1.Exploring Therapeutic Mechanism of Xihuang Wan for Hyperplasia of Mammary Glands Based on Network Pharmacology, Molecular Docking, and Cell Experiments
Junliang WANG ; Peihua LIANG ; Xueli MA ; Juanxia SUN ; Tao HAN ; Yongmei LAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):41-49
ObjectiveTo study the therapeutic mechanism of Xihuang Wan for hyperplasia of mammary glands based on network pharmacology, molecular docking, and cell experiments. MethodsThe active ingredients and targets of Xihuang Wan were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and A Bioinformatics Annotation daTabase for Molecular mechANism of Traditional Chinese Medicine (BATMAN-TCM) and supplemented by searching against PubChem and Swiss Target Prediction. The targets of differential metabolites in tissues and urine were obtained from previous metabolomics studies through PubChem and Swiss Target Prediction. GeneCards, Online Mendelian Inheritance in Man (OMIM), PharmGKB, Therapeutic Target Database (TTD), Drunbank were searched for the targets of hyperplasia of mammary glands. After the common targets were obtained via Veeny2.1.0, the STRING database was used to analyze the protein-protein interactions, and Cytoscape was used for the core target analysis and visualization. Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) were employed for enrichment analysis. Molecular docking was carried out in Autodock, and cell experiments were conducted to verify the prediction results. In the cell experiments, estradiol and progesterone (E2+P) were used to intervene in human mammary epithelial/MCF-10A cells, and thus the MCF-10A cell proliferation model was established. The cells were then treated with Xihuang Wan-medicated serum. The cell counting kit-8 (CCK-8) was used to measure the cell proliferation, and flow cytometry was used to detect apoptosis. The mRNA and protein levels of key factors in MCF-10A cells were determined by real-time PCR and Western blot, respectively. ResultsThe results of network pharmacology showed that 90 active ingredients and 316 common targets were obtained, from which 20 core targets and 38 corresponding active ingredients were screened out. The results of GO and KEGG enrichment analyses showed that Xihuang Wan exerted effect against hyperplasia of mammary glands by regulating a variety of biological processes, which may be related to protein kinase B (Akt)-related molecular functions, estrogen signaling pathway, prolactin signaling pathway and other biological processes. The results of molecular docking showed that estrogen receptor 1 (ESR1), serine/threonine kinase 1 (Akt1), non-receptor tyrosine kinase (SRC), and signal transducer and activator of transcription 3 (STAT3) all had strong binding activity with the nine active ingredients, suggesting that Xihuang Wan exert the effect through the ESR1/SRC/phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway and the Janus kinase (JAK)/STAT3 signaling pathway. The results of cell experiments showed that E2+P intervention in MCF-10A cells promoted the proliferation of MCF-10A cells (P<0.05), while the Xihuang Wan-medicated serum inhibited the proliferation of MCF-10A cells exposed to E2+P (P<0.05). Flow cytometry showed that the Xihuang Wan-medicated serum promoted the apoptosis of MCF-10A cells exposed to E2+P (P<0.01). The results of Real-time PCR showed that the Xihuang Wan-medicated serum down-regulated the mRNA levels of PI3K, Akt, JAK2, and STAT3 in MCF-10A cells treated with E2+P (P<0.01). The results of Western blot showed that the Xihuang Wan-medicated serum inhibited the expression of p-PI3K/PI3K, p-Akt/Akt, p-JAK2/JAK2, and p-STAT3/STAT3 in MCF-10A cells treated with E2+P (P<0.05). ConclusionXihuang Wan may exert the effect against hyperplasia of mammary glands by inhibiting the proliferation and promoting the apoptosis of MCF-10A cells, which may related to the inhibition of the activation of PI3K/Akt and JAK2/STAT3 signaling pathways.
2.Mechanisms of Jiangtang No. 3 Prescription in Alleviating Adipose Tissue Insulin Resistance in Diabetic Rats via TLR4/NF-κB/NLRP3 Signaling Pathway-mediated Inflammation
Tongxun WANG ; Lantian LIU ; Runqi LI ; Haoxiang LI ; Yi ZHAO ; Tian TIAN ; Rufeng MA ; Sihua GAO ; Dandan ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):109-117
ObjectiveTo observe the effects of Jiangtang No. 3 prescription on inflammatory pathways and insulin resistance-related indicators in rats with type 2 diabetes mellitus (T2DM), and to elucidate its molecular mechanism in combating diabetes. MethodsA T2DM rat model was established using a high-fat diet combined with intraperitoneal injection of streptozotocin (STZ). Successfully modeled rats were randomly assigned to the model group, metformin group, and low-, medium-, and high-dose Jiangtang No. 3 prescription groups, and a normal group was also set. Daily gavage was administered for 8 weeks as follows: metformin at 0.1 g·kg-1·d-1, Jiangtang No. 3 prescription granules at 1.62, 3.24, 6.48 g·kg-1·d-1 for the respective dose groups, and sterile water for the normal and model groups. Rat body weight, fasting blood glucose (FBG), oral glucose tolerance test (OGTT), and insulin tolerance test (ITT) were measured. After drug intervention, enzyme-linked immunosorbent assay (ELISA) was used to determine serum levels of total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL), high-density lipoprotein cholesterol (HDL), non-esterified fatty acids (NEFA), interleukin (IL)-1β, IL-18, and insulin (INS). Hematoxylin-eosin (HE) staining was used to observe morphological changes in adipose tissue. Real-time quantitative PCR was used to detect the mRNA expression of Toll-like receptor 4 (TLR4), nuclear factor-κB (NF-κB), NOD-like receptor protein 3 (NLRP3), Caspase-1, IL-1β, IL-18, and gasdermin D (GSDMD) in adipose tissue. Western blot was used to measure the corresponding protein expression levels. ResultsCompared with the model group, Jiangtang No. 3 prescription groups exhibited significantly increased body weight (P<0.05, P<0.01), significantly reduced FBG (P<0.05, P<0.01), significant reductions in TC, TG, NEFA, and LDL (P<0.05, P<0.01), and a significant increase in HDL (P<0.01). Serum levels of inflammatory mediators IL-1β and IL-18 were significantly decreased (P<0.01), the homeostatic model assessment of insulin resistance (HOMA-IR) index was significantly reduced (P<0.05, P<0.01), and adipose tissue pathology was improved. The protein expression levels of TLR4, NF-κB, NLRP3, Caspase-1, IL-1β, IL-18, and GSDMD were markedly decreased (P<0.05, P<0.01), and the mRNA expression levels of these indicators were also significantly downregulated (P<0.05, P<0.01). Some effects were superior to those of the positive control drug metformin, and certain indicators exhibited dose-dependent improvements. ConclusionT2DM rats display significant inflammatory responses, disordered glucose and lipid metabolism, and insulin resistance. Jiangtang No. 3 prescription effectively suppresses inflammatory mediators, improves glucose and lipid metabolism and insulin resistance, and ameliorates pathological changes in adipose tissue. Its mechanism may be related to the regulation of the TLR4/NF-κB/NLRP3 signaling pathway in visceral adipose tissue, thereby influencing downstream inflammatory mediators.
3.Ancient and Modern Documentation of Classic Formula Sangjuyin
Xiaofang WANG ; Lyuyuan LIANG ; Jialei CAO ; Ziming XU ; Wangju ZHOU ; Yiping WANG ; Yujie CHANG ; Ruiting SU ; Yihan LI ; Jingwen LI ; Bingqi WEI ; Bingxiang MA ; Wenli SHI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):200-207
Sangjuyin, as a pungent and cooling agent with precise therapeutic effect, is a classic pungent formula for cooling relief of the epidermis, which is highly respected by medical practitioners. This formula is from the Wenbing Tiaobian written by WU Jutong in the Qing dynasty, on the basis of which subsequent medical practitioners have made additions and subtractions to apply it. The authors used the bibliometric method to systematically organize the medical books from the Qing dynasty and the Republic of China and modern literature to analyze the composition, concoction, decoction, efficacy, and previous and modern application of Sangjuyin. After examination, the drug base of this formula is basically clear. Armeniacae Semen Amarum is the dried mature seeds of Armeniaca vulgaris, family Rosaceae. Forsythiae Fructus is the dried fruit of Forsythia suspensa, family Mulleinaceae. Menthae Haplocalycis Herba is the dried above-ground part of Mentha haplocalyx, family Labiatae. Mori Folium is the dried leaves of Morus alba, family Moraceae. Chrysanthemi Flos is the dried head of Chrysanthemum morifolium, family Asteraceae. Platycodonis Radix is the dried root of Eryngium grandiflorum, family Eryngium. Glycyrrhizae Radix et Rhizoma is the dried root and rhizome of Glycyrrhiza uralensis of the Leguminosae family, and Phragmitis Rhizoma is the fresh or dried rhizome of Phragmites communis of the Gramineae family. It is recommended that the eight drugs be used in raw form as medicine. The dosage and method of decoction were converted into a modern single dosage of 7.46 g Armeniacae Semen Amarum, 5.60 g Forsythiae Fructus, 2.98 g Menthae Haplocalycis Herba, 9.33 g Mori Folium, 3.73 g Chrysanthemi Flos, 7.46 g Platycodonis Radix, 2.98 g Glycyrrhizae Radix et Rhizoma, and 11.19 g Phragmitis Rhizoma, with 400 mL water added, and the solution was boiled to obtain 200 mL, taken twice a day. Sangjuyin has the efficacy of dispersing wind and clearing heat, promoting lung and relieving cough, and it is used for treating the initial onset of wind-warmth and the evidence of evil spirits in the lungs and collaterals. Modern research has shown that Sangjuyin is often used in the treatment of cough, pneumonia, rhinitis, and other respiratory diseases, and the results of this study provide a reference for the later development of Sangjuyin.
4.Effects of total flavonoids from Carthamus tinctorius L. on hepatic stellate cell activation based on miRNA-204/NUAK1/Hippo signaling axis
Mingqi LI ; Xiaolu ZHAO ; Chenlu ZHANG ; Yinghe WANG ; Yuehong MA
China Pharmacy 2026;37(3):311-316
OBJECTIVE To investigate the effects of total flavonoids from Carthamus tinctorius L. (TFCTL) on hepatic stellate cell (HSC) activation based on the microRNA (miRNA)-204/NUAK family SNF1-like kinase 1 (NUAK1)/Hippo signaling axis, thereby elucidating the potential mechanism underlying their antifibrotic effects. METHODS The HSC-T6 cells were divided into control group, model group, TFCTL low-concentration group (20 μg/mL), TFCTL medium-concentration group (40 μg/mL), and TFCTL high-concentration group (60 μg/mL). Except for control group, the remaining groups were treated with 5 ng/mL of transforming growth factor-β to induce the activation of hepatic stellate cells, followed by the addition of corresponding drug solutions/culture medium and incubation for 24 hours. Cell apoptosis was assessed, the expression levels of α-smooth muscle actin (α-SMA), type Ⅰ collagen (Collagen Ⅰ) and proteins associated with the Hippo/Yes-associated protein (YAP) pathway [YAP, large tumor suppressor kinase 1 (LATS1), and mammalian STE20-like kinase 1 (MST1)] were detected. Additionally, cell transfection was used to investigate the activity of the miRNA-204/NUAK1/Hippo signaling axis at both the genetic and protein levels. RESULTS After intervention with TFCTL, the apoptosis rate of HSC-T6 cells and the protein expressions of MST1 (except for the TFCTL high-concentration group) and LATS1 were significantly increased (P<0.05), while the protein expressions of α-SMA, CollagenⅠ, and YAP (except for the TFCTL medium-concentration group) were significantly decreased (P<0.05). Further results from cell transfection experiments revealed that after transfection with miRNA-204 mimics, the mRNA it’s protein expressions of α-SMA, CollagenⅠ, NUAK1, and YAP in HSC-T6 cells were significantly decreased (P<0.05), while the mRNA and protein expressions of LATS1 and the mRNA expression of MST1 were significantly increased (P<0.05). Conversely, the results were opposite following transfection with miRNA-204 inhibitors. CONCLUSIONS TFCTL can exert anti-hepatic fibrosis effects by up-regulating the expression of miRNA-204, thereby down- regulating the expressions of NUAK1, inactivating the Hippo/YAP pathway, which in turn suppresses the activation of HSC and promotes their apoptosis.
5.Construction of an Evaluation System for Traditional Chinese Medicine Appropriate Technologies Based on Evidence-Based Medicine
Jixing WANG ; Zhiyi ZHOU ; Sisi LIN ; Dieyu MA ; Zhizhen ZHOU ; Conghua JI
Journal of Traditional Chinese Medicine 2026;67(4):355-359
Based on the theory of evidence-based medicine (EBM), this paper systematically constructed a multi-dimensional evaluation framework for traditional Chinese medicine (TCM) appropriate technologies, encompassing three core dimensions including evidence, practitioner, and patient. For the current practical challenges in the promotion of TCM technologies such as lack of high-quality evidence, inconsistent operational standards, and varying patient acceptance, the paper proposed the integration of published literature evidence and real-world research data to construct a scientific and applicable evaluation pathway. Regarding the evidence dimension, it emphasizes syste-matic assessment of effectiveness, safety, and economic efficiency, introducing methods like the target trial emulation framework to enhance evidence quality; for the practitioner dimension, it suggests developing multi-aspects competency evaluation tools based on educational background, training assessment, and clinical outcomes; for the patient dimension, it recommends designing patient acceptance assessment tools by considering factors such as technical characte-ristics, expected efficacy, patient-practitioner interaction, and the availability of alternative treatments. The purpose of the above measures is to provide methodological support for the standardized popularization and precision application of TCM appropriate technologies.
6.Impacts of wearing orthokeratology lenses on ocular surface and meibomian gland of children and adolescents in high-altitude areas
Duqin WANG ; Xiaomei MA ; Xueying LIU
International Eye Science 2026;26(3):489-492
AIM: To explore the impacts of wearing orthokeratology lenses on the ocular surface and meibomian gland of children and adolescents in high-altitude areas.METHODS: Retrospective study. Myopic children and adolescents who visited at the ophthalmology department and received orthokeratology lens from June 2023 to December 2023 were selected as orthokeratology group, and those who visited and wore regular spectacle group during the same period were selected as regular spectacle group. Then the ocular surface disease index(OSDI)score, tear meniscus height, non-invasive breakup time(NIBUT)[first NIBUT(fNIBUT), average NIBUT(avNIBUT)], and meibomian gland score were compared between the two groups before wearing lenses, and at 1, 3, 6, and 12 mo after wearing lenses.RESULTS: A total of 80 children(160 eyes)with myopia were included in this study. Among them, 40 cases(80 eyes)were in the orthokeratology lens group, with 25 males and 15 females and an average age of 11.33±2.76 years old, and 40 cases(80 eyes)were in the regular spectacle group, with 23 males and 17 females and an average age of 11.58±2.94 years old. The OSDI scores and tear meniscus heights showed no clear difference between two groups at various time points before and after wearing glasses(all P>0.05). The fNIBUT and avNIBUT of the orthokeratology lens group decreased compared with the regular spectacle group at 6 and 12 mo after wearing lenses, and meibomian gland score of the orthokeratology group was higher than that of the regular spectacle group at 12 mo after wearing lenses(all P<0.001). Among them, the fNIBUT and avNIBUT of the orthokeratology lenses group at 3, 6, and 12 mo after wearing glasses were lower than those before wearing glasses(all P<0.05). Moreover, the meibomian gland scores of the orthokeratology lenses group at 12 mo after wearing glasses were higher than those before wearing glasses, and 1, 3, and 6 mo after wearing glasses(all P<0.05), while there was no statistical difference in fNIBUT, avNIBUT, and meibomian gland score at each time point in the regular spectacle group(all P>0.05).CONCLUSION: Long term wearing of orthokeratology lenses can shorten the NIBUT of myopic children and adolescents in high-altitude areas, and have a certain impact on their meibomian gland function.
7.Influence of pterygium thickness and area on corneal refractive status
Xiaodong CHENG ; Jie WANG ; Song GAO ; Yanhong LU ; Yanbo MA ; Xinming CUI ; Xihui CHEN
International Eye Science 2026;26(1):152-156
AIM: To investigate the influence of pterygium thickness and area on corneal refractive status.METHODS: Prospective longitudinal study. A total of 60 cases(60 eyes)of pterygium patients admitted to our hospital from January 2024 to September 2024 were randomly selected. All patients underwent pterygium excision combined with pedicle conjunctival flap transplantation for treatment. Optical coherence tomography(OCT)was used to measure the preoperative thickness of patient's pterygium, and a digital slit lamp microscope was used to measure the area of pterygium. The corneal refractive status(degree of corneal astigmatism and average curvature)and changes in uncorrected visual acuity of patients before surgery, 1 d, 1, and 3 mo after surgery were compared. The relationship between preoperative thickness and area of pterygium in patients and corneal refractive status indicators at different postoperative time points were analyzed, and Logistic regression was used to analyze the impact of pterygium thickness and area on postoperative visual improvement in patients.RESULTS: All patients completed follow-up after surgery for 3 mo. At 3 mo after surgery, visual acuity improved in 21 eyes(35%). The results of bivariate Pearson correlation analysis showed that the thickness and area of pterygium positively correlated with the degree of corneal astigmatism and uncorrected visual acuity before surgery and 1 d, 1, and 3 mo after surgery(all P<0.05), and negatively correlated with the average corneal curvature before surgery and 1 d, 1, and 3 mo after surgery(all P<0.05). Logistic regression analysis showed that the thickness and area of pterygium before surgery, high degree of corneal astigmatism, and low uncorrected visual acuity(large LogMAR value)were all risk factors for poor postoperative visual improvement in patients(OR>1, P<0.05). The large average corneal curvature before surgery was a protective factor for poor postoperative visual improvement in patients(OR<1, P<0.05).CONCLUSION: The increase in thickness and area of pterygium can, to some extent, improve corneal astigmatism, reduce the average curvature of the cornea, and affect postoperative visual recovery.
8.Three-dimensional displacement and risk factors of midshaft clavicle fractures treated with titanium elastic intramedullary nailing
Junwei ZHANG ; Lingling CHEN ; Zhenyuan MA ; Weizhi NIE ; Chaohui LI ; Haitao WANG ; Laibao DUAN ; Jinyong HOU ; Hongzheng BI
Chinese Journal of Tissue Engineering Research 2026;30(2):269-277
BACKGROUND:Titanium elastic intramedullary nailing for the treatment of significantly displaced midshaft clavicle fractures has the characteristics of minimally invasive and elastic fixation.The displacement of the fracture is closely related to the later function.However,there are few studies on the three-dimensional displacement analysis of the fracture ends before surgery and after intramedullary fixation such as titanium elastic intramedullary nailing.OBJECTIVE:To explore the three-dimensional displacement of fracture ends after midshaft clavicle fracture and fixation with titanium elastic intramedullary nails,and to analyze the risk factors.METHODS:A total of 91 patients with midshaft clavicle fracture(fracture end shortening ≥15 mm)admitted to Wendeng Orthopedic Hospital of Shandong Province from April 2019 to April 2024 were selected,including 57 males and 34 females,aged(51.73±10.21)years old.All patients received closed reduction and internal fixation with titanium elastic intramedullary nail.CT scans of the affected clavicle were performed before and on the first day after surgery.The CT data were imported into Mimics software for modeling.The length of the clavicle,lateral displacement of the fracture end,and rotation of the distal end of the fracture along the X,Y,and Z axes were measured and recorded before and after surgery.Pearson correlation coefficient was used for correlation analysis of various parameters,and generalized linear regression was used to evaluate risk factors.RESULTS AND CONCLUSION:(1)Preoperatively,the variable that increased the risk of lateral displacement was the number of comminuted bone fragments,the variable that increased the risk of shortening displacement was male patients,and the variable that increased the risk of Z-axis rotation was the left limb.Shortening displacement was significantly positively correlated with lateral displacement(r=0.715,P<0.001);shortening displacement was significantly positively correlated with X-axis rotation displacement and Y-axis rotation displacement(r=0.265,P=0.028;r=0.303,P=0.011);lateral displacement was significantly positively correlated with Y-axis rotation and Z-axis rotation(r=0.258,P=0.032;r=0.250,P=0.038);X-axis rotation was significantly positively correlated with Y-axis rotation(r=0.382,P=0.001),and Z-axis rotation was significantly positively correlated with Y-axis rotation(r=0.280,P=0.020).(2)Postoperatively:The number of scapula fractures and comminuted bone fragments were variables that increased the risk of postoperative shortening and lateral displacement:Preoperative X-,Y-,and Z-axis rotation displacements were risk variables that increased postoperative X-,Y-,and Z-axis rotation displacements,respectively.Postoperative lateral displacement was significantly positively correlated with postoperative shortening and displacement(r=0.584,P=0.000),and postoperative lateral displacement was also significantly positively correlated with postoperative Y axis rotation and Z axis rotation(r=0.360,P=0.002;r=0.250,P=0.038).Postoperative Y axis rotation was significantly positively correlated with postoperative Z axis rotation(r=0.248,P=0.040).(3)The results showed that the three-dimensional displacement of the clavicle end before and after surgery was affected by many factors,especially the number of comminuted bone fragments,scapula fractures,gender,and original rotation displacement.At the same time,there were complex correlations between various displacements,especially the correlation between shortening displacement and lateral displacement was the strongest.
9.Expression and significance of tumor necrosis factor alpha,nuclear factor kappaB and ionized calcium binding adaptor molecule-1 in the hippocampus of mice with aortic dissection
Hong MA ; Xueling DING ; Qi WANG ; Hui LYU ; Albusm ASYA ; Xinyi CHENG ; Xiang MA
Chinese Journal of Tissue Engineering Research 2026;30(4):858-863
BACKGROUND:Hippocampal injury caused by aortic coarctation has been poorly studied,and combined detection of tumor necrosis factor α,nuclear factor κB and ionized calcium binding adaptor molecule-1 expression in aortic dissection has not been reported.OBJECTIVE:To observe histomorphologic changes in the hippocampus of a mouse model of aortic dissection and investigate the expression and significance of tumor necrosis factor alpha,nuclear factor kappaB and ionized calcium binding adaptor molecule-1 in the hippocampus of aortic dissection mice.METHODS:Sixteen healthy 3-week-old male C57BL/6 mice were randomly divided into two groups:control group and aortic dissection group,with eight mice in each group.In the aortic dissection group,mice were given β-aminopropionitrile monofumarate as drinking water for 4 weeks,and the angiotensin Ⅱ microinfiltration pump was then implanted to establish an animal model of aortic dissection.Mice in the control group were given normal diet and water.After the model was established,the maximum diameter of the ascending aorta was measured,hematoxylin-eosin staining and EVG staining were performed to evaluate the model formation rate,and the levels of inflammatory factors tumor necrosis factor α and interleukin 6 in serum were detected by enzyme-linked immunosorbent assay.The hippocampus was dissected and stained with hematoxylin-eosin to observe the pathological changes of the hippocampus in brain sections.The protein expression of tumor necrosis factor α,nuclear factor κB and ionized calcium binding adaptor molecule-1 was detected by western blot analysis.RESULTS AND CONCLUSION:(1)Compared with the control group,the maximum diameter of the ascending aorta in the aortic dissection group was significantly enlarged.(2)Hematoxylin-eosin staining of the aorta showed obvious thickening of the middle aorta and destruction and disorder of the aortic wall structure in mice.Neurons in the CA1 and CA3 regions of mice were sparsely arranged,reduced in size,and showed pyknosis with deeply stained nuclei.(3)Serum levels of inflammatory factors tumor necrosis factor α and interleukin 6 were increased in the aortic dissection group compared with the control group(P<0.01).(4)The expression levels of tumor necrosis factor α,nuclear factor κB,phosphorylated nuclear factor κB,and ionized calcium binding adaptor molecule-1 in the hippocampus were increased in the aortic dissection group compared with the control group(P<0.05).To conclude,microglial activation and increased expression of tumor necrosis factor α and nuclear factor κB may be involved in hippocampal neuron injury in aortic dissection mice.
10.Yinchenhao Tang Regulates Pyroptosis to Intervene in Cholestatic Liver Injury
Linlin WANG ; Zhengwang ZHU ; Jinghan ZHAO ; Ruixue MA ; Bing WANG ; Pingsheng ZHU ; Mingsan MIAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(1):55-62
ObjectiveTo explore the mechanism by which Yinchenhao Tang intervenes in α-naphthylisothiocyanate (ANIT)-induced cholestatic liver injury by regulating the Takeda G-protein-coupled receptor 5(TGR5)/NOD-like receptor protein 3(NLRP3)/cysteine aspartate-specific protease-1 (Caspase-1) pyroptosis signaling pathway. MethodsForty male Wistar rats were randomly assigned into blank, model, ursodeoxycholic acid, and Yinchenhao Tang groups. Except the blank group, other groups were treated with ANIT dissolved in olive oil for the modeling of cholestatic liver injury. Ursodeoxycholic acid (0.1 g·kg-1) and Yinchenhao Tang (9.23 g·kg-1) were administered by gavage. The blank group and the model group were administrated with the same amount of pure water, once a day for 3 days. The blood and liver tissue samples were collected, and the serum levels of liver function indicators were measured by an automatic biochemical analyzer. Hematoxylin-eosin staining was employed to observe the pathological changes of the liver. The levels of interleukin (IL)-1β and IL-18 in the liver tissue were determined by ELISA. The mRNA levels of IL-1β, IL-18, TGR5, NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), Caspase-1, and GSDMD in the liver tissue were assessed by Real-time PCR. The protein levels of TGR5, NLRP3, ASC, Caspase-1, and GSDMD in the liver tissue were determined by Western blot. ResultsCompared with the blank group, the model group showed elevated levels of alanine amino-transferase (ALT), aspartate transferase (AST), alkaline phosphatase (ALP), total bile acid (TBA), and total bilirubin (TBil) in the serum (P<0.01), inflammatory cell infiltration, hepatocyte swelling, and bile duct epithelial cell proliferation in the liver, raised levels of IL-1β and IL-18 in the liver tissue (P<0.01), down-regulated mRNA and protein levels of TGR5 (P<0.01), up-regulated mRNA levels of IL-18 (P<0.01), ASC (P<0.01), Caspase-1 (P<0.01), GSDMD (P<0.01), IL-1β (P<0.05), and NLRP3 (P<0.05), and up-regulated protein levels of NLRP3 (P<0.01), ASC (P<0.01), Caspase-1 (P<0.01), and GSDMD (P<0.05). Compared with the model group, the ursodeoxycholic acid group showed declined levels of AST (P<0.01), TBA (P<0.01), TBil (P<0.01), and ALT (P<0.05) in the serum, lowered levels of IL-1β and IL-18 in the liver tissue (P<0.01), down-regulated mRNA levels of NLRP3 (P<0.01), Caspase-1 (P<0.01), GSDMD (P<0.01), IL-1β (P<0.05), IL-18 (P<0.05), and ASC (P<0.05), up-regulated mRNA and protein levels of TGR5 (P<0.05), and down-regulated protein levels of NLRP3, ASC, Caspase-1, and GSDMD (P<0.05). Compared with the model group, the Yinchenhao Tang group showed lowered levels of ALT, AST, ALP, TBA, and TBil in the serum (P<0.01), declined levels of IL-1β and IL-18 in the liver tissue (P<0.01), down-regulated mRNA levels of IL-1β (P<0.01), NLRP3 (P<0.01), ASC (P<0.01), Caspase-1 (P<0.01), GSDMD (P<0.01), and IL-18 (P<0.05), up-regulated mRNA and protein levels of TGR5 (P<0.01), and down-regulated protein levels of Caspase-1 and GSDMD (P<0.05). The liver tissue of the administration groups showed reduced infiltration of inflammatory cells, reduced swelling of hepatocytes, and alleviated proliferation of bile duct epithelial cells. ConclusionYinchenhao Tang can ameliorate ANIT-induced cholestatic liver injury by regulating the hepatocyte pyroptosis mediated by the TGR5/NLRP3/Caspase-1 signaling pathway.

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