1.Safety of 3D printed titanium alloy bone trabecular cup prosthesis combined with modified Kidney Tonifying and Blood Activating Decoction for elderly hip arthroplasty
Jiangjing WANG ; Na ZHAO ; Xiaona HU ; Na ZHAO
Chinese Journal of Tissue Engineering Research 2026;30(3):612-619
BACKGROUND:The titanium alloy bone trabecular bone socket cup prosthesis printed by 3D technology has superior biological characteristics,which helps to promote osteogenic differentiation of stem cells.The Kidney Tonifying and Blood Activating Decoction has the functions of tonifying the kidneys,strengthening tendons,promoting blood circulation,and relieving pain.However,the application of the combination of the two in elderly hip arthroplasty is rarely reported.OBJECTIVE:To evaluate the application value and safety of 3D printed titanium alloy bone trabecular cup prosthesis combined with modified Kidney Tonifying and Blood Activating Decoction in elderly hip arthroplasty.METHODS:A total of 200 elderly patients who received hip arthroplasty in Cangzhou Hospital of Integrated Traditional Chinese and Western Medicine from January 2022 to January 2024 were selected and randomly divided into control group(n=100)and trial group(n=100).The control group underwent Pinnaele cup prosthesis hip arthroplasty combined with modified Kidney Tonifying and Blood Activating Decoction,while the trial group underwent 3D printed titanium alloy bone trabecular cup prosthesis hip arthroplasty combined with modified Kidney Tonifying and Blood Activating Decoction.The medication was taken continuously for 1 week before hip arthroplasty and for 4 weeks after arthroplasty.The differences in pain,swelling,hemorheology,inflammatory factors,bone metabolism,intra-articular stress,and hip abduction muscle strength were compared between the two groups to evaluate the clinical efficacy and safety of the two treatment options.RESULTS AND CONCLUSION:(1)Four weeks after surgery,the pain visual analog scale score,limb swelling,whole blood viscosity,and plasma viscosity of the trial group were lower than those of the control group(P<0.05).The interleukin-6 level of the trial group was lower than that of the control group,and the interleukin-10 level was higher than that of the control group(P<0.05).The levels of type Ⅰ collagen carboxyl terminal peptide β specific sequence and tartrate-resistant acid phosphatase 5b in the trial group were lower than those in the control group,while the levels of osteocalcin,bone-specific phosphatase and typeⅠ procollagen amino-terminal propeptide were higher than those in the control group(P<0.05).The intra-articular stress and hip abduction muscle strength in the trial group were higher than that of the control group(P<0.05).(2)Three months after surgery,the excellent and good treatment rate in the trial group was higher than that in the control group,and the incidence of safety evaluation events such as postoperative incision hematoma and infection was lower than that in the control group(P<0.05).(3)It suggests that 3D printed titanium alloy trabecular acetabular cup prosthesis combined with modified Kidney Tonifying and Blood Activating Decoction can improve the blood rheology of elderly patients with hip arthroplasty,inhibit inflammatory response,reduce postoperative pain and swelling,and enhance bone metabolism,thereby promoting postoperative hip joint function recovery,and reducing the incidence of adverse events,with high safety.
2.WANG Yaoxian's Experience in Treating Diabetic Kidney Disease from the Perspective of Spleen and Stomach:Based on the Theory of "Internal Heat Leading to Concretions"
Bo ZHANG ; Yuxin HU ; Cong ZHAO ; Jiale ZHANG ; Weimin JIANG ; Chang YU ; Yang LIU ; Liqiao SUN ; Weiwei SUN ;
Journal of Traditional Chinese Medicine 2026;67(5):482-486
This paper summarizes Professor WANG Yaoxian's experience in treating diabetic kidney disease (DKD) from the perspective of spleen and stomach based on the "internal heat leading to concretions" theory. It is considered that internal heat leading to concretions constitutes the core pathogenesis of DKD, with the spleen and stomach serving as the source of internal heat; therefore, treatment should be based on regulating the spleen and stomach. In the early stage of DKD, dysfunction of the spleen and stomach leads to the initial generation of internal heat. Common syndrome patterns include gastrointestinal heat accumulation and constrained heat in the liver and stomach, for which modified Gegen Qinlian Decoction (葛根芩连汤) can be used to clear heat bind while modified Dachaihu Decoction (大柴胡汤) is used to clear stomach and soothe liver, respectively. In the middle stage of DKD, weakness of the spleen and stomach results in the initial formation of concretions and conglomerations. Common patterns include spleen deficiency with prevalence of dampness and deficiency of both the spleen and kidney. Treatment emphasizes strengthening the spleen and resolving dampness, raising yang and boosting the stomach with modified Shengyang Yiwei Decoction (升阳益胃汤), or supplementing spleen and boosting kidney, dissipating bind and dispe-ring concretions with modified Shenqi Dihuang Decoction (参芪地黄汤), respectively. In the late stage of DKD, it is characterized by spleen and stomach depletion, and rampant accumulation of turbidity and toxin, and the common syndrome patterns are damp-turbidity obstruction in the middle jiao (焦) and spleen-kidney yang deficiency. Treatment aims to remove turbidity and harmonize the stomach, or to warm the kidney and strengthen the spleen while elimina-ting turbidity, using modified Dahuang Gancao Decoction(大黄甘草汤) and Jupi Zhuru Decoction (橘皮竹茹汤) or modified Baoyuan Decoction (保元汤) and Lizhong Decoction (理中汤), respectively. In clinical practice, appropriate formulas and medications are flexibly selected according to specific syndromes.
3.Protective effect and mechanism of genistein on etoposide-induced chondrocyte senescence
Jinhong WANG ; Tianyu CHEN ; Lifang MAO ; Yingjie ZHAO ; Renpeng ZHOU ; Wei HU ; Chao LU
Acta Universitatis Medicinalis Anhui 2026;61(4):636-643
ObjectiveTo investigate the protective effect of genistein (Gen) on etoposide-induced chondrocyte senescence and its underlying mechanism. MethodsThe C28/I2 cell line was treated with different concentrations of Gen and etoposide, and the cell viability was detected by the CCK-8 assay. The senescence model of C28/I2 chondrocytes was induced by etoposide, with Gen intervention. Senescence-associated β-galactosidase (SA-β-gal) staining was performed to detect the senescence-positive rate and staining characteristics of chondrocytes. The expressions of peroxiredoxin 6 (Prdx6), cyclin-dependent kinaseto clarify the functional necessity of Prdx6. ResultsCompared with the etoposide group, the C28/I2 chondrocyte viability significantly increased (P<0.01), the expression ofsenescence-associated proteins p21 and p16 decreased (P<0.01, P<0.05), the expression of senescence-associated genes p21 and p16 reduced (both P<0.01), the fluorescence intensity of senescence-associated proteins p21 and p16 was diminished (P<0.05, P<0.01), and the proportion of SA-β-gal-positive cells decreased (P<0.01) in the Gen+etoposide group. Compared with the Control group, the expression of Prdx6 was downregulated in the etoposide group (P<0.05). Compared with the etoposide group, the expression of Prdx6 was upregulated in the Gen+etoposide group (P<0.01). Compared with the Control group, the GPx activity significantly decreased in the si-Prdx6 group (P<0.01). Furthermore, compared with the si-Prdx6 group, the GPx activity increased in the si-Prdx6+Gen group (P<0.05). Molecular docking results revealed that Gen formed hydrogen bond interactions with the active site of Prdx6. After Prdx6 knockdown, the expression of senescence-associated genes p21 and p16 and the fluorescence intensity of senescence-associated proteins p21 and p16 both increased in the Gen+etoposide+si-Prdx6 group (both P<0.01). ConclusionGen can inhibit etoposide-induced senescence of C28/I2 chondrocytes by upregulating the expression of Prdx6. This study provides potential drug targets and experimental basis for the prevention and treatment of chondrocyte senescence-related diseases.
4.Effect Difference and Mechanisms of Zishenwan Against Chronic Prostatitis Before and After Salt-processing of Anemarrhenae Rhizoma and Phellodendri Chinensis Cortex by Integrating Network Pharmacology and Metabolomics
Shangling ZHAO ; Xiao MENG ; Sirui LI ; Rui TAN ; Changjiang HU ; Lingying YU ; Zhimin CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(13):177-187
ObjectiveThis paper aims to systematically reveal the effect difference and mechanisms of Zishenwan against chronic prostatitis (CP) before and after salt-processing of Anemarrhenae rhizoma and Phellodendri chinensis cortex based on an integrated strategy of ultra-high performance liquid chromatography-quadrupole-orbitrap mass spectrometry (UPLC-Q-Orbitrap-MS/MS), network pharmacology, and serum metabolomics. MethodsZishenwan samples before and after salt-processing of Anemarrhenae rhizoma and Phellodendri chinensis cortex were extracted by alcohol-water dual extraction. The chemical components of each sample were detected by UPLC-Q-Orbitrap-MS/MS, and differential components were screened by multivariate statistical analysis. Network pharmacology analysis was performed based on the identified chemical components of Zishenwan to construct a protein-protein interaction (PPI) network of "component, target, and pathway", and the core components, targets, and pathways of Zishenwan against CP were screened. Forty-two male Sprague-Dawley (SD) rats were randomly divided into a blank group, a model group, a Qianliekang group (1.54 g·kg-1), low- and high-dose raw Zishenwan groups (1.8, 5.4 g·kg-1), and low- and high-dose salt-processed Zishenwan groups (1.8, 5.4 g·kg-1). The CP rat model was established by intraprostatic injection of carrageenan. After one week of recovery, the rats were administered the corresponding drugs for 21 days, while those in the blank group and model group received the same volume of normal saline. After the experiment, serum and tissue samples were collected to evaluate pharmacodynamic indicators including organ indices, histopathology, and inflammatory factors in serum. Subsequently, untargeted serum metabolomics technology was used to analyze metabolite changes and perform pathway enrichment analysis. The network pharmacology was used to construct a network of "differential metabolite, reaction, enzyme, and gene". ResultsA total of 76 components were identified in raw and salt-processed Zishenwan, and 34 differential components were screened by multivariate statistical analysis. Among them, the contents of 14 components, including berberine, berberrubine, and phellodendrine, increased after salt-processing, while the contents of 20 components, such as neomangiferin, decreased. The 28 active components and 185 potential targets were screened out by network pharmacology. The core components included berberine, phellodendrine, magnoflorine, and jatrorrhizine, and the core targets included signal transducer and activator of transcription 3 (STAT3), protein kinase B1 (Akt1), and transcription factor AP-1 (JUN). These targets were significantly enriched in pro-inflammatory signaling pathways such as phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) and mitogen-activated protein kinase (MAPK). Compared with the model group, all Zishenwan administration groups showed decreased prostate index, reduced levels of interleukin (IL)-1β, IL-18, and B-cell lymphoma-2 (Bcl-2) in serum (P<0.05, P<0.01), as well as varying degrees of alleviation in histopathological damage. At the same dose, compared with the raw Zishenwan groups, the salt-processed Zishenwan groups showed lower prostate index, pathological scores, and IL-1β, IL-18, and Bcl-2 levels in serum, but the differences were not statistically significant. Metabolomics reveals that 38 differential metabolites were reversed after salt-processed Zishenwan administration. Both raw and salt-processed Zishenwan regulated pathways such as β-alanine metabolism and tryptophan metabolism. In addition to the common regulated pathways, the salt-processed group specifically regulated pantothenate and coenzyme A biosynthesis, pyrimidine metabolism, and arginine and proline metabolism. The intersecting pathways between network pharmacology and metabolomics were tryptophan metabolism and arginine and proline metabolism, with overlapping targets including monoamine oxidase A (MAOA) and arginase 1 (ARG1). ConclusionThe increased contents of components such as berberine and phellodendrine in salt-processed Zishenwan may enhance its therapeutic effect on CP by inhibiting the PI3K/Akt and MAPK signaling pathways, along with multi-target regulation of tryptophan, arginine, and pantothenate metabolism pathways to comprehensively regulate inflammatory and immune responses.
5.Sleep Traits and Malignant Risk of Pulmonary Nodules: Evidence Triangulation From Questionnaire, Cohort, and Mendelian Randomization
Xiangyu CHEN ; Yiqiao XUE ; Mengqing LIU ; Yile HU ; Weizuo LIANG ; Hanqing LIU ; Yizheng WANG ; Mingfang ZHAO
Medical Journal of Peking Union Medical College Hospital 2026;17(3):663-676
To investigate the association between sleep-related phenotypes and the risk of malignancy in pulmonary nodules, and to provide complementary evidence from a general population cohort and genetic analyses. This study comprised three parts. Part 1 was a cross-sectional study that consecutively enrolled patients with imaging-confirmed pulmonary nodules at the First Hospital of China Medical University from November 2024 to December 2025. Nine sleep domains were constructed using items from the Pittsburgh sleep quality index (PSQI), with domain severity coded on a 0-6 scale according to the frequency of occurrence. Benign or malignant status of pulmonary nodules was determined based on pathological results or clinical follow-up. Multivariable Logistic regression models with progressive adjustment were constructed. Stratified, interaction, and dose-response analyses (including categorical grouping and restricted cubic splines) were performed focusing on the insomnia symptom domain to explore the association between sleep-related phenotypes and the risk of malignant pulmonary nodules. Part 2 was a prospective cohort study using the China Health and Retirement Longitudinal Study (CHARLS) to investigate the association between sleep duration and incident lung cancer risk in the general population. Part 3 comprised genetic causality analyses, including two-sample Mendelian randomization (MR) and linkage disequilibrium score regression (LDSC), using data from the OpenGWAS database, to assess whether directionally consistent genetic association signals exist between sleep-related phenotypes and lung cancer risk. In the cross-sectional study, a total of 800 patients with pulmonary nodules were included, of whom 288 (36.0%) were in the malignant group. In the continuous-variable main model fully adjusted for baseline confounders, all nine sleep domains, imaging findings, and depression and anxiety status, the severity of the insomnia symptom domain showed a positive association signal with the risk of malignant pulmonary nodules (fully adjusted model: per 1-point increase, In patients with pulmonary nodules, an association signal exists between insomnia-related symptoms and the risk of malignancy, but the dose-response relationship remains unclear. The CHARLS cohort and genetic analyses provide supplementary directional clues for the above associations, albeit with limited statistical strength and result consistency. Definitive conclusions regarding the association between sleep phenotypes and the risk of malignant pulmonary nodules require further validation in prospective studies.
6.Consideration of Health Economics Evidence in Clinical Practice Guidelines: Methods and Steps
Dongrui PENG ; Qi ZHOU ; Xufei LUO ; Zijun WANG ; Hui LIU ; Junxian ZHAO ; Jinghong HUANG ; Hongyu HU ; Xin XING ; Jing WU ; Shitong XIE ; Xiaohui WANG ; Yaolong CHEN
Medical Journal of Peking Union Medical College Hospital 2026;17(3):862-870
Health economics evidence plays an important role in linking clinical value evidence with health resource allocation decisions in the development of clinical practice guidelines. It can not only effectively balance clinical effectiveness and economic feasibility but also avoid forming "idealized" recommendations that are detached from the affordability of the healthcare system or the burden-bearing capacity of patients. To promote guideline developers to use health economics evidence more standardizedly and fully, this paper conducts an in-depth analysis of the current application status, existing challenges, access channels, and application processes of health economics evidence in current guidelines, and on this basis, puts forward considerations and suggestions for strengthening and standardizing the application of health economics evidence in China's clinical practice guidelines.
7.Safety of 3D printed titanium alloy bone trabecular cup prosthesis combined with modified Kidney Tonifying and Blood Activating Decoction for elderly hip arthroplasty
Jiangjing WANG ; Na ZHAO ; Xiaona HU ; Na ZHAO
Chinese Journal of Tissue Engineering Research 2026;30(3):612-619
BACKGROUND:The titanium alloy bone trabecular bone socket cup prosthesis printed by 3D technology has superior biological characteristics,which helps to promote osteogenic differentiation of stem cells.The Kidney Tonifying and Blood Activating Decoction has the functions of tonifying the kidneys,strengthening tendons,promoting blood circulation,and relieving pain.However,the application of the combination of the two in elderly hip arthroplasty is rarely reported.OBJECTIVE:To evaluate the application value and safety of 3D printed titanium alloy bone trabecular cup prosthesis combined with modified Kidney Tonifying and Blood Activating Decoction in elderly hip arthroplasty.METHODS:A total of 200 elderly patients who received hip arthroplasty in Cangzhou Hospital of Integrated Traditional Chinese and Western Medicine from January 2022 to January 2024 were selected and randomly divided into control group(n=100)and trial group(n=100).The control group underwent Pinnaele cup prosthesis hip arthroplasty combined with modified Kidney Tonifying and Blood Activating Decoction,while the trial group underwent 3D printed titanium alloy bone trabecular cup prosthesis hip arthroplasty combined with modified Kidney Tonifying and Blood Activating Decoction.The medication was taken continuously for 1 week before hip arthroplasty and for 4 weeks after arthroplasty.The differences in pain,swelling,hemorheology,inflammatory factors,bone metabolism,intra-articular stress,and hip abduction muscle strength were compared between the two groups to evaluate the clinical efficacy and safety of the two treatment options.RESULTS AND CONCLUSION:(1)Four weeks after surgery,the pain visual analog scale score,limb swelling,whole blood viscosity,and plasma viscosity of the trial group were lower than those of the control group(P<0.05).The interleukin-6 level of the trial group was lower than that of the control group,and the interleukin-10 level was higher than that of the control group(P<0.05).The levels of type Ⅰ collagen carboxyl terminal peptide β specific sequence and tartrate-resistant acid phosphatase 5b in the trial group were lower than those in the control group,while the levels of osteocalcin,bone-specific phosphatase and typeⅠ procollagen amino-terminal propeptide were higher than those in the control group(P<0.05).The intra-articular stress and hip abduction muscle strength in the trial group were higher than that of the control group(P<0.05).(2)Three months after surgery,the excellent and good treatment rate in the trial group was higher than that in the control group,and the incidence of safety evaluation events such as postoperative incision hematoma and infection was lower than that in the control group(P<0.05).(3)It suggests that 3D printed titanium alloy trabecular acetabular cup prosthesis combined with modified Kidney Tonifying and Blood Activating Decoction can improve the blood rheology of elderly patients with hip arthroplasty,inhibit inflammatory response,reduce postoperative pain and swelling,and enhance bone metabolism,thereby promoting postoperative hip joint function recovery,and reducing the incidence of adverse events,with high safety.
8.Effects of human umbilical cord blood platelet-rich plasma,mononuclear cells,and mesenchymal stem cells in repairing thin endometrium in rats
Yanli MU ; Anchun HU ; Wenchi XU ; Panpan CHEN ; Hao CHEN ; Shuyun ZHAO ; Guanyou HUANG ; Xiaojuan CHEN
Chinese Journal of Tissue Engineering Research 2026;30(1):78-92
BACKGROUND:Research has found that human umbilical cord blood platelet rich plasma and human umbilical cord blood derived mesenchymal stem cells have certain therapeutic effects on thin endometrium.However,there are currently no reports on the study of human umbilical cord blood mononuclear cells on thin endometrium,and there is a lack of relevant research comparing the three.OBJECTIVE:To explore the effects and mechanisms of human umbilical cord blood platelet rich plasma,monocytes,and mesenchymal stem cells in repairing thin endometrium in rats.METHODS:Sixty female SPF grade SD rats were randomly divided into sham operation group,model group,human umbilical cord blood platelet rich plasma group,human umbilical cord blood mononuclear cell group,and human umbilical cord blood derived mesenchymal stem cell group,with 12 rats in each group.The sham operation group received 0.5 mL physiological saline injection into the uterine horn,followed by 0.5 mL of PBS infusion after 5 minutes;The model group,human umbilical cord blood platelet rich plasma group,human umbilical cord blood mononuclear cell group,and human umbilical cord blood derived mesenchymal stem cell group were injected with 0.5 mL of 95%ethanol by volume.After 5 minutes,the remaining ethanol was aspirated and washed twice with physiological saline.Then,0.5 mL of PBS,human umbilical cord blood platelet rich plasma,human umbilical cord blood mononuclear cells(1×107 cells/mL),and human umbilical cord blood derived mesenchymal stem cells(1×107 cells/mL)were perfused separately.During the third normal estrus cycle after reperfusion,organs,tissues and serum were collected for testing of relevant indicators.RESULTS AND CONCLUSION:(1)The macroscopic view of uterine tissue,hematoxylin eosin staining and Masson staining results:the sham operation group had intact structure,moderate endometrial thickness,and clear vascular structure.Compared with the sham operation group,the model group showed uterine atrophy,incomplete structure,significantly reduced endometrial thickness and glandular quantity,disordered vascular structure,and increased fibrosis.Compared with the model group,after treatment with human umbilical cord blood derivatives,the size,structure,and endometrial thickness of the uterus were restored(all P<0.01),and fibrosis was reduced,with the most significant recovery observed in the human umbilical cord blood mononuclear cell group.The increase in glandular quantity was most significant in the human umbilical cord blood platelet rich plasma group(P<0.000 1).(2)The immunohistochemistry and immunofluorescence results of uterine tissue showed that compared with the sham operation group,the expression levels of cell proliferation related indicators such as keratin 9 and vimentin,endometrial receptivity related indicators such as leukemia inhibitory factor and integrin αyβ3,platelet endothelial cell adhesion molecule,basic fibroblast growth factor,and vascular endothelial growth factor were all reduced in the model group(all P<0.05).Compared with the model group,the above indicators were significantly increased after treatment with human umbilical cord blood derivatives.Comparison of human umbilical cord blood derivatives among groups showed that keratin 9 and vascular endothelial growth factor protein:human umbilical cord blood mononuclear cell group>human umbilical cord blood derived mesenchymal stem cell group>human umbilical cord blood platelet rich plasma group;Wave shaped protein and leukemia inhibitory factor protein:human umbilical cord blood derived mesenchymal stem cell group>human umbilical cord blood mononuclear cell group>human umbilical cord blood platelet rich plasma group;Integrin αyβ3 protein:human umbilical cord blood platelet rich plasma group>human umbilical cord blood derived mesenchymal stem cell group>human umbilical cord blood mononuclear cell group;Platelet endothelial cell adhesion molecule protein:human umbilical cord blood platelet rich plasma group>human umbilical cord blood mononuclear cell group>human umbilical cord blood derived mesenchymal stem cell group;Basic fibroblast growth factor protein:human umbilical cord blood mononuclear cell group>human umbilical cord blood platelet rich plasma group>human umbilical cord blood derived mesenchymal stem cell group.(3)Western blot analysis showed that compared with the sham operation group,the protein levels of interleukin-6,interleukin-1β,and tumor necrosis factor alpha in the model group were significantly increased(all P<0.001),and their expression levels decreased after treatment(all P<0.05).(4)ELISA assay showed that compared with the sham operation group,the model group had lower levels of anti Mullerian hormone,estradiol,and progesterone,and increased levels of follicle stimulating hormone and luteinizing hormone(except for luteinizing hormone,all P<0.05).After treatment,there was a certain degree of recovery in the levels of sex hormones and anti Mullerian hormones.(5)Fertility experiments showed that compared with the sham operation group,the model group had an increase in conception time and a significant decrease in litter size(all P<0.05).After treatment with human umbilical cord blood derivatives,the litter size of all three groups increased(P<0.05),and no significant differences were found between the groups.This study preliminarily reveals that human umbilical cord blood mononuclear cells have a certain therapeutic effect on thin endometrium,and human umbilical cord blood platelet rich plasma,human umbilical cord blood mononuclear cells,and human umbilical cord blood derived mesenchymal stem cells have different advantages and differences in improving endometrial regeneration function,endometrial receptivity,angiogenesis,inflammation regulation,and improving pregnancy outcomes in thin endometrium.
9.Mechanisms of Mahuang Lianqiao Chixiaodoutang in Improving Obesity-type Polycystic Ovary Syndrome in Rats Based on PI3K/Akt Signaling Pathway
Shiwei HU ; Biran ZHU ; Jinrong ZHANG ; Luyao RUAN ; Ji KUANG ; Jianghuan HUA ; Zhe LIU ; Yanyue YAO ; Ji WANG ; Min ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):21-31
ObjectiveTo investigate the mechanisms by which Mahuang Lianqiao Chixiaodoutang (MLC) improves obesity-type polycystic ovary syndrome (PCOS) through the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway. MethodsThirty-six female Sprague-Dawley (SD) rats were randomly divided into a blank control group (Con) and an obesity-type PCOS model preparation group. The model was induced by gavage with letrozole (1 mg·kg-1) combined with a high-fat diet (HFD). After model establishment, the obesity-type PCOS model preparation group was further divided into the model group (Mod, normal saline), metformin group (Met, 0.3 g·kg-1), low-dose MLC group (MLC-L, 4.3 g·kg-1), medium-dose MLC group (MLC-M, 8.6 g·kg-1), and high-dose MLC group (MLC-H, 17.2 g·kg-1). Active components of MLC and targets of obesity-type PCOS were screened from databases, a protein-protein interaction (PPI) network was constructed, and gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was performed. The gut microbiota structure was analyzed based on 16S rRNA sequencing and correlated with network pharmacology pathways. Body weight and estrous cycle were dynamically monitored. Ovarian morphology was observed by hematoxylin-eosin (HE) staining. Cell apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). Enzyme-linked immunosorbent assay (ELISA) was used to detect levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), anti-Müllerian hormone (AMH), testosterone (T), and estradiol (E2). Western blot was used to detect the protein expression levels of phosphorylated PI3K/PI3K (p-PI3K/PI3K), phosphorylated Akt/Akt (p-Akt/Akt), B-cell lymphoma-2 (Bcl-2), and Bcl-2-associated X protein (Bax). ResultsNetwork pharmacology screening identified 124 active components of MLC and 408 overlapping targets between the herbal formula and the disease. Core targets such as Akt1 and Bcl-2 were revealed. As indicated by 16S rRNA sequencing, the abundances of Lachnospiraceae, Lachnoclostridium, and Dorea were increased in the MLC groups (P<0.05), while the abundance of Veillonella was decreased (P<0.05). KEGG correlation analysis integrating network pharmacology and gut microbiota data showed significant enrichment of the PI3K/Akt signaling pathway. Animal experiments showed that, compared with the Mod group, body weight decreased to normal levels in the Met, MLC-M, and MLC-H groups. The estrous cycle became regular. The number of corpora lutea increased and cystic follicles decreased. Serum levels of T, FSH, and LH/FSH were reduced (P<0.05, P<0.01), while the E2 level was increased (P<0.01). Ovarian cell apoptosis was reduced (P<0.01), and the protein expression levels of p-PI3K/PI3K, p-Akt/Akt, and Bcl-2 in ovarian tissue were significantly increased, whereas Bax protein expression was significantly decreased (P<0.05, P<0.01). ConclusionMLC can regulate gut microbiota structure, effectively improve ovarian pathology in rats with obesity-type PCOS, and inhibit ovarian granulosa cell apoptosis. The mechanism may be associated with upregulation of the PI3K/Akt signaling pathway.
10.Mechanisms of Mahuang Lianqiao Chixiaodoutang in Improving Obesity-type Polycystic Ovary Syndrome in Rats Based on PI3K/Akt Signaling Pathway
Shiwei HU ; Biran ZHU ; Jinrong ZHANG ; Luyao RUAN ; Ji KUANG ; Jianghuan HUA ; Zhe LIU ; Yanyue YAO ; Ji WANG ; Min ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):21-31
ObjectiveTo investigate the mechanisms by which Mahuang Lianqiao Chixiaodoutang (MLC) improves obesity-type polycystic ovary syndrome (PCOS) through the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway. MethodsThirty-six female Sprague-Dawley (SD) rats were randomly divided into a blank control group (Con) and an obesity-type PCOS model preparation group. The model was induced by gavage with letrozole (1 mg·kg-1) combined with a high-fat diet (HFD). After model establishment, the obesity-type PCOS model preparation group was further divided into the model group (Mod, normal saline), metformin group (Met, 0.3 g·kg-1), low-dose MLC group (MLC-L, 4.3 g·kg-1), medium-dose MLC group (MLC-M, 8.6 g·kg-1), and high-dose MLC group (MLC-H, 17.2 g·kg-1). Active components of MLC and targets of obesity-type PCOS were screened from databases, a protein-protein interaction (PPI) network was constructed, and gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was performed. The gut microbiota structure was analyzed based on 16S rRNA sequencing and correlated with network pharmacology pathways. Body weight and estrous cycle were dynamically monitored. Ovarian morphology was observed by hematoxylin-eosin (HE) staining. Cell apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). Enzyme-linked immunosorbent assay (ELISA) was used to detect levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), anti-Müllerian hormone (AMH), testosterone (T), and estradiol (E2). Western blot was used to detect the protein expression levels of phosphorylated PI3K/PI3K (p-PI3K/PI3K), phosphorylated Akt/Akt (p-Akt/Akt), B-cell lymphoma-2 (Bcl-2), and Bcl-2-associated X protein (Bax). ResultsNetwork pharmacology screening identified 124 active components of MLC and 408 overlapping targets between the herbal formula and the disease. Core targets such as Akt1 and Bcl-2 were revealed. As indicated by 16S rRNA sequencing, the abundances of Lachnospiraceae, Lachnoclostridium, and Dorea were increased in the MLC groups (P<0.05), while the abundance of Veillonella was decreased (P<0.05). KEGG correlation analysis integrating network pharmacology and gut microbiota data showed significant enrichment of the PI3K/Akt signaling pathway. Animal experiments showed that, compared with the Mod group, body weight decreased to normal levels in the Met, MLC-M, and MLC-H groups. The estrous cycle became regular. The number of corpora lutea increased and cystic follicles decreased. Serum levels of T, FSH, and LH/FSH were reduced (P<0.05, P<0.01), while the E2 level was increased (P<0.01). Ovarian cell apoptosis was reduced (P<0.01), and the protein expression levels of p-PI3K/PI3K, p-Akt/Akt, and Bcl-2 in ovarian tissue were significantly increased, whereas Bax protein expression was significantly decreased (P<0.05, P<0.01). ConclusionMLC can regulate gut microbiota structure, effectively improve ovarian pathology in rats with obesity-type PCOS, and inhibit ovarian granulosa cell apoptosis. The mechanism may be associated with upregulation of the PI3K/Akt signaling pathway.

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