1.Efficacy and Mechanism of Lutongning Granules in Treatment of Trigeminal Neuralgia Induced by Injection of Talc into Infraorbital Foramen of Model Rats Based on P2X7R-mediated Neuroinflammation
Qiyue SUN ; Shuran LI ; Shuangrong GAO ; Shanshan GUO ; Zihan GENG ; Lei BAO ; Ronghua ZHAO ; Jingsheng ZHANG ; Bo PANG ; Yingli XU ; Yu ZHANG ; Shan CAO ; Yaxin WANG ; Xiaolan CUI ; Bing HAN ; Jing SUN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(15):56-63
ObjectiveTo evaluate the effectiveness of Lutongning granules in the treatment of trigeminal neuralgia in animal models and study its mechanism of action, so as to provide laboratory data support for the clinical application of Lutongning granules and precise treatment. MethodMale SD rats were randomly divided into normal group, model group, carbamazepine group (0.06 g·kg-1·d-1), high-dose Lutongning group (2.70 g·kg-1·d-1), and low-dose Lutongning group (1.35 g·kg-1·d-1) according to the stratified basic mechanical pain thresholds, with 10 rats in each group. A trigeminal neuralgia model of rats was prepared by injecting 30% talc suspension into the infraorbital foramen area of the rat. The drug groups were administered 10 mL·kg-1 of drugs by gavage after 2 h of modeling. The normal group and the model group were administered distilled water by gavage under the same conditions once a day for 10 consecutive days. Von Frey brushes were used to determine the mechanical pain threshold of rats. A fully automated blood and body fluid analyzer was employed to detect the blood routine of rats. Hematoxylin and eosin (HE) staining was utilized to detect the pathological changes in the trigeminal ganglion and medulla oblongata tissue. Transmission electron microscopy was used to scan the ultrastructure of the medulla oblongata tissue. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of inflammatory factors interleukin (IL)-1, IL-6, IL-8, tumor necrosis factor (TNF)-α, neuropeptide substance P, and β-endorphins (β-EP) in the serum of rats, and Western blot was used to detect the protein expression levels of IL-1β, purinergic receptor P2X7 (P2X7R), and phosphorylated p38 mitogen-activated protein kinase (p-p38 MAPK). ResultCompared with that in the normal group, the pain threshold of rats in the model group was significantly lower (P<0.01). The absolute value of neutrophils (NEUT#) and the percentage of neutrophils (NEUT) were significantly improved, and the percentage of lymphocytes (LYMPH) was significantly reduced (P<0.01). The serum levels of IL-1, IL-6, IL-8, and TNF-α were significantly increased (P<0.01). SP content in brain tissue was significantly increased, and β-EP content was significantly decreased (P<0.01). The relative protein expression of IL-1β, P2X7R, and p-p38 MAPK was significantly increased (P<0.05). HE staining and transmission electron microscopy results of medulla oblongata tissue revealed neuronal degeneration, mild proliferation of microglial cells, reduction in the number of myelinated nerves, and obvious demyelination. The trigeminal nerve fibers of rats were disarranged, and some nerve fibers showed vacuolization. Axons were swollen, and Schwann cells proliferated. Demyelination was observed. Compared with the model group, each administration group significantly increased the pain threshold of rats (P<0.05, P<0.01), reduced NEUT# and NEUT, and elevated LYMPH (P<0.05, P<0.01). The administration group significantly decreased the levels of IL-1, IL-6, IL-8, and TNF-α in serum and SP in brain tissue (P<0.01) and increased the level of β-EP (P<0.01). They significantly down-regulated the protein expression of IL-1β, P2X7R, and p-p38 MAPK(P<0.05, P<0.01) and significantly ameliorated the pathological changes in medulla oblongata tissue and trigeminal nerves of rats. ConclusionLutongning Granules had significant therapeutic effects on trigeminal neuralgia induced by injection of talc into the infraorbital foramen of model rats, and the mechanism may be related to amelioration of P2X7R-mediated neuroinflammation and inhibition of demyelination of myelinated nerves.
2.Hypericin inhibits the expression of NLRP3 in microglia of Parkinson's disease mice and alleviates the damage of DA-ergic neurons
Li-Shan FAN ; Jia ZHANG ; Si-Xiang NIU ; Qi XIAO ; Hui-Jie FAN ; Lei XU ; Li-Xia YANG ; Lu JIA ; Shao-Chen QIN ; Bao-Guo XIAO ; Cun-Gen MA ; Zhi CHAI
The Chinese Journal of Clinical Pharmacology 2024;40(17):2523-2527
Objective To observe the intervention effect of hypericin(HYP)on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced Parkinson's disease(PD)mice model and its mechanism.Methods Thirty C57BL/6 mice were randomly divided into normal,model and experimental groups with 10 mice per group.PD mouse model was established after 7 days of intraperitoneal injection of MPTP,and drug intervention was carried out from the first day of modeling.Normal group and model group were intraperitoneally injected with 500 μL·kg·d-1 0.9%NaCl.The experimental group was intraperitoneally injected with 25 mg·kg·d-1 HYP.The three groups of rats were given the drug once each time for 14 days.The expression levels of tyrosine hydroxylase(TH),Nod-like receptor thermal protein domain protein 3(NLRP3)and ionized calcium binding adapter molecule 1(Iba1)in the striatum of nigra were detected by Western blot.Results The climbing time of normal,model and experimental groups was(5.35±0.43),(9.71±1.19)and(8.07±0.34)s;suspension scores were(2.92±0.15),(1.38±0.28)and(1.96±0.28)points;the relative expression levels of TH protein were 1.04±0.06,0.51±0.09 and 0.75±0.07;the relative expression levels of NLRP3 protein were 0.51±0.03,1.00±0.04 and 0.77±0.06;the relative expression levels of Iba1 protein were 0.68±0.10,1.30±0.28 and 0.89±0.05,respectively.The above indexes in the model group were statistically significant compared with the experimental group and the normal group(all P<0.01).Conclusion HYP plays a therapeutic role in PD by inhibiting the expression of NLRP3 inflammasome in PD mice.
3.Effect of Influenza A Virus on BEAS-2B in Human Lung Epithelial Cells and Intervention Effect of Shufeng Jiedu Capsule-containing Serum
Shan CAO ; Zihan GENG ; Lei BAO ; Yingli XU ; Bo PANG ; Jingsheng ZHANG ; Yu ZHANG ; Mengping CHEN ; Yaxin WANG ; Ronghua ZHAO ; Shanshan GUO ; Xiaolan CUI
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(23):90-97
ObjectiveTo observe the effect of Shufeng Jiedu capsule (SFJD)-containing serum on human lung epithelial cells infected by influenza A virus, and investigate the protective effect of the drug on the cells and the potential antiviral effect. MethodThe SFJD-containing serum was prepared and used to treat human lung epithelial cells (BEAS-2B) cultured in vitro. The viability of cells treated with different concentrations of SFJD-containing serum was measured by the cell counting kit-8 (CCK-8), and the optimal concentration of SFJD-containing serum was screened for subsequent experiments. BEAS-2B cells were classified into normal control, virus infection, and SFJD-containing serum groups, and the CCK-8 method was used to detect the survival rate of BEAS-2B cells after virus infection and drug administration. The expression of influenza virus nucleic acid in the cells of each group was determined, and the apoptosis of cells in different groups was observed by fluorescence microscopy. Real-time PCR was employed to determine the mRNA levels of influenza virus nucleoprotein (NP), Toll-like receptor 4 (TLR4), and myeloid differentiation primary response gene 88 (MyD88) in each group of cells. The immunofluorescence assay was used to detect the fluorescence intensities of TLR4, MyD88, and phosphorylated nuclear factor-κB (p-NF-κB) in lung epithelial cells. ResultCompared with that in the control group (normal serum), the cell survival rates in the blank serum and the SFJD-containing serum (5%, 10%, and 20%) groups were 100.00%±0.00%, 89.05%±4.80%, 87.13%±5.90%, 93.83%±6.03%, and 99.33%±3.39%, respectively (P<0.01). The SFJD-containing serum of 20% was selected as the optimal treatment for subsequent experiments. Compared with the normal control group, the virus infection group showed reduced cell survival rate (P<0.01), and the reduction was increased by the SFJD-containing serum (P<0.01). Compared with the virus infection group, SFJD-containing serum reduced the virus load (P<0.01) to decrease apoptosis. Compared with the normal control group, the virus infection group showed up-regulated mRNA levels of NP, TLR4, and MyD88 (P<0.01), and the up-regulation was down-regulated by the SFJD-containing serum (P<0.05, P<0.01). The fluorescence intensities of TLR4, MyD88, and p-NF-κB proteins in the cells increased after virus infection compared with those in the normal control (P<0.05, P<0.01), and they were decreased after administration with the SFJD-containing serum (P<0.05). ConclusionThe SFJD-containing serum can inhibit influenza virus in vitro by increasing the survival rate, reducing the apoptosis, and down-regulating the protein levels of TLR4, MyD88, and p-NF-κB in BEAS-2B cells.
4.Identification and anti-inflammatory activity of chemical constituents and a pair of new monoterpenoid enantiomers from the fruits of Litsea cubeba
Mei-lin LU ; Wan-feng HUANG ; Yu-ming HE ; Bao-lin WANG ; Fu-hong YUAN ; Ting ZHANG ; Qi-ming PAN ; Xin-ya XU ; Jia HE ; Shan HAN ; Qin-qin WANG ; Shi-lin YANG ; Hong-wei GAO
Acta Pharmaceutica Sinica 2024;59(5):1348-1356
Eighteen compounds were isolated from the methanol extract of the fruits of
5.Tujia medicine Toddalia asiatica improves synovial pannus in rats with collagen-induced arthritis through the PI3K/Akt signaling pathway
Shan XIANG ; Zongxing ZHANG ; Lu JIANG ; Daozhong LIU ; Weiyi LI ; Zhuoma BAO ; Rui TIAN ; Dan CHENG ; Lin YUAN
Journal of Southern Medical University 2024;44(8):1582-1588
Objective To investigate the therapeutic mechanism of Tujia medicine Toddalia asiatica alcohol extract(TAAE)for synovial pannus formation in rats with college-induced arthritis(CIA).Methods Sixty male SD rats were randomized into normal control group,CIA model group,TGT group,3 TAAE treatment groups at low,medium and high doses(n=10).Except for those in the normal control group,all the rats were subjected to CIA modeling using a secondary immunization method and treatment with saline,TGT or TAAE by gavage once daily for 35 days.The severity of arthritis was assessed using arthritis index(AI)score,and knee joint synovium pathologies were examined with HE staining.Serum levels of TNF-α,IL-6,and IL-1β were detected with ELISA;the protein expressions of PI3K,Akt,p-PI3K,p-Akt,VEGF,endostatin,HIF-1α,MMP1,MMP3,and MMP9 in knee joint synovial tissues were determined using Western blotting,and the mRNA expressions of TNF-α,IL-6,IL-1β,VEGF,HIF-1α,PI3K,and Akt were detected with RT-PCR.Results Treatment of CIA rat models with TAAE and TGT significantly alleviated paw swelling,lowered AI scores,and reduced knee joint pathology,neoangiogenesis,and serum levels of inflammatory factors.TAAE treatment obviously increased endostatin protein expression,downregulated p-PI3K,p-Akt,MMP1,MMP3,MMP9,VEGF,and HIF-1α proteins,and reduced TNF-α,IL-6,IL-1β,PI3K,Akt,VEGF,and HIF-1α mRNA levels in the synovial tissues,and these changes were comparable between high-dose TAAE group and TGT group.Conclusion TAAE can improve joint symptoms and inhibit synovial pannus formation in CIA rats by regulating the expressions of HIF-1α,VEGF,endostatin,MMP1,MMP3,and MMP9 via the PI3K/Akt signalling pathway.
6.Standardized operational protocol for the China Human Brain Bank Consortium(2nd edition)
Xue WANG ; Zhen CHEN ; Juan-Li WU ; Nai-Li WANG ; Di ZHANG ; Juan DU ; Liang YU ; Wan-Ru DUAN ; Peng-Hao LIU ; Han-Lin ZHANG ; Can HUANG ; Yue-Shan PIAO ; Ke-Qing ZHU ; Ai-Min BAO ; Jing ZHANG ; Yi SHEN ; Chao MA ; Wen-Ying QIU ; Xiao-Jing QIAN
Acta Anatomica Sinica 2024;55(6):734-745
Human brain banks use a standardized protocol to collect,process and store post-mortem human brains and related tissues,along with relevant clinical information,and to provide the tissue samples and data as a resource to foster neuroscience research according to a standardized operating protocols(SOP).Human brain bank serves as the foundation for neuroscience research and the diagnosis of neurological disorders,highlighting the crucial rule of ensuring the consistency of standardized quality for brain tissue samples.The first version of SOP in 2017 was published by the China Human Brain Bank Consortium.As members increases from different regions in China,a revised SOP was drafted by experts from the China Human Brain Bank Consortium to meet the growing demands for neuroscience research.The revised SOP places a strong emphasis on ethical standards,incorporates neuropathological evaluation of brain regions,and provides clarity on spinal cord sampling and pathological assessment.Notable enhancements in this updated version of the SOP include reinforced ethical guidelines,inclusion of matching controls in recruitment,and expansion of brain regions to be sampled for neuropathological evaluation.
7.Tujia medicine Toddalia asiatica improves synovial pannus in rats with collagen-induced arthritis through the PI3K/Akt signaling pathway
Shan XIANG ; Zongxing ZHANG ; Lu JIANG ; Daozhong LIU ; Weiyi LI ; Zhuoma BAO ; Rui TIAN ; Dan CHENG ; Lin YUAN
Journal of Southern Medical University 2024;44(8):1582-1588
Objective To investigate the therapeutic mechanism of Tujia medicine Toddalia asiatica alcohol extract(TAAE)for synovial pannus formation in rats with college-induced arthritis(CIA).Methods Sixty male SD rats were randomized into normal control group,CIA model group,TGT group,3 TAAE treatment groups at low,medium and high doses(n=10).Except for those in the normal control group,all the rats were subjected to CIA modeling using a secondary immunization method and treatment with saline,TGT or TAAE by gavage once daily for 35 days.The severity of arthritis was assessed using arthritis index(AI)score,and knee joint synovium pathologies were examined with HE staining.Serum levels of TNF-α,IL-6,and IL-1β were detected with ELISA;the protein expressions of PI3K,Akt,p-PI3K,p-Akt,VEGF,endostatin,HIF-1α,MMP1,MMP3,and MMP9 in knee joint synovial tissues were determined using Western blotting,and the mRNA expressions of TNF-α,IL-6,IL-1β,VEGF,HIF-1α,PI3K,and Akt were detected with RT-PCR.Results Treatment of CIA rat models with TAAE and TGT significantly alleviated paw swelling,lowered AI scores,and reduced knee joint pathology,neoangiogenesis,and serum levels of inflammatory factors.TAAE treatment obviously increased endostatin protein expression,downregulated p-PI3K,p-Akt,MMP1,MMP3,MMP9,VEGF,and HIF-1α proteins,and reduced TNF-α,IL-6,IL-1β,PI3K,Akt,VEGF,and HIF-1α mRNA levels in the synovial tissues,and these changes were comparable between high-dose TAAE group and TGT group.Conclusion TAAE can improve joint symptoms and inhibit synovial pannus formation in CIA rats by regulating the expressions of HIF-1α,VEGF,endostatin,MMP1,MMP3,and MMP9 via the PI3K/Akt signalling pathway.
8.Renal tubular epithelial cell quality control mechanisms as therapeutic targets in renal fibrosis
Bao YINI ; Shan QIYUAN ; Lu KEDA ; Yang QIAO ; Liang YING ; Kuang HAODAN ; Wang LU ; Hao MIN ; Peng MENGYUN ; Zhang SHUOSHENG ; Cao GANG
Journal of Pharmaceutical Analysis 2024;14(8):1099-1109
Renal fibrosis is a devastating consequence of progressive chronic kidney disease,representing a major public health challenge worldwide.The underlying mechanisms in the pathogenesis of renal fibrosis remain unclear,and effective treatments are still lacking.Renal tubular epithelial cells(RTECs)maintain kidney function,and their dysfunction has emerged as a critical contributor to renal fibrosis.Cellular quality control comprises several components,including telomere homeostasis,ubiquitin-proteasome system(UPS),autophagy,mitochondrial homeostasis(mitophagy and mitochondrial metabolism),endoplasmic reticulum(ER,unfolded protein response),and lysosomes.Failures in the cellular quality control of RTECs,including DNA,protein,and organelle damage,exert profibrotic functions by leading to senescence,defective autophagy,ER stress,mitochondrial and lysosomal dysfunction,apoptosis,fibro-blast activation,and immune cell recruitment.In this review,we summarize recent advances in un-derstanding the role of quality control components and intercellular crosstalk networks in RTECs,within the context of renal fibrosis.
9.Status quo and influencing factors of postnatal sense of security in patients with hypertensive disorder complicating pregnancy
Lulu BAO ; Chunhua WANG ; Chunjian SHAN ; Xiangdi ZHANG ; Congshan PU ; Mengtian JI
Chinese Journal of Modern Nursing 2024;30(10):1311-1317
Objective:To explore the status quo and influencing factors of postnatal sense of security in patients with hypertensive disorder complicating pregnancy.Methods:This study was a cross-sectional study. Using the convenience sampling method, a total of 215 patients with hypertensive disorder complicating pregnancy and their spouses who gave birth in Women's Hospital of Nanjing Medical University from December 2022 to May 2023 were selected as the research objects. The general data questionnaire, Parents' Postnatal Sense of Security (PPSS), Ego-Resiliency Scale (ERS) and Perceived Social Support Scale (PSSS) were used to investigate them. Multiple linear regression analyses were used to explore the factors influencing postpartum security in patients with hypertensive disorders complicating pregnancy.Results:A total of 215 questionnaires were sent out in this study, and 202 were effectively collected, with an effective recovery rate of 94.0%. The total score of PPSS maternal version for 202 patients with hypertensive disorder complicating pregnancy was (59.32±6.81), while the total score of PPSS paternal version for spouses was (41.28±5.24). The results of multiple linear regression analysis showed that the number of antenatal examinations accompanied by family members, the effect of postpartum blood pressure control, the current state of mother and child, postnatal sense of security of spouse, ego-resiliency and perceived social support were the influencing factors of postnatal sense of security in patients with hypertensive disorder complicating pregnancy ( P<0.05) . Conclusions:The postnatal sense of security in patients with hypertensive disorder complicating pregnancy is at a medium high level. Medical personnel should focus on patients with hypertensive disorder complicating pregnancy who are accompanied by their family members less times, poor postpartum blood pressure control and postpartum separation from mother and child, providing them with targeted clinical care to improve their postnatal sense of security.
10.Pterostilbene inhibits the growth of esophageal squamous cell carcinoma by targeting PPARα signaling pathway and inducing ferroptosis
Yi YANG ; Wen-Jie SHI ; Shan LI ; Yue ZHANG ; Yuan-Qian MIN ; Bao-Ping LU
Chinese Pharmacological Bulletin 2024;40(12):2354-2360
Aim To study the molecular mechanism of pterostilbene(PTS)inhibiting the growth of esophage-al squamous cell carcinoma(ESCC).Methods Soft agar assay was used to detect the effect of PTS on the anchored independent growth of KYSE150.TMT-la-beled quantitative proteomics analysis was used to ana-lyze the influence of PTS on the proteome of KYSE150.Then the differentially expressed proteins(DEPs)enrichment was analyzed by GO and KEGG,and signaling pathway interactions were analyzed by STRING database.The molecular docking model of PTS and PPARα was established by computer.Trans-mission electron microscopy was used to observe the in-fluence of PTS on the morphology change of KYSE150.Western blot analysis the effects of PTS on PPARα sig-naling pathway and ferroptosis related proteins expres-sion.Results PTS inhibited the anchorage-independ-ent growth capability of KYSE150.A total of 249 DEPs were identified by proteomic analysis,including 175 up-regulated proteins and 74 down-regulated pro-teins.The DEPs enrichment analysis showed that PPAR signaling pathway was related to unsaturated fat-ty acid synthesis,pyruvate metabolism and other meta-bolic signaling pathways.PTS caused the reduction of mitochondrial volume and mitochondrial cristae of KYSE150.PTS inhibited the expression of PPARα sig-naling pathway and ferroptosis related proteins.Con-clusion PTS induced the ferroptosis of ESCC by in-hibiting PPARα signaling pathway.

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