1.α-ketoglutarate ameliorated arsenic-induced hepatic lipid deposition in offspring via PI3K/AKT signaling pathway
Shuangrui BAO ; Hongyan WU ; Ying SUN ; Tong ZHAN ; Qian YANG ; Xinru LIANG ; Zhiyan WAN ; Wenyi CHEN ; Cheng ZHANG
Acta Universitatis Medicinalis Anhui 2026;61(2):225-231
ObjectiveTo investigate the protective effect of α-ketoglutarate (α-KG) on hepatic lipid deposition in offspring caused by arsenic exposure during pregnancy. Methods8-week-old institute of cancer research (ICR) mice were mated in a ratio of 2∶1 between females and males, and the detection of vaginal plugs confirmed pregnant. A total of 32 pregnant mice were randomly divided into four groups: control group, arsenic group, α-KG group, arsenic+α-KG group. On gestational day 0-16 (GD0-GD16), the arsenic and arsenic+α-KG groups were exposed to sodium arsenite (NaAsO2 ,15 mg/L) in drinking water everyday, and the α-KG and arsenic+α-KG groups were gavaged with α-KG (2 g/kg) everyday. On GD16, pregnant mice were euthanized to collect fetal liver, and fetal body weight and crown-rump length were measured. Gene expression differences between the control group and the arsenic group were analyzed by transcriptome. The total triglycerides (TGs) and subtypes in fetal liver were detected by liquid chromatography tandem mass spectrometry (LC-MS/MS). Oil red O staining was used to observe the histopathological changes in the liver. Quantitative polymerase chain reaction (qPCR) was used to detect the expression level of genes related to lipid synthesis, transport, and degradation, and phosphatidylinositol 3' -kinase/ protein kinase B (PI3K/AKT) in the liver of fetus. ResultsTranscriptomics analysis showed that 2 144 genes were downregulated and 1 675 genes were upregulated in the arsenic exposed fetal liver; body weight and crown-rump length were reduced (PTuKey<0.05); the level of hepatic TGs was elevated in arsenic group (PTuKey<0.05); oil-red O staining showed a significant increase in lipid droplets in arsenic group (PTuKey<0.01); the expression of lipid synthesis-related genes were significantly upregulated (PTuKey<0.05); the expression of β-oxidation-related genes and lipid degradation-related genes were downregulated (PTuKey<0.05); the expression of PI3K, AKT decreased(PTuKey<0.05). Compared with the arsenic group, the body weight and crown-rump length of fetus increased in the arsenic+α-KG group (PTuKey<0.05); the level of hepatic TGs decreased in the arsenic+α-KG group (PTuKey<0.05); oil red O staining showed lipid droplets significantly decreased (PTuKey<0.01); the expression of lipid synthesis-related genes were downregulated (PTuKey<0.05), the expression of β-oxidation-related genes and lipid degradation-related genes were upregulated (PTuKey<0.05); the expression levels of PI3K and AKT increased (PTuKey<0.05). Conclusionα-KG alleviated hepatic lipid deposition in offspring exposed to arsenic during pregnancy through activating PI3K/AKT signaling pathway.
2.The Role and Molecular Mechanism of N⁶-methyladenosine Modification in Spermatogenesis
Shi-Qi MENG ; Wen-Ting LU ; Xu CHENG ; Fan YANG ; Chang-Min NIU ; Ying ZHEGN
Progress in Biochemistry and Biophysics 2026;53(5):1297-1312
Spermatogenesis is a highly ordered and spatiotemporally regulated developmental process in the male reproductive system, during which spermatogonial stem cells (SSCs), supported by the seminiferous tubule microenvironment, sequentially undergo mitosis, meiosis, and spermiogenesis to ultimately generate structurally intact spermatozoa. This complex process is accompanied by extensive transcriptional reprogramming, chromatin remodeling, and finely tuned post-transcriptional regulation. Precise control of RNA fate is therefore essential for maintaining the continuity and fidelity of spermatogenesis, and its disruption represents a major molecular basis of male infertility. N6-methyladenosine (m6A), the most abundant internal RNA modification in eukaryotes, has emerged as a critical regulator of post-transcriptional gene expression. m6A methyltransferases (“writers”) catalyze the addition of a methyl group to the N6 position of adenosine, m6A demethylases (“erasers”) remove the modification, and m6A-binding proteins (“readers”) recognize m6A-modified transcripts. Through the coordinated actions of these factors, m6A regulates transcript fate at multiple levels, including RNA splicing, nuclear export, stability, translation, and decay. Emerging evidence indicates that m6A-mediated regulation is essential across multiple stages of spermatogenesis, including SSC self-renewal and differentiation, meiotic progression, maintenance of chromosomal stability, and sperm morphogenesis. Beyond its intrinsic functions in germ cells, m6A also contributes to the regulation of the testicular microenvironment. In sertoli cells, m6A is involved in maintaining blood-testis barrier integrity, RNA processing, and paracrine signaling, thereby providing structural and metabolic support for germ cell development. In Leydig cells, m6A regulates steroidogenesis, particularly testosterone synthesis, and participates in cellular stress responses and metabolic homeostasis. Through these mechanisms, m6A indirectly influences spermatogenesis by modulating the functional state of testicular somatic cells, highlighting an integrated regulatory mode that combines cell-intrinsic and microenvironment-mediated effects. Notably, distinct classes of m6A regulators exhibit pronounced stage-specific functions and coordinated division of labor, collectively forming a multilayered and dynamic regulatory network. Writers often display dosage- and temporal window-dependent effects; erasers contribute to stage-specific demethylation and functional compensation; while readers function through a “switch-buffer” dual-layer architecture, and RNA-binding proteins (RBPs) participate in substrate selection and post-transcriptional regulation. Importantly, emerging evidence suggests that some m6A-related proteins can function through noncanonical mechanisms independent of m6A recognition, such as intrinsic RNA-binding activity, helicase function, or ribonucleoprotein complex assembly, thereby expanding the functional landscape of the m6A regulatory system. Dysregulation of m6A machinery can lead to multiple spermatogenic defects, including impaired SSC self-renewal, meiotic arrest, abnormal chromatin remodeling, and defective sperm formation, ultimately resulting in male infertility. Despite substantial advances, several critical questions remain unresolved, including the distinction between m6A-dependent and -independent mechanisms, the spatiotemporal dynamics of m6A modifications at single-cell resolution, and the coordination and antagonism among different regulatory factors. In this review, we systematically summarize the dual regulation of spermatogenesis by germ cell-intrinsic mechanisms and the testicular microenvironment, and delineate the molecular mechanisms and stage-specific functions of the dynamic m6A regulatory network. We further discuss the current limitations in the field and propose feasible experimental strategies for future investigation. Collectively, this work aims to provide a comprehensive framework for understanding the epitranscriptomic regulation of spermatogenesis and to offer theoretical insights into the pathogenesis and clinical management of male infertility.
3.The Role and Molecular Mechanism of N⁶-methyladenosine Modification in Spermatogenesis
Shi-Qi MENG ; Wen-Ting LU ; Xu CHENG ; Fan YANG ; Chang-Min NIU ; Ying ZHEGN
Progress in Biochemistry and Biophysics 2026;53(5):1297-1312
Spermatogenesis is a highly ordered and spatiotemporally regulated developmental process in the male reproductive system, during which spermatogonial stem cells (SSCs), supported by the seminiferous tubule microenvironment, sequentially undergo mitosis, meiosis, and spermiogenesis to ultimately generate structurally intact spermatozoa. This complex process is accompanied by extensive transcriptional reprogramming, chromatin remodeling, and finely tuned post-transcriptional regulation. Precise control of RNA fate is therefore essential for maintaining the continuity and fidelity of spermatogenesis, and its disruption represents a major molecular basis of male infertility. N6-methyladenosine (m6A), the most abundant internal RNA modification in eukaryotes, has emerged as a critical regulator of post-transcriptional gene expression. m6A methyltransferases (“writers”) catalyze the addition of a methyl group to the N6 position of adenosine, m6A demethylases (“erasers”) remove the modification, and m6A-binding proteins (“readers”) recognize m6A-modified transcripts. Through the coordinated actions of these factors, m6A regulates transcript fate at multiple levels, including RNA splicing, nuclear export, stability, translation, and decay. Emerging evidence indicates that m6A-mediated regulation is essential across multiple stages of spermatogenesis, including SSC self-renewal and differentiation, meiotic progression, maintenance of chromosomal stability, and sperm morphogenesis. Beyond its intrinsic functions in germ cells, m6A also contributes to the regulation of the testicular microenvironment. In sertoli cells, m6A is involved in maintaining blood-testis barrier integrity, RNA processing, and paracrine signaling, thereby providing structural and metabolic support for germ cell development. In Leydig cells, m6A regulates steroidogenesis, particularly testosterone synthesis, and participates in cellular stress responses and metabolic homeostasis. Through these mechanisms, m6A indirectly influences spermatogenesis by modulating the functional state of testicular somatic cells, highlighting an integrated regulatory mode that combines cell-intrinsic and microenvironment-mediated effects. Notably, distinct classes of m6A regulators exhibit pronounced stage-specific functions and coordinated division of labor, collectively forming a multilayered and dynamic regulatory network. Writers often display dosage- and temporal window-dependent effects; erasers contribute to stage-specific demethylation and functional compensation; while readers function through a “switch-buffer” dual-layer architecture, and RNA-binding proteins (RBPs) participate in substrate selection and post-transcriptional regulation. Importantly, emerging evidence suggests that some m6A-related proteins can function through noncanonical mechanisms independent of m6A recognition, such as intrinsic RNA-binding activity, helicase function, or ribonucleoprotein complex assembly, thereby expanding the functional landscape of the m6A regulatory system. Dysregulation of m6A machinery can lead to multiple spermatogenic defects, including impaired SSC self-renewal, meiotic arrest, abnormal chromatin remodeling, and defective sperm formation, ultimately resulting in male infertility. Despite substantial advances, several critical questions remain unresolved, including the distinction between m6A-dependent and -independent mechanisms, the spatiotemporal dynamics of m6A modifications at single-cell resolution, and the coordination and antagonism among different regulatory factors. In this review, we systematically summarize the dual regulation of spermatogenesis by germ cell-intrinsic mechanisms and the testicular microenvironment, and delineate the molecular mechanisms and stage-specific functions of the dynamic m6A regulatory network. We further discuss the current limitations in the field and propose feasible experimental strategies for future investigation. Collectively, this work aims to provide a comprehensive framework for understanding the epitranscriptomic regulation of spermatogenesis and to offer theoretical insights into the pathogenesis and clinical management of male infertility.
4.Discussion on processing time for Polygonatum kingianum based on analysis of correlation between sugar components and color changes
GUO Hong ; YAO Rui ; LI Zhe ; FAN Jing ; WANG Ying ; GUO Xiaohan ; CHEN Jia ; DUAN Baozhong ; YANG Jianbo ; JING Wenguang ; CHENG Xianlong ; WEI Feng
Drug Standards of China 2026;27(1):0083-0091
Objective: To investigate the correlation between color parameters (L*, a*, b*, Eab*) and the contents of reducing sugars, total polysaccharides, total oligosaccharides, as well as four saccharides (fructose, glucose, sucrose, and kestose) during the nine cycles of steaming and sun-drying processing of Polygonatum kingianum, and to preliminarily explore the optimal processing duration.
Methods: The color changes were objectively evaluated using a colorimeter. The anthrone-sulfuric acid method was employed to determine total polysaccharides and oligosaccharides. The 3,5-dinitrosalicylic acid (DNS) colorimetric method was used to measure total reducing sugar content. High-performance liquid chromatography coupled with charged aerosol detection (HPLC-CAD) was applied for quantitative analysis of fructose, glucose, sucrose, and kestose. Multivariate statistical analysis was conducted to assess samples from different processing stages.
Results: Significant variations in color and component contents were observed across processing stages. The herbal pieces progressively darkened with increased processing cycles: brightness (L*) and total color difference (Eab*) initially decreased then stabilized, while a* (red-green) and b* (yellow-blue) values first increased then declined. Total polysaccharides and oligosaccharides showed overall decreasing trends, whereas reducing sugars initially increased before stabilizing. Fructose and glucose levels rose continuously, while sucrose and kestose decreased progressively, becoming undetectable after five cycles.
Conclusion: The chromatic alteration and saccharide composition of P. kingianum showed significant correlation with processing duration. Both total color difference (Eab*) and reducing sugar content stabilized after four processing cycles (12 hours), suggesting that four cycles of steaming and sun-drying may represent the optimal processing duration.
5.Progress of Research on Advanced Non-Small Cell Lung Cancer with HER-2 Mutation
Liang ZHANG ; Changliang YANG ; Peidong LI ; Ying CHENG
Cancer Research on Prevention and Treatment 2025;52(2):87-92
Anti-tumor drug research and development in non-small cell lung cancer (NSCLC) is rapidly developing, and the clinical application of high-throughput sequencing technology is also becoming widespread. Accordingly, researchers are focusing on human epidermal growth factor receptor-2 (HER-2) gene as a rare target of NSCLC, and a series of exploratory studies has been performed. Traditional chemotherapy and immunotherapy are unsatisfactory in the HER-2 mutant population, whereas the survival improvement of anti-HER-2 monoclonal antibodies and pan-HER inhibitors is limited. The development of antibody drug conjugate (ADC) ushers in a turning point for HER-2-mutated NSCLC, and new ADC drugs represented by trastuzumab deruxtecan are making a breakthrough. It opens up a new era of precision therapy for advanced HER-2-mutated NSCLC. Additionally, novel HER-2 inhibitors show very encouraging initial efficacy and safety, and clinical trials are ongoing. This review focuses on the latest progress of research on HER-2-mutated NSCLC.
6.Efficacy of Differential Dosage of Pueraria in Gegen Qinliantang on Acute Enteritis Model in Mice
Ruiying ZHANG ; Ping WANG ; Di ZHANG ; Hongfa CHENG ; Ying ZHANG ; Zhu DENG ; Hui FENG ; Min LIU ; Yang TANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):197-204
ObjectiveTo investigate whether there are differences in the efficacy of Gegen Qinliantang with different contents of Puerariae Lobatae Radix on the acute enteritis (AE) model mice and provide a scientific basis for the interpretation of Gegen Qinliantang in the treatment of "Xie Re Li". MethodsA total of 112 male BALB/c mice were randomly divided into a blank group,model group,single Puerariae Lobatae Radix group,non-Puerariae Lobatae Radix group,regular dose Gegen Qinliantang group (regular dose group),half-dose Puerariae Lobatae Radix group,and doubled-dose Puerariae Lobatae Radix group, with 16 mice in each group. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of the colon tissue. Western blot was employed to detect the expression of ZO-1 (a protein in the tight junction) and Occludin in the colon tissue, as well as the changes of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β). ResultsCompared with the blank group,the DAI scores of the mice in the model group were significantly higher (P<0.05),and the histopathological sections of their colon tissues showed mucosal damage,glandular atrophy,disordered arrangement,and a large number of inflammatory cells infiltration,and the expression of ZO-1 and Occludin proteins in their colon tissues was significantly down-regulated (P<0.05,P<0.01). The expression of inflammatory factors TNF-α and IL-1β was significantly up-regulated (P<0.05,P<0.01). Compared with the model group,the DAI scores of mice in all dosing groups decreased significantly (P<0.05),with the most significant effect in the regular dose group. After 7 d of drug administration,the regular dose group had the best impact on the repair of colonic mucosa in the AE mouse model. The regular dose group significantly down-regulated the expression of TNF-α (P<0.05) and significantly up-regulated the expression of ZO-1 protein (P<0.05). The doubled-dose Puerariae Lobatae Radix group significantly down-regulated the expression of IL-1β protein (P<0.01),and there was no significant difference between all dosing groups and the model group in terms of the expression of Occludin protein. After 14 d of drug administration,the best effect on the repair of colonic mucosa in the AE mouse model was observed in the doubled dose Puerariae Lobatae Radix group. All groups except the non-Puerariae Lobatae Radix group significantly down-regulated the expression of TNF-α (P<0.01). Meanwhile,the regular dose group and doubled-dose Puerariae Lobatae Radix group significantly elevated the expression level of Occludin protein (P<0.01). The doubled-dose Puerariae Lobatae Radix group also significantly inhibited the expression of IL-1β protein (P<0.05) and up-regulated ZO-1 protein expression (P<0.05). ConclusionGegen Qinliantang can reduce the pathological damage of colon tissue, protect the barrier function and structure of intestinal epithelial cells, and reduce the expression of inflammatory factors, so as to achieve the therapeutic effect of AE model mice. When comparing the therapeutic efficacy of Gegen Qinliantang containing different Gegen contents, Gegen Qinliantang with the proportion of the original formula of Zhongjing was the most effective in AE model mice.
7.Efficacy of Differential Dosage of Pueraria in Gegen Qinliantang on Acute Enteritis Model in Mice
Ruiying ZHANG ; Ping WANG ; Di ZHANG ; Hongfa CHENG ; Ying ZHANG ; Zhu DENG ; Hui FENG ; Min LIU ; Yang TANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):197-204
ObjectiveTo investigate whether there are differences in the efficacy of Gegen Qinliantang with different contents of Puerariae Lobatae Radix on the acute enteritis (AE) model mice and provide a scientific basis for the interpretation of Gegen Qinliantang in the treatment of "Xie Re Li". MethodsA total of 112 male BALB/c mice were randomly divided into a blank group,model group,single Puerariae Lobatae Radix group,non-Puerariae Lobatae Radix group,regular dose Gegen Qinliantang group (regular dose group),half-dose Puerariae Lobatae Radix group,and doubled-dose Puerariae Lobatae Radix group, with 16 mice in each group. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of the colon tissue. Western blot was employed to detect the expression of ZO-1 (a protein in the tight junction) and Occludin in the colon tissue, as well as the changes of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β). ResultsCompared with the blank group,the DAI scores of the mice in the model group were significantly higher (P<0.05),and the histopathological sections of their colon tissues showed mucosal damage,glandular atrophy,disordered arrangement,and a large number of inflammatory cells infiltration,and the expression of ZO-1 and Occludin proteins in their colon tissues was significantly down-regulated (P<0.05,P<0.01). The expression of inflammatory factors TNF-α and IL-1β was significantly up-regulated (P<0.05,P<0.01). Compared with the model group,the DAI scores of mice in all dosing groups decreased significantly (P<0.05),with the most significant effect in the regular dose group. After 7 d of drug administration,the regular dose group had the best impact on the repair of colonic mucosa in the AE mouse model. The regular dose group significantly down-regulated the expression of TNF-α (P<0.05) and significantly up-regulated the expression of ZO-1 protein (P<0.05). The doubled-dose Puerariae Lobatae Radix group significantly down-regulated the expression of IL-1β protein (P<0.01),and there was no significant difference between all dosing groups and the model group in terms of the expression of Occludin protein. After 14 d of drug administration,the best effect on the repair of colonic mucosa in the AE mouse model was observed in the doubled dose Puerariae Lobatae Radix group. All groups except the non-Puerariae Lobatae Radix group significantly down-regulated the expression of TNF-α (P<0.01). Meanwhile,the regular dose group and doubled-dose Puerariae Lobatae Radix group significantly elevated the expression level of Occludin protein (P<0.01). The doubled-dose Puerariae Lobatae Radix group also significantly inhibited the expression of IL-1β protein (P<0.05) and up-regulated ZO-1 protein expression (P<0.05). ConclusionGegen Qinliantang can reduce the pathological damage of colon tissue, protect the barrier function and structure of intestinal epithelial cells, and reduce the expression of inflammatory factors, so as to achieve the therapeutic effect of AE model mice. When comparing the therapeutic efficacy of Gegen Qinliantang containing different Gegen contents, Gegen Qinliantang with the proportion of the original formula of Zhongjing was the most effective in AE model mice.
8.Biosynthesis of ganoderic acid and its derivatives.
Hong-Yan SONG ; Wan YANG ; Li-Wei LIU ; Xia-Ying CHENG ; Dong-Feng YANG ; Zong-Qi YANG
China Journal of Chinese Materia Medica 2025;50(5):1155-1163
Ganoderic acid is a class of lanostane-type triterpenoids found in Ganoderma species, and is one of the most important pharmacologically active components in G. lucidum, exhibiting antioxidant, anti-neuropsychiatric, anti-tumor, and immune-enhancing properties. The content of ganoderic acid in G. lucidum is very low, and the traditional extraction process is complex, yielding minimal amounts at high cost. The biosynthetic pathway of G. lucidum triterpenoids(GLTs), including the synthesis of different structural forms of ganoderic acid from lanosterol, as well as the molecular regulatory mechanisms involving key regulatory enzyme genes and their functions, are not yet fully understood. With the continuous development of synthetic biology technologies, there has been a deeper understanding of the biosynthesis and metabolic regulation pathways of ganoderic acid and its derivatives at the molecular level. Research has explored the key regulatory enzyme genes related to ganoderic acid biosynthesis and their functions. Moreover, through the optimization of synthetic biology and culture conditions, large-scale production and preparation of GLTs at the cellular level have been achieved. This paper reviews and analyzes the latest research progress on the biosynthesis pathways and metabolic regulation of GLTs, focusing on the configuration of ganoderic acid and its derivatives, the biosynthetic pathways, key enzyme genes, transcription factors related to ganoderic acid biosynthesis, signal transduction mechanisms, and factors affecting triterpenoid biotransformation. This review is expected to provide a theoretical basis and technical reference for improving the efficient production of triterpenoid pharmacological components and the exploitation and utilization of G. lucidum resources.
Triterpenes/chemistry*
;
Reishi/chemistry*
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Biosynthetic Pathways
;
Lanosterol
9.Effect of medicinal parts and harvest seasons on nature-flavor correlation of plant-based Chinese materia medica.
Qi-Ao MA ; Guang YANG ; Hong-Chao WANG ; Ying LI ; Meng CHENG ; Tie-Lin WANG ; Kai SUN ; Xiu-Lian CHI
China Journal of Chinese Materia Medica 2025;50(15):4228-4237
This study selected 6 529 plant-based Chinese materia medica(PCMM) from Chinese Materia Medica as research subjects and applied a random permutation test to explore the overall correlation characteristics between nature and flavor, as well as the correlation characteristics after distinguishing different medicinal parts and harvest seasons. The results showed that the overall correlation characteristics between nature and flavor in PCMM were significantly associated in the following pairs: cold and bitter, cool and bitter, cool and astringent, cool and light, neutral and sweet, neutral and astringent, neutral and light, neutral and sour, hot and pungent, and warm and pungent. When analyzing the data by distinguishing medicinal parts and/or harvest seasons, new correlation patterns emerged, characterized by the disappearance of some significant correlations and the emergence of new ones. When analyzing by medicinal parts alone, significant correlations were found in the following cases: cold and light in leaves, cold and salty in barks, cool and sweet in fruits and seeds, neutral and pungent in whole herbs, neutral and salty in stems, and warm and salty in flowers. However, no significant correlations were found between cool and bitter in stems and other types of herbs, cool and astringent in fruits, seeds, flowers, and other types of herbs, cool and light in leaves, fruits, seeds, barks, flowers and other types of herbs, neutral and sweet in barks, neutral and astringent in whole herbs and stems, neutral and light in leaves, fruits, seeds, and flowers, neutral and sour in whole herbs, stems, barks, flowers, and other types of herbs, and hot and pungent in whole herbs, stems, flowers, and other types of herbs. When analyzing by harvest season alone, significant correlations were found in the following cases: cold and salty, and cool and sour in herbs harvested in winter, and neutral and salty in herbs harvested year-round. However, no significant correlation was found between cool and light in herbs harvested in winter. When considering both medicinal parts and harvest seasons, compared to the independent influence of medicinal parts, 14 new significant correlations emerged(e.g., the correlation between cool and bitter in stems harvested in spring), while 53 previously significant correlations disappeared(e.g., the correlation between cool and bitter in barks harvested in summer). Compared to the independent influence of harvest seasons, 11 new significant correlations appeared(e.g., the correlation between cold and light in barks harvested in autumn), while 50 previously significant correlations disappeared(e.g., the correlation between hot and pungent in leaves harvested in winter). This study is the first to reveal the influence of medicinal parts and harvest seasons on the correlation between nature and flavor in PCMM, which highlights that these two factors can interact and jointly affect nature-flavor correlations. Further research is needed to explore the underlying mechanisms. This study provides a deeper understanding of the inherent scientific connotations of herbal properties and offers a theoretical foundation for the cultivation and harvesting of PCMM.
Seasons
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Plants, Medicinal/growth & development*
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Drugs, Chinese Herbal/chemistry*
;
Taste
10.Mechanism of Chaijin Jieyu Anshen Formula in regulating synaptic damage in nucleus accumbens neurons of rats with insomnia complicated with depression through TREM2/C1q axis.
Ying-Juan TANG ; Jia-Cheng DAI ; Song YANG ; Xiao-Shi YU ; Yao ZHANG ; Hai-Long SU ; Zhi-Yuan LIU ; Zi-Xuan XIANG ; Jun-Cheng LIU ; Hai-Xia HE ; Jian LIU ; Yuan-Shan HAN ; Yu-Hong WANG ; Man-Shu ZOU
China Journal of Chinese Materia Medica 2025;50(16):4538-4545
This study aims to investigate the effect of Chaijin Jieyu Anshen Formula on the neuroinflammation of rats with insomnia complicated with depression through the regulation of triggering receptor expressed on myeloid cells 2(TREM2)/complement protein C1q signaling pathway. Rats were randomly divided into a normal group, a model group, a positive drug group, as well as a high, medium, and low-dose groups of Chaijin Jieyu Anshen Formula, with 10 rats in each group. Except for the normal group, the other groups were injected with p-chlorophenylalanine and exposed to chronic unpredictable mild stress to establish the rat model of insomnia complicated with depression. The sucrose preference experiment, open field experiment, and water maze test were performed to evaluate the depression in rats. Enzyme-linked immunosorbent assay was employed to detect serum 5-hydroxytryptamine(5-HT), dopamine(DA), and norepinephrine(NE) levels. Hematoxylin and eosin staining and Nissl staining were used to observe the damage in nucleus accumbens neurons. Western blot and immunofluorescence were performed to detect TREM2, C1q, postsynaptic density 95(PSD-95), and synaptophysin 1(SYN1) expressions in rat nucleus accumbens, respectively. Golgi-Cox staining was utilized to observe the synaptic spine density of nucleus accumbens neurons. The results show that, compared with the model group, Chaijin Jieyu Anshen Formula can significantly increase the sucrose preference as well as the distance and number of voluntary activities, shorten the immobility time in forced swimming test and the successful incubation period of positioning navigation, and prolong the stay time of space exploration in the target quadrant test. The serum 5-HT, DA, and NE contents in the model group are significantly lower than those in the normal group, with the above contents significantly increased after the intervention of Chaijin Jieyu Anshen Formula. In addition, Chaijin Jieyu Anshen Formula can alleviate pathological damages such as swelling and loose arrangement of tissue cells in the nucleus accumbens, while increasing the Nissl body numbers. Chaijin Jieyu Anshen Formula can improve synaptic damage in the nucleus accumbens and increase the synaptic spine density. Compared to the normal group, the expression of C1q protein was significantly higher in the model group, while the expression of TREM2 protein was significantly lower. Compared to the model group, the intervention with Chaijin Jieyu Anshen Formula significantly downregulated the expression of C1q protein and significantly upregulated the expression of TREM2. Compared with the model group, the PSD-95 and SYN1 fluorescence intensity is significantly increased in the groups receiving different doses of Chaijin Jieyu Anshen Formula. In summary, Chaijin Jieyu Anshen Formula can reduce the C1q protein expression, relieve the TREM2 inhibition, and promote the synapse-related proteins PSD-95 and SNY1 expression. Chaijin Jieyu Anshen Formula improves synaptic injury of the nucleus accumbens neurons, thereby treating insomnia complicated with depression.
Animals
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Male
;
Rats
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Nucleus Accumbens/metabolism*
;
Drugs, Chinese Herbal/administration & dosage*
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Depression/complications*
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Membrane Glycoproteins/genetics*
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Rats, Sprague-Dawley
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Sleep Initiation and Maintenance Disorders/complications*
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Neurons/metabolism*
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Receptors, Immunologic/genetics*
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Signal Transduction/drug effects*
;
Synapses/metabolism*

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