1.Effect of Gynostemma pentaphyllum Alcohol Extract on Glucose and Lipid Metabolism Disorders in db/db Mice Based on Transcriptomics and Gut Microbiota
Yifei ZHU ; Lei DING ; Wei LIU ; Yahui SUN ; Lingling QIN ; Lili WU ; Tonghua LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):80-89
ObjectiveTo investigate the efficacy and underlying mechanisms of Gynostemma pentaphyllum alcohol extract in improving glucose and lipid metabolism disorders in db/db mice through transcriptomics and gut microbiota analysis. MethodsEighteen db/db mice were randomly assigned to the model(DM) group, metformin(MET) group, and G. pentaphyllum alcohol extract(GP) group, with six mice in each group, based on stratification of fasting blood glucose and body weight. An additional six db/m mice were selected as the normal control(NC) group. Mice in the NC and DM groups were administered deionized water (10 mL·kg-1) daily. The MET group received metformin (0.195 g·kg-1) by gavage. The GP group was treated with G. pentaphyllum alcohol extract (3.9 g·kg-1) by gavage for six weeks. Fasting blood glucose was measured every two weeks. After six weeks of intervention, serum levels of total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatinine (CREA), and blood urea nitrogen (BUN) were assessed. Enzyme-linked immunosorbent assay (ELISA) was used to measure insulin (FINS), adiponectin (ADP), and tumor necrosis factor-α (TNF-α). Hematoxylin-eosin (HE) staining was used to observe liver histomorphology, periodic acid-Schiff (PAS) staining was employed to assess hepatic glycogen synthesis, and Oil Red O staining was used to detect hepatic lipid deposition. Liver transcriptomic data were used to identify differentially expressed genes in the liver and conduct enrichment analysis. Real-time PCR was employed to verify the expression levels of adiponectin gene (Adipoq), peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α), AMP-activated protein kinase (AMPK), peroxisome proliferator-activated receptor α (PPARα), glucokinase (GCK), forkhead box (Fox)O1, FoxO3, phosphoenolpyruvate carboxykinase (PEPCK), and glucose-6-phosphatase (G6PC). Metagenomic sequencing was conducted to analyze changes in gut microbiota composition. ResultsCompared with the NC group, the DM group exhibited significantly elevated fasting blood glucose (P<0.01), serum AST, ALT, TC, TG, LDL-C, and HDL-C (P<0.01). FINS, homeostatic model assessment for insulin resistance (HOMA-IR), and the inflammatory cytokine TNF-α were significantly increased (P<0.01), while ADP was significantly decreased (P<0.05). Histological analysis confirmed severe hepatic steatosis and excessive lipid accumulation in the DM group, along with markedly reduced glycogen synthesis. Compared with the DM group, the GP group showed significantly decreased fasting blood glucose (P<0.01), reduced serum TC, LDL-C, and HDL-C levels (P<0.05), significantly decreased serum TG and AST levels (P<0.01), significantly reduced FINS, HOMA-IR, and TNF-α levels (P<0.01), and significantly increased ADP (P<0.01). Hepatic steatosis and lipid deposition were significantly alleviated, while glycogen synthesis was markedly enhanced. Transcriptomic differential and enrichment analyses suggested that the mechanisms by which G. pentaphyllum alcohol extract improved hepatic glucose and lipid metabolism in db/db mice may involve regulation of the AMPK and FoxO signaling pathways. Real-time PCR results confirmed that expression of PGC-1α, PEPCK, G6PC, FoxO1, and FoxO3 was significantly downregulated following treatment with G. pentaphyllum alcohol extract (P<0.05, P<0.01), whereas mRNA expression of Adipoq, PPARα, GCK, and AMPK was significantly upregulated (P<0.05, P<0.01). Metagenomic analysis showed that the relative abundance of Lactobacillus, Alistipes, and Akkermansia species was higher in the GP group than in the DM group. ConclusionG. pentaphyllum alcohol extract may improve glucose and lipid metabolism disorders in db/db mice by regulating the hepatic AMPK/PPARα pathway to suppress lipid deposition and alleviate hepatic steatosis, by inhibiting gluconeogenesis through the AMPK/PGC-1α and FoxO pathways to lower fasting blood glucose, and by increasing the abundance of beneficial gut bacteria such as Lactobacillus, Alistipes, and Akkermansia to restore gut microbiota balance.
2.Effect of Gynostemma pentaphyllum Alcohol Extract on Glucose and Lipid Metabolism Disorders in db/db Mice Based on Transcriptomics and Gut Microbiota
Yifei ZHU ; Lei DING ; Wei LIU ; Yahui SUN ; Lingling QIN ; Lili WU ; Tonghua LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):80-89
ObjectiveTo investigate the efficacy and underlying mechanisms of Gynostemma pentaphyllum alcohol extract in improving glucose and lipid metabolism disorders in db/db mice through transcriptomics and gut microbiota analysis. MethodsEighteen db/db mice were randomly assigned to the model(DM) group, metformin(MET) group, and G. pentaphyllum alcohol extract(GP) group, with six mice in each group, based on stratification of fasting blood glucose and body weight. An additional six db/m mice were selected as the normal control(NC) group. Mice in the NC and DM groups were administered deionized water (10 mL·kg-1) daily. The MET group received metformin (0.195 g·kg-1) by gavage. The GP group was treated with G. pentaphyllum alcohol extract (3.9 g·kg-1) by gavage for six weeks. Fasting blood glucose was measured every two weeks. After six weeks of intervention, serum levels of total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatinine (CREA), and blood urea nitrogen (BUN) were assessed. Enzyme-linked immunosorbent assay (ELISA) was used to measure insulin (FINS), adiponectin (ADP), and tumor necrosis factor-α (TNF-α). Hematoxylin-eosin (HE) staining was used to observe liver histomorphology, periodic acid-Schiff (PAS) staining was employed to assess hepatic glycogen synthesis, and Oil Red O staining was used to detect hepatic lipid deposition. Liver transcriptomic data were used to identify differentially expressed genes in the liver and conduct enrichment analysis. Real-time PCR was employed to verify the expression levels of adiponectin gene (Adipoq), peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α), AMP-activated protein kinase (AMPK), peroxisome proliferator-activated receptor α (PPARα), glucokinase (GCK), forkhead box (Fox)O1, FoxO3, phosphoenolpyruvate carboxykinase (PEPCK), and glucose-6-phosphatase (G6PC). Metagenomic sequencing was conducted to analyze changes in gut microbiota composition. ResultsCompared with the NC group, the DM group exhibited significantly elevated fasting blood glucose (P<0.01), serum AST, ALT, TC, TG, LDL-C, and HDL-C (P<0.01). FINS, homeostatic model assessment for insulin resistance (HOMA-IR), and the inflammatory cytokine TNF-α were significantly increased (P<0.01), while ADP was significantly decreased (P<0.05). Histological analysis confirmed severe hepatic steatosis and excessive lipid accumulation in the DM group, along with markedly reduced glycogen synthesis. Compared with the DM group, the GP group showed significantly decreased fasting blood glucose (P<0.01), reduced serum TC, LDL-C, and HDL-C levels (P<0.05), significantly decreased serum TG and AST levels (P<0.01), significantly reduced FINS, HOMA-IR, and TNF-α levels (P<0.01), and significantly increased ADP (P<0.01). Hepatic steatosis and lipid deposition were significantly alleviated, while glycogen synthesis was markedly enhanced. Transcriptomic differential and enrichment analyses suggested that the mechanisms by which G. pentaphyllum alcohol extract improved hepatic glucose and lipid metabolism in db/db mice may involve regulation of the AMPK and FoxO signaling pathways. Real-time PCR results confirmed that expression of PGC-1α, PEPCK, G6PC, FoxO1, and FoxO3 was significantly downregulated following treatment with G. pentaphyllum alcohol extract (P<0.05, P<0.01), whereas mRNA expression of Adipoq, PPARα, GCK, and AMPK was significantly upregulated (P<0.05, P<0.01). Metagenomic analysis showed that the relative abundance of Lactobacillus, Alistipes, and Akkermansia species was higher in the GP group than in the DM group. ConclusionG. pentaphyllum alcohol extract may improve glucose and lipid metabolism disorders in db/db mice by regulating the hepatic AMPK/PPARα pathway to suppress lipid deposition and alleviate hepatic steatosis, by inhibiting gluconeogenesis through the AMPK/PGC-1α and FoxO pathways to lower fasting blood glucose, and by increasing the abundance of beneficial gut bacteria such as Lactobacillus, Alistipes, and Akkermansia to restore gut microbiota balance.
3.Allogeneic intrastromal lenticule implantation combined with corneal collagen cross-linking for moderate to advanced keratoconus
Jing ZHANG ; Jie HOU ; Yahui DONG ; Yulin LEI ; Yafei XU ; Fangfang SUN
International Eye Science 2025;25(9):1517-1522
AIM: To evaluate the safety and efficacy of allogeneic intrastromal lenticule implantation combined with corneal collagen cross-linking(CXL)in patients with moderate to advanced keratoconus.METHODS: A retrospective case series analysis was conducted. A total of 19 patients(20 eyes)with moderate to advanced keratoconus who underwent combined allogeneic intrastromal lenticule implantation and CXL at the Jinan Mingshui Eye Hospital from June 2021 to December 2023 were included. The uncorrected distance visual acuity(UCVA), thinnest corneal thickness, central corneal epithelial thickness, anterior corneal flat keratometry(Kf), steep keratometry(Ks), and mean keratometry(Km), as well as the first applanation time(A1T), the first applanation length(A1L), the velocity during the first applanation moment(VIN), the second applanation time(A2T), the second applanation length(A2L), the velocity during the second applanation moment(VOUT), highest concavity time(HCT), highest concavity radius(HCR), peak distance(PD), deformation amplitude(DA), stiffness parameter at first applanation(SP-A1), integrated radius(IR), central corneal thickness(CCT), intraocular pressure(IOP), corneal thickness-corrected IOP, biomechanically intraocular pressure IOP(bIOP), and corneal thickness variation rate(ARTH)were compared between the two groups before surgery and at 1 wk, 1, 3 and 6 mo after surgery.RESULTS: All patients successfully completed the surgery without any intraoperative complications. No significant differences were observed between pre-operative and post-operative measurements for UCVA or the corneal biomechanical parameters, including A1L, A2L, PD, A1T, A2T, VIN, VOUT, DA, IOP, and bIOP(all P>0.05). Significant differences were found between pre-operative and post-operative values for corneal thinnest point thickness, central corneal epithelial thickness, Kf, Ks, Km, and the corneal biomechanical parameters, including HCT, HCR, SP-A1, ARTH, IR, and CCT(all P<0.05). The anterior corneal curvature demonstrated an initial increase followed by a decrease post-operatively. Furthermore, significant differences were observed between pre-operative and post-operative values for HCT, HCR, SP-A1, ARTH, IR, and CCT(all P<0.005).CONCLUSION: Allogenic intrastromal lenticule implantation combined with corneal collagen cross-linking demonstrates favorable safety and stability in treating moderate-to-advanced keratoconus. This combined procedure effectively increases corneal thickness and rigidity, resulting in corneas that are more resistant to deformation postoperatively.
4.Shugan Jianpi Yangxin Prescription Regulates Orexin-A and Its Receptors in Mouse Model of Insomnia
Zhen WANG ; Jie ZHANG ; Feixiang LIU ; Jiangyan SUN ; Yahui GAO ; Yuxin ZHENG ; Min ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(10):54-61
ObjectiveTo investigate the interventional effects of Shugan Jianpi Yangxin prescription on the expression of orexin-A (OXA), orexin-1 receptor (OX1R), and orexin-2 receptor (OX2R) in the mouse model of insomnia. MethodThe mouse model of insomnia was established by intraperitoneal injection of DL-4-chlorophenylalanine (PCPA). Fifty BALB/c mice were randomized into a blank group, a model group, an eszopiclone (0.13 mg·kg-1) group, and low- and high-dose (8.4 and 33.6 g·kg-1, respectively) Shugan Jianpi Yangxin prescription groups and treated with the corresponding drugs for 14 consecutive days. The weight changes of mice were monitored, and Morris water maze and pentobarbital-induced sleep tests were conducted. Immunohistochemistry (IHC) was employed to examine the expression of OXA in the hypothalamus. Enzyme-linked immunosorbent assay was used to measure the levels of OXA and 5-hydroxytryptamine (5-HT) in the hypothalamus, serum, and spleen. Real-time fluorescence quantitative polymerase chain reaction was employed to determine the mRNA levels of OXA, OX1R, and OX2R in the hypothalamus. ResultCompared with the blank group, the model group had decreased body weight (P<0.01), increased escape latency (P<0.01), increased sleep latency (P<0.01), shortened sleep duration (P<0.01), elevated OXA level and lowered 5-HT level in the hypothalamus, serum, and spleen (P<0.05), and up-regulated mRNA levels of OXA, OX1R, and OX2R in the hypothalamus (P<0.01). Compared with the model group, the low- and high-dose groups of Shugan Jianpi Yangxin prescription showed increased body weight (P<0.05, P<0.01), shortened escape latency (P<0.05), shortened sleep latency and prolonged sleep duration (P<0.01), and lowered OXA level and elevated 5-HT level in the hypothalamus, serum, and spleen (P<0.05, P<0.01). Moreover, the two doses of Shugan Jianpi Yangxin prescription down-regulated the mRNA levels of OXA, OX1R, and OX2R in the hypothalamus (P<0.01). ConclusionShugan Jianpi Yangxin prescription exerts sedative and hypnotic effects in mice by increasing the content of 5-HT in the brain and inhibiting the expression of OXA and its receptors in the hypothalamus.
5.Electrocardiographic characteristics and their correlation with indicators of disease severity in patients with chronic pulmonary artery stenosis
Mingjun DENG ; Yahui SUN ; Yao MI ; Kaiyu JIANG ; Aqian WANG ; Hongling SU ; Yunshan CAO
Chinese Journal of General Practitioners 2024;23(2):146-152
Objective:To analyze the electrocardiographic characteristics of patients with chronic pulmonary artery stenosis (PAS), and to explore their relationship with disease severity indicators.Methods:The study was a retrospective case-series analysis. Patients with chronic PAS admitted to Gansu Provincial Hospital from January 2018 to July 2021 were enrolled. The clinical data and the results of electrocardiography, transthoracic echocardiography, right cardiac catheterization, N-terminal B-type natriuretic peptide (NT-proBNP) measurement and 6-min walking distance test of patients were analyzed. The linear regression model or logistic regression model was used to analyze the relationship between electrocardiographic characteristics and the disease severity in patients with chronic PAS.Results:Sixty-three patients aged (62.1±9.7) years including 43 females (68.3%) were enrolled in the study. Among them, 62 patients (98.4%) had (R 1+S Ⅲ)-(S Ⅰ+R Ⅲ)<1.5 mV, and no patients had V 5lead R: S ratio to V 1 lead R: S<0.04 and V 6 lead R: S ratio<0.4. There were 55 patients (87.3%), with flat or inverted T-waves in V 1, and 10 patients (15.9%) with flat or inverted T-waves in all precordial leads (V 1-V 6). There were 18 patients (28.6%) with flat or inverted T-waves in inferior leads (Ⅱ, Ⅲ, aVF). Multiple liner regression analysis showed that Max R V1, 2+Max S I, aVL-S V1 combined with the number of flat or inverted T-waves in limb leads was independently correlated with atrial area ( R2=0.290, P=0.002); R V1+S V5 was independently correlated with right ventricular area ( R2=0.257, P=0.001); R peak V 1 combined with the number of flat or inverted T waves in precordial leads was independently correlated with tricuspid annular plane systolic excursion ( R2=0.407, P<0.001); (R 1+S Ⅲ)-(S Ⅰ+R Ⅲ) combined with the number of flat or inverted T waves in precordial leads was independently correlated with NT-proBNP ( R2=0.504, P<0.001); Max R V1, 2+Max S I, aVL-S V1 were independently correlated with right atrial pressure ( R2=0.803, P=0.036); (R 1+S Ⅲ)-(S Ⅰ+R Ⅲ) were independently correlated with mean pulmonary artery pressure ( R2=0.302, P<0.001); R aVRcombined with the number of flat or inverted T-waves in precordial leads was independently correlated with cardiac index ( R2=0.173, P=0.003); (R 1+S Ⅲ)-(S Ⅰ+R Ⅲ) was independently correlated with pulmonary vascular resistance ( R2=0.173, P=0.002); R peak V 1 combined with the number of flat or inverted T-waves in precordial leads was independently correlated with mixed vein oxygen saturation ( R2=0.302, P<0.001). Conclusion:The vast majority of patients with chronic PAS have (R 1+S Ⅲ)-(S Ⅰ+R Ⅲ)<1.5 mV and flat or inverted T-wave in V 1 lead, and some characteristic electrocardiographic manifestations are correlated with indicators of disease severity.
6.Model informed precision medicine of Chinese herbal medicines formulas-A multi-scale mechanistic intelligent model
Qian YUANYUAN ; Wang XITING ; Cai LULU ; Han JIANGXUE ; Huang ZHU ; Lou YAHUI ; Zhang BINGYUE ; Wang YANJIE ; Sun XIAONING ; Zhang YAN ; Zhu AISONG
Journal of Pharmaceutical Analysis 2024;14(4):585-600
Recent trends suggest that Chinese herbal medicine formulas(CHM formulas)are promising treatments for complex diseases.To characterize the precise syndromes,precise diseases and precise targets of the precise targets between complex diseases and CHM formulas,we developed an artificial intelligence-based quantitative predictive algorithm(DeepTCM).DeepTCM has gone through multilevel model cali-bration and validation against a comprehensive set of herb and disease data so that it accurately captures the complex cellular signaling,molecular and theoretical levels of traditional Chinese medicine(TCM).As an example,our model simulated the optimal CHM formulas for the treatment of coronary heart disease(CHD)with depression,and through model sensitivity analysis,we calculated the balanced scoring of the formulas.Furthermore,we constructed a biological knowledge graph representing interactions by associating herb-target and gene-disease interactions.Finally,we experimentally confirmed the thera-peutic effect and pharmacological mechanism of a novel model-predicted intervention in humans and mice.This novel multiscale model opened up a new avenue to combine"disease syndrome"and"macro micro"system modeling to facilitate translational research in CHM formulas.
7.Stratified management for cardiovascular diseases risk in community population based on China-PAR
Xue BAI ; Ying NIE ; Shihuan LUO ; Xinhuan ZHANG ; Ronghui XIA ; Yahui SUN ; Yuntao LU
Chinese Journal of General Practitioners 2023;22(3):271-277
Objective:To evaluate the effect of stratified management of cardiovascular diseases risk in community population based on China-PAR.Methods:It was a single arm study. Beijing Jiaotong University faculty and staff who participated in annual health check-up from 2019 to 2021 and met the inclusion/exclusion criteria were enrolled in the study. The general data, physical examination and laboratory test results, including age, residence region, waist circumference, total cholesterol, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C),blood pressure (BP), taking antihypertensive drugs,diabetes, family history of cardiovascular diseases,were collected. Participants were stratified according to China-PAR assessment model and then stratified management was carried out. For low-risk populations, family doctors provided online guidance or outpatient follow-up if necessary after their first interview. For middle and high-risk groups, outpatient and telephone follow-up were arranged in addition to online guidance. Relevant examinations were completed and drug treatment or adjustment were given by doctors when necessary. Frequency of outpatient follow-up for middle and high-risk groups was different and patients in these two groups were scored again at the end of 2-year follow-up.Results:A total 284 participants were enrolled,197 participants (69.4%) were males with a age of (46.9±8.8) years. Among them, 205 participants (72.2%) were in low-risk group, including 136 males (66.3%), and their 10-year risk of cardiovascular diseases was (2.5±0.1)%; 59 participants (20.8%) were in middle-risk group, including 43 males (72.9%), and their 10-year risk of cardiovascular diseases was (7.1±0.2)%;20 participants (7.0%) were in high-risk group,including 18 males (90.0%) and their 10-year risk of cardiovascular diseases was (14.0±1.1)%. After 2 years follow-up, the proportion of dietary imbalances and alcohol drinking, waist circumference, blood pressure, fasting glucose levels and risk score decreased significantly in high risk group ( P<0.05). The proportion of dietary imbalances, waist circumference, blood pressure, total cholesterol, and LDL-C levels decreased significantly in medium risk group ( P<0.05). In high-risk group, 2 participants (10.0%) converted to low-risk, 8 participants (40.0%) converted to middle-risk. In middle-risk group, 5 participants (8.5%) converted to low-risk and 7 participants (11.9%) converted to high-risk. Conclusion:The risk factors and risk stratification of cardiovascular disease in community population can be improved by stratified management based on China-PAR risk assessment model.
8.Prevalence of subclinical thyroid disease in the faculty and staff of a university and their affecting factors
Shihuan LUO ; Ying NIE ; Xinhuan ZHANG ; Xue BAI ; Yahui SUN ; Lingwei KONG ; Yifei WANG
Chinese Journal of General Practitioners 2023;22(10):1032-1037
Objective:To investigate the status quo of subclinical thyroid diseases in the faculty and staff of a university and to explore their affecting factors.Methods:A total of 4 219 faculty and staff members who met the exclusion criteria and underwent the health examination in the Community Health Service Center of Beijing Jiaotong University in 2021 were enrolled in the study. General clinical data and laboratory findings of the enrolled subjects were collected. According to the upper and low reference range of thyroid stimulating hormone (TSH) in our laboratory (0.35-5.5 μIU/ml), subjects were classified into subclinical hyperthyroidism group, subclinical hypothyroidism group and normal thyroidism group. The association of gender, age and body mass index (BMI), as well as the metabolic indices with the prevalence of subclinical thyroid disease was analyzed.Results:The prevalence rates of subclinical hypothyroidism and subclinical hyperthyroidism were 4.10% (173/4 219) and 0.69% (29/4 219), respectively. The prevalence of subclinical thyroid diseases in females was higher than that in males(5.90% (77/2 101) vs. 3.66%(125/2 018),χ 2=11.58, P<0.05); there was a significant difference in prevalence among different age groups(χ 2=39.49, P<0.05)and the prevalence increased with the age. There were significant differences in levels of total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), diastolic blood pressure(DBP), fasting blood glucose (FBG), free triiodothyronine (FT 3), free thyroxine (FT 4), thyroglobulin antibody (TGAb) and thyroid peroxidase antibody (TPOAb) among three groups ( P<0.05). TSH levels were positively correlated with the age ( r=0.58, P<0.001), and levels of TG ( r=0.66, P<0.001), TC ( r=0.67, P<0.001), LDL-C ( r=0.62, P<0.001), TPOAb ( r=0.78, P<0.001), TGAb ( r=0.77, P<0.001); was negatively correlated with FBG ( r=-0.50, P<0.001). Conclusion:The prevalence of subclinical thyroid diseases among faculty and staff of the studied university is relatively high, and it is related to gender, age, thyroid antibodies, blood glucose and lipid levels.
9.Platycodon grandiflorus polysaccharide regulates colonic immunity through mesenteric lymphatic circulation to attenuate ulcerative colitis.
Yang LIU ; Yahui DONG ; Wei SHEN ; Jiahui DU ; Quanwei SUN ; Ye YANG ; Dengke YIN
Chinese Journal of Natural Medicines (English Ed.) 2023;21(4):263-278
Platycodon grandiflorus polysaccharide (PGP) is one of the main components of P. grandiflorus, but the mechanism of its anti-inflammatory effect has not been fully elucidated. The aim of this study was to evaluate the therapeutic effect of PGP on mice with dextran sodium sulfate (DSS)-induced ulcerative colitis (UC) and explore the underlying mechanisms. The results showed that PGP treatment inhibited the weight loss of DSS-induced UC mice, increased colon length, and reduced DAI, spleen index, and pathological damage within the colon. PGP also reduced the levels of pro-inflammatory cytokines and inhibited the enhancement of oxidative stress and MPO activity. Meanwhile, PGP restored the levels of Th1, Th2, Th17, and Treg cell-related cytokines and transcription factors in the colon to regulate colonic immunity. Further studies revealed that PGP regulated the balance of colonic immune cells through mesenteric lymphatic circulation. Taken together, PGP exerts anti-inflammatory and anti-oxidant effect and regulates colonic immunity to attenuate DSS-induced UC through mesenteric lymphatic circulation.
Animals
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Mice
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Colitis, Ulcerative/drug therapy*
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Platycodon
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Colon/pathology*
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Cytokines
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Anti-Inflammatory Agents/therapeutic use*
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Polysaccharides/therapeutic use*
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Dextran Sulfate
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Disease Models, Animal
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Colitis/chemically induced*
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Mice, Inbred C57BL
10.Aurantii Fructus Immaturus, Atractylodis Macrocephalae Rhizoma, and Their Combination Treat Colonic Motility Disorders in Rats with Slow Transit Constipation via PINK1/Parkin Signaling Pathway-mediated Mitophagy
Xiaopeng WANG ; Huiju YANG ; Mingming SUN ; Jing LIU ; Bensheng WU ; Yinzi YUE ; Yuanyuan QIN ; Yinghui CHEN ; Lie TIAN ; Yan LI ; Yahui WANG ; Shuai YAN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(13):45-53
ObjectiveTo observe the effects of Aurantii Fructus Immaturus, Atractylodis Macrocephalae Rhizoma, and their combination on slow transit constipation via PTEN-induced putative kinase 1 (PINK1)/Parkin pathway-mediated mitophagy. MethodFifty-six male SD rats were randomly assigned into normal group, model group, natural recovery group, Aurantii Fructus Immaturus group, Atractylodis Macrocephalae Rhizoma group, Aurantii Fructus Immaturus combined with Atractylodis Macrocephalae Rhizoma group, and mosapride group, with 8 rats in each group. Slow transit constipation model was established by gavage with loperamide (3 mg·kg-1·d-1) for 14 days in other groups except the normal group. After successful modeling, except that the model group was continuously induced by loperamide, the normal group and the natural recovery group were administrated with 0.9% normal saline by gavage, and the rats in the Aurantii Fructus Immaturus (1.35 g·kg-1·d-1) group, the Atractylodis Macrocephalae Rhizoma (2.7 g·kg-1·d-1) group, the Aurantii Fructus Immaturus combined with Atractylodis Macrocephalae Rhizoma (4.05 g·kg-1·d-1) group, and the mosapride (1.56 mg·kg-1·d-1) group were administrated with corresponding drugs by gavage for 7 days. The amount of feces, fecal water content, and intestinal propulsion rate of rats were determined. The pathological changes of the colon were evaluated by hematoxylin-eosin (HE) staining and Alcian blue-periodic acid-Schiff (AB-PAS) staining. The activity of respiratory chain complex and the ultrastructure of the colon tissue were determined by ultraviolet spectrophotometry and observed by transmission electron microscopy, respectively. Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) was employed to determine the mRNA levels of PINK1, Parkin, and p62, and Western blot to determine the protein levels of microtubule-associated protein 1 light chain 3 (LC3), PINK1, and Parkin. ResultCompared with the normal group, the model group and the natural recovery group showed decreases in the amount of feces, fecal water content, intestinal propulsion rate (P<0.05,P<0.01), and activities of mitochondrial respiratory chain complexes Ⅱ, Ⅲ, and Ⅳ in the colon tissue (P<0.05,P<0.01). Further, the mRNA levels of PINK1 and Parkin and the protein levels of PINK1, Parkin, and LC3 were up-regulated (P<0.01) and the mRNA level of p62 was down-regulated in the model group (P<0.05) and the natural recovery group. Compared with the model group and the natural recovery group, the Aurantii Fructus Immaturus combined with Atractylodis Macrocephalae Rhizoma group showed increased amount of feces, fecal water content, intestinal propulsion rate, and activities of mitochondrial respiratory chain complexes Ⅱ, Ⅲ, and Ⅳ (P<0.05,P<0.01). Moreover, the combination meliorated the degree of mitochondrial swelling in the colon tissue, down-regulated the mRNA levels of PINK1 and Parkin and the protein levels of PINK1, Parkin, and LC3 (P<0.05,P<0.01), and up-regulated the mRNA level of p62 (P<0.05). ConclusionAurantii Fructus Immaturus and Atractylodis Macrocephalae Rhizoma, and their combination may remedy the colonic motility disorders in rats with slow transit constipation by blocking PINK1/Parkin signaling pathway to inhibit the excessive mitophagy in interstitial cells of Cajal in the colon tissue.

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