1.Genetic analysis and reproductive intervention for 46 Chinese pedigrees affected with Hereditary multiple exostoses.
Lilan SU ; Xiao HU ; Jing DAI ; Zhengxing WAN ; Duo YI ; Shuangfei LI ; Liang HU ; Yueqiu TAN ; Fei GONG ; Ge LIN ; Guangxiu LU ; Qianjun ZHANG ; Juan DU ; Wenbin HE
Chinese Journal of Medical Genetics 2026;43(4):253-258
OBJECTIVE:
To explore the genetic etiology of 46 Chinese pedigrees affected with Hereditary multiple exostoses (HME) and provide genetic counseling and reproductive intervention.
METHODS:
Whole-exome sequencing and Sanger sequencing were carried out on 87 patients from the 46 pedigrees to analyze the variants of EXT1 and EXT2 genes. Pathogenicity of the variants was assessed based on the guidelines from the American College of Medical Genetics and Genomics and Association for Molecular Pathology (ACMG/AMP). Prenatal diagnosis and preimplantation genetic testing (PGT) were provided for couples with identified pathogenic mutations. This study was approved by the Medical Ethics Committee of the hospital (Ethics No.: LL-SC-SG-2014-010).
RESULTS:
In total 17 and 22 pathogenic variants were respectively identified in the EXT1 and EXT2 genes, among which 5 EXT1 and 12 EXT2 variants were unreported previously. Three patients with no family history were found to harbor de novo variants of the EXT1 gene. Twenty nine couples had opted for PGT or underwent prenatal diagnosis following natural conception, and 17 healthy babies were born.
CONCLUSION
This study has clarified the genetic etiology of 45 HME pedigrees and identified 17 novel variants, which has enriched the mutational spectrum of the EXT1 and EXT2 genes. Reproductive intervention through PGT and prenatal diagnosis have prevented the recurrence of HME in these families.
Humans
;
Female
;
Male
;
Pedigree
;
Exostoses, Multiple Hereditary/diagnosis*
;
N-Acetylglucosaminyltransferases/genetics*
;
Adult
;
Exostosin 1
;
Asian People/genetics*
;
Genetic Testing
;
Exostosin 2
;
Mutation
;
China
;
Prenatal Diagnosis
;
Pregnancy
;
Genetic Counseling
;
Preimplantation Diagnosis
;
Exome Sequencing
;
East Asian People
2.Mechanism of Yigan huayu formula in alleviating liver fibrosis based on proteomics
Conghui WANG ; Guiping MA ; Longzhu WANG ; Fenping LU ; Yanfang LI ; Qiuhan GE ; Shiping HU
China Pharmacy 2026;37(9):1155-1160
OBJECTIVE To investigate the effects and mechanism of Yigan huayu formula in alleviating liver fibrosis in mice. METHODS Mice were randomly divided into blank group (normal saline), model group (normal saline), Yigan huayu formula low- and high-dose groups (28.98, 57.96 g/kg, calculated by crude drug), with 8 mice in each group. Except for the blank group, the liver fibrosis model was induced by intraperitoneal injection of 15%CCl 4 -olive oil solution. From the third week, the mice received the medicine/normal saline intragastrically, once a day, for 4 consecutive weeks. After the last medication, liver indexes were calculated, the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum, as well as the hydroxyproline (HYP) content in liver tissue, were measured. Liver histopathology was evaluated. Differentially expressed proteins (DEPs) in liver tissue were analyzed based on proteomics, followed by bioinfo rmatics analysis. The expressions of core DEPs were validated using Western blot (WB) and immunohistochemistry (IHC) methods. RESULTS Compared with the blank group, the model group showed significantly elevated liver indexes, serum activities of ALT and AST, and hepatic HYP content ( P <0.05), along with obvious pathological damage and collagen deposition. Compared with the model group, the above indexes of mice in the Yigan huayu formula high-dose group were decreased significantly ( P <0.05), with marked improvement in liver pathological damage and collagen deposition. Proteomics identified 210 DEPs between the model group and Yigan huayu formula high-dose group. DEPs were significantly enriched in extracellular matrix (ECM)-receptor interaction and lipid metabolism pathways. WB and IHC confirmed that Yigan huayu formula could significantly inhibit the abnormally elevated expressions of collagen type Ⅳ alpha1 chain (COL4A1), secreted protein acidic and rich in cysteine (SPARC), vitronectin (VTN) and laminin subunit alpha5 (LAMA5) in liver tissue of mice ( P <0.05). CONCLUSIONS Yigan huayu formula may exert anti-hepatic fibrosis effects by inhibiting the expressions of proteins such as COL4A1, LAMA5, SPARC, and VTN, thereby blocking the ECM-receptor interaction signaling pathway, and subsequently suppressing excessive ECM deposition and basement membrane remodeling.
3.Effect and mechanism of the azo-podophyllotoxin derivative SU056 in a mouse model of carbon tetrachloride-induced liver fibrosis
Qichao GE ; Rui CHEN ; Yufei YANG ; Yuecheng GUO ; Dihanjing ZHANG ; Hui DONG ; Lungen LU
Journal of Clinical Hepatology 2026;42(6):1310-1320
ObjectiveTo investigate the effect of SU056, an azo-podophyllotoxin derivative, on carbon tetrachloride (CCl4)-induced liver fibrosis in mice and related mechanisms of action. MethodsA total of 12 mice were randomly divided into control group, model group (CCl4+normal saline), and treatment group (CCl4+SU056), with 4 mice in each group. Mice were given intraperitoneal injection of CCl4 to establish a model of liver fibrosis, and during the middle stage of modeling, the mice in the treatment group were given daily intraperitoneal injection of SU056. Liver histopathological injury, collagen deposition, and liver function were assessed based on HE staining, Masson staining, Sirius Red staining, the content of hydroxyproline in liver tissue, and the serum levels of alanine aminotransferase and aspartate aminotransferase, and immunofluorescence assay was used to measure the expression levels of smooth muscle actin α (α-SMA), collagen type Ⅰ, and Y-box binding protein 1 (YB1). The human hepatic stellate cell (HSC) line LX-2 and primary mouse HSC were used, and CCK-8 assay was used to measure cell proliferation; Transwell assay was used to observe cell migration; quantitative reverse transcription-polymerase chain reaction and Western Blot were used to measure the expression levels of collagen type Ⅰ, collagen type Ⅲ, YB1, phosphorylated mammalian target of rapamycin (mTOR), and phosphorylated S6K, so as to validate the function of the YB1/mTOR signaling axis. The one-way or two-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsIn the mouse model of liver fibrosis induced by CCl4, compared with the model group, the treatment group had significant alleviation of inflammatory cell infiltration, collagen deposition, and pseudolobule formation in liver tissue and significant reductions in the serum levels of alanine aminotransferase and aspartate aminotransferase and the content of hydroxyproline in liver tissue (all P<0.01). Immunofluorescence assay showed that SU056 significantly inhibited the abnormal high expression of α-SMA, collagen type I, and YB1 in liver tissue (all P<0.01). In vitro experiments showed that SU056 inhibited the transforming growth factor-β1-induced proliferation of LX-2 cells (P<0.01), the migration of LX-2 cells (P<0.05), and the transcriptional up-regulation of collagen type Ⅰ and collagen type Ⅲ (all P<0.05) in a dose-dependent manner, and SU056 could inhibit the spontaneous activation of primary HSC in vitro. Mechanistic studies revealed that transforming growth factor-β1 simultaneously upregulated the expression levels of YB1, phosphorylated mTOR, and phosphorylated S6K in LX-2 cells, and treatment with SU056 (10 and 20 µmol/L) could downregulate the protein expression levels of collagen type I, YB1, phosphorylated mTOR, and phosphorylated S6K. Specific knockdown of YB1 or administration of the mTOR inhibitor rapamycin exerted a similar effect as SU056. SU056 also inhibited the co-upregulation of α-SMA and phosphorylated mTOR in liver tissue of model mice (P<0.01). ConclusionSU056 can effectively inhibit HSC activation, proliferation, migration, and extracellular matrix production both in vivo and in vitro and thus delay the progression of liver fibrosis, by disrupting the YB1/mTOR positive feedback signaling axis.
4.Circadian mechanisms underlying cardiometabolic dysfunction induced by chronic PM2.5 exposure
Wenqing ZHANG ; Biao WU ; Jianshu GUO ; Dongxia FAN ; Ge WANG ; Lu YU ; Chihang ZHANG ; Xianying LIAO ; Xihao DU ; Yuquan XIE ; Jinzhuo ZHAO
Journal of Environmental and Occupational Medicine 2026;43(8):926-935
Background Long-term exposure to ambient fine particulate matter (PM2.5) is a significant risk factor for cardiometabolic disorders. However, the mechanisms of its interaction with the endogenous circadian system remain incompletely understood. Objective To investigate whether chronic PM2.5 exposure interferes with the rhythmic expression of the cardiac circadian clock, thereby disrupting downstream antioxidant defenses and metabolic homeostasis, and ultimately driving cardiometabolic dysfunction. Methods Seventy-two male C57BL/6 mice were randomly divided into a PM2.5 exposure group (PM group) and a filtered air control group (FA group). Whole-body exposure was conducted for 8 weeks in a meteorological environmental animal exposure system. Samples were collected at six distinct zeitgeber time (ZT) points post-exposure. The 24 h ambulatory blood pressure and serum lipid profiles were monitored. Rhythm parameters were derived via cosinor analysis to compare differences in Midline statistic of rhythm (Mesor), amplitude, and phase between the two groups. The rhythmic expression of core circadian clock genes and antioxidant genes in the myocardium was detected by quantitative polymerase chain reaction (qPCR). Myocardial reactive oxygen species (ROS) levels and downstream pathway protein expression were analyzed by immunofluorescence and Western blot (WB), respectively. The expression changes of the clock gene retinoic acid receptor-related orphan receptor α (RORα) were assessed at both the mRNA and protein levels. Finally, Spearman correlation analysis was used to explore the relationships among myocardial RORα expression, lipid profiles, and oxidative stress indicators. Results Compared to the FA group, mice in the PM group exhibited a blunted circadian rhythm in blood pressure, characterized by sustained elevation throughout the day. Chronic PM2.5 exposure showed a significant interaction with ZT on systolic blood pressure (SBP), diastolic blood pressure (DBP), and mean arterial pressure (MAP) (F-interaction=9.11, 5.70, and 6.02, respectively; P<0.05), as well as on serum triglycerides (TG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) (F-interaction=16.32, 11.12, 15.39, and 28.09, respectively; P<0.05). Cosinor analysis further revealed that the Mesor values of T-CHO, TG, and LDL-C were significantly increased (P<0.05), while that of HDL-C was significantly decreased in the PM group (P<0.05). The oscillation amplitudes of SBP, DBP, and MAP showed a decreasing trend, whereas those of TG and LDL-C were significantly increased (P<0.05). Furthermore, SBP, T-CHO, and HDL-C all exhibited a significant phase delay (P<0.05). Mechanistically, PM2.5 exposure significantly suppressed the expression of the positive circadian regulator RORα in the myocardium, leading to disordered rhythmic expression of core clock genes (Bmal1, Clock, Per1/2, and Cry1/2). This exposure also inhibited the rhythmic expression of antioxidant genes (GPX1, SOD2, and CAT), resulting in increased ROS generation and elevated expression of calcium/calmodulin-dependent protein kinase II (CaMKII) and reduced nicotinamide adenine dinucleotide phosphate (NADPH) proteins. Correlation analysis further revealed that myocardial RORα expression level was negatively correlated with T-CHO, TG, and LDL-C (r=−0.55, −0.63, and −0.51, respectively; P<0.001), and positively correlated with HDL-C (r=0.37, P=0.010), and antioxidant genes GPX1, SOD2, and CAT expression (r=0.34, 0.35, and 0.56, respectively; P < 0.001). Conclusion Chronic PM2.5 exposure induces cardiometabolic dysfunction by suppressing myocardial RORα expression. This suppression disrupts the cardiac circadian clock and the diurnal balance of oxidative stress, triggering oxidative damage and elevating expression of CaMKII/NADPH pathway proteins. Collectively, these alterations precipitate the loss of cardiac metabolic rhythms and subsequent functional impairment.
5.An excerpt of EASL-AASLD Delphi consensus statement on surrogate endpoints and real-world evidence in primary biliary cholangitis (2026)
Qichao GE ; Xiaobo CAI ; Lungen LU
Journal of Clinical Hepatology 2026;42(8):1807-1814
In 2026, the European Association for the Study of the Liver and the American Association for the Study of Liver Diseases jointly published a Delphi consensus statement on surrogate endpoints and real-world evidence in primary biliary cholangitis. This consensus proposes 16 statements and 42 recommendations centering on the following three most critical issues in the development of new drugs for primary biliary cholangitis: whether liver biochemical parameters and noninvasive assessments of liver fibrosis can be used as surrogate endpoints for clinical outcomes; how real-world data and real-world evidence can complement or support confirmatory studies in a manner compliant with regulatory requirements; how patient-reported outcomes can be standardized for the assessment of quality of life, pruritus, and fatigue. This article provides an excerpt and expert interpretation of the background, methodological framework, core statements, recommendations, and key figures and tables of this consensus, in order to provide a reference for the diagnosis and treatment of primary biliary cholangitis, drug evaluation, real-world study design, and the development of patient-centered clinical endpoints in China.
6.Optimization of polysaccharide extraction from Abelmoschus manihot flowers using composite enzymes and a study on the activities of the polysaccharides
Ge LIU ; Cheng LUO ; Huifang CHAI ; Yan LI ; Tian LU
China Pharmacy 2026;37(18):2369-2375
OBJECTIVE To optimize the extraction process of polysaccharides from Abelmoschus manihot flowers, and to investigate the antioxidant, moisturizing and hygroscopicity activities of the polysaccharides.METHODS The optimal ratio of composite enzymes was determined by orthogonal experiments, and the extraction process of polysaccharides was optimized by single-factor experiments and Box-Behnken response surface methodology, and validated. The scavenging rates of 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radicals, hydroxyl free radicals, and superoxide anion free radicals, as well as the total reducing ability of the polysaccharides were investigated by the ABTS method, Fenton-salicylic acid colorimetric method, pyrogallol autoxidation method, and potassium ferricyanide reduction method, respectively. The moisturizing and hygroscopicity activities of the polysaccharides were evaluated by the weighing method.RESULTS The optimal ratio of composite enzymes was 136 000 U of papain, 125 000 U of pectinase,and 5 000 U of cellulase. The optimal extraction process was as follows: enzymatic hydrolysis time of 2.8 h, enzymatic hydrolysis temperature of 57.50 ℃, pH of the sodium dihydrogen phosphate-disodium hydrogen phosphate buffer was 5.5, and solid-to-liquid ratio of 1∶40. The results of three experiments showed that the average polysaccharides extraction rate of 23.595%, with an RSD of 0.90%. The scavenging rates of ABTS free radicals, hydroxyl free radicals, superoxide anion free radicals, and total reducing ability were 81.759%, 87.110%, 54.244%, and 1.129 (the mass concentrations of polysaccharides were 10、 5、 10, and 10 mg/mL, respectively), respectively. At relative humidity of 43% for 12 h, the moisturizing rate and hygroscopicity rate of polysaccharides were 99.387% and 100.478%, respectively; at relative humidity of 83% for 12 h, they were 99.764% and 100.455%, respectively.CONCLUSIONS The optimal extraction process of polysaccharides from A.manihot flowers based on the composite enzymes is stable and feasible; the polysaccharides obtained under this process have good antioxidant, moisturizing, and hygroscopicity activities.
7.Effects of Electrothermal Acupuncture on the HIF-1α/S1P/S1PR3 Pathway and Macrophage Polarization in Epididymal White Adipose Tissue of Obese Model Mice
Xiaobo GE ; Yuanyuan LI ; Jiangyan WEI ; Limin NIE ; Xin YANG ; Quan MIAO ; Mina WANG ; Yawei XI ; Luopeng ZHAO ; Yine SONG ; Xinru SUN ; Lu LIU ; Bin LI
Journal of Traditional Chinese Medicine 2026;67(20):2227-2235
ObjectiveTo investigate the mechanism underlying the weight loss effect of electrothermal acupuncture (ETA) in diet-induced obese (DIO) model mice based on the hypoxia-inducible factor-1α (HIF-1α)/sphingosine-1-phosphate (S1P)sphingosine-1-phosphate receptor 3 (S1PR3) signaling pathway-mediated macrophage polarization. MethodsAn obesity mouse model was established by continuously feeding at a high-fat diet (HFD) for 8 weeks. Twenty successfully modeled obese mice were randomly divided into a model group and an electrothermal acupuncture (ETA) group with 10 mice per group, both of which continued to receive the HFD till week 12. Another 10 mice were assigned to the normal group and fed a standard chow diet continuously for 12 weeks. Starting from week 9 of the experiment, mice in the ETA group received electrothermal acupuncture stimulation at Zhongwan (CV12), bilateral Tianshu (ST25), and bilateral Zusanli (ST36) for 30 minutes per session, with the instrument current set at (40±0.5)mA and a constant temperature of (40±0.5)℃. Mice in the normal group and the model group were restrained in the same manner for 30 minutes without acupuncture. All interventions were performed once every other day for a total of 4 weeks. Ten hours after the final intervention, the body weight, waist circumference, and body length of mice in each group were measured, and Lee's index was calculated. Serum biochemical analysis was performed to assess fasting blood glucose (FBG), total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), and creatinine (Cr). Serum leptin (LEP) and adiponectin (ADPN) levels, as well as sphingosine-1-phosphate (S1P) levels in epididymal white adipose tissue (eWAT), were detected by ELISA. Histomorphological changes in eWAT were monitored via HE staining. Flow cytometry was used to analyze macrophage subtype-related indexes in eWAT, including the total proportion of macrophages, the proportion of M1 macrophages, the proportion of M2 macrophages, and the M1/M2 ratio. Antibody array technology was used to determine the levels of differentially expressed inflammatory factors in eWAT between the model and blank groups, as well as between the ETA and model groups. Western blot and qPCR were used to detect the protein and mRNA expression levels of HIF-1α and S1PR3 in the eWAT, respectively. ResultsCompared to the normal group, the model group showed significantly increased body weight, Lee's index, waist circumference, and FBG, TC, TG, HDL-C, LDL-C, AST, and ALT levels, upgraded LEP and S1P levels, decreased ADPN level and ADPN/LEP ratio, elevated proportion of M1 macrophages and the M1/M2 ratio, reduced proportion of M2 macrophages, and upregulated protein and mRNA expression levels of HIF-1α and S1PR3 (P<0.05 or P<0.01). With the exception of HDL-C, all the abovementioned indicators in the ETA group were significantly improved compared to the model group (P<0.05 or P<0.01). Analysis of differentially expressed inflammatory cytokines revealed that, compared to the normal group, the levels of interleukin-1β (IL-1β), interferon-γ (IFN-γ), interleukin-6 (IL-6), and interleukin-12p70 (IL-12p70) in the eWAT of the model group were elevated; conversely, the levels of interleukin-17 (IL-17), interleukin-5 (IL-5), IL-6, and interleukin-23 (IL-23) in the eWAT of the ETA group decreased compared to the model group (P<0.05 or P<0.01). ConclusionETA exerts a weight-reducing effect in obese model mice, and the mechanism may involve inhibition of the HIF-1α/S1P/S1PR3 signaling pathway activation, which shifts macrophage polarization from the M1 to M2 phenotype, thereby alleviating chronic inflammation.
8.The core factors and application of multiple linear regression fusion model in evaluating the risk of myopia in preschool children
Qingjin WANG ; Cangxia ZHANG ; Hanxiao GE ; Yue MA ; Jie LU
International Eye Science 2026;26(11):2018-2025
AIM: To construct an assessment model for the risk of myopia in preschool children and evaluate its clinical value.METHODS: A retrospective analysis was conducted on the data of preschool children who established refractive profiles in our hospital from January 2020 to December 2024. The eye with lower refractive error was selected. The right eye was chosen in cases where the refractive error of both eyes was identical. The data were divided into a training set(n=385)and a validation set(n=165)in a 7∶3 ratio. The data of the children were collected in detail, and the children's spherical equivalent(SE)was used as the dependent variable. The multiple linear regression model was used to screen the indicators that affected the biological indicators related to myopia in preschool children.Subsequently, the children in the training set were divided into a myopia group and a normal vision group based on whether myopia occurred(SE≤-0.50 D can be diagnosed as myopia according to refractive power assessment). Subsequently, the influencing factors selected by the multiple linear regression model were included as predictive variables, and a risk assessment equation was constructed using Logistic regression. The discrimination, calibration and clinical application value of the assessment model were verified through the receiver operating characteristic(ROC)curve, Hosmer-Lemeshow test and decision curve.RESULTS: This study included 550 preschool children(550 eyes). The training set comprised 385 children(385 eyes; 190 boys and 195 girls), and the validation set comprised 165 children(165 eyes; 82 boys and 83 girls). Among the children in the training set, the myopia group included 34 children(34 eyes; 16 boys and 18 girls), and the non-myopic group included 351 children(351 eyes; 174 boys and 177 girls). Multiple linear regression analysis showed that the ratio of axial length(AL)to corneal curvature radius(CR), subfoveal choroidal thickness(SFCT), anterior chamber depth(ACD), age, parental myopia history, and outdoor activity time were risk factors for myopia in preschool children(all P<0.05). Based on the above risk factors, a Logistic regression risk assessment model was constructed, and the evaluation model Logit(P)was obtained as 0.894×AL/CR-1.312×SFCT+1.182×ACD+2.063×age+0.763×parental myopia history-5.868. The effectiveness of the myopia risk assessment model for preschool children based on the training set was evaluated. The results showed that the area under the ROC curve(AUC)was 0.939(95%CI: 0.912-0.965), the H-L test was χ2=1.487, P=0.993, and the decision curve indicated that within the threshold range of 0.03-0.81 and 0.89-0.97, the net return rate was greater than 0. This suggested that the model had good discrimination, calibration, and clinical application value in evaluating the myopia risk and actual risk of preschool children. Internal validation was conducted in an independent validation set, and the results showed that the AUC of the ROC curve was 0.967(95%CI: 0.919-1.000), the H-L test was χ2=3.818, P=0.873, and the decision curve indicated that within the threshold range of 0.01-0.99, the net return rate was greater than 0, further suggesting that the model assessment had good consistency with the actual risk.CONCLUSION: Age, parental history of myopia, AL/CR, SFCT, and ACD are risk factors for myopia in preschool children. The Logistic regression risk assessment model constructed based on these factors has high evaluation value and can effectively evaluate the probability of myopia occurrence in individuals, providing reference for individualized prevention of myopia risk in preschool children.
9.Summary of the best evidence for the management of gastrointestinal graft-versus-host disease after hematopoietic stem cell transplantation
Fangchen GU ; Yongqin GE ; Huijuan MEI ; Yin LU ; Xiaming ZHU
Chinese Journal of Blood Transfusion 2026;39(5):610-618
Objective: To summarize the best evidence for the management of patients with gastrointestinal graft-versushost disease after hematopoietic stem cell transplantation, and provide evidence-based references for clinical nursing work. Methods: A systematic search was conducted for relevant evidence on the management of patients with gastrointestinal graft-versus-host disease after hematopoietic stem cell transplantation from domestic and foreign databases, as well as websites of blood and bone marrow transplantation related societies. The search period was from the establishment of the database until February 2026. Results: After screening, a total of 19 articles were included, encompassing 3 clinical decisions, 1 recommended practice, 4 guidelines, 7 expert consensuses, and 4 evidence summaries. Twenty-five pieces of best evidence were summarized across 8 aspects: multidisciplinary collaboration, influencing factors, assessment and monitoring, diagnostic differentiation, symptom management, dietary nutrition, medication guidance, and health education. Conclusion: Summarizing the relevant evidence on the management of patients with gastrointestinal graft-versus-host disease after hematopoietic stem cell transplantation can provide evidence-based support for clinical medical staff, and it is recommended to apply it in combination with clinical practice and patient wishes.
10.Fufang Kangjiaolv Capsules Treat Anxiety in Rat Model of Chronic Restraint Stress via Microbiota-gut-brain Axis
Wenxin FAN ; Tingyue JIANG ; Yu WANG ; Ge ZHANG ; Yifan LU ; Mengmeng LIU ; Jiayuan LI ; Renzhi MA ; Jinli SHI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):95-107
ObjectiveTo observe the intervention effect of Fufang Kangjiaolv capsules on anxiety-like behaviors in the rat model of chronic restraint stress (CRS) and explore the mechanism underlying the anti-anxiety effect via the microbiota-gut-brain axis. MethodsRats were assigned into blank, model, positive drug (diazepam, 1 mg·kg-1), and low-, medium-, and high-dose (0.75, 1.5, 3 g·kg-1, respectively) Fufang Kangjiaolv capsules groups. After 14 days of administration, the elevated plus maze test, open field test, light and dark box test, and marble burying test were performed. Hematoxylin-eosin staining was employed to observe the pathological changes in the hippocampus and colon of rats, and Nissl staining was conducted to observe the damage of hippocampal neurons. The gut microbiota was analyzed by 16S rRNA gene sequencing. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was employed to determine the mRNA levels of zonula occludens-1 (ZO-1) and occludin in the colon of rats. The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) in the colon, serum, and hippocampus were determined by enzyme-linked immunosorbent assay. Western blot was employed to determine the protein levels of ZO-1, occludin, nuclear factor-κB p65 (NF-κB p65) in the colon tissue and NF-κB p65 and brain-derived neurotrophic factor (BDNF) in the hippocampal tissue. ResultsCompared with the blank group, the model group showed reductions in the time and frequency ratio of rats entering the elevated plus maze, the time and frequency of rats entering the central area of the open field, the time of entering the open box, the times of passing through the light and dark box, and the number of unburied beads (P<0.05, P<0.01). Compared with the model group, Fufang Kangjiaolv capsules ameliorated the anxiety of the model rats to varying degrees, and the high-dose group had the best effect, with increases in the proportions of time and frequency of rats entering the open arm in the elevated plus maze (P<0.05), the number of rats entering the central area in the open field (P<0.05), the time of entering the open box, the times of passing through the light and dark boxes, and the number of unburied beads (P<0.01). Moreover, the high-dose group showed alleviated pathological damage of hippocampal neurons and colon. The results of 16S rRNA gene sequencing showed that the model group had increased relative abundance of Firmicutes, Deferribacterota, Romboutsia, and Phascolarctobacterium, while it had decreased relative abundance of Bavcteroidota and Lactobacillus. The drug administration groups showed increased relative abundance of Bavcteroidota, Bacteroides, norank f norank o Clostridia UCG-014, and Blautia and decreased relative abundance of Firmicutes and Deferribacterota. Compared with the blank group, the model group showed down-regulated protein and mRNA levels of ZO-1 and occludin in the colon (P<0.01), elevated levels of TNF-α, IL-6, and IL-β in the colon, serum, and hippocampus (P<0.01), up-regulated protein level of NF-κB p65 in the colon and hippocampus (P<0.01), and down-regulated protein level of BDNF in the hippocampus (P<0.05). Compared with the model group, high-dose Fufang Kangjiaolv capsules up-regulated the mRNA levels of ZO-1 and occludin in the colon (P<0.01), lowered the levels of TNF-α, IL-6, and IL-β in the colon, serum, and hippocampus (P<0.01), up-regulated the protein levels of ZO-1 (P<0.01) and occludin (P<0.05) in the colon, down-regulated the protein level of NF-κB p65 in the colon and hippocampus (P<0.05), and up-regulated the protein level of BDNF in the hippocampus. ConclusionFufang Kangjiaolv capsules can reduce the anxiety-like behaviors in the rat model of CRS by regulating the gut microbiota disturbance, up-regulating the expression of tight junction proteins in the colon, repairing intestinal mucosal mechanical barrier, and down-regulating NF-κB/BDNF signaling pathway, thereby reducing peripheral and central inflammation. This study proves the hypothesis that Fufang Kangjiaolv capsules play an anti-anxiety role via the microbiota-gut-brain axis, providing a new idea for further research.

Result Analysis
Print
Save
E-mail