triterpenoids were more abundant in Poriae Cutis, all four types of triterpenoids were higher in Fushenmu, and type Ⅰ, Ⅱ, and Ⅳ triterpenoids were higher in wild Fushen. A total of 12 common differential chemical constituents were screened, including serine, guanosine, gallic acid, 2-octenal, maltotriose, trametenolic acid, dehydroeburicoic acid, dehydrotrametenolic acid, poricoic acid A, poricoic acid B, poricoic acid E and G, but the relative contents of them varied significantly among different medicinal materials. ConclusionAmong the five P. cocos medicinal materials, the types of constituents are generally similar, but their relative contents differed significantly among these medicinal materials, especially in the distribution of triterpenoids. The integration of UPLC-Q-Orbitrap-MS, MDF and GNPS can provide a reference for the rapid qualitative analysis of other Chinese medicines.

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