To investigate the protective effects and underlying mechanisms of Babao Dan (BBD) against doxorubicin (Dox)-induced cardiomyopathy (DIC). A zebrafish model of DIC alongside an in vitro model of cardiomyocyte injury induced by Dox was utilized to elucidate the protective effects of BBD against DIC. The zebrafish and cell models were divided into control, Dox, and BBD (low-, medium-, and high-dose) plus Dox treatment groups. Reactive oxygen species (ROS) and malondialdehyde (MDA) content were determined by commercial kits. Nrf2/GPX4 signaling activity was assessed by Western blot and RT-qPCR. A comprehensive network pharmacology analysis was conducted to explore the potential mechanisms of action of BBD on DIC. Compared with Dox group, BBD treatment significantly reduced ROS levels and decreased MDA content in zebrafish (P<0.01), indicating ameliorated oxidative stress, which contributed to a reduction in Dox-induced cardiac injury. Notably, BBD was found to inhibit cardiomyocyte ferroptosis by restoring mitochondrial function in vitro. Network pharmacology analysis identified 4 core network genes, including Nrf2, which is pivotal in linking oxidative stress with ferroptosis, along with 6 key active components of BBD. Molecular docking and dynamics simulations demonstrated that these active components exhibit a strong binding affinity with the identified core targets. BBD effectively activates the Nrf2/GPX4 signaling pathway, thereby inhibiting cardiomyocyte ferroptosis and mitigating DIC. These insights provide a theoretical foundation for the future exploration of BBD as a preventive and therapeutic agent against Dox-related cardiotoxicity.
To elucidate whether Danggui Sini Decoction (DGSD) relieves sciatica by downregulating transient receptor potential (TRP) channels involved in peripheral sensitization. After conducting chronic constriction injury of sciatic nerve (CCI) operation, 32 SD rats, randomly assigned to the sham, CCI 7d, CCI 14d, and CCI 21d groups (n=8 per group), were used to observe the course of sciatica. mRNA sequencing of dorsal root ganglia (DRG) was conducted on rats in the sham and CCI 21d groups. Another 32 rats were divided into the sham, CCI, mecobalamin, and DGSD groups, 8 in each group. mRNA sequencing analysis and single-cell RNA-sequencing (scRNA-seq) data analysis were conducted. Behavioural tests were conducted to evaluate the therapeutic effects of DGSD. The levels of TRP vanilloid (TRPV)1–4, TRP ankyrin 1 (TRPA1), TRP melastatin 8 (TRPM8), Ras homolog family member A (RhoA), and phosphorylated p38 mitogen-activated protein kinase (p-p38MAPK) were measured via Western blot, and the serum levels of C-C motif chemokine ligand 2 and 5 (CCL2, CCL5), substance P (SP), and prostaglandin E2 (PGE2) were conducted via ELISA. scRNA-seq and mRNA sequencing analysis revealed that inflammatory mediator regulation of TRP channels played a role in the development of sciatica. CCI rats exhibited cold hyperalgesia from day 4 to 21, increased the DRG protein expression levels of TRP channels, including TRPV1–4, TRPA1, TRPM8, RhoA, p-p38MAPK, CCL2, CCL5, SP, and PGE2. Treatment with DGSD decreased the serum levels of CCL2, CCL5, SP, and PGE2, and reduced the levels of RhoA, p-p38MAPK, TRPV1–4, TRPA1, and TRPM8 in DRG (P<0.05). CCI increased the levels of TRP channels and inflammatory factors, inducing cold hyperalgesia, whereas DGSD relieved pain via inhibiting peripheral sensitization to relieve cold hyperalgesia by reducing the inflammatory factors and TRP.
To determine whether gypenosides (GPs) induce preadipocyte browning via famesoid X receptor (FXR)/Rubicon-mediated autophagy inhibition and ameliorate obesity-related metabolic disorders in mice. In vitro, differentiated 3T3-L1 preadipocytes were treated with 1–6 µ g/mL GPs. Browning markers [uncoupling protein 1 (UCP1), peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1 α), PR domain containing 16 (Prdm16)], autophagy markers [microtubule-associated protein 1 light chain 3 form II/form I (LC3II/LC3I), Beclin-1, recombinant autophagy related protein 7 (ATG7), p62], and FXR/Rubicon expression were assessed by Western blot and qPCR. FXR knockdown was performed using shRNA lentivirus. Autophagic flux was evaluated using an mRFP-GFP-LC3 tandem reporter. In vivo, high-fat diet-induced obese mice were randomly divided into 5 groups using a random number sequence (n=6 per group): control, model, low- and high-dose GPs (300, 600 mg/kg per day by oral gavage, respectively), and sitagliptin (SIT, 30 mg/kg daily) groups. All treatments were administered daily for 12 weeks. Metabolic parameters, adipocyte histology, and protein expression in inguinal white adipose tissue were then analyzed. GPs (3–6 µ g/mL) significantly upregulated UCP1, PGC-1α, and Prdm16 protein levels and reduced the LC3II/LC3I ratio compared with the differentiation medium control (all P<0.01). FXR and Rubicon expression were increased by GPs (P<0.01). FXR knockdown abolished these effects (P<0.01). In obese mice, GPs (600 mg/kg) reduced body weight, serum triglycerides, total cholesterol, and fasting blood glucose, decreased adipocyte size, and increased UCP1, PGC-1α, Prdm16, FXR, and Rubicon while decreasing LC3II/LC3I, Beclin-1, and ATG7 compared with the model group (P<0.05). GPs activate FXR/Rubicon signal pathway to inhibit autophagy, inducing preadipocyte browning and offering a novel herbal-based anti-metabolic diseases strategy.
To evaluate the renoprotective effects and underlying molecular mechanisms of sodium danshensu (SDSS) in renal ischemia–reperfusion (RIR) induced acute kidney injury. Network pharmacology, Weighted Gene Co-expression Network Analysis, single-cell RNA sequencing and machine learning algorithms were integrated to systematically identify SDSS’s potential therapeutic targets in RIR. Molecular docking was also employed to confirm key target interactions. Target validation was performed both in vivo and in vitro. In vivo, a rat RIR model was established by occluding bilateral renal arteries, and SDSS (20 mg•kg−1•d−1) was administered intraperitoneally for 28 d. Renal injury was evaluated by HE and TTC stainings, while the expression of signal transducer and activator of transcription 3 (STAT3), phosphorylated STAT3 (p-STAT3), BCL2 associated X (BAX), and caspase 3 (CASP3) in kidney tissues was detected via immunofluorescence, Western blot, and qRT-PCR. In vitro, hypoxia/reoxygenation-treated HK-2 cells were used to assess the protective effect of SDSS (13 µg/mL), with cell viability and apoptosis measured by cell counting kit-8 and flow cytometry, respectively, and STAT3 phosphorylation levels examined by Western blot, further confirming the involvement of the STAT3 pathway. The results of network pharmacology and multiomics analysis identified STAT3, P-STAT3, RELA proto-oncogene (RELA), C-C motif chemokine ligand 2 (CCL2), tumor necrosis factor (TNF), BAX, and CASP3 as therapeutic targets involved in inflammation, apoptosis and immune regulation during RIR progression. More importantly, SDSS significantly reduced the infarction volume and preserved tissue integrity in vivo (P<0.01). RIR upregulated STAT3, BAX, and CASP3 expressions, which SDSS treatment effectively reversed both in vivo and in vitro (P<0.01). SDSS alleviates RIR injury by suppressing inflammatory and apoptotic signaling, primarily through downregulation of STAT3, BAX, and CASP3, providing new insights into its therapeutic mechanism and potential clinical application.
To investigate the therapeutic effects of Yinchenhao Decoction (YCHD) on a mouse model of autoimmune hepatitis (AIH) induced by cytochrome P450 2D6 adenovirus (Ad-CYP2D6), and reveal its potential molecular mechanisms through integrated transcriptomic and proteomic analyses. AIH model was established via tail vein injection of 100 µL Ad-CYP2D6 (1 × 109 PFU/mL) to C57BL/6 mice. Seven days post-injection, the mice were stratified by weight and randomly divided into 3 groups (n=6–8 per group), including the normal control, AIH model, and YCHD treatment groups. YCHD treatment group received daily oral gavage of YCHD at a dosage of 10 mL/kg for 2 consecutive weeks, while the normal and model groups received an equivalent volume of normal saline. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured and anti-liver kidney microsomal-1 (LKM-1) autoantibody was detected by ELISA. Liver histopathology was assessed using HE staining. RNA sequencing (RNA-seq) and tandem mass tag (TMT) proteomics were employed to analyze the differential gene and protein expressions in liver tissues, with validation via qPCR and Western blot. Compared with the normal control group, the model group showed significantly increased liver and spleen indices and markedly elevated serum ALT and AST levels, which were markedly reduced following YCHD treatment (P<0.05 or P<0.01). HE staining revealed prominent inflammatory cell infiltration and hepatocyte nuclear loss in the model group; these pathological alterations were significantly ameliorated in the YCHD treatment group. Serum level of specific autoantibody LKM-1 was increased in the model group and significantly reduced following YCHD treatment (P<0.05). Integrated transcriptomic and proteomic analyses demonstrated signal transducer and activator of transcription 1 (Stat1), guanylate-binding protein 3 (Gbp3), and T-cell-specific guanosine triphosphatase 2 (Tgtp2) were significantly upregulated in the model group and were significantly downregulated after YCHD treatment (P<0.05 or P<0.01). YCHD effectively alleviates liver injury in Ad-CYP2D6-induced AIH mice. Its underlying mechanism may be associated with the suppression of interferon-β cellular response pathway mediated by Stat1, Gbp3, and Tgtp2.
To investigate the specific mechanism by which Ruanjian Sanjie Formula (RJSJ) inhibits the occurrence and metastasis of lung cancer. This study developed an in vivo model using H460 cells in nude mice to investigate postoperative lung cancer metastasis. Two groups received either RJSJ (0.2 mL) or saline to assess RJSJ’s anti-cancer effects, including disease-free survival duration, recurrence-free metastasis rate, number of pulmonary metastases, as well as weight and coefficient of major organs. An in vitro study was conducted to evaluate the effects of RJSJ at concentrations ranging from 0 to 500 µg/mL on the proliferation, migration, and invasion capabilities of H460 and H1975 cell lines. These effects were assessed using the CCK-8 assay for cell proliferation, the wound healing assay for cell migration, and the Transwell assay for cell invasion. RJSJ’s chemical profile was analyzed via UPLC-Q-TOF-MS, and its mechanism was explored through network pharmacology. Specific microRNAs and their target genes were identified and analyzed using bioinformatics, qRT-PCR, and Western blot to examine RJSJ’s impact on metastasis-related genes and proteins during epithelial-mesenchymal transition (EMT). Virtual docking and molecular simulations were used to evaluate interactions between RJSJ compounds and microRNAs. RJSJ could inhibit the proliferation, migration, and invasion of lung cancer cells in vitro (P<0.05 or P<0.01), and it had also been shown to prevent lung cancer recurrence and metastasis in vivo (P<0.05); it contains 47 chemical compounds that act on 191 common targets, including 36 core targets. It reduced cancer cell proliferation and invasion, downregulated miR-182-5p, thereby regulating its target gene EPAS1, and regulated EMT-related markers by decreasing the expression of N-cadherin, MMP2, MMP9, Snail, and Twist, while increasing the expression of E-cadherin. Chlorogenic acid was identified as the main anti-tumor component due to its strong binding affinity to nucleic acids. RJSJ inhibits lung cancer occurrence and metastasis both in vivo and in vitro. Mechanistically, RJSJ suppresses the EMT process by downregulating miR-182-5p and inhibiting EPAS1 expression, thereby restraining lung cancer malignant progression, with chlorogenic acid identified as a key active component.
To determine whether radiation directly compromises AT2 cell stemness through oxidative and DNA damage, and to evaluate whether Compound Kushen Injection (CKI) protects AT2 cells, restores alveolar regeneration, and mitigates radiation-induced pulmonary fibrosis. Mice were randomly assigned to a control, irradiated model, or CKI-treated group. CKI (4 mL·kg−1·d−1) was administered intraperitoneally from 3 days before irradiation until the designated endpoints. A thoracic irradiation plus left pneumonectomy model was established to mimic clinical scenarios requiring postoperative lung regeneration. Lung function, histopathology, and compensatory growth of the accessory lobe were assessed. AT2 cells proliferation, differentiation, and transitional Krt8+ accumulation were evaluated using lineage tracing in Sftpc-CreER and Rosa26-ZsGreen mice. Oxidative stress and DNA damage were examined by immunofluorescence. Transcriptomic profiling of sorted lineage-labeled AT2 cells was conducted to identify radiation- and CKI-responsive genes. Radiation caused marked oxidative stress and γ H2AX-defined DNA damage in AT2 cells, accompanied by suppression of Sod1 and Parp1 and increased Il6 and Tgfb1 expression (P<0.01). These epithelial-intrinsic injuries led to reduced AT2 proliferation, impaired AT2-to-AT1 differentiation, and long-term accumulation of Krt8+ transitional cells. Functionally, irradiation significantly impaired compensatory lung regeneration after pneumonectomy and promoted fibrotic remodeling. CKI markedly alleviated ROS accumulation and DNA damage in AT2 cells, restored Sod1 and Parp1 expression, suppressed inflammatory and profibrotic signaling, rescued AT2 proliferative and differentiative capacities, reduced transitional Krt8+ cell accumulation, and improved both alveolar regeneration and fibrosis (P<0.05). CKI preserves epithelial integrity by reducing oxidative and DNA damage, restoring AT2 stemness, and enhancing alveolar repair. These findings provide mechanistic support for CKI as a potential epithelial-directed therapeutic strategy for preventing or treating radiation-induced lung fibrosis.
To explore the effectiveness and safety of Chinese medicine (CM) treatment based on pattern identification (PI) in elderly patients (⩾ 60 years) with severe community-acquired pneumonia (CAP). This post hoc analysis was based on a multicenter, randomized, placebo-controlled trial. Elderly patients with severe CAP were randomly assigned to receive either conventional medicine plus CM placebo (conventional group) or conventional medicine combined with CM treatment based on PI (combination group) for a 28-d treatment period. The primary outcome was the rate of treatment failure. Secondary outcomes included 28- and 90-d mortality, time to clinical stability, rate of clinical stability, length of hospital stay, and adverse events. A total of 128 elderly patients with severe CAP were included in the intention-to-treat population, comprising 67 patients in the combination group and 61 in the conventional group. The rate of treatment failure was lower in the combination group than in the conventional group (20.9
To investigate the mechanisms of Modified Shenqi Pill (MSQP) in the treatment of allergic rhinitis comorbid with asthma (ARCA). According to a random number table, 30 BALB/c mice were divided into a control group, an ARCA group, a MSQP low-dose group (MSQP-L, 24.6 g/kg), a MSQP high-dose group (MSQP-H, 49.2 g/kg), and a dexamethasone (DXMS) group (1 mg/kg), with 6 mice in each group. The ARCA mouse model was established using ovalbumin (OVA). MSQP and DXMS were administered by gavage for 10 d. The airway allergic symptom scores, serum OVA-specific immunoglobulin E (OVA-sIgE), histopathological changes in nasal and lung tissues were evaluated. Differential protein expression profiles in lung tissues were detected by proteomics. Functional enrichment analysis of differential proteins was performed using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. Activation of the JAK2/STAT1 signaling pathway was assessed by immunohistochemistry (IHC) and Western blot. Activation of B lymphocytes was detected by CD19 and MHC II immunofluorescence (IF) double staining. The proportions of Th1, Th2, and Th17 cells in lung and spleen tissues were analyzed by flow cytometry. Compared with the ARCA group, MSQP significantly decreased airway allergic symptom scores and serum OVA-sIgE levels, and reduced eosinophil infiltration and goblet cell hyperplasia in nasal and lung tissues (P<0.05 or P<0.01). GO and KEGG analyses revealed that differential proteins between MSQP and ARCA groups were primarily associated with immune regulation, involving signaling pathways including JAK/STAT, MHC II-mediated antigen presentation, Th1/Th2 differentiation, and Th17 differentiation. Western blot and IHC results showed that MSQP-H reduced the expressions of JAK2, p-JAK2, STAT1, p-STAT1 in nasal and lung tissues (P<0.01). IF results showed that MSQP-H reduced the numbers of CD19+ and MHC II+ positive cells, and CD19+MHC II+ double-positive cells in nasal and lung tissues (P<0.01). Flow cytometry results showed that MSQP-H changed the proportional relationship among Th1, Th2, and Th17 cells in lung and spleen tissues (P<0.05 or P<0.01). MSQP improved airway allergic symptoms and inflammatory responses in ARCA mice, possibly by inhibiting the activation of JAK2/STAT1 signaling pathway and B lymphocytes, and correcting imbalances in Th1/Th2 and Th17 differentiation.
To investigate the therapeutic mechanisms of Eucommia ulmoides Oliv. (EC) in a collagen-induced arthritis (CIA) rat model. The CIA model was established by immunizing rats with type II collagen. Wistar rats were assigned to the normal group (n=6), and the remaining model rats were further divided by body weight into the CIA model, EC, and Iguratimod (IGU) positive control groups (n=6 per group). EC (4 g/kg per day) and IGU (4.5 mg/kg daily) was administered orally for 4 weeks. Arthritis severity was evaluated by measuring hind paw volume. Histopathological analysis of ankle joints was performed using hematoxylin and eosin (HE) and safranin O/fast green. Serum levels of procollagen type I N-terminal propeptide (PINP), cross-linked carboxy-terminal telopeptide of type I collagen (ICTP), osteocalcin (BGP), β-crosslaps (β-CTX), tumor necrosis factor (TNF-α), interleukin (IL)-1β, and CTNNB1 (β-catenin) were measured by ELISA. Serum nitric oxide (NO) levels were measured using a nitrate reductase assay. Transcriptomic sequencing and RT-qPCR were performed to assess mRNA expression, and Western blot was used to assess protein expression of Wnt2 and β-catenin in the spleen. Compared with the CIA group, EC significantly alleviated arthritis severity and paw swelling (P<0.05) and reduced synovial inflammation and cartilage damage. Serum analysis showed that EC decreased TNF-α, IL-1β, NO, ICTP, and β-CTX, while increasing PINP and BGP (P<0.05 or P<0.01), and also reduced CTNNB1 (P<0.01) compared with the CIA group. In the spleen, EC reduced mRNA expressions of IL-1β, IL-6, and TNF-α (P<0.05 or P<0.01), upregulated IL-10, and decreased β-catenin protein (P<0.05); Wnt2 mRNA showed a non-significant downward trend compared with the CIA group. In the ankle joints, EC significantly reduced mRNA expressions of Wnt2 and β-catenin (P<0.05 or P<0.01). EC may alleviate inflammation and bone destruction in RA by regulating the Wnt/β-catenin signaling pathway, suggesting its potential as a therapeutic agent for RA.
To investigate whether hyperoside (HYP), the main active ingredients of Wuzi Yanzong Pill, can reduce oxidative stress (OS) damage and treat Parkinson’s disease (PD) by activating the nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway. MPTP-induced PD mouse model was established and animals were divided into 5 groups by random number table method: control group, MPTP group (15, 20, and 30 mg/kg on days 1, 2 and 3–7, respectively), and MPTP+HYP low-, medium-, and high-dose groups (25, 50, and 100 mg·kg−1·d−1, respectively). Behavioral tests were conducted and protein expression levels of tyrosine hydroxylase (TH), Nrf2, HO-1, B-cell lymphoma 2 (Bcl-2), and Bcl-2-associated X protein (Bax) were analyzed using Western blot. Glutathione (GSH), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT) levels were measured by ELISA. Molecular docking and dynamics simulations were performed on HYP and Nrf2, and neuronal apoptosis was detected by TUNEL assay. Multivariate statistical analysis was used to investigate the correlation between HYP’s therapeutic effect on PD and OS via the Nrf2/HO-1 pathway. In the SH-SY5Y cell model (MPP+ and ML385), cell viability and toxicity were assessed with cell counting kit-8 and lactate dehydrogenase assays, and protein expression levels and OS indicatars were also measured. In animal experiments, HYP intervention improved motor abilities, increased TH, Nrf2, HO-1, and Bcl-2 expressions, and reduced Bax expression compared to the MPTP group (P<0.05 or P<0.01). It also elevated GSH, GSH-Px, SOD, and CAT levels, decreased MDA, and reduced neuronal apoptosis (P<0.05 or P<0.01). Molecular docking and dynamics showed HYP binds effectively to Nrf2. Multivariate analysis revealed a strong correlation between HYP’s therapeutic effect on PD and OS via the Nrf2/HO-1 pathway. In cell experiments, ML385 inhibited Nrf2/HO-1 activation by HYP (P<0.05 or P<0.01), diminishing its effect on OS and apoptosis. HYP can reduce OS and apoptosis by activating the Nrf2/HO-1 pathway, thereby exerting a therapeutic effect in PD.
OBJECTIVE:To elucidate the therapeutic potential and mechanisms of Fuzi (Aconiti Lateralis Radix Praeparata) against intrahepatic cholangiocarcinoma (ICC). METHODS:Fuzi-related targets were obtained from TCMSP, HERB, and ETCM databases, and ICC-related differentially expressed genes were retrieved from Gene Expression Omnibus. Common hub targets were identified using network pharmacology and machine learning, followed by functional enrichment and immune infiltration analyses. Effects of aconitine, a major alkaloid of Fuzi, were validated in RBE cells using qPCR, Western blot, Transwell assays, and flow cytometry. RESULTS:Thirty-six common genes were identified, mainly enriched in metabolic and cancer-related pathways. Topoisomerase II alpha (TOP2A) and alpha1A-adrenergic receptor (ADRA1A) were identified as hub genes with high diagnostic value (area under curve>0.98). High TOP2A expression was linked to poor disease-free survival. Several immune cells, including regulatory T cells, showed altered infiltration in ICC and correlated with hub gene expression. Aconitine inhibited the proliferation, migration, and invasion of RBE cells in a dose- and time-dependent manner (P<0.05 or P<0.01). It also induced G1 phase arrest and apoptosis, accompanied by modulation of Bcl-2 and Bax expression, and downregulation of TOP2A. TOP2A knockdown similarly reduced migration and invasion and altered Bcl-2/Bax expression, but had limited effects on apoptosis rate or cell cycle distribution. CONCLUSION:TOP2A was identified as Fuzi's potential therapeutic target in ICC, where its compound aconitine exerts antitumor effects by downregulating TOP2A, providing novel mechanistic insights and targeted therapy potential.
To evaluate the efficacy and safety of acupuncture combined with estazolam for hemodialysis patients with insomnia. This prospective, randomized controlled trial was conducted from August 15, 2022 to February 22, 2024. By the random number table method, 110 end-stage renal-disease undergoing hemodialysis patients with insomnia were randomly divided into 2 groups to receive 1 mg/d of estazolam (control group, 55 cases) or acupuncture combined with 1 mg/d of estazolam (intervention group, 55 cases) for 4 weeks, and followed up for another 4 weeks. All patients received their standard treatments and common medications. The Pittsburgh Sleep Quality Index (PSQI), Hamilton Anxiety Scale (HAMA), and Chinese Medicine Syndrome Score of Insomnia (CMSSI) were the primary outcomes measured at baseline and at 2, 4, and 8 weeks, as well as clinical effectiveness before and after treatment; the second outcomes were the sleep stucture parameters assessed before and after treatment. Adverse effects were also assessed. At 2, 4, and 8 weeks post-treatment, the scores of PSQI, HAMA, and CMSSI in the intervention group were lower than those in the control group (all P<0.01). At 4 weeks, the intervention group also had a significantly higher clinical effective rate than that in the control group (81.82
To evaluate the long-term efficacy of Chinese herbal medicine (CHM) in enhancing reproductive outcomes and reducing recurrence rates among endometriosis-associated infertility women infertile women. This study is a retrospective analysis of a randomized, double-blind, placebo-controlled clinical trial implemented from May 2014 to April 2018. The study included patients with endometriosis-associated infertility who had undergone standardized laparoscopic surgery. After surgery, the participants were randomly assigned to 2 groups: the intervention group received a CHM treatment regimen, while the control group was given placebo therapy. Both groups were subjected to the same blinding procedures and received treatment for a period of 6 months. Five years after surgery, the researchers conducted follow-up with the participants and analysed the data on pregnancy outcomes and recurrence of endometriosis. In this trial, a total of 202 patients with endometriosis-associated infertility (101 per group) were followed up at 5 years post-surgery for clinical pregnancy, live birth, and recurrence outcomes (101 patients per group). According to our data, pregnancies occurred in 83 (82.18
To evaluate the therapeutic potential of Chinese medicine prescription Gancao Ganjiang Decoction (Licorice and Dried Ginger Decoction, LDGD) against renal cell carcinoma (RCC) and to elucidate its underlying mechanisms. A mouse RCC model was established in BALB/c mice by subcutaneous injection of Renca cells. The effects of LDGD (5, 10, and 15 g/kg), cisplatin (3 mg/kg), and their combination (10 g/kg LDGD + 3 mg/kg cisplatin) on tumor growth were assessed. Ki-67 expression in tumor tissue was detected by immunohistochemistry. The anti-proliferative effects of LDGD and its 7 blood-absorbed constituents were evaluated on 769-P and 786-O human RCC cell lines using MTT and colony formation assays. Network pharmacology was conducted to predict core targets and signaling pathways. Molecular docking was performed to forecast interactions between active components of LDGD and key proteins in related signaling pathways. Western blot was used to validate phosphorylation levels of the key proteins. LDGD and cisplatin inhibited mouse RCC growth in vivo. The combination of LDGD with cisplatin enhanced the tumor inhibition rate as compared with cisplatin alone. Ki-67 expression was reduced following treatments. LDGD and its blood-absorbed constituents inhibited RCC cell viability and proliferation in a time- and dose-dependent manner in vitro. Network pharmacology showed that PI3K-AKT and JAK-STAT pathways implicated in the anti-RCC effect of LDGD and its blood-absorbed constituents, and AKT1 and STAT3 were potential targets. Molecular docking suggested that isoliquiritigenin and 6-shogaol had strong binding affinity for AKT1 and STAT3. Western blot showed that LDGD suppressed phosphorylation of AKT and STAT3 in 769-P cells, while inhibited phosphorylation of STAT3 protein in 786-O cells. Isoliquiritigenin inhibited phosphorylation of AKT and STAT3 proteins in both 769-P and 786-O cells, and 6-shogaol inhibited phosphorylation of STAT3 protein in 786-O cells. LDGD exerts significant anti-tumor effects against RCC, possibly via suppression of AKT and STAT3 phosphorylation.
