1.Mechanism of Modified Erxian Decoction Regulating Perimenopausal Syndrome via SIRT1/Kisspeptin/GnRH Signaling Pathway
Ruiyu HUANG ; Fang LEI ; Wuchaonan LIU ; Jingjing YANG ; Qianru ZENG ; Shengping LUO ; Yanling CHEN ; Mengge ZHANG ; Fanshun SHEN ; Yihui DENG ; Dingxiang LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(22):51-62
ObjectiveTo explore the regulation of hypothalamic-pituitary-gonadal (HPG) axis by modified Erxian decoction in rats with perimenopausal syndrome (PMS) and to further analyze the expression of proteins related to the silent information regulator 1 (SIRT1)/hypothalamic kisspeptin (Kisspeptin)/gonadotropin-releasing hormone (GnRH) signaling pathway in the arcuate nucleus region (ARC) of the hypothalamus, so as to reveal the potential target of action and molecular biological mechanism of modified Erxian decoction for the treatment of perimenopausal syndrome. MethodsAn animal model was established via the incomplete castration method, with successful modeling confirmed by the exfoliated cervical cell smear method. The 48 rats were divided into six groups based on the randomization principle after successful modeling, including a sham operation group, a model group, an estradiol valerate group (0.09 mg∙kg-1∙d-1), high-, medium-, and low-dose modified Erxian decoction groups (7.614, 3.807,1.903 5 g∙kg-1∙d-1), with 8 rats in each group. The estradiol valerate group and the high-, medium- and low-dose modified Erxian decoction groups were continuously administered by gavage for 28 days, and the indicators were detected 24 hours after the last administration. Body weights and uterine indices were measured. The pathological changes of the uterus were observed by hematoxylin-eosin (HE) staining. Enzyme-linked immunosorbent assay (ELISA) was performed to measure the levels of estradiol (E2), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and gonadotropin-releasing hormone (GnRH). Real-time quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to determine the expression levels of SIRT1, Kisspeptin, kisspeptin receptor (GPR54), and GnRH in the ARC region of the hypothalamus and gonadotropin-releasing hormone receptor (GnRH-R) in pituitary. ResultsCompared with the sham operation group, rats in the model group had a significantly increased body weight (P0.01), reduced wet weight and index of uterus (P0.01), endometrial thinning or atrophy, glandular atrophy, and a decreasing number of glands. Additionally, serum levels of E2 and the expression of SIRT1 in the ARC region of the hypothalamus significantly decreased (P0.01). Serum levels of FSH, LH, and GnRH, the expression of Kisspeptin, GPR54, and GnRH in the ARC region of the hypothalamus, and GnRH-R in pituitary significantly increased (P0.01). Compared with the model group, the estradiol valerate group and the high-, medium-dose modified Erxian decoction groups had significantly reduced body weight, serum levels of FSH, LH, and GnRH, and expression of Kisspeptin, GPR54, and GnRH in the ARC region of the hypothalamus and GnRH-R in pituitary (P0.05, P0.01) and significantly increased wet weight and index of uterus, serum level of E2, and expression of SIRT1 in the ARC region of the hypothalamus (P0.05, P0.01). In addition, they showed thickened endometrium, increased number of endometrial glands, and improved glandular atrophy. ConclusionModified Erxian decoction regulates the function of the HPG axis through multi-targets, and its mechanism of action may be related to the up-regulation of the expression of SIRT1 in the ARC region of the hypothalamus, the inhibition of the over-activation of the Kisspeptin/GnRH signaling pathway, the regulation of the expression of GnRH-R in the pituitary, the restoration of secretion balance of gonadotropins, and the elevation of the estrogen level. This study provides an experimental basis for the interpretation of the scientific connotation of modified Erxian decoction in the treatment of perimenopausal syndrome and a theoretical reference for the development of a novel therapeutic strategy based on the SIRT1/Kisspeptin/GnRH pathway.
2.Effect of Zuogui Jiangtang Tongmai Prescription on Inflammatory Injury of Human Umbilical Vein Endothelial Cells Induced by High Glucose and LPS Based on GPR43/β-arrestin-2/IκBα/NF-κB Pathway
Lanyu PENG ; Jingxin YAO ; Yujia LI ; Dingxiang LI ; Xun LIU ; Yihui DENG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(3):64-74
ObjectiveTo investigate the effects and mechanism of Zuogui Jiangtang Tongmai prescription (ZJTP) on human umbilical vein endothelial cells (HUVECs) damaged by high glucose combined with lipopolysaccharide (LPS). MethodThe survival rate of cells was determined by cell counting kit-8 (CCK-8), and the level of tumor necrosis factor-α (TNF-α) was determined by enzyme-linked immunosorbent assay (ELISA) to determine the optimal injury concentration and action time of LPS, as well as the optimal action concentration of ZJTP drug-containing serum. HUVECs were divided into a blank control group, a model group, a ZJTP drug-containing serum group, and an SCFA mixed liquid group. ELISA was used to detect the level of endothelin-1 (ET-1), nitric oxide (NO), interleukin-1β (IL-1β), interleukin-6 (IL-6), and TNF-α. Western blot was performed to detect the protein expression of G protein-coupled receptor43 (GPR43), β-suppressor protein-2 (β-arrestin-2), nuclear factor-κB suppressor α (IκBα), and nuclear factor κB p65 (NF-κB p65). The nucleation of NF-κB p65 was observed by immunofluorescence staining (IF). The role of GPR43 in the regulation of inflammatory injury was observed by means of small interfering ribonucleic acid (siRNA). The cells after intervention were divided into an empty carrier group, a ZJTP drug-containing serum group, a Si-GPR43 group, and a Si-GPR43 + ZJTP drug-containing serum group. The content of IL-1β, IL-6, and TNF-α was detected by ELISA. The protein expression of pathways was detected by Western blot. IF was used to observe the nucleation of NF-κB p65. ResultThe optimal molding condition was 1 mg·L-1 LPS for 24 h. The optimal drug intervention condition was 5% ZJTP drug-containing serum for 24 h. Compared with the blank control group, the content of ET-1 in the model group was significantly increased, and the content of NO was significantly decreased (P<0.01). The levels of inflammatory factors were significantly increased (P<0.01). The expressions of GPR43 and IκBα were significantly decreased, while the protein expressions of β-arrestin-2 and NF-κB p65 were significantly increased (P<0.01). NF-κB p65 protein was transferred from the extranuclear to the intranuclear (P<0.01). Compared with the model group, the content of ET-1 in the ZJTP drug-containing serum group was decreased, and the content of NO was increased (P<0.05). The levels of inflammatory factors decreased (P<0.05). The protein expressions of GPR43 and IκBα were increased, while the expressions of β-arrestin-2 and NF-κB p65 were decreased (P<0.05). The amount of NF-κB p65 transferred from the intranuclear to the extranuclear decreased (P<0.01). The mechanism study showed that compared with the Si-GPR43 group, the content of IL-1β, IL-6, and TNF-α were significantly decreased after treatment with ZJTP drug-containing serum (P<0.01). The protein expressions of GPR43 and IκBα were significantly increased (P<0.01), while the protein expressions of β-arrestin-2 and NF-κB p65 were significantly decreased (P<0.01). The amount of NF-κB p65 transferred from the extranuclear to the intranuclear decreased (P<0.01). ConclusionZJTP has a protective effect on HUVECs with high glucose and LPS-induced inflammatory injury, which may be related to the regulation of GPR43/β-arrestin-2/IκBα/NF-κB pathway.
3.Mechanism of Xumingtang in Gu Jin Lu Yan for Treatment of Ischemic Stroke Based on HIF-1α/NLRP3 Pathway-mediated Pyroptosis
Yuzhe CAI ; Dingxiang LI ; Zheng LUO ; Yixuan LIU ; Jingjing YANG ; Qin WU ; Yanan ZHANG ; Jing CHEN ; Yihui DENG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(17):9-17
ObjectiveTo investigate the mechanism of Xumingtang in Gu Jin Lu Yan (《古今录验》) in regulating cell pyroptosis through the hypoxia-inducible factor-1α (HIF-1α)/NOD-like receptor pyrin domain-containing protein 3 (NLRP3) pathway in ischemic stroke (IS). MethodSD rats were randomly divided into a sham operation group, a model group, low- and high-dose Xumingtang groups, and a metformin group, with 20 rats in each group. Oral administration was performed for 3 days, and tissue samples were collected. Differential messenger RNA (mRNA) was screened using high-throughput sequencing, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed on key differentially expressed genes. The modified neurological severity score (mNSS) and 2,3,5-triphenyltetrazolium chloride (TTC) staining were used to evaluate the effect of brain infarction. Hematoxylin-eosin (HE) staining was used for pathological morphological observation of brain tissue. Enzyme-linked immunosorbent assay (ELISA) was used to compare the levels of interleukin-1β (IL-1β) and interleukin-18 (IL-18) in the ischemic cortical region. Double staining immunohistochemistry was used to detect the co-localization of HIF-1α and NLRP3. Real-time quantitative polymerase chain reaction (PCR) was performed to detect the mRNA expression of NLRP3, HIF-1α, Caspase-1 (CASP-1), and gasdermin D (GSDMD). Western blot was used to detect the protein expression of HIF-1α, NLRP3, CASP-1, and GSDMD. ResultA total of 5 705 differentially expressed genes (2 733 downregulated and 2 972 upregulated) were obtained by mRNA sequencing. After conversion to homologous genes and intersection with the pyroptosis gene set, 95 key differentially expressed pyroptosis genes were obtained. Compared with the sham operation group, the model group showed significantly increased mNSS scores, larger brain infarction areas (P<0.01), diverse neuronal morphology, disordered arrangement, widened cell gaps, significantly increased levels of IL-1β and IL-18 in the ischemic cortical region (P<0.01), enhanced co-localization fluorescence intensity, and significantly increased mRNA and protein expression levels of HIF-1α, NLRP3, CASP-1, and GSDMD (P<0.01). Compared with the model group, the high-dose Xumingtang group showed the most significant improvement in neurological function scores and brain infarction areas (P<0.01). The neuronal integrity and arrangement were more complete, and the cell gaps were narrower in all groups with drug treatment, with significantly reduced co-localization fluorescence intensity. Xumingtang could reduce the levels of IL-1β, IL-18, and the mRNA and protein expression of HIF-1α, NLRP3, CASP-1, and GSDMD (P<0.05, P<0.01), with the high-dose Xumingtang group showing the most significant effect (P<0.01). ConclusionXumingtang in Gu Jin Lu Yan can inhibit cell pyroptosis and promote neurological function recovery after IS, which may be related to the inhibition of the HIF-1α/NLRP3 pathway.
4.Experimental study on the improvement of inflammatory response in rats with cerebral ischemia by Chinese ultrasound drug permeation electrotherapy device
Jingjing YANG ; Zheng LUO ; Yuzhe CAI ; Yixuan LIU ; Dingxiang LI ; Yihui DENG
International Journal of Traditional Chinese Medicine 2023;45(1):54-59
Objective:To observe the effects of Traditional Chinese Medicine (TCM)ultrasound drug permeation electrotherapy device on the inflammatory response of rats with cerebral ischemia, and to provide an experimental basis for the clinical application of TCM ultrasound drug permeation electrotherapy device in the treatment of cerebral ischemia.Methods:A total of 72 SD rats were randomly divided into sham-operation group (12 rats) and modeling group (60 rats). The middle cerebral artery occlusion (MCAO) model was prepared by thread embolism in the model group. The rats were divided into model group, Chinese medicine tablet group, blank tablet + TCM ultrasound drug permeation electrotherapy group (hereinafter referred to as "blank tablet + electrotherapy group"), Chinese medicine tablet + TCM ultrasound drug permeation electrotherapy group (hereinafter referred to as "Chinese medicine tablet + electrotherapy group") and butylphthalide group according to the random number table method, with 12 rats in each group. The corresponding treatment was given continuously for 7 days. The neurological function was scored using Longa method evaluation criteria; TTC staining was used to observe the infarct volume and calculate the percentage of infarct volume; HE staining was used to observe the cell morphology of cortical area in each group of rats; ELISA was used to detect the serum TNF-α and IL-1β levels in each group of rats; TLR4, MyD88 and NF-κBp65 protein expressions in hippocampal tissue of each group of rats on the infarct side were detected by Western blot method.Results:Compared with the model group, the neurological function scores of rats in the blank tablet + electrotherapy group, the herbal tablet + electrotherapy group, and the butylphthalein group significantly decreased ( P<0.05), the percentage of cerebral infarct volume significantly decreased ( P<0.05), the contents of serum TNF-α and IL-1β significantly decreased ( P<0.05), and the expressions of TLR4 (0.42±0.07, 0.31±0.07, 0.19±0.04 vs. 0.68±0.14), MyD88 (0.39±0.12, 0.30±0.07, 0.23±0.11 vs. 0.67±0.10), NF-κBp65 (0.32±0.03, 0.27±0.02, 0.17±0.03 vs. 0.57±0.12) protein in hippocampal tissue significantly decreased ( P<0.05). Conclusion:The TCM ultrasound drug permeation electrotherapy device can inhibit TLR4, MyD88, NF-κBp65 protein expressions and reduce the release of serum inflammatory factors TNF-α and IL-1β, thus exerting cerebral ischemic protective effects.
5.The application in patients with chronic heart failure based on the "Internet plus" three entities double track interactive continuity nursing
Haiyan XIONG ; Tingting HU ; Meifang LENG ; Wen JIN ; Jun LIU ; Dingxiang CAI
Chinese Journal of Practical Nursing 2018;34(34):2641-2647
Objective To explore the application effect of Interactive continuing nursing intervention model with three subjects and two tracks,which is based on internet and applied to chronic heart failure patients.Methods During January 1 to December 30,2017,110 chronic heart failure patients,who were going to discharge from the Second People's Hospital of Guangdong Province,were selected for this research.Participants were equally divided into control group and observation group with random number table with 55 cases each.Participants were adopted conventional medication and traditional care,while patients in control group were given extra interactive continuing nursing intervention with three subjects and two tracks,which was relied on internet-based cardiovascular disease management platform.Comparing patients' scores of life quality,self-care ability,anxiety and depression degree before intervention and that in 6 months after intervention with Minnesota Living with Heart Failure Questionnaire (MLHFQ),Self Care of Heart Failure Index (SCHFI),Self-rating Anxiety Scale(SAS)and Hamilton Depression Scale (HAMD) respectively.Moreover,6 minute walk test was conducted,simultaneously,the incidence of cardiovascular events was calculated respectively before intervention,3 months after intervention and 6 months after intervention.Results It showed that total scores of MLHFQ,SCHFI,SAS and HAMD for patients in observation group before intervention were (49.05 ± 4.39),(101.57 ±21.04),(58.3 ± 5.2) and (40.1 ± 3.5) points respectively while that for patients in observation group after 6 months intervention were (43.03 ± 6.85),(123.17 ± 18.23),(42.3 ± 4.2) and (24.4 ± 4.3) points respectively.The difference was statistically significant (t =5.49-20.60,all P <0.01).In comparison,intervention effect of observation group was better than that of control group,although,the SAS scores of the control group were statistically significant.Furthermore,after intervention,the difference of MLHFQ,SCHFI,SAS and HAMD scores were statistically significant(t =3.48-17.79,all P <0.01).Moreover,6 months after intervention,6 minute walk test evaluation showed that there was statistical significance between observation group and control group (~=8.11,P <0.05).Besides,The death rate in observation group was lower than that in control group after 6 months intervention (~=6.30,P <0.05).Meanwhile,readmission rate after 3 months and 6 months indicates that there was statistical significance between observation group and control group(x2=5.25,6.30,P <0.05).Conclusions Interactive nursing intervention model with three subjects and two tracks which based on internet is able to enhance chronic heart failure patients' self-care ability and life quality,decrease the incidence of cardiovascular events and negative emotion of patients.
6.Nursing experience of vacuum sealing drainage applied for plastic surgery post skin flap necrosis
Jia GUO ; Yuhong LI ; Shasha LI ; Jiantao SHANG ; Li LI ; Dingxiang LIU
Chinese Journal of Modern Nursing 2016;22(13):1875-1877
Objective To summarize nursing methods of vacuum sealing drainage (VSD)for plastic surgery patients with skin flap necrosis.Methods VSD was applied to 4 plastic surgery patients with skin flap necrosis in Middle Second Ward of Plastic Surgery Hospital,Chinese Academy of Medical Sciences.The effectiveness of VSD was retrospectively summarized and analyzed.Results Two patients with intercostalartery perforator flap had been treated with VSD for two cycles.There were massive fresh granulation tissue characterized with fine granules,abundant capillaries and no edema.The area of wound had decreased to 2 cm × 2 cm.Moreover,the wound recovered well after skin-graft repairing. One patient with ectatic sensate paraumbilical flap had been treated with VSD for one cycle.The area of wound had decreased to 1 cm ×1 cm and recovered after four times dressing change.The last case of patient with scapular flap had been treated with VSD for 3 cycles and the area wound had decreased to 2 cm ×3 cm.The wound recovered favorably after skin-graft repairing.There were no systemic or local toxicity,allergic reaction and complications in all four cases. Conclusions Application of VSD,which is worth of clinical expansion,is a simple-handling and effective treatment for plastic surgery patients with skin flap necrosis after debridement.

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