1.Trends in incidence and mortality of prostate cancer in Ningbo City from 2011 to 2023
ZHANG Qun ; WANG Yong ; CHEN Jieping ; BAO Kaifang ; FENG Yueyi ; Wang Xiaoli
Journal of Preventive Medicine 2025;37(1):46-50
Objective:
To analyze the trends in incidence and mortality of prostate cancer in Ningbo City, Zhejiang Province, so as to provide insights into the prevention and control of prostate cancer.
Methods:
Data of the incidence and mortality of prostate cancer in Ningbo City from 2011 to 2023 were collected through Ningbo Chronic Disease Collaborative Management System. The incidence and mortality of prostate cancer were calculated and standardized by the data from the sixth national population census in 2010 and the Segi's world standard population in 1960. The trends in incidence and mortality of prostate cancer were evaluated using average annual percent change (AAPC).
Results:
A total of 15 411 cases of prostate cancer were reported in Ningbo City from 2011 to 2023, and the crude incidence, Chinese-standardized incidence and world-standardized incidence were 39.62/105, 22.18/105 and 16.49/105, respectively, showing upward trends (AAPC=14.782%, 10.390% and 10.608%, all P<0.05). The Chinese-standardized incidence of prostate cancer was higher in urban areas than in rural areas, and both showed upward trends (25.14/105 vs. 19.44/105; AAPC=9.057% and 14.272%, both P<0.05). The crude incidence of prostate cancer in the groups aged 50-<60 years, 60-<70 years, 70-<80 years and ≥80 years showed upward trends (AAPC=11.657%, 14.031%, 10.734% and 5.300%, all P<0.05). A total of 3 739 deaths were reported, and the crude mortality, Chinese-standardized mortality and world-standardized mortality were 9.66/105, 5.23/105 and 3.71/105, respectively, showing upward trends (AAPC=8.458%, 3.620% and 3.602%, all P<0.05). The Chinese-standardized mortality of prostate cancer was higher in urban areas than in rural areas, and both showed upward trends (5.35/105 vs. 5.13/105; AAPC=3.183% and 3.962%, both P<0.05). The crude mortality of prostate cancer the groups ageds ≥80 years showed an upward trend (AAPC=7.482%, P<0.05).
Conclusions
From 2011 to 2023, the incidence and mortality of prostate cancer in Ningbo City showed upward trends. Special attention should be paid to urban residents, and prostate cancer screening should be strengthened among males aged 50 years and older.
2.Modified Ditan Tang Regulates Biorhythm-related Genes in Rat Model of Non-alcoholic Fatty Liver Disease
Zhiwen PANG ; Yu LIU ; Nan SONG ; Jie WANG ; Jingxuan ZHU ; Zhen HUA ; Yupeng PEI ; Qun WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):115-124
ObjectiveTo investigate the effects of modified Ditan tang on genes related to the transcription-translation feedback loop (TTFL) of biorhythm in the rat model of non-alcoholic fatty liver disease (NAFLD) and its mechanism for prevention and treatment of NAFLD. MethodsSixty-five healthy SPF male SD rats were randomly assigned into blank (n=20), model (n=15), and low-, medium-, and high-dose (2.68, 5.36, and 10.72 g·kg-1·d-1, respectively) modified Ditan tang (n=10) groups. Other groups except the blank group were fed a high-fat diet for 12 weeks. The modified Ditan tang groups were treated with the decoction at corresponding doses by gavage, and the blank and model groups were treated with an equal volume of normal saline from the 9th week for 4 weeks. The levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) in the serum were measured by an automatic biochemical analyzer. TG and non-esterified fatty acid (NEFA) assay kits were used to measure the levels of TG and NEFA in the liver. The pathological changes in the hypothalamus and liver were observed by hematoxylin-eosin staining, and the lipid deposition in the liver was observed by oil red O staining. The levels of brain-muscle ARNT-like protein 1 (BMAL1/ARNTL) in the hypothalamus and liver were determined by immunohistochemical staining. The mRNA and protein levels of BMAL1, circadian locomotor output cycles kaput (CLOCK), period circadian clock 2 (PER2), and cryptochrome1 (Cry1) in the hypothalamus and liver were determined by Real-time PCR and Western blot, respectively. ResultsCompared with the blank group, the model group showed elevated levels of TG, TC, LDL-C, AST, and ALT (P<0.01) and a lowered level of HDL-C (P<0.05) in the serum, elevated levels of TG and NEFA in the liver (P<0.01), pyknosis and deep staining of hypothalamic neuron cells, and a large number of vacuoles in the brain area. In addition, the model group showed lipid deposition in the liver, up-regulated mRNA and protein levels of CLOCK and BMAL1 (P<0.01), and down-regulated mRNA and protein levels of Cry1 and PER2 (P<0.01) in the hypothalamus and liver. Compared with the model group, all the three modified Ditan tang groups showed lowered levels of TG, TC, LDL-C, ALT, and AST (P<0.05, P<0.01) and an elevated level of HDL-C (P<0.05) in the serum, and lowered levels of TG and NEFA (P<0.05, P<0.01) in the liver. Furthermore, the three groups showed alleviated pyknosis and deep staining of hypothalamic neuron cells, reduced lipid deposition in the liver, down-regulated mRNA and protein levels of CLOCK and BMAL1 (P<0.05, P<0.01), and up-regulated mRNA and protein levels of Cry1 and PER2 (P<0.05, P<0.01) in the hypothalamus and liver. ConclusionModified Ditan tang can reduce lipid deposition in the liver and regulate the expression of CLOCK, BMAL1, Cry1, and PER2 in the TTFL of NAFLD rats.
3.Prevalence of latent tuberculosis infection among freshmen in middle schools and their willingness of preventive treatment in Yunyan District of Guiyang
PENG Xiaowei, ZHANG Jiangping, LUO Peng, CHEN Siyin, XU Mengqun, WANG Qun
Chinese Journal of School Health 2025;46(1):129-133
Objective:
To investigate the status of latent tuberculosis infection(LTBI)among freshmen in middle schools and their willingness of preventive treatment in Yunyan District of Guiyang, so as to provide a reference for the prevention and control of tuberculosis in schools.
Methods:
The tuberculin skin test (TST) and X-ray scans were used to screen the TB infection of 13 915 freshmen in middle schools in Yunyan District of Guiyang in 2023, and a questionnaire survey on the willingness of accepting tuberculosis preventive treatment was conducted to LTBI patients. The χ 2 test and Fisher exact probability was conducted for the comparison of the rates among the groups.
Results:
Among the freshmen screened, the detection rate of LTBI was 3.29%. There were statistically significant difference in LTBI rates among freshmen of different genders (boys:2.87%, girls:3.81%), age groups (12-15 years old:3.31%, 16-17 years old:3.92%, 18-20 years old:1.91%), and school stages (junior high school:3.52%, ordinary high school:5.96%, vocational high school:2.29%)( χ 2=9.59, 13.08, 54.30, P <0.01). A total of 356 LTBI freshmen completed questionnaire survey,and 299(83.99%) were willing to accept tuberculosis preventive treatment, but the actual number of LIBI freshman who underwent preventive treatment was zero. Those LTBI who had received Bacille Calmette Guérin(BCG) vaccine(86.97%) was higher in the reporting rates of being willing to accept preventive treatment than that of LTBI who had not received BCG vaccine( 75.79 %),the differences were statistically significant( χ 2=6.48, P <0.05). The main reasons for refusing preventive treatment was worry about adverse drug reactions(80.70%), social acceptance and the support of social institutions were needed most(85.96%).
Conclusions
The LTBI rate among freshmen in Yunyan District of GuiYang is higher. Although the freshmen with LTBI have a higher willingness to accept preventive treatment, however, no one has undergone preventive treatment. Corresponding measures need to be taken for improving the preventive treatment rate of LIBI freshmen.
4.Comprehensive Application of AHP-CRITIC Hybrid Weighting Method, Grey Correlation Analysis and BP-ANN in Optimization of Extraction Process of Qizhi Prescription
Qun LAN ; Yi CHENG ; Zian LI ; Bingyu WU ; Jinyu WANG ; Dewen LIU ; Yan TONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(8):176-186
ObjectiveBased on analytic hierarchy process(AHP)-criteria importance through intercriteria correlation(CRITIC) hybrid weighting method, grey relational analysis and backpropagation artificial neural network(BP-ANN), to optimize the water extraction process of Qizhi prescription, so as to provide an experimental basis for optimization of the preparation process of this prescription and the establishment of quality standards. MethodsL9(34) orthogonal test was employed, and the AHP-CRITIC hybrid weighting method was utilized to determine the weight coefficients of the quality fractions of various components, including astragaloside Ⅳ, polygalaxanthone Ⅲ, calycosin-7-O-β-D-glucoside, tenuifolin, and 3,6′-disinapoylsucrose, as well as the dry extract yield. The comprehensive score of each factor level combination in the orthogonal test were calculated as evaluation indicator to select the optimal extraction process parameters. The effects of extraction times, extraction time, and solvent dosage on the aqueous extraction process of the formula were investigated through intuitive analysis, variance analysis, and grey relational analysis. Meanwhile, a BP-ANN model was established to reverse-predict the optimal extraction process parameters of Qizhi prescription, and the optimized process parameters were validated. ResultsThe weight coefficients of the five index components(astragaloside Ⅳ, tenuifolin, calycosin-7-O-β-D-glucoside, polygalaxanthone Ⅲ, and 3,6′-disinapoylsucrose) and dry extract yield were 25.7%, 20.82%, 16.41%, 12.45%, 15.96% and 8.67%, respectively. The optimized extraction process parameters were extracted 3 times with 8, 6, 6 times the amount of water, each time for 1 h. The network prediction results of BP-ANN test samples were consistent with the orthogonal test results, and the mean square error(MSE) of the predicted and measured values of the network was <1%. The water extraction process of Qizhi prescription analyzed and predicted by relevant mathematical models was stable and feasible, which could effectively improve the extraction efficiency of the active ingredients of Astragali Radix and Polygalae Radix, and the average comprehensive score of the validation test was 90.85 with the relative standard deviation(RSD) of 1.55%. ConclusionThis study establishes a water extraction process for compound Qizhi granules, and the optimized extraction process can effectively improve the extraction efficiency of active ingredients, which provides useful references for the optimization of preparation process and the establishment of quality standards for other clinical experience formulas.
5.Effect of Yiqi Yangyin Jiedu Huayu prescription on intestinal metabolites in liver cirrhosis with minimal hepatic encephalopathy: A study based on metabolomics
Tingting JIANG ; Qun ZHANG ; Xianbo WANG ; Yuyong JIANG ; Xiaoli LIU ; Hao YU ; Zhiyun YANG
Journal of Clinical Hepatology 2025;41(3):469-477
ObjectiveTo investigate the effect of Yiqi Yangyin Jiedu Huayu prescription on minimal hepatic encephalopathy in liver cirrhosis based on intestinal metabolomics. MethodsA total of 11 patients with liver cirrhosis who were hospitalized in Beijing Ditan Hospital, Capital Medical University, from March to May 2024, and were diagnosed with MHE based on psychometric hepatic encephalopathy score were enrolled as subjects, and 11 healthy family members of the patients were enrolled as control group. Fecal samples were collected for metabolomics analysis from the control group and the patients with MHE before and after treatment with Yiqi Yangyin Jiedu Huayu prescription, and a population cohort study was conducted to investigate the effect of Yiqi Yangyin Jiedu Huayu prescription on intestinal metabolism of patients with MHE. The Fisher’s exact test was used for categorical data between two groups; the independent samples t-test was used for comparison of normally distributed continuous data between two groups, the paired t-test was used for comparision before and after treatment within the same group, and the Wilcoxon rank-sum test was used for comparison of non-normally distributed continuous data between two groups. ResultsA total of 29 differentially expressed metabolites were detected between the MHE group and the control group, mainly amino acids, organic acids, organic amines, carbohydrates, fatty acids, and vitamins, and there were 12 upregulated metabolites and 17 downregulated metabolites in the MHE group, which were mainly enriched in the metabolic pathways of ornithine, branched-chain amino acid, and aromatic amino acid. After the treatment with Yiqi Yangyin Jiedu Huayu prescription, 80 differentially expressed metabolites were detected in the patients with MHE, mainly carbohydrates, organic acids, and amino acids, and there were 56 upregulated metabolites and 24 downregulated metabolites, which were mainly enriched in the metabolic pathways of ornithine, branched-chain amino acid, and aromatic amino acid. ConclusionYiqi Yangyin Jiedu Huayu prescription can exert a therapeutic effect on patients with MHE by regulating intestinal metabolism.
6.Effect of intestinal flora and metabolites on the development and progression of acute-on-chronic liver failure
Ke SHI ; Qun ZHANG ; Xianbo WANG ; Ying FENG
Journal of Clinical Hepatology 2025;41(3):568-573
Acute-on-chronic liver failure (ACLF) is a dangerous disease with severe conditions, rapid progression, and high short-term mortality. Intestinal flora and metabolites are closely associated with the development, progression, and pathogenesis of ACLF. During the development of ACLF, the destruction of intestinal integrity and the dysregulation of intestinal microecology and its metabolites mediate immune disturbance and thus aggravate systemic inflammatory response. This article elaborates on the role of intestinal flora and metabolites in the development and progression of ACLF and related therapeutic strategies.
7.Exploring Molecular Mechanism of Gypenoside L against Ovarian Cancer Based on Ferroptosis Pathway Mediated by Mature-tRNA-Asp-GTC/ATF3-LPCAT3
Jingxuan ZHU ; Jiao ZHAO ; Qun WANG ; Xiaofei SUN ; Jiaxin WANG ; Hongda ZHANG ; Nan SONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):107-117
ObjectiveTo investigate the role of mature-tRNA-Asp-GTC and pre-tRNA-Arg-TCT in the ferroptosis phenotype of ovarian cancer (OC) cells and the regulatory mechanism of gypenoside L (Gyp-L) on mature-tRNA-Asp-GTC and pre-tRNA-Arg-TCT in OC cells. MethodsThe proliferation of human ovarian adenocarcinoma OVCAR3 cells was detected by cell counting kit-8 (CCK-8) assay, and the half-maximal inhibitory concentration (IC50) values of cisplatin (DDP), Gyp-L, and DDP in the presence of Gyp-L were calculated to determine the intervention concentration for subsequent experiments. Cell cloning assay and scratch assay reflected the proliferation and migration ability of OVCAR3 cells. PANDORA-seq small RNA sequencing was used to detect the differentially expressed transfer RNA-derived small RNAs (tsRNAs) in the cells after Gyp-L intervention, and the corresponding target genes of the tsRNAs were found by the RNAhybrid software. Malondialdehyde (MDA), glutathione (GSH), and lipid peroxide (LPO) levels were measured by colorimetry or enzyme linked immunosorbent assay (ELISA) method, Fe2+ content by FerroOrange fluorescent probe, and reactive oxygen species (ROS) content by DCFH-DA fluorescent probe to reflect the occurrence of ferroptosis in OVCAR3 cells. OVCAR3 cells were divided into a control group, a 50 µmol·L-1 Gyp-L group, and a 100 µmol·L-1 Gyp-L group. Quantitative real-time polymerase chain reaction (PCR) was performed to detect the expression of mature-tRNA-Asp-GTC, mature-tRNA-Leu-CAA, mature-mt_tRNA-Tyr-GTA_5_end, mature-tRNA-Val-CAC, mature-mt_tRNA-Glu-TTC, pre-tRNA-Arg-TCT, mature-tRNA-Asn-GTT, hydroxymethylbilane synthase (HMBS), Wnt, β-catenin, glutathione peroxidase 4 (GPX4), Kelch-like ECH-associated protein 1 (KEAP1), nuclear factor erythroid 2-related factor 2 (Nrf2), activating transcription factor 3 (ATF3), cystine/glutamate antiporter xCT, lysophosphatidylcholine acyltransferase 3 (LPCAT3), and arachidonate 15-lipoxygenase (ALOX15). Western blot was performed to detect the expression of HMBS, Wnt, β-catenin, GPX4, KEAP1, Nrf2, ATF3, xCT, LPCAT3, and ALOX15 proteins. ResultsThe 50 µmol·L-1 Gyp-L, 100 µmol·L-1 Gyp-L, DDP, 50 µmol·L-1 Gyp-L+DDP, and 100 µmol·L-1 Gyp-L+DDP groups showed significantly inhibited proliferation and migration of OVCAR3 cells (P<0.05) and exacerbated cell ferroptosis as reflected by the increase in the content of ROS, MDA, LPO, and Fe2+, as well as a decrease in the content of GSH (P<0.05). Compared with the control group, Gyp-L effectively interfered with the expression of 25 tsRNAs in OVCAR3 cells (P<0.05, |log2Fc|>1). Pre-tRNA-Arg-TCT/HMBS/Wnt/β-catenin/GPX4, pre-tRNA-Arg-TCT/KEAP1/NRF2/xCT, mature-tRNA-Asp-GTC/ATF3/KEAP1/NRF2/xCT, and mature-tRNA-Asp-GTC/LPCAT3/ALOX15 axial expression was significantly aberrant after Gyp-L intervention (P<0.05). ConclusionThe pre-tRNA-Arg-TCT/HMBS/Wnt/β-catenin/GPX4, pre-tRNA-Arg-TCT/KEAP1/Nrf2/xCT, mature-tRNA-Asp-GTC/ATF3/KEAP1/Nrf2/xCT, and mature-tRNA-Asp-GTC/LPCAT3/ALOX15 signaling pathways are involved in OC development. Gyp-L inhibits OC development by activating OVCAR3 cell ferroptosis onset mainly through the mature-tRNA-Asp-GTC/ATF3/KEAP1/Nrf2/xCT and mature-tRNA-Asp-GTC/LPCAT3/ALOX15 signaling axes.
8.Oxylipidomics Combined with Transcriptomics Reveals Mechanism of Jianpi Huogu Prescription in Treating Steroid-induced Osteonecrosis of Femoral Head in Rats
Lili WANG ; Qun LI ; Zhixing HU ; Qianqian YAN ; Liting XU ; Xiaoxiao WANG ; Chunyan ZHU ; Yanqiong ZHANG ; Weiheng CHEN ; Haijun HE ; Chunfang LIU ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):190-199
ObjectiveTo unveil the mechanism of Jianpi Huogu prescription (JPHGP) in ameliorating the dyslipidemia of steroid-induced osteonecrosis of the femur head (SONFH) by oxylipidomics combined with transcriptomics. MethodsSixty SD rats were assigned into normal, model, low-, medium-, and high-dose (2.5, 5, 10 g·kg-1, respectively) JPHGP, and Jiangushengwan (1.53 g·kg-1) groups. Lipopolysaccharide was injected into the tail vein at a dose of 20 μg·kg-1 on days 1 and 2, and methylprednisolone sodium succinate was injected at a dose of 40 mg·kg-1 into the buttock muscle on days 3 to 5. The normal group received an equal volume of normal saline. Drug administration by gavage began 4 weeks after the last injection, and samples were taken after administration for 8 weeks. Hematoxylin-eosin staining was conducted to reveal the histopathological changes of the femoral head, and the number of adipocytes, the rate of empty bone lacunae, and the trabecular area were calculated. Micro-computed tomography was used for revealing the histological and histomorphometrical changes of the femoral head. Enzyme-linked immunosorbent assay was employed to measure the serum levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), apolipoprotein A1 (ApoA1), and apolipoprotein B (ApoB). At the same time, the femoral head was collected for oxylipidomic and transcriptomic detection. The differential metabolites and differential genes were enriched and analyzed, and the target genes regulating lipid metabolism were predicted. The predicted target proteins were further verified by molecular docking, immunohistochemistry, and Western blot. ResultsCompared with the normal group, the model group showcased thinning of the femoral head, trabecular fracture, karyopyknosis, subchondral cystic degeneration, increases in the number of adipocytes and the rate of empty bone lacunae (P<0.01), a reduction in the trabecular area (P<0.01), decreases in BMD, Tb.Th, Tb.N, and BV/TV, and increases in Tb.Sp and BS/BV (P<0.01). Compared with the model group, the JPHGP groups showed no obvious thinning of the femoral head or subchondroidal cystic degeneration. The high- and medium-dose JPHGP groups presented declines in the number of adipocytes and the rate of empty bone lacunae, an increase in the trabecular area (P<0.05, P<0.01), rises in BMD, Tb.Th, Tb.N, and BV/TV, and decreases in Tb.Sp and BS/BV (P<0.05, P<0.01). Compared with the normal group, the model group showcased raised serum levels of TG, TC, LDL, and ApoB and lowered serum levels of HDL and ApoA1 (P<0.01). Compared with the model group, the JPHGP groups had lowered serum levels of TG, TC, LDL, and ApoB (P<0.05, P<0.01) and a risen serum level of ApoA1 (P<0.05, P<0.01). Moreover, the serum level of HDL in the high-dose JPHGP group increased (P<0.01). A total of 19 different metabolites of disease set and drug set were screened out by oxylipidomics of the femoral head, and 119 core genes with restored expression were detected by transcriptomics. The enriched pathways were mainly concentrated in inflammation, lipids, apoptosis, and osteoclast differentiation. Molecular docking, immunohistochemistry, and Western blot results showed that compared with the normal group, the model group displayed increased content of 5-lipoxygenase (5-LO) and peroxisome proliferator-activated receptor γ (PPARγ) in the femoral head (P<0.01). Compared with the model group, medium- and high-dose JPHGP reduced the content of 5-LO and PPARγ (P<0.05, P<0.01). ConclusionJPHGP can restore the levels of oxidized lipid metabolites by regulating the 5-LO-PPARγ axis to treat SONFH in rats. Relevant studies provide experimental evidence for the efficacy mechanism of JPHGP in the treatment of SONFH.
9.Exploring Molecular Mechanism of Gypenoside L against Ovarian Cancer Based on Ferroptosis Pathway Mediated by Mature-tRNA-Asp-GTC/ATF3-LPCAT3
Jingxuan ZHU ; Jiao ZHAO ; Qun WANG ; Xiaofei SUN ; Jiaxin WANG ; Hongda ZHANG ; Nan SONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):107-117
ObjectiveTo investigate the role of mature-tRNA-Asp-GTC and pre-tRNA-Arg-TCT in the ferroptosis phenotype of ovarian cancer (OC) cells and the regulatory mechanism of gypenoside L (Gyp-L) on mature-tRNA-Asp-GTC and pre-tRNA-Arg-TCT in OC cells. MethodsThe proliferation of human ovarian adenocarcinoma OVCAR3 cells was detected by cell counting kit-8 (CCK-8) assay, and the half-maximal inhibitory concentration (IC50) values of cisplatin (DDP), Gyp-L, and DDP in the presence of Gyp-L were calculated to determine the intervention concentration for subsequent experiments. Cell cloning assay and scratch assay reflected the proliferation and migration ability of OVCAR3 cells. PANDORA-seq small RNA sequencing was used to detect the differentially expressed transfer RNA-derived small RNAs (tsRNAs) in the cells after Gyp-L intervention, and the corresponding target genes of the tsRNAs were found by the RNAhybrid software. Malondialdehyde (MDA), glutathione (GSH), and lipid peroxide (LPO) levels were measured by colorimetry or enzyme linked immunosorbent assay (ELISA) method, Fe2+ content by FerroOrange fluorescent probe, and reactive oxygen species (ROS) content by DCFH-DA fluorescent probe to reflect the occurrence of ferroptosis in OVCAR3 cells. OVCAR3 cells were divided into a control group, a 50 µmol·L-1 Gyp-L group, and a 100 µmol·L-1 Gyp-L group. Quantitative real-time polymerase chain reaction (PCR) was performed to detect the expression of mature-tRNA-Asp-GTC, mature-tRNA-Leu-CAA, mature-mt_tRNA-Tyr-GTA_5_end, mature-tRNA-Val-CAC, mature-mt_tRNA-Glu-TTC, pre-tRNA-Arg-TCT, mature-tRNA-Asn-GTT, hydroxymethylbilane synthase (HMBS), Wnt, β-catenin, glutathione peroxidase 4 (GPX4), Kelch-like ECH-associated protein 1 (KEAP1), nuclear factor erythroid 2-related factor 2 (Nrf2), activating transcription factor 3 (ATF3), cystine/glutamate antiporter xCT, lysophosphatidylcholine acyltransferase 3 (LPCAT3), and arachidonate 15-lipoxygenase (ALOX15). Western blot was performed to detect the expression of HMBS, Wnt, β-catenin, GPX4, KEAP1, Nrf2, ATF3, xCT, LPCAT3, and ALOX15 proteins. ResultsThe 50 µmol·L-1 Gyp-L, 100 µmol·L-1 Gyp-L, DDP, 50 µmol·L-1 Gyp-L+DDP, and 100 µmol·L-1 Gyp-L+DDP groups showed significantly inhibited proliferation and migration of OVCAR3 cells (P<0.05) and exacerbated cell ferroptosis as reflected by the increase in the content of ROS, MDA, LPO, and Fe2+, as well as a decrease in the content of GSH (P<0.05). Compared with the control group, Gyp-L effectively interfered with the expression of 25 tsRNAs in OVCAR3 cells (P<0.05, |log2Fc|>1). Pre-tRNA-Arg-TCT/HMBS/Wnt/β-catenin/GPX4, pre-tRNA-Arg-TCT/KEAP1/NRF2/xCT, mature-tRNA-Asp-GTC/ATF3/KEAP1/NRF2/xCT, and mature-tRNA-Asp-GTC/LPCAT3/ALOX15 axial expression was significantly aberrant after Gyp-L intervention (P<0.05). ConclusionThe pre-tRNA-Arg-TCT/HMBS/Wnt/β-catenin/GPX4, pre-tRNA-Arg-TCT/KEAP1/Nrf2/xCT, mature-tRNA-Asp-GTC/ATF3/KEAP1/Nrf2/xCT, and mature-tRNA-Asp-GTC/LPCAT3/ALOX15 signaling pathways are involved in OC development. Gyp-L inhibits OC development by activating OVCAR3 cell ferroptosis onset mainly through the mature-tRNA-Asp-GTC/ATF3/KEAP1/Nrf2/xCT and mature-tRNA-Asp-GTC/LPCAT3/ALOX15 signaling axes.
10.Oxylipidomics Combined with Transcriptomics Reveals Mechanism of Jianpi Huogu Prescription in Treating Steroid-induced Osteonecrosis of Femoral Head in Rats
Lili WANG ; Qun LI ; Zhixing HU ; Qianqian YAN ; Liting XU ; Xiaoxiao WANG ; Chunyan ZHU ; Yanqiong ZHANG ; Weiheng CHEN ; Haijun HE ; Chunfang LIU ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):190-199
ObjectiveTo unveil the mechanism of Jianpi Huogu prescription (JPHGP) in ameliorating the dyslipidemia of steroid-induced osteonecrosis of the femur head (SONFH) by oxylipidomics combined with transcriptomics. MethodsSixty SD rats were assigned into normal, model, low-, medium-, and high-dose (2.5, 5, 10 g·kg-1, respectively) JPHGP, and Jiangushengwan (1.53 g·kg-1) groups. Lipopolysaccharide was injected into the tail vein at a dose of 20 μg·kg-1 on days 1 and 2, and methylprednisolone sodium succinate was injected at a dose of 40 mg·kg-1 into the buttock muscle on days 3 to 5. The normal group received an equal volume of normal saline. Drug administration by gavage began 4 weeks after the last injection, and samples were taken after administration for 8 weeks. Hematoxylin-eosin staining was conducted to reveal the histopathological changes of the femoral head, and the number of adipocytes, the rate of empty bone lacunae, and the trabecular area were calculated. Micro-computed tomography was used for revealing the histological and histomorphometrical changes of the femoral head. Enzyme-linked immunosorbent assay was employed to measure the serum levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), apolipoprotein A1 (ApoA1), and apolipoprotein B (ApoB). At the same time, the femoral head was collected for oxylipidomic and transcriptomic detection. The differential metabolites and differential genes were enriched and analyzed, and the target genes regulating lipid metabolism were predicted. The predicted target proteins were further verified by molecular docking, immunohistochemistry, and Western blot. ResultsCompared with the normal group, the model group showcased thinning of the femoral head, trabecular fracture, karyopyknosis, subchondral cystic degeneration, increases in the number of adipocytes and the rate of empty bone lacunae (P<0.01), a reduction in the trabecular area (P<0.01), decreases in BMD, Tb.Th, Tb.N, and BV/TV, and increases in Tb.Sp and BS/BV (P<0.01). Compared with the model group, the JPHGP groups showed no obvious thinning of the femoral head or subchondroidal cystic degeneration. The high- and medium-dose JPHGP groups presented declines in the number of adipocytes and the rate of empty bone lacunae, an increase in the trabecular area (P<0.05, P<0.01), rises in BMD, Tb.Th, Tb.N, and BV/TV, and decreases in Tb.Sp and BS/BV (P<0.05, P<0.01). Compared with the normal group, the model group showcased raised serum levels of TG, TC, LDL, and ApoB and lowered serum levels of HDL and ApoA1 (P<0.01). Compared with the model group, the JPHGP groups had lowered serum levels of TG, TC, LDL, and ApoB (P<0.05, P<0.01) and a risen serum level of ApoA1 (P<0.05, P<0.01). Moreover, the serum level of HDL in the high-dose JPHGP group increased (P<0.01). A total of 19 different metabolites of disease set and drug set were screened out by oxylipidomics of the femoral head, and 119 core genes with restored expression were detected by transcriptomics. The enriched pathways were mainly concentrated in inflammation, lipids, apoptosis, and osteoclast differentiation. Molecular docking, immunohistochemistry, and Western blot results showed that compared with the normal group, the model group displayed increased content of 5-lipoxygenase (5-LO) and peroxisome proliferator-activated receptor γ (PPARγ) in the femoral head (P<0.01). Compared with the model group, medium- and high-dose JPHGP reduced the content of 5-LO and PPARγ (P<0.05, P<0.01). ConclusionJPHGP can restore the levels of oxidized lipid metabolites by regulating the 5-LO-PPARγ axis to treat SONFH in rats. Relevant studies provide experimental evidence for the efficacy mechanism of JPHGP in the treatment of SONFH.


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