1.Mechanism study of SIRT3 alleviating oxidative-stress injury in renal tubular cells by promoting mitochondrial biogenesis via regulating mitochondrial redox balance
Yaojun LIU ; Jun ZHOU ; Jing LIU ; Yunfei SHAN ; Huhai ZHANG ; Pan XIE ; Liying ZOU ; Lingyu RAN ; Huanping LONG ; Lunli XIANG ; Hong HUANG ; Hongwen ZHAO
Organ Transplantation 2026;17(1):86-94
Objective To elucidate the molecular mechanism of sirtuin-3 (SIRT3) in regulating mitochondrial biogenesis in human renal tubular epithelial cells. Methods Cells were stimulated with different concentrations of H2O2 and divided into four groups: control (NC), 50 μmol/L H2O2, 110 μmol/L H2O2 and 150 μmol/L H2O2. SIRT3 protein expression was then measured. SIRT3 was knocked down with siRNA, and cells were further assigned to five groups: control (NC), negative-control siRNA (NCsi), SIRT3-siRNA (siSIRT3), NCsi+H2O2, and siSIRT3+H2O2. After 24 h, cellular adenosine triphosphate (ATP) and mitochondrial superoxide anion (O2•−) levels were determined, together with mitochondrial expression of SIRT3, peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α), nuclear respiratory factor 1 (NRF1), mitochondrial transcription factor A (TFAM), superoxide dismutase 2 (SOD2), acetylated-SOD2 and adenosine monophosphate activated protein kinase α1 (AMPKα1). Results The 110 and 150 μmol/L H2O2 decreased SIRT3 protein (both P<0.05). ATP and mitochondrial O2•− did not differ between NC and NCsi groups (both P>0.05). Compared to the NCsi group, the siSIRT3 group exhibited elevated O2•− level, decreased SIRT3 protein and increased expression levels of SOD2 and acetylated SOD2 protein (all P<0.05). Compared to the NCsi group, the NCsi+H2O2 group exhibited decreased cellular ATP levels, elevated mitochondrial O2•− levels, and reduced protein expression levels of SIRT3, SOD2, TFAM, AMPKα1, PGC-1α and NRF1 (all P<0.05). Compared with the siSIRT3 group, the siSIRT3+H2O2 group showed a decrease in cellular ATP levels, an increase in mitochondrial O2•− levels, a decrease in SIRT3, SOD2, TFAM, AMPKα1, PGC-1α and NRF1 protein expression levels and a decrease in acetylated SOD2 protein expression levels (all P<0.05). Compared with the NCsi+H2O2 group, the siSIRT3+H2O2 group showed a decrease in cellular ATP levels, an increase in mitochondrial O2•− levels, a decrease in SIRT3, AMPKα1, PGC-1α and NRF1, TFAM protein expression levels, and an increase in SOD2 and acetylated SOD2 protein expression levels (all P<0.05). Conclusions SIRT3 promotes mitochondrial biogenesis in tubular epithelial cells via the AMPK/PGC-1α/NRF1/TFAM axis, representing a key mechanism through which SIRT3 ameliorates oxidative stress-induced mitochondrial dysfunction.
2.Effect of Anmeidan in Ameliorating Neuronal Synaptic Structural and Functional Impairment in Aged Sleep Deprivation Model via EphA4/ephrinA3 Signaling Pathway
Junlu ZHANG ; Guangjing XIE ; Ping WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(6):36-45
ObjectiveTo investigate the effects of Anmeidan (AMD) on protein expression of the ephrin type-A receptor 4 (EphA4)/ephrinA3 signaling pathway and synaptic structural function in an aged sleep deprivation model. MethodsSeventy-two 18-month-old aged mice were randomly divided into a blank group, a model group, AMD high-, medium-, and low-dose groups (26.26, 13.13, 6.565 g·kg-1·d-1, respectively), and a melatonin group (1.3 mg·kg-1·d-1), with 12 mice in each group. Cognitive function was assessed using the novel object recognition test. Hematoxylin-eosin (HE) staining was used to observe cell number and morphology in hippocampal tissues, and Nissl staining was performed to examine cellular structure and quantify Nissl bodies. Transmission electron microscopy was used to observe synaptic ultrastructure, with emphasis on changes in synaptic morphology and structure. Western blot was employed to detect the expression levels of EphA4, ephrinA3, brain-derived neurotrophic factor (BDNF), glutamate aspartate transporter (GLAST), glutamate transporter-1 (GLT-1), growth-associated protein 43 (GAP43), postsynaptic density protein 95 (PSD95), and synaptophysin (SYN) in hippocampal tissues. Immunofluorescence double labeling was performed to co-stain EphA4 and ephrinA3 with glial fibrillary acidic protein (GFAP) and neuronal nuclei antigen (NeuN), respectively, to observe the colocalization of target proteins with neurons and astrocytes. ResultsCompared with the blank group, the model group exhibited increased exploration time of familiar objects (P<0.01), while exploration time of novel objects and the recognition index were decreased (P<0.01). The number of neurons in the CA1, CA3, and dentate gyrus (DG) regions of the hippocampus was reduced, Nissl bodies were decreased, and synaptic structures were damaged. Protein expression levels of BDNF, GLAST, GLT-1, GAP43, PSD95, and SYN in hippocampal tissues were decreased, whereas the expression levels of EphA4, ephrinA3, and GFAP were increased. Compared with the model group, the AMD low-, medium-, and high-dose groups and the melatonin group showed increased exploration time of novel objects and higher novel object recognition indices (P<0.01), along with significantly reduced exploration time of familiar objects (P<0.01). Neuronal damage in the CA1 and DG regions was ameliorated, the number of Nissl bodies in the CA1 region was increased, and organelle and synaptic structural damage was alleviated. Protein expression levels of BDNF, GLAST, GLT-1, GAP43, PSD95, and SYN were increased, and protein expression levels of EphA4, ephrinA3, and GFAP were decreased (P<0.05,P<0.01). ConclusionAMD can regulate protein expression of the EphA4/ephrinA3 signaling pathway in an aged sleep deprivation model, enhance synaptic protein expression, and improve neuronal synaptic damage.
3.Expert Consensus on Clinical Application of Qidong Yixin Oral Liquid
Changkuan FU ; Xiaochang MA ; Mingjun ZHU ; Yue DENG ; Hongxu LIU ; Mingxue ZHANG ; Ying CHEN ; Yan ZHOU ; Ling ZHANG ; Jianhua FU ; Wei YANG ; Yu'er HU ; Ming CHEN ; Yanming XIE ; Yuanyuan LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):147-158
The prescription of Qidong Yixin oral liquid is derived from the experience of national medical master Ren Jixue in treating viral myocarditis (VMC). It has the functions of tonifying Qi, nourishing the heart,calming the mind, and relieving palpitations. It is used to treat VMC and angina pectoris of coronary heart disease caused by deficiency of both Qi and Yin. However,the understanding of its efficacy evidence, advantageous aspects, dosage and administration, and medication safety remains insufficient in clinical practice. Therefore,the development of the Expert Consensus on the Clinical Application of Qidong Yixin Oral Liquid (hereinafter referred to as consensus) was initiated. Consensus strictly followed the process and methods of the expert consensus on the clinical application of Chinese patent medicines of the China Association of Chinese Medicine,successively completing multiple tasks such as the consensus project initiation,determination of clinical problems,evidence search and evaluation,formation of recommendation opinions and consensus suggestions,solicitation of opinions,peer review, submission for review and release, and so on. Consensus formed a total of 10 recommendation opinions and 12 consensus suggestions,clarifying the clinical positioning,efficacy advantages,syndrome differentiation,dosage and administration,combination therapy,timing of medication,adverse reactions,contraindications, and precautions of Qidong Yixin oral liquid,indicating that it has good clinical advantages and safety in the treatment of VMC and angina pectoris of coronary heart disease,providing norms and references for physicians to safely and rationally apply Qidong Yixin oral liquid. Consensus was reviewed and approved for release by the Standardization Office of the China Association of Chinese Medicine on December 23, 2024. Standard number:GSCACM-376-2024.
4.Polygonati Odorati Rhizoma Polysaccharides Ameliorate Hyperlipidemia in Mice by Regulating Gut Microbiota
Jingchen XIE ; Qianqian LIU ; Suhui XIONG ; Zhimin ZHANG ; Yuexin LIU ; Ping WU ; Duanfang LIAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):84-92
ObjectiveTo explore the effect of Polygonati Odorati Rhizoma polysaccharides on hyperlipidemia in mice by modulating the gut microbiota. MethodsNinety male C57BL/6J mice were randomized into the following groups (n=15): control, model, simvastatin, low- (100 mg·kg-1), medium- (200 mg·kg-1), and high-dose (400 mg·kg-1) Polygonati Odorati Rhizoma polysaccharides groups. Other groups except the control group were fed with a high-fat diet for the modeling of hyperlipidemia, and drug interventions lasted for 12 weeks. Serum levels of total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) were measured by an automatic biochemical analyzer. The pathological changes in the liver and epididymal fat were observed by hematoxylin-eosin staining, and lipid accumulation in the liver was assessed by oil red O staining. The gut microbiota was analyzed by 16S rRNA gene sequencing. ResultsCompared with the control group, the model group exhibited an increase in body weight (P<0.01), along with marked elevations in serum levels of TC, TG, and LDL-C (P<0.05,P<0.01). Furthermore, the model group showcased increase in the liver index and epididymal fat coefficient (P<0.05), increased liver fat accumulation, enlargement of adipocytes in the epididymal fat, decreases in both alpha and beta diversity of the gut microbiota, and an increase in the relative abundance of Allobaculum (P<0.01). Compared with the model group, Polygonati Odorati Rhizoma polysaccharides suppressed the increase in body weight (P<0.01), lowered the serum levels of TC, TG, and LDL-C (P<0.05,P<0.01), reduced the liver index and epididymal fat coefficient (P<0.05), alleviated liver fat accumulation, and decreased the size of adipocytes in the epididymal fat. Furthermore, it enhanced the alpha and beta diversity of the gut microbiota in mice, reduced the relative abundance of Allobaculum, Erysipelotrichaceae, and Clostridium (P<0.01), and increased the relative abundance of Akkermansia and Blautia (P<0.01). ConclusionPolygonati Odorati Rhizoma polysaccharides can ameliorate hyperlipidemia induced by a high-fat diet in mice by regulating the diversity and composition of the gut microbiota.
5.Improvement mechanism of Pangshi antai zhixue decoction on spontaneous abortion with heat syndrome by regulating NLRP3 inflammasome
Liya MA ; Xingfei WU ; Liujun WU ; Yanduo SHEN ; Bingheng XIE ; Jiale ZHANG ; Jinhao HAO ; Meng YU ; Yumiko NAKAYAMA ; Minghao ZHANG ; Dawei ZHANG
China Pharmacy 2025;36(1):37-43
OBJECTIVE To investigate the mechanism of Pangshi antai zhixue decoction in the improvement of spontaneous abortion with heat syndrome by regulating the NOD-like receptor protein 3 (NLRP3) inflammasome. METHODS The binding activities of 13 main components in Pangshi antai zhixue decoction with NLRP3, apoptosis-associated speck-like protein , containing a CARD (ASC), and caspase-1 precursor (pro- No.20-21ZY1053) caspase-1) were predicted by molecular docking. Sixty 1-day-old pregnant rats were randomly divided into normal group, model group, dexamethasone group (0.002 g/kg), and Pangshi antai zhixue decoction low-, medium-, and high-dose groups (11.025, 22.05, 44.10 g/kg), with 10 rats in each group. Each group was given distilled water/corresponding medicinal solution intragastrically, once a day, for 12 consecutive days. Except for normal group, other groups were given traditional Chinese medicine for warming yang and mifepristone to establish a model of spontaneous abortion with heat syndrome. 24 h after the last medication, serum levels of triiodothyronine (T3), thyroxine (T4), interleukin-2 (IL-2), IL-4, IL-6, IL-10, and interferon-γ (IFN-γ) were all detected; the abortion rate and uterine coefficient were calculated; the pathological morphology of the pregnant uterus was observed; protein expressions of NLRP3, ASC and caspase-1 were detected. RESULTS The molecular docking results showed that the binding energies of 13 main components of Pangshi antai zhixue decoction with NLRP3, ASC, and pro-caspase-1 were all less than -5 kJ/moL. The animal experiment results showed that compared with normal group, the uterine coefficient and serum levels of IL-4, IL-6 and IL-10 were decreased significantly in model group (P<0.05); the abortion rate and serum levels of T3, T4, IL-2 and IFN-γ as well as protein expressions of NLRP3, ASC and caspase-1 were increased significantly (P<0.05); there were abortion lesions in the pregnant endometrium. Compared with the model group, most of the quantitative indicators mentioned above were significantly reversed in Pangshi antai zhixue decoction groups (P<0.05), and the endometrial miscarriage lesions in pregnancy were improved to varying degrees. CONCLUSIONS Pangshi antai zhixue decoction influences the immune balance between mother and fetus by regulating the formation of NLRP3 inflammasome, down-regulating pro-inflammatory cytokines such as IFN-γ and IL-2, and up-regulating anti-inflammatory cytokines such as IL-4, IL-6 and IL-10, thereby improving spontaneous abortion with heat syndrome.
6.Quality analysis of Pinghuo tea standard decoction
Xinmei ZHAO ; Hongyang SANG ; Chunjing YANG ; Jingwei LEI ; Haiyan GONG ; Caixia XIE ; Chunya ZHANG ; Haohan DUAN ; Hao YU
China Pharmacy 2025;36(1):71-78
OBJECTIVE To establish the fingerprint of Pinghuo tea standard decoction and a method for determination of multi-component to clarify the transfer relationship of quantities and quality from pieces and standard decoction. METHODS Fifteen batches of Pinghuo tea standard decoction were prepared and the extract rate was determined; the fingerprint of the preparation was established by using high-performance liquid chromatography(HPLC); the similarity evaluation and the determination of common peaks were performed, and chemometric analysis was performed; the same method was used to determine the content of indicator components and the transfer rate was calculated. The chromatographic column was Venusil C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid solution (gradient elution); the column temperature was 30 ℃, and the detection wavelengths were 238 nm (0-37 min, 85-102 min) and 330 nm (37-85 min) at a flow rate of 1.0 mL/min with an injection volume of 10 μL. RESULTS The similarity of HPLC fingerprints for 15 batches of Pinghuo tea standard decoction was not lower than 0.968. A total of 24 common peaks were calibrated and 9 peaks were recognized, which were as follows neochlorogenic acid (peak 3), chlorogenic acid (peak 6), geniposide (peak 9), glycyrrhizin (peak 10), galuteolin (peak 11), isochlorogenic acid A (peak 14), luteolin (peak 21), kaempferol (peak 23) and glycyrrhizic acid (peak 24). Cluster analysis, principal component analysis and orthogonal partial least squares discriminant analysis showed consistent results, all of which could classify the 15 batches of samples into three categories. The linear range of indicator components in 15 batches of Pinghuo tea standard decoction, such as geniposide, luteolin, isochlorogenic acid A, glycyrrhizin, and glycyrrhizic acid, were 0.020 580-0.411 600, 0.001 617-0.080 850, 0.006 076-0.607 600, 0.005 125-0.071 740, and 0.017 288-0.432 200 mg/mL, respectively; RSDs of precision, repeatability, stability and recovery rate tests were all not higher than 4% (n=6). The mass fractions ranged 3.227 9-10.002 2, 0.297 4-0.554 6, 3.350 1-6.159 6, 0.720 6-1.073 3, 2.003 1-3.030 1 mg/g; transfer rates from the pieces and standard decoction were 19.762 8%-35.840 5%, 12.123 3%-21.254 0%, 46.097 2%-82.869 4%, 58.708 8%-91.629 6%, 39.114 3%-63.710 6%. The transfer rates of the extract from 15 batches of Pinghuo tea standard decoction ranged from 61.15%-84.68%. CONCLUSIONS Established HPLC fingerprint and content determination methods in this study are simple and accurate, which can provide reference for the quantitative value transfer study, quality control, clinical application and the development of subsequent formulations of Pinghuo tea standard decoction.
7.A case analysis of multiple immune-related adverse event induced by toripalimab
Zhen FENG ; Yuzhen ZHANG ; Cheng XIE
China Pharmacy 2025;36(1):96-100
OBJECTIVE To explore the clinical features, diagnosis, treatment and management measures of multiple immune- related adverse event (irAE) after toripalimab treatment, in order to provide reference for the clinical management of similar cases. METHODS A retrospective analysis was conducted on a post-operative bladder cancer patient who developed various irAEs after toripalimab treatment. The patient’s medical history, laboratory tests, imaging studies, treatment course and outcomes were detailed. The Naranjo scale was used to assess the relationship between the irAEs and toripalimab. RESULTS The patient developed irAEs after receiving toripalimab, such as immune-related myocarditis, liver dysfunction, and myasthenia. According to the Naranjo scale evaluation, it was assessed that these adverse events were related to the administration of toripalimab. After high- dose corticosteroid pulse therapy and immunoglobulin immunomodulation, the patient’s symptoms were effectively relieved, with a improved trend in various physiological indicators and significant improvement in the degree of system/organ damage. CONCLUSIONS Toripalimab may lead to multiple irAEs, including immune-related myocarditis, liver injury, and myasthenia. Early recognition, timely diagnosis, and appropriate treatment of irAE are crucial for improving patient’s prognosis.
8.Ideas of Traditional Chinese Medicine Treatment of Pancreatic Endocrine and Exocrine Co-Morbidities from the Attributes of Zang-Fu Organs of Pancreas
Yulin LENG ; Jiacheng YIN ; Xianglong LI ; Jiahong ZHANG ; Yi SU ; Hong GAO ; Chunguang XIE ; Xiaoxu FU
Journal of Traditional Chinese Medicine 2025;66(2):145-149
Based on advancements in modern medical research regarding the intricate connection between the endocrine and exocrine functions of the pancreas, as well as the relationship between pancreatic functions and traditional Chinese medicine (TCM) spleen system, this paper discussed the categorization of the pancreas. It is proposed that the pancreas is neither a true zang organ nor a fu organ, but possessed the attributes of an extraordinary fu-organ and can be classified under the spleen. The spleen governs transportation and transformation, ascent of the clear and dispersion of essence, which encompasses the endocrine and exocrine functions, and pancreatic enzymes and glucose-regulating hormones form the material basis for the spleen's function of dispersing essence. Diseases of the pancreas exhibit characteristics of both zang-organ deficiency and fu-organ excess, so treatment should simultaneously supplement zang-organ disease and regulate fu-organ disease when pancreas showing endocrine and exocrine co-morbidities, with focus on restoring the pancreas (spleen)'s dispersing essence function. Therapeutic strategies include supplementing spleen qi, nourishing spleen yin to strengthen spleen earth, unblocking spleen collaterals, raising spleen yang, and removing spleen turbidity to support the spleen's dispersing essence function, so as to replenish the essential qi of zang-fu organs, ensure their distribution throughout the body, and improve the endocrine and exocrine functions of the pancreas.
9.Yishen Tongluo Prescription Ameliorates Oxidative Stress Injury in Mouse Model of Diabetic Kidney Disease via Nrf2/HO-1/NQO1 Signaling Pathway
Yifei ZHANG ; Xuehui BAI ; Zijing CAO ; Zeyu ZHANG ; Jingyi TANG ; Junyu XI ; Shujiao ZHANG ; Shuaixing ZHANG ; Yiran XIE ; Yuqi WU ; Zhongjie LIU ; Weijing LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):41-51
ObjectiveTo investigate the effect and mechanism of Yishen Tongluo prescription in protecting mice from oxidative stress injury in diabetic kidney disease (DKD) via the nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1)/NAD(P)H quinone oxidoreductase 1 (NQO1) signaling pathway. MethodsSpecific pathogen-free (SPF) male C57BL/6 mice were assigned into a control group (n=10) and a modeling group (n=50). The DKD model was established by intraperitoneal injection of streptozotocin. The mice in the modeling group were randomized into a model group, a semaglutide (40 μg·kg-1) group, and high-, medium-, and low-dose (18.2, 9.1, 4.55 g·kg-1, respectively) Yishen Tongluo prescription groups, with 10 mice in each group. The treatment lasted for 12 weeks. Blood glucose and 24-h urine protein levels were measured, and the kidney index (KI) was calculated. Serum levels of creatinine (SCr), blood urea nitrogen (BUN), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were assessed. The pathological changes in the renal tissue were evaluated by hematoxylin-eosin, periodic acid-Schiff, periodic acid-silver methenamine, and Masson’s trichrome staining. Enzyme-linked immunosorbent assay kits were used to measure the levels of β2-microglobulin (β2-MG), neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule-1 (KIM-1), liver fatty acid-binding protein (L-FABP), nitric oxide synthase (NOS), glutathione (GSH), total antioxidant capacity (T-AOC), and 8-hydroxy-2'-deoxyguanosine (8-OHdG). Immunohistochemical staining was performed to examine the expression of Kelch-like ECH-associated protein 1 (Keap1) and malondialdehyde (MDA). Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot were employed to determine the mRNA and protein levels, respectively, of factors in the Nrf2/HO-1/NQO1 signaling pathway. ResultsCompared with the control group, the DKD model group showed rises in blood glucose, 24-h urine protein, KI, SCr, BUN, and ALT levels, along with glomerular hypertrophy, renal tubular dilation, thickened basement membrane, mesangial expansion, and collagen deposition. Additionally, the model group showed elevated levels of β2-MG, NGAL, KIM-1, L-FABP, NOS, and 8-OHdG, lowered levels of GSH and T-AOC, up-regulated expression of MDA and Keap1, and down-regulated expression of Nrf2, HO-1, NQO1, and glutamate-cysteine ligase catalytic subunit (GCLC) (P<0.05). Compared with the model group, the semaglutide group and the medium- and high-dose Yishen Tongluo prescription groups showed reductions in blood glucose, 24-h urine protein, KI, SCr, BUN, and ALT levels, along with alleviated pathological injuries in the renal tissue. In addition, the three groups showed lowered levels of β2-MG, NGAL, KIM-1, L-FABP, NOS, and 8-OHdG, elevated levels of GSH and T-AOC, down-regulated expression of MDA and Keap1, and up-regulated expression of Nrf2, HO-1, NQO1, and GCLC (P<0.05). ConclusionYishen Tongluo prescription exerts renoprotective effects in the mouse model of DKD by modulating the Nrf2/HO-1/NQO1 signaling pathway, mitigating oxidative stress, and reducing renal tubular injuries.
10.Dipsacus asper Treats Alzheimer's Disease in Caenorhabditis elegans by Regulating PPARα/TFEB Pathway
Mengmeng WANG ; Jianping ZHAO ; Limin WU ; Shuang CHU ; Yanli HUANG ; Zhenghao CUI ; Yiran SUN ; Pan WANG ; Hui WANG ; Zhenqiang ZHANG ; Zhishen XIE
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):104-114
ObjectiveTo investigate the anti-Alzheimer's disease (AD) effect of Dipsacus asper(DA) in the Caenorhabditis elegans model, and decipher the underlying mechanism via the peroxisome proliferator-activated receptor α (PPARα)/transcription factor EB (TFEB) pathway. MethodsFirst, transgenic AD C. elegans individuals were assigned into the blank control, model, positive control (WY14643, 20 µmol·L-1), and low-, medium-, and high-dose (100, 200, and 400 mg·L-1, respectively) DA groups. The amyloid β-42 (Aβ42) formation in the muscle cells, the paralysis time, and the deposition of amyloid β-protein (Aβ) in the head were detected. The lysosomal autophagy in the BV2 cell model was examined by Rluc-LC3wt/G120A. The expression levels of lysosomal autophagy-related proteins LC3Ⅱ, LC3I, LAMP2, and TFEB were detected by Western blot. Real-time quantitative polymerase chain reaction (Real-time PCR) was employed to determine the mRNA levels of autophagy-related genes beclin1 and Atg5 and lysosome-related genes LAMP2 and CLN2 downstream of PPARα/TFEB. A reporter gene assay was used to detect the transcriptional activities of PPARα and TFEB. Immunofluorescence was used to detect the fluorescence intensity of PPARα, and the active components of the ethanol extract of DA were identified by UPLC-MS. RCSB PDB, Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), and Autodock were used to analyze the binding between the active components and PPARα-ligand-binding domain (LBD). ResultsCompared with the model group, the positive control group and 200 and 400 mg·L-1 DA groups showed prolonged paralysis time (P<0.05), and all the treatment groups showed decreased Aβ deposition in the head (P<0.01). DA within the concentration range of 50-500 mg·L-1 did not affect the viability of BV2 cells. In addition, DA enhanced the autophagy flux (P<0.05), up-regulated the mRNA levels of beclin1, Atg5, LAMP2, and CLN2 (P<0.05, P<0.01), promoted the nuclear translocation of TFEB (P<0.05), increased LAMP2 expression and autophagy flux (P<0.05, P<0.01), and enhanced the transcriptional activities of PPARα and TFEB (P<0.01). The positive control group and 200 and 400 mg·L-1 DA groups showed enhanced fluorescence intensity of PPARα in the BV2 nucleus (P<0.01). UPLC-MS detected nine known compounds of DA, from which 8 active components of DA were screened out. The docking results suggested that a variety of components in DA could bind to PPARα-LBD and form stable hydrogen bonds. ConclusionDA may reduce the pathological changes in AD by regulating the PPARα-TFEB pathway.

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