1.Treatment of Hyperuricemia and Gouty Arthritis by Buyang Huanwu Tongfeng Decoction via Inhibition of PPAR-γ/NF-κB/AGEs/RAGE Pathway Based on Network Pharmacology
Yue CAO ; Wanmei YAO ; Tao YANG ; Man YANG ; Ruimin JIA ; Rongrong LU ; Xue FENG ; Biwang LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):182-192
ObjectiveThis paper aims to investigate the potential molecular biological mechanism of Buyang Huanwu Tongfeng decoction in treating hyperuricemia and gouty arthritis by network pharmacology and molecular docking technology and preliminarily verify the mechanism through animal experiments. MethodsThe active ingredients and targets in the Buyang Huanwu Tongfeng decoction were obtained by the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and ETCM databases. The DisGeNET and GeneCards databases were utilized to acquire disease targets associated with hyperuricemia and gouty arthritis. These disease targets were then intersected with drug targets to identify key targets. The R language ClusterProfiler package and Python were employed for conducting gene ontology(GO) enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG) enrichment analysis. The regulatory network diagram of the drug-key target-function-pathway was visualized using Cytoscape 3.9.1 software, and the protein-protein interaction (PPI) network for key targets was depicted. Finally, the hub gene was determined through topological analysis. Auto Dock, PyMOL, and other software were used for molecular docking to explore the possible therapeutic mechanism of Buyang Huanwu Tongfeng decoction for hyperuricemia and gouty arthritis. In animal experiments, a composite rat model of hyperuricemia induced by intraperitoneal injection of oteracil potassium combined with gouty arthritis induced by the modified Coderre method was established. Through hematoxylin-eosin(HE) staining, uric acid test, enzyme linked immunosorbent assay(ELISA), Western blot, and real-time polymerase chain reaction(Real-time PCR), the molecular mechanism and key targets of Buyang Huanwu Tongfeng decoction for treating hyperuricemia and gouty arthritis were observed. ResultsAfter screening and removing duplicate values, 76 active ingredients and 15 key targets were finally obtained. GO enrichment analysis yielded that the treatment of hyperuricemia and gouty arthritis with Buyang Huanwu Tongfeng decoction was significantly associated with acute inflammatory response, astrocyte activation, regulation of interleukin (IL)-8 production, nuclear receptor activity, and binding of growth factor receptor. KEGG pathway enrichment analysis obtained that the key target genes were significantly associated with the IL-17 signaling pathway, advanced glycosylation end/receptor of advanced glycation endproducts(AGE/RAGE) signaling pathway, anti-inflammatory, and other pathways. PPI network indicated that albumin(ALB), peroxisome proliferator-activated receptor-γ (PPAR-γ), IL-6, IL-1β, and C-reactive protein(CRP) were the key protein targets. The molecular docking results showed that ALB had the strongest binding force with beta-carotene (β-carotene). Biochemical results showed that blood uric acid decreased in the Buyang Huanwu Tongfeng decoction groups. HE staining results showed that the low-dose (7.76 g·kg-1·d-1), medium-dose (15.53 g·kg-1·d-1), and high-dose (31.05 g·kg-1·d-1) groups of Buyang Huanwu Tongfeng decoction had different degrees of remission, and the remission of the high-dose group was the most obvious. Fibroblastic tissue hyperplasia in synovial joints accompanied with inflammatory cell infiltration, as well as inflammatory cell infiltration in renal tissue of the high-dose group was significantly reduced, followed by the medium-dose and low-dose groups, and the expression of ALB, PPAR-γ, IL-6, IL-1β, and CRP was down-regulated to different degrees. ConclusionBy regulating the targets such as ALB, PPAR-γ, IL-6, IL-1β, and CRP, inhibiting the PPAR-γ/nuclear transcription factor (NF)-κB pathway, and reducing AGEs/RAGE-mediated inflammation, Buyang Huanwu Tongfeng decoction exerts anti-inflammatory and analgesic effects and activates blood circulation and diuresis in the treatment of hyperuricemia and gouty arthritis.
2.Effect of Modified Shibaotang on Serum Sex Hormone Levels in Patients with Male Late-onset Hypogonadism of Kidney Essence Deficiency Syndrome Complicated with Diabetes Mellitus
Yi SHAN ; Shaokang CHEN ; Zhenfu SHI ; Haifeng XU ; Yi LU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):227-233
ObjectiveTo observe the effect of modified Shibaotang on serum sex hormone levels in patients with male late-onset hypogonadism of kidney essence deficiency syndrome complicated with diabetes mellitus. MethodsA total of 60 patients with male late-onset hypogonadism of kidney essence deficiency syndrome complicated with diabetes mellitus,who met the inclusion criteria and were admitted to Yinchuan Hospital of Traditional Chinese Medicine from October 2022 to October 2023,were selected and randomly divided into an observation group and a control group,with 30 patients in each group. Both groups continued their original treatments,including blood glucose lowering and blood lipid regulation. The observation group was treated with modified Shibaotang,while the control group was treated with testosterone undecanoate capsules. The treatment lasted for 12 weeks. The changes in traditional Chinese medicine (TCM)syndrome scores,partial androgen deficiency in aging males (PADAM)symptom scores,glucose metabolism indexes [fasting plasma glucose (FPG),2-hour postprandial glucose (2 h PG),glycosylated hemoglobin (HbA1c)],and serum sex hormone indexes [sex hormone-binding globulin (SHBG),free testosterone (FT),total testosterone (TT),prolactin (PRL),luteinizing hormone (LH),follicle stimulating hormone (FSH),estrogen (E2)] were compared between the two groups before and after treatment. Safety was also evaluated. Results(1)Clinical efficacy comparison:After treatment,the clinical efficacy in both groups was similar,and there was no statistically significant difference between the two groups. (2)TCM syndrome score and PADAM symptom score comparison:After treatment,both groups showed a significant reduction in TCM syndrome scores and PADAM symptom scores (P<0.01),and the observation group showed a significantly greater reduction compared to the control group (P<0.05). (3)Glucose metabolism indexes comparison:After treatment,the levels of FPG,2 h PG,and HbA1c were significantly reduced in both groups (P<0.01),and there was no statistically significant difference between the two groups regarding FPG,2 h PG,and HbA1c levels after treatment. (4)Serum sex hormone indexes comparison:After treatment,the levels of FT,TT,PRL,LH,and FSH were significantly increased in both groups (P<0.01),while E2 levels were significantly decreased (P<0.01). There was no statistically significant difference between the two groups in the levels of FT,TT,PRL,LH,FSH,and E2 after treatment. There was also no significant difference in SHBG levels within or between the groups before and after treatment. During the clinical observation,neither group exhibited any obvious adverse reactions. ConclusionModified Shibaotang can significantly improve the clinical symptoms of male late-onset hypogonadism of kidney essence deficiency syndrome complicated with diabetes mellitus,reduce blood glucose,and increase sex hormone levels. The mechanism may involve the inhibition of aromatase transformation in adipocytes,promotion of GnRH production,and regulation of the hypothalamic-pituitary-gonadal axis function.
3.Comparison of myopia progression before and after discontinuation of low-concentration atropine in children wearing orthokeratology lenses
Yaozeng WANG ; Pan LI ; Jin WANG ; Zikang LU
International Eye Science 2025;25(1):134-139
AIM: To compare the progression of myopia in children wearing orthokeratology lenses combined with low-concentration atropine before and after drug withdrawal, to determine the rebound effect of drug withdrawal in orthokeratology lens wearers, and to analyze its causes based on changes in pupil diameter.METHODS:A prospective case-control study was conducted to collect 80 children with myopia who were treated with orthokeratology lenses combined with 0.01% atropine ophthalmic gel at the Xi'an No.1 Hospital from January to June 2022. One year later, they were divided into a drug withdrawal group(Group A, 40 cases)and a continuous medication group(Group B, 40 cases)based on whether they stopped taking the medication. The progression of myopia before and after drug withdrawal was observed by analyzing changes in axial length(AL)and spherical equivalent(SE)in the group A within 1 a before and after drug withdrawal. The changes in AL, pupil diameter(PD), and SE were compared between the group A and group B within 2 a, and the correlation between PD and AL growth was analyzed.RESULTS:In the group A, the AL increased by 0.17±0.23 and 0.29±0.18 mm at 0.5 and 1 a after drug withdrawal, respectively, which were both greater than before drug withdrawal(t=5.318, 2.983, both P<0.001). There was no statistically significant difference in SE growth between the two time points(P>0.05). There were no statistically significant differences in AL and PD between the group A and group B at baseline, 0.5 and 1 a during combined medication(all P>0.05). At 1.5 a, the AL growth of the group A was greater than that of the group B(0.32±0.27 mm vs 0.26±0.20 mm, t=7.363, P<0.001), and the PD was smaller than that of the group B(3.60±1.25 mm vs 4.12±1.92 mm, t=-7.541, P<0.001). At 2 a, the AL growth of the group A was greater than that of the group B(0.44±0.18 mm vs 0.32±0.14 mm, t=5.709, P<0.001), and the PD was smaller than that of the group B(3.67±1.31 mm vs 4.02±1.67 mm, t=-4.281, P<0.001). Correlation analysis showed a negative correlation between PD and AL growth at 0.5 and 1 a follow-ups over 2 a(R2=-0.156, -0.190, both P<0.001).CONCLUSION: After stopping low-concentration atropine in children wearing orthokeratology lenses, AL increased more rapidly than before drug withdrawal, PD decreased, and SE changed little. Compared with continuous medication, discontinuation of medication led to faster progression of AL with little change in diopter, and the larger the PD during orthokeratology lens wear, the slower the progression of AL.
4.Quantitative analysis on microvasculature in the optic disc area of patients with unilateral branch retinal vein occlusion
Jia SUN ; Jian LIU ; Peng YAN ; Nan LU ; Zhiming SHAN ; Dongni YANG
International Eye Science 2025;25(1):152-156
AIM: To observe the changes of retinal nerve fiber layer(RNFL)thickness and radial peripheral capillary(RPC)density in patients with unilateral branch retinal vein occlusion(BRVO), and further analyze the correlation between RPC density and RNFL thickness.METHODS: Observational study. Totally 37 patients with unilateral BRVO diagnosed at the ophthalmology department of First Hospital of Qinhuangdao from October 2020 to January 2022 were selected, the 37 affected eyes were the unilateral BRVO group, and 37 fellow healthy eyes were the contralateral unaffected group, and 35 healthy individuals(35 right eyes were selected)without ocular diseases during the same period were selected as the normal control group. The best corrected visual acuity, intraocular pressure, anterior segment, fundus and optical coherence tomography angiography(OCTA)were examined in both eyes of all BRVO patients and healthy individuals. The central macular thickness(CMT), the RNFL thickness, and the optic disc-AV crossing distance(DAVD)were measured by built-in software of the OCTA equipment. The optimized U-net algorithm was used to eliminate the large blood vessels, and then the RPC density was calculated. The CMT, RNFL thickness and RPC density were compared among the three groups. And the correlations of the RPC density with the CMT, RNFL thickness, and the DAVD were investigated.RESULTS: Compared with the contralateral unaffected group and the normal control group, the CMT and the RNFL thickness were significantly thickened in the unilateral BRVO group(all P<0.05); there were no statistical differences in the CMT and the RNFL thickness between the contralateral unaffected group and the normal control group(all P>0.05). The RPC density in the unilateral BRVO group increased compared with the contralateral unaffected group and decreased compared with the normal control group, but there was no statistically difference(all P>0.05). However, the RPC density in the contralateral unaffected group decreased compared with the normal control group(P<0.05). The RPC density in the unilateral BRVO group was not correlated with the CMT(P=0.960), but positively correlated with the RNFL thickness(r=0.401, P=0.014)and negatively correlated with the DAVD(r=-0.339, P=0.040).CONCLUSION: The RNFL thickened significantly and the RPC density did not change significantly in the optic disc area of BRVO patients. The RPC density is positively correlated with the RNFL thickness, indicating that the RNFL thickness can be used as a monitoring indicator to analyze and study the damage degree of the RPC density.
5.Intense pulsed light combined with Yuyin Runmu formula fumigation and meibomian gland massage for the treatment of meibomian gland dysfunction-related dry eye
Maodan RAN ; Lu JIN ; Gang REN
International Eye Science 2025;25(1):157-161
AIM: To investigate the effect of intense pulsed light(IPL)combined with Yuyin Runmu formula fumigation and meibomian gland massage on the treatment of patients with meibomian gland dysfunction(MGD)-related dry eye.METHODS: Prospectively selected 198 cases(396 eyes)of MGD-related dry eye patients admitted to our hospital from November 2021 to November 2023, and they were randomly divided into 99 cases(198 eyes)in control group treated with fumigation of Yuyin Runmu formula and meibomian gland massage, and 99 cases(198 eyes)in observation group treated with combined IPL on the basis of the control group. The efficacy of the two groups was compared, as well as the changes in the levels of ocular indexes [tear film break-up time(BUT), Schirmer I test(SⅠt)], visual quality [objective scattering index(OSI), Strehl ratio(SR), and modulation transfer function(MTF)], lipid layer thickness(LLT)of the tear film, and changes in tear fluid levels of inflammatory factors [tumour necrosis factor-α(TNF-α)and transforming growth factor-beta 1(TGF-β1)].RESULTS: All the patients completely received the treatment and follow-up. The levels of BUT, SⅠt, SR, MTF, and LLT increased and the levels of OSI, TNF-α, and TGF-β1 decreased in the two groups at 2 mo after treatment(all P<0.001), and the observation group was more favourable(all P<0.001).CONCLUSION: IPL combined with Yuyin Runmu formula fumigation and meibomian gland massage is effective in treating MGD-related dry eye, improving patients' ocular parameters, visual quality, and LLT, and decreasing the levels of inflammatory factors in the tear fluid.
6.Wendantang Regulates Energy Metabolism in Treatment of Myocardial Ischemia via SIRT3/PGC-1α Pathway
Xinjun ZHANG ; Zhiqiang XIAO ; Jia LU ; Wenliang DUN ; Ning GU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):1-8
ObjectiveTo investigate the mechanism by which Wendantang regulates the silent information regulator 3 (SIRT3)/peroxisome proliferator-activated receptor-gamma coactivator-1α (PGC-1α) pathway to influence energy metabolism and thereby prevent and treat myocardial ischemia (MI) in a rat model of hyperlipidemia (HL). MethodsThirty SD rats were randomly assigned into five groups: control, model, low-dose (3.702 g·kg-1·d-1) Wendantang, high-dose (7.404 g·kg-1·d-1) Wendantang, and positive control (trimetazidine, 0.006 g·kg-1·d-1), with six rats in each group. The control group was fed normally, while the other groups were fed with a high-fat diet for six weeks for the modeling of HL. Subsequently, the drug intervention groups were administrated with corresponding drugs by gavage, and the control and model groups received an equivalent volume of normal saline for 14 days. One hour after the last gavage, the other groups except the control group were injected intraperitoneally with posterior pituitary hormone (30 U·kg-1) to induce MI. Electrocardiography (ECG) was employed to detect changes in the electrocardiogram. Hematoxylin-eosin staining was performed to observe cardiac pathological changes. Enzyme-linked immunosorbent assay was employed to measure the serum levels of cardiac troponin I(cTnI), myoglobin (MYO), and creatine kinase-MB (CK-MB). Colorimetry was used to determine the levels of total cholesterol (TC) and triglycerides (TG) in the serum and ATP, malondialdehyde (MDA), and superoxide dismutase (SOD) in the myocardial tissue. Western blot was employed to determine the protein levels of SIRT3, PGC-1α, adenosine monophosphate-activated protein kinase (AMPK), and phosphorylated AMPK (p-AMPK) in the myocardial tissue. Real-time PCR was employed to measure the mRNA levels of SIRT3, PGC-1α, and AMPKα in the myocardial tissue. ResultsCompared with the control group, the model group showed significant J-point deviation and elevation in the ECG image, increased heart rate, disarrangement of myocardial fibers with unclear boundaries, elevated levels of CK-MB, cTnI, MYO, TC, and TG (P<0.05, P<0.01), declined levels of SOD and ATP (P<0.01), down-regulated mRNA levels of SIRT3, PGC-1α, and AMPK (P<0.05), and down-regulated protein levels of SIRT3, PGC-1α, and p-AMPK (P<0.05). Compared with the model group, the low-dose and high-dose Wendantang groups and the trimetazidine group showed inhibited J-point deviation and elevation in the ECG image, slowed heart rate, reduced inflammatory cell infiltration, alleviated disarrangement of myocardial fibers, declined levels of CK-MB, cTnI, MYO, TC, and TG (P<0.05, P<0.01), elevated level of SOD (P<0.01), up-regulated mRNA levels of SIRT3, PGC-1α, and AMPK (P<0.05, P<0.01) and up-regulated protein levels of SIRT3, PGC-1α, and p-AMPK (P<0.05, P<0.01). ConclusionWendantang can effectively intervene in HL-associated MI in rats by reducing oxidative stress in myocardial cells, alleviating lipid metabolism disorders, and improving myocardial energy metabolism via the SIRT3/PGC-1α signaling pathway.
7.Effect of Yiqi Wenyang Huoxue Lishui Components on Cardiac Function and Mitochondrial Energy Metabolism in CHF Rats
Hui GAO ; Zeqi YANG ; Xin LIU ; Fan GAO ; Yangyang HAN ; Aiyangzi LU ; Xingchao LIU ; Qiuhong GUO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):27-36
ObjectiveTo investigate the effects of Yiqi Wenyang Huoxue Lishui components on the cardiac function and mitochondrial energy metabolism in the rat model of chronic heart failure (CHF) and explore the underlying mechanism. MethodsThe rat model of CHF was prepared by transverse aortic constriction (TAC). Eight of the 50 SD rats were randomly selected as the sham group, and the remaining 42 underwent TAC surgery. The 24 SD rats successfully modeled were randomized into model, trimetazidine (6.3 mg·kg-1), and Yiqi Wenyang Huoxue Lishui components (60 mg·kg-1 total saponins of Astragali Radix, 10 mg·kg-1 total phenolic acids of Salviae Miltiorrhizae Radix et Rhizoma, 190 mg·kg-1 aqueous extract of Lepidii Semen, and 100 mg·kg-1 cinnamaldehyde) groups. The rats were administrated with corresponding agents by gavage, and those in the sham and model groups were administrated with the same amount of normal saline at a dose of 10 mL·kg-1 for 8 weeks. Echocardiography was used to examine the cardiac function in rats. Enzyme-linked immunosorbent assay was employed to determine the serum levels of N-terminal pro-B-type natriuretic peptide (NT-ProBNP), hypersensitive troponin(cTnI), creatine kinase (CK), lactate dehydrogenase (LD), free fatty acids (FFA), superoxide dismutase (SOD), and malondialdehyde (MDA). The colorimetric assay was employed to measure the levels of adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP) in the myocardial tissue. The pathological changes in the myocardial tissue were observed by hematoxylin-eosin staining and Masson staining. The Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities in the myocardial tissue were determined by the colorimetric assay. The ultrastructural changes of myocardial mitochondria were observed by transmission electron microscopy. Western blot was employed to determine the protein levels of ATP synthase subunit delta (ATP5D), glucose transporter 4 (GLUT4), and carnitine palmitoyltransferase-1 (CPT-1). The mitochondrial complex assay kits were used to determine the activities of mitochondrial complexes Ⅰ, Ⅱ, Ⅲ, and Ⅳ. ResultsCompared with the sham group, the model group showed a loosening arrangement of cardiac fibers, fracture and necrosis of partial cardiac fibers, inflammatory cells in necrotic areas, massive blue fibrotic tissue in the myocardial interstitium, increased collagen fiber area and myocardial fibrosis, destroyed mitochondria, myofibril disarrangement, sparse myofilaments, and fractured and reduced cristae. In addition, the rats in the model group showed declined ejection fraction (EF) and fractional shortening (FS), risen left ventricular end-diastolic diameter (LVIDd), left ventricular end-systolic diameter (LVIDs), left ventricular end-diastolic posterior wall thickness (LVPWd), left ventricular end-systolic posterior wall thickness (LVPWs), left ventricular end-diastolic volume (LVVOLd), and left ventricular end-systolic volume (LVVOLs), elevated levels of NT-ProBNP, cTnI, CK, MDA, FFA, and LD, lowered level of SOD, down-regulated protein levels of GLUT4 and CPT-1, decreased activities of Na+-K+-ATPase, Ca2+-Mg2+-ATPase, and respiratory complexes Ⅰ-Ⅳ, and declined levels of ATP5D, ATP, ADP, and AMP (P<0.05, P<0.01). Compared with the model group, the Yiqi Wenyang Huoxue Lishui components and trimetazidine groups showed alleviated pathological damage of the mitochondria and mycardial tissue, risen EF and FS, declined LVIDd, LVIDs, LVPWd, LVPWs, LVVOLd, and LVVOLs, lowered levels of NT-ProBNP, cTnI, CK, MDA, FFA, and LD, elevated level of SOD, up-regulated protein levels of GLUT4 and CPT-1, increased activities of Na+-K+-ATPase, Ca2+-Mg2+-ATPase, and respiratory complexes Ⅰ-Ⅳ, and elevated levels of ATP5D, ATP, ADP, and AMP (P<0.05, P<0.01). ConclusionYiqi Wenyang Huoxue Lishui components can improve the cardiac function, reduce myocardial injury, regulate glucose and lipid metabolism, optimize the utilization of substrates, and alleviate the damage of mitochondrial structure and function, thus improving the energy metabolism of the myocardium in the rat model of CHF.
8.Compound Glycyrrhizin Tablets Ameliorate Liver Injury Induced by Tripterygium Glycosides Tablet by Regulating Cholesterol Metabolism
Xiaotong FU ; Chunyu CAO ; Chun LI ; Chenna LU ; Ting LIU ; Yifei YANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):46-55
ObjectiveTo investigate the mechanism of liver injury induced by tripterygium glycosides tablets (TG) and the molecular mechanism of compound glycyrrhizin tablets (CG) in alleviating the abnormalities of cholesterol metabolism caused by TG via cholesterol metabolism. MethodsAccording to the body weights, male Sprague-Dawley (SD) rats were randomly grouped as follows: control (pure water), low-dose TG (TG-L, 189.0 mg·kg-1·d-1), high-dose TG (TG-H, 472.5 mg·kg-1·d-1), TG-L+CG (189.0 mg·kg-1·d-1 TG + 20.25 mg·kg-1·d-1 CG), and TG-H+CG (472.5 mg·kg-1·d-1 TG + 20.25 mg·kg-1·d-1 CG), with 6 rats in each group. Rats were administrated with corresponding drugs once daily for 3 weeks. At the end of the last administration, the mRNA and protein levels of liver X receptor-alpha (LXR-α), low-density lipoprotein receptor (LDLR), adenosine triphosphate-binding cassette transporter A1 (ABCA1), adenosine triphosphate-binding cassette transporter G1 (ABCG1), 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR), cholesterol 7α-hydroxylase (CYP7A1), cholesterol 12α-hydroxylase (CYP8B1), and sterol 27-hydroxylase (CYP27A1) in the liver tissue were determined by Real-time PCR and Western blotting, respectively. The level of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoAR), a regulatory enzyme of cholesterol synthesis, was measured by enzyme-linked immunosorbent assay (ELISA). HepG2 cells were used to observe the effect of TG on the cell proliferation in vitro. Specifically, HepG2 cells were grouped as follows: Low-dose TG (TG-l, 15 mg·L-1), medium-dose TG (TG-m, 45 mg·L-1), high-dose TG (TG-h, 135 mg·L-1), fenofibrate (FB, 10 μmol·L-1), CG extract, TG-h+FB (135 mg·L-1 TG + 10 μmol·L-1 FB), TG-m+FB (45 mg·L-1 TG + 10 μmol·L-1 FB), TG-l+FB (15 mg·L-1 TG + 10 μmol·L-1 FB), TG-h+CG (135 mg·L-1 TG + 60 μmol·L-1 CG), TG-m+CG (45 mg·L-1 TG + 60 μmol·L-1 CG), and TG-l+CG (15 mg·L-1 TG + 60 μmol·L-1 CG). The mRNA and protein levels of LXR-α, ABCG1, LDLR, CYP7A1, CYP8B1, and CYP27A1 in HepG2 cells were determined by Real-time PCR and Western blotting, respectively. ResultsThe rat experiment showed that compared with the control group, the TG-H group showed down-regulated mRNA levels of CYP7A1, CYP8B1, and CYP27A1 in the liver tissue (P<0.05, P<0.01), which were up-regulated by the application of CG (P<0.05, P<0.01), and the TG-H+CG group showed up-regulated mRNA level of LDLR (P<0.01). Compared with the control group, the TG-L and TG-H groups showed down-regulated protein levels of LDLR, CYP7A1, and CYP8B1 in the liver tissue (P<0.05, P<0.01). In addition, the protein levels of ABCG1 and LXR-α were down-regulated in the TG-H and TG-L groups, respectively (P<0.05). Compared with TG alone, TG+CG up-regulated the protein levels of ABCG1 and LDLR (P<0.05, P<0.01), and the protein levels of CYP7A1 and CYP8B1 in the TG-H+CG group were up-regulated (P<0.05, P<0.01). The cell experiment showed that compared with the control group, the TG-h group presented up-regulated mRNA level of LXR-α (P<0.01), and the TG-m and TG-h groups showcased down-regulated mRNA levels of LDLR and CYP7A1 (P<0.01) and up-regulated mRNA level of CYP27A1 (P<0.01) in HepG2 cells. The combination of CG with TG restored the above changes (P<0.01). Western blotting results showed that compared with the control group, the TG-m and TG-h groups showed down-regulated protein levels of LXR-α, ABCG1, LDLR, CYP7A1, CYP8B1, and CYP27A1 in HepG2 cells (P<0.01). Compared with the TG-h group, the TG-h+CG group showed up-regulated protein level of LDLR (P<0.05). Compared with the TG-m group, the TG-m+CG group showcased up-regulated protein levels of LDLR, ABCG1, CYP7A1, and CYP27A1 (P<0.05, P<0.01). ConclusionThe administration of TG at 189.0, 472.5 mg·kg-1 for 3 weeks could modulate the signaling pathways associated with cholesterol efflux, endocytosis, and cholesterol biotransformation in hepatocytes, leading to the accumulation of cholesterol and subsequent liver injury in rats. CG could ameliorate the liver injury induced by lipid metabolism disorders caused by TG by up-regulating the expression of LXR-α, LDLR, ABCG1, CYP7A1, CYP8B1, and CYP27A1 to promote cholesterol biotransformation.
9.Effect of Wulao Qisun Prescription on Proliferation and Osteogenic Differentiation of AS Fibroblasts by Regulating Wnt/β-catenin Signaling Pathway
Juanjuan YANG ; Ping CHEN ; Haidong WANG ; Zhendong WANG ; Haolin LI ; Zhimin ZHANG ; Yuping YANG ; Weigang CHENG ; Jin SU ; Jingjing SONG ; Dongsheng LU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):67-73
ObjectiveTo investigate the effect and underlying mechanism of the Wulao Qisun prescription on pathological new bone formation in ankylosing spondylitis (AS). MethodsSynovial fibroblasts were isolated from the hip joints of AS patients and observed under a microscope to assess cell morphology. The cells were identified using immunofluorescence staining. The isolated AS fibroblasts were divided into blank group, low drug-containing serum group, medium drug-containing serum group, high drug-containing serum group, and positive drug group. After drug intervention, cell proliferation was measured using the cell counting kit-8 (CCK-8) assay to observe fibroblast growth and determine the optimal intervention time. Alkaline phosphatase (ALP) activity was measured using the alkaline phosphatase assay. Protein expression of osteocalcin (OCN), osteopontin (OPN), and runt-related transcription factor 2 (Runx2) was detected by Western blot. The mRNA expression levels of Wnt5a, β-catenin, and Dickkopf-1 (DKK-1) were measured by real-time quantitative polymerase chain reaction (Real-time PCR). ResultsCompared with the blank group, each drug-containing serum group of Wulao Qisun prescription and the positive drug group inhibited the proliferation of AS fibroblasts and reduced ALP expression (P<0.01). Compared with the blank group, the low drug-containing serum group of Wulao Qisun prescription downregulated β-catenin mRNA expression (P<0.05). The medium and high drug-containing serum groups and the positive drug group significantly downregulated Wnt5a and β-catenin mRNA expression (P<0.05, P<0.01), with the positive drug group showing the most pronounced effect (P<0.01). The high drug-containing serum group and the positive drug group significantly upregulated DKK-1 mRNA expression (P<0.01). Compared with the blank group, the low drug-containing serum group of Wulao Qisun prescription inhibited the expression of OPN and Runx2 proteins (P<0.05, P<0.01), while the medium and high drug-containing serum groups and the positive drug group inhibited the expression of OCN, OPN, and Runx2 proteins (P<0.05, P<0.01). ConclusionThe Wulao Qisun prescription can inhibit the proliferation and osteogenic differentiation of AS fibroblasts, thereby delaying the formation of pathological new bone in AS. The possible mechanism involves the regulation of Wnt/β-catenin-related gene expression, further inhibiting the transcription of downstream target genes.
10.Herbal Textual Research on Arcae Concha in Famous Classical Formulas
Yiqin ZHANG ; Yixue ZHUANG ; Yinan LU ; Yanning CHEN ; Yichong CHEN ; Shuiyu XU ; Zhilai ZHAN ; Chengzi YANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):208-218
In this paper, the name, origin, producing area, harvesting, processing and functional indications of Arcae Concha were systematically combed and verified by consulting the ancient and modern literature, in order to provide a basis for the development of famous classical formulas containing Arcae Concha. Arcae Concha was first recorded in the name of Han in Bencao Shiyi, but later, due to the influence of LI Shizhen's error of combining Han item with Kuiha in the Ming dynasty, there were aliases such as Kuilu and Fulao, and Yizong Bidu began to include Walengzi as its correct name and has been used ever since. The textual descriptions and illustrations of the medicinal materials of Arcae Concha contained in the materia medica of the past generations were consistent with the modern Arca inflata, A. subcrenata and A. granosa. In ancient times, there were medicinal records of two parts of shell and meat, but now the shell is used as medicine, and the meat is mostly edible. In ancient times, Zhejiang, Shandong, Guangdong and Guangxi were the main producing areas, and Zhejiang was the best. It is now believed that A. inflata is mostly distributed in the northern part of the Huanghai Sea, A. granosa is mostly distributed in the coastal areas south of Shandong Peninsula in China, and A. subcrenata is widely distributed in the coastal areas of China. Its quality is better in a complete, white, no residual meat and sand. In ancient times, there was no clear harvesting period, and the processing was mainly based on vinegar quenching after calcination or powdering of calcined shell, but now the harvesting period is autumn and winter. After harvesting, it is directly washed and crushed for raw use or processed by calcined method. The records of the medicinal materials in the past dynasties on the properties of Arcae Concha were mainly warm, sweet, salty and mild, and it is now believed that Arcae Concha is salty in taste and mild in nature. In ancient times, it was believed that Arcae Concha were mainly used for coldness in the heart and abdomen, coldness in the waist and spine, benefiting the five internal organs, strengthening the stomach. Nowadays, it is believed that Arcae Concha can eliminate phlegm and remove blood stasis, soften the hardness and dissipate the lumps, produce acid and relieve pain. It can be used in the treatment of stubborn phlegm, gall tumor, scrofula and other symptoms. In conclusion, it is suggested that for the famous classical formulas containing Arcae Concha, the corresponding methods should be selected according to the processing requirements of the drug in the formulas, while those without processing requirements can be determined according to the functional position of the products.

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