1.Current status of dental caries among primary and middle school students in Xinjiang Production and Construction Corps
CHEN Hui ; MIAO Shanshan ; LIU Xianfeng ; ZHANG Hui
Journal of Preventive Medicine 2026;38(1):102-106
Objective:
To understand the prevalence of dental caries and its influencing factors among primary and middle school students in the Xinjiang Production and Construction Corps (hereinafter referred to as "the Corps"), so as to provide a reference for formulating targeted prevention strategies and promoting oral health of primary and middle school students.
Methods:
Primary and middle school students in the Corps were selected as survey subjects by a multi-stage stratified random cluster sampling method in September 2023. Basic information and dietary behaviors were collected through questionnaire surveys, and dental caries status was examined by oral health technicians. Multivariable logistic regression model was used to analyze the influencing factors of dental caries among primary and middle school students.
Results:
A total of 80 370 primary and middle school students were investigated, including 40 582 males (50.49%) and 39 788 females (49.51%). There were 37 608 primary school students (46.79%), 34 612 junior high school students (43.07%), and 8 150 senior high school students (10.14%). There were 26 669 students with dental caries, with a prevalence rate of 33.18%. Multivariable logistic regression analysis showed that the risk of dental caries was higher among female students (OR=1.170, 95%CI: 1.136-1.206), students in suburban counties (OR=1.212, 95%CI: 1.166-1.258), boarding students (OR=1.306, 95%CI: 1.257-1.357), and those with a frequency of fried food intake ≥1 time per day (OR=1.175, 95%CI: 1.084-1.273). Conversely, the risk of dental caries was lower among middle school students (OR=0.542, 95%CI: 0.524-0.560), high school students (OR=0.661, 95%CI: 0.620-0.705), and those with a frequency of vegetable intake ≥1 time per day (1 time per day, OR=0.900, 95%CI: 0.838-0.967), (≥2 time per day, OR=0.879, 95%CI: 0.819-0.944), and those who sometimes ate breakfast (OR=0.907, 95%CI: 0.874-0.942).
Conclusions
The prevalence of dental caries among primary and middle school students in the Corps is relatively high, and is influenced by various factors such as gender, school stage, area, boarding status, and dietary behaviors. It is suggested to strengthen oral health knowledge education among students, conduct regular oral health examinations, and improve the overall level of oral health.
2.Mechanism of miR-21 targeting inhibition of the PTEN/AKT/mTOR pathway in ameliorating chronic renal fibrosis in mice
Jiao QI ; Shanshan XU ; Qige QI ; Yan MENG ; Jianrong ZHAO ; Liying ZHANG
Acta Universitatis Medicinalis Anhui 2026;61(2):217-224
ObjectiveTo investigate the mechanism through which miR‑21 improves chronic renal fibrosis in mice via targeted modulation of the phosphatase and tensin homolog (PTEN)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway. MethodsThirty‑two chronic kidney disease model mice were randomly divided into four groups (n=8 each group): model group, miR‑21 overexpression group, miR‑21 inhibition group, and miR‑21 inhibition + MK‑2206 group. Eight healthy mice were included as the control group. The miR‑21 overexpression, miR‑21 inhibition, and miR‑21 inhibition + MK‑2206 groups received tail‑vein injections of lentivirus (50 μL, 1×10⁸ TU per mouse) once weekly for three weeks. The control and model groups were injected with an equal volume of empty vector (LV‑NC). The miR‑21 inhibition + MK‑2206 group additionally received gavage of the AKT/mTOR pathway inhibitor MK‑2206 (480 mg/kg) once weekly for three weeks. The expressions of miR‑21, 24 h urinary protein, serum creatinine (Scr), blood urea nitrogen (BUN), and renal tissue levels of collagen Ⅰ, collagen Ⅲ, α‑smooth muscle actin (α‑SMA), and PTEN protein, as well as p‑AKT/AKT and p‑mTOR/mTOR ratios, were compared among groups. HE staining was used to observe pathological changes in renal tissue, and Masson staining was used to observe the degree of renal fibrosis. A dual‑luciferase assay was performed to verify the targeting relationship between miR‑21 and PTEN. ResultsCompared with the model group, miR‑21 expression in renal tissue increased in the miR‑21 overexpression group (P<0.05) and decreased in the miR‑21 inhibition group (P<0.05). Compared with the model group, the miR‑21 overexpression group showed increased 24 h urinary protein, Scr, BUN, and renal tissue expression of collagen Ⅰ, collagen Ⅲ, and α‑SMA (all P<0.05), while these indicators decreased in the miR‑21 inhibition group (P<0.05). Compared with the miR‑21 inhibition group, the miR‑21 inhibition + MK‑2206 group exhibited lower 24‑h urinary protein, Scr, BUN, and renal tissue expression of Collagen Ⅰ, Collagen Ⅲ, and α‑SMA (all P<0.05). Compared with the model group, the miR‑21 overexpression group showed decreased PTEN protein expression (P<0.05) and increased p‑AKT/AKT and p‑mTOR/mTOR ratios (P<0.05), while the miR‑21 inhibition group showed increased PTEN expression (P<0.05) and decreased p‑AKT/AKT and p‑mTOR/mTOR ratios (P<0.05). Compared with the miR‑21 inhibition group, the miR‑21 inhibition + MK‑2206 group had lower p‑AKT/AKT and p‑mTOR/mTOR ratios (P<0.05), with no significant difference in PTEN protein expression. HE and Masson staining showed normal kidney structure and almost no fibrosis in the control group. The model group exhibited glomerular enlargement, capillary loop adhesion, and focal fibrosis. The miR-21 overexpression group showed severe destruction of glomerular structure, accompanied by extensive fibrosis and renal tubular atrophy. The pathological changes and degree of fibrosis were alleviated in the miR-21 inhibition group. The miR-21 inhibition + MK-2206 group showed only mild pathological changes and mild fibrosis, with the interstitium being largely normal. Compared with PTEN-WT + NC mimics 1, the relative luciferase activity in the PTEN-WT + miR-21 mimics group decreased (P<0.001). There was no statistically significant difference in relative luciferase activity between PTEN-WT + NC mimics group and PTEN-MUT + miR-21 mimics group. ConclusionmiR‑21 may improve renal function indicators and alleviate renal fibrosis in chronic kidney disease mice via targeted modulation of PTEN and subsequently inhibiting the AKT/mTOR pathway.
3.Research on the correlation between Ddit3-Trib3-Akt signaling pathway and spermatogenesis in rats based on the testicular tissue co-culture system
Yan LI ; Shanshan LIU ; Lin GAO ; Lingyi KONG ; Xia YUN ; Yan ZHANG ; Taodi LIU
Acta Universitatis Medicinalis Anhui 2026;61(1):91-97
ObjectiveTo verify the association between the Ddit3-Trib3-Akt signaling pathway and rat spermatogenesis by constructing an in vitro co-culture system of testis. MethodsTesticular tissue blocks from 20-25-day-old male rats were placed in an in vitro culture system, and the culture medium was replaced every 2 to 3 days. PCR was used to verify the expression of marker genes of various spermatogenic cells. RNA interference technology was employed to verify the correlation between the Ddit3-Trib3-Akt signaling pathway and rat spermatogenesis. ResultsThe co-culture system could be continuously cultured for more than 2.5 months in vitro. RT-PCR showed that specific marker genes of spermatogonia, spermatocyte and spermoblast were expressed. The RNA and protein expression of Trib3 and Akt changed after the knocking down of Ddit3 and Trib3, respectively. It demonstrated the existence of Ddit3-Trib3-Akt signaling pathway in rat spermatogenesis. ConclusionThe culture time of more than 2.5 months indicates that the culture system can temporarily maintain the proliferation and differentiation of stem cells, and simultaneously maintain and stabilize spermatogenesis in a simple system. The successful validation of the Ddit3-Trib3-Akt signaling pathway also confirms that this culture system can be used to study possible molecular mechanisms of spermatogenesis in vitro.
4.Effect of Xiaoqinglong Decoction (小青龙汤) on Th2 Immune Responses and Nasal Mucosal TSLP/OX40L Pathway in Allergic Rhinitis Model Rats
Yimeng CHEN ; Yuye CHEN ; Guangchun YU ; Bei CHEN ; Jianwei ZHANG ; Shanshan DING ; Xiaoting YANG ; Baifan YU ; Yating CAI ; Xuejuan LIN ; Mengting ZHANG
Journal of Traditional Chinese Medicine 2026;67(9):994-1002
ObjectiveTo explore the potential mechanism of Xiaoqinglong Decoction (小青龙汤, XD) in the treatment of allergic rhinitis. MethodsForty-five rats were randomly assigned to a control group, a model group, a loratadine group, low-, medium- and high-dose XD groups, and low-, medium- and high-dose Mahuang Decoction and Cang'erzi Powder (麻黄汤合苍耳子散, MDCP) groups. Except for the control group, rats were administered with ovalbumin (OVA) and aluminum hydroxide via intraperitoneal injection for 14 days to establish an allergic rhinitis model. After the 14th-day injection, nasal stimulation was continued with 20 μl of 10% OVA solution to maintain the model. Rats in the control group and the model group received 10 ml/(kg·d) of saline, whereas those in the loratadine group were administered with 0.9 mg/(kg·d) of loratadine. The low-, medium- and high-dose XD groups were administered XD at the dose of 2.7, 5.4, and 10.8 g/(kg·d), respectively. The low-, medium- and high-dose MDCP groups were administered MDCP at the dose of 2.43, 4.86, and 9.72 g/(kg·d), respectively. All treatments were administered by gavage once daily for 7 consecutive days. One hour after the final gavage, nasal symptom scores were recorded for all group of rats. The next day, serum levels of immunoglobulin E (IgE), interleukin-4 (IL-4), and interleukin-13 (IL-13) were measured. HE staining was used to observe the pathological morphology of the nasal mucosal tissue. Quantitative reverse transcription PCR (RT-qPCR) and Western Blot were performed to assess mRNA and protein expression of thymic stromal lymphopoietin (TSLP) and OX40 ligand (OX40L) in the nasal mucosa. ResultsCompared to the control group, total nasal symptom score in the model group significantly increased (P<0.01). HE staining revealed disrupted and adhered cilia, thickened basement membranes, and extensive inflammatory cell infiltration in the nasal mucosa. Serum levels of total IgE, IL-4, and IL-13, as well as TSLP and OX40L mRNA and protein expression in the nasal mucosa, were significantly elevated in the model group (P<0.05 or P<0.01). Compared to the model group, the total nasal symptom scores in all drug intervention groups were significantly reduced; the serum total IgE levels in the loratadine group, the low- and medium-dose XD groups, and the low- and high-dose MDCP groups were significantly reduced; and the serum levels of IL-4 and IL-13 in the high-dose XD group and the high-dose MDCP group decreased (P<0.05 or P<0.01). Nasal mucosal structure was improved. Except for the low-dose MDCP group, all other intervention groups showed a significant reduction in TSLP and OX40L mRNA expression in the nasal mucosa (P<0.01). All doses of XD and the medium- and high-dose MDCP groups significantly decreased the protein levels of TSLP and OX40L (P<0.05). The medium-dose XD group exhibited more improvement of nasal symptom scores and greater suppression of expression of TSLP and OX40L mRNA, and TSLP protein levels compared to the loratadine group (P<0.05). ConclusionXD may protect nasal mucosa of rats and alleviate allergic rhinitis by suppressing the TSLP/OX40L pathway, thereby attenuating Th2-mediated immune responses.
5.Effect of Xiaoqinglong Decoction (小青龙汤) on Th2 Immune Responses and Nasal Mucosal TSLP/OX40L Pathway in Allergic Rhinitis Model Rats
Yimeng CHEN ; Yuye CHEN ; Guangchun YU ; Bei CHEN ; Jianwei ZHANG ; Shanshan DING ; Xiaoting YANG ; Baifan YU ; Yating CAI ; Xuejuan LIN ; Mengting ZHANG
Journal of Traditional Chinese Medicine 2026;67(9):994-1002
ObjectiveTo explore the potential mechanism of Xiaoqinglong Decoction (小青龙汤, XD) in the treatment of allergic rhinitis. MethodsForty-five rats were randomly assigned to a control group, a model group, a loratadine group, low-, medium- and high-dose XD groups, and low-, medium- and high-dose Mahuang Decoction and Cang'erzi Powder (麻黄汤合苍耳子散, MDCP) groups. Except for the control group, rats were administered with ovalbumin (OVA) and aluminum hydroxide via intraperitoneal injection for 14 days to establish an allergic rhinitis model. After the 14th-day injection, nasal stimulation was continued with 20 μl of 10% OVA solution to maintain the model. Rats in the control group and the model group received 10 ml/(kg·d) of saline, whereas those in the loratadine group were administered with 0.9 mg/(kg·d) of loratadine. The low-, medium- and high-dose XD groups were administered XD at the dose of 2.7, 5.4, and 10.8 g/(kg·d), respectively. The low-, medium- and high-dose MDCP groups were administered MDCP at the dose of 2.43, 4.86, and 9.72 g/(kg·d), respectively. All treatments were administered by gavage once daily for 7 consecutive days. One hour after the final gavage, nasal symptom scores were recorded for all group of rats. The next day, serum levels of immunoglobulin E (IgE), interleukin-4 (IL-4), and interleukin-13 (IL-13) were measured. HE staining was used to observe the pathological morphology of the nasal mucosal tissue. Quantitative reverse transcription PCR (RT-qPCR) and Western Blot were performed to assess mRNA and protein expression of thymic stromal lymphopoietin (TSLP) and OX40 ligand (OX40L) in the nasal mucosa. ResultsCompared to the control group, total nasal symptom score in the model group significantly increased (P<0.01). HE staining revealed disrupted and adhered cilia, thickened basement membranes, and extensive inflammatory cell infiltration in the nasal mucosa. Serum levels of total IgE, IL-4, and IL-13, as well as TSLP and OX40L mRNA and protein expression in the nasal mucosa, were significantly elevated in the model group (P<0.05 or P<0.01). Compared to the model group, the total nasal symptom scores in all drug intervention groups were significantly reduced; the serum total IgE levels in the loratadine group, the low- and medium-dose XD groups, and the low- and high-dose MDCP groups were significantly reduced; and the serum levels of IL-4 and IL-13 in the high-dose XD group and the high-dose MDCP group decreased (P<0.05 or P<0.01). Nasal mucosal structure was improved. Except for the low-dose MDCP group, all other intervention groups showed a significant reduction in TSLP and OX40L mRNA expression in the nasal mucosa (P<0.01). All doses of XD and the medium- and high-dose MDCP groups significantly decreased the protein levels of TSLP and OX40L (P<0.05). The medium-dose XD group exhibited more improvement of nasal symptom scores and greater suppression of expression of TSLP and OX40L mRNA, and TSLP protein levels compared to the loratadine group (P<0.05). ConclusionXD may protect nasal mucosa of rats and alleviate allergic rhinitis by suppressing the TSLP/OX40L pathway, thereby attenuating Th2-mediated immune responses.
6.Research progress on Ecoflex in the field of stomatology
BI Huimin ; CHEN Jianhang ; ZHANG Jingxin ; YANG Maohua ; DENG Shuangshan ; SU Yingyue ; GAO Shanshan
Journal of Prevention and Treatment for Stomatological Diseases 2026;34(5):507-517
Ecoflex is a commercial designation for elastomers developed based on the principles of environmental sustainability and flexibility. Various manufacturers offer different types of Ecoflex products with distinct compositions and functions. Among these, the platinum-catalyzed silicone rubber Ecoflex series has demonstrated considerable applicability in various fields of oral medicine due to its excellent flexibility, biocompatibility, stability across a wide temperature range, and tunable mechanical properties. In tissue engineering, it can simulate the mechanical behavior of oral mucosa, and is used in cleft lip surgical training models and preoperative evaluation for temporal bone defect reconstruction. In the field of wearable devices, leveraging its encapsulation protection and flexible characteristics, highly sensitive sensors constructed from Ecoflex can monitor signals such as oral bite force and masticatory muscle activity, thereby aiding in the diagnosis of temporomandibular joint disorders and postoperative evaluation of cleft lip and palate. Moreover, when combined with bio-waste materials, it promotes the functionalization and sustainability of oral wearable devices.In drug delivery systems, its conformability and controlled-release capability address challenges in localized oral drug administration. Designs such as flexible microneedles and temperature-responsive composite systems provide precise solutions for treating periodontitis and oral ulcers. In minimally invasive surgical instruments, its softness enables the development of soft robots and magnetically controlled microfluidic valves, enhancing surgical safety and precision. In the field of oral rehabilitation, Ecoflex soft liner materials, inspired by the suction cup structure of octopus tentacles, improve denture retention. Their low modulus reduces mucosal irritation, ensuring both comfort and durability. Although Ecoflex shows great potential in biomedical applications, it still faces certain challenges, particularly regarding long-term stability after implantation, mechanical fatigue resistance, and microbial colonization, which require further investigation. In the future, with advancements in 3D printing technology, Ecoflex is expected to achieve more precise clinical translation across multiple fields and drive innovation in intelligent biomaterials.
7.Research progress on strategies for toxicity reduction and efficacy enhancement of triptolide
Xiaoqing ZHENG ; Ying DING ; Shanshan XU ; Long WANG ; Shanshan HAN ; Yaping XING ; Meng ZHANG ; Wenhao LI
China Pharmacy 2026;37(11):1496-1501
Triptolide (TP), the core active component of the traditional Chinese medicine Tripterygium wilfordii , exhibits remarkable pharmacological activities including anti-inflammatory, immunosuppressive and anti-tumor effects, and holds broad application prospects in the treatment of major diseases such as autoimmune diseases and malignant tumors. However, TP has a narrow therapeutic window and causes multi-organ toxicities including liver, kidney and reproductive toxicities, which severely restrict its safe clinical application and new drug development. Therefore, toxicity reduction and efficacy enhancement has become a core scientific problem urgently to be solved in this field. This paper systematically reviews the four core strategies for TP toxicity reduction and efficacy enhancement, including structural modification, dosage form improvement, herbal compatibility, and external therapies of traditional Chinese medicine. Among them, structural modification optimizes the toxic and efficacy characteristics of TP from the molecular structure level, with typica l derivatives including (5 R )-5-hydroxy triptolide, ZT01, PG490-88, etc. Dosage form modification achieves toxicity reduction and efficacy enhancement via targeted and sustained-controlled drug release of diverse delivery systems. It includes triptolide preparations such as nanoparticles, liposomes, microemulsion gels and liquid crystals, possessing favorable clinical transformation potential. The herbal compatibility and external therapies of traditional Chinese medicine conform to the holistic view of traditional Chinese medicine and have a profound clinical application foundation, but their mechanisms of action are insufficiently elucidated, and they lack unified standardized specifications and high-quality evidence-based proof. In the future, we should rely on multi-omics technology to elucidate the toxic and efficacy mechanisms, integrate technologies to optimize preparations, improve the evaluation system and promote clinical transformation.
8.Intravitreal Conbercept for macular edema secondary to non-ischemic retinal vein occlusion
Min YANG ; Shanshan LI ; Shuang LIU ; Dawei ZHANG
International Eye Science 2026;26(7):1147-1151
AIM:To observe the clinical efficacy of intravitreal injection of conbercept in the treatment of macular edema secondary to non-ischemic retinal vein occlusion(RVO).METHODS: Single center retrospective study. ME patients secondary to non-ischemic RVO admitted to the hospital from January 2023 to March 2024 were selected, and were divided into central retinal vein occlusion(CRVO)group and branch retinal vein occlusion(BRVO)group according to the location of obstruction. All patients were treated with intravitreal injection of conbercept once a month for 3 mo. The best corrected visual acuity(BCVA), macular foveal thickness(CMT), superficial capillary density(SVD), deep capillary density(DVD), and foveal avascular zone(FAZ)area were recorded before and after treatment(with 3 injections per course)at 1, 3, and 6 mo.RESULTS:This study included a total of 120 ME secondary to non-ischemic RVO patients(128 eyes), who were divided into CRVO group(51 cases, 56 eyes, 31 males, 20 females, mean age 61.39±10.32 y)and BRVO group(69 cases, 72 eyes, 41 males, 28 females, mean age 61.48±10.41 y)based on the location of obstruction. There was no significant difference in general data between the two groups before treatment(both P>0.05). After 1, 3, and 6 mo of treatment, both groups showed improvement in BCVA, CMT, SVD, and DVD compared to before treatment(all P<0.05). BCVA in the BRVO group was better than that in the CRVO group at all time points after treatment(all P<0.05), while there was no difference in CMT, SVD, and DVD between the two groups(all P>0.05); There was no significant difference in FAZ area between the two groups before and after treatment(both P>0.05). Follow up for 6 mo showed no significant difference in the incidence of complications between the two groups of patients(both P>0.05), but there was a significant difference in the recurrence rate(P<0.05).CONCLUSION: The first intravitreal injection of conbercept is effective in treating macular edema caused by non-ischemic CRVO and BRVO, improving visual function, reducing macular edema, and repairing retinal structure and blood flow perfusion. Notably, the recovery of visual function and improvement of capillary density are more significant in BRVO patients.
9.Therapeutic Effect of Cranial Painkiller Pills' Extract Powder in Treatment of Trigeminal Neuralgia Induced by Injection of Talci Pulvis into Infraorbital Foramen of Model Rats Based on OTULIN-regulated Neuroinflammation
Shuran LI ; Xinwei WANG ; Jing SUN ; Dan XIE ; Ronghua ZHAO ; Lei BAO ; Zihan GENG ; Qiyue SUN ; Jingsheng ZHANG ; Yaxin WANG ; Xihe CUI ; Xinying LI ; Bing HAN ; Tianjiao LU ; Xiaolan CUI ; Liying LIU ; Shanshan GUO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):21-28
ObjectiveThis paper aims to verify the therapeutic effect of Cranial Painkiller pills' extract powder prepared by the new process on the rat's trigeminal neuralgia model caused by infraorbital injection of Talci Pulvis, evaluate its potential clinical application value, and compare the therapeutic effect with that of Cranial Painkiller granules, so as to provide data support for the application of the Cranial Painkiller pills' extract powder and precise treatment. MethodsThe rat's trigeminal neuralgia model was constructed by infraorbital injection of Talci Pulvis, and the rats were randomly divided into the normal group, model group, carbamazepine group (60 mg·kg-1), Cranial Painkiller granules group (2.70 g·kg-1), and low, medium, and high dosage groups of Cranial Painkiller pills' extract powder (1.35, 2.70, 5.40 g·kg-1) according to the basal mechanical pain thresholds, and there were 10 rats in each group. The drug was administered by gavage to each group 2 h after modeling, and distilled water was given by gavage to the normal and model groups under the same conditions once a day for 10 d. Von Frey brushes were used to measure mechanical pain thresholds in rats. Hematoxylin-eosin (HE) staining was used to detect pathological changes in the trigeminal ganglion, and enzyme-linked immunosorbent assay (ELISA) was used to detect the inflammatory factors interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-α (TNF-α) levels in rat serum, as well as neuropeptide substance P (SP) and β-endorphin (β-EP) levels in rat brain tissue. Western blot technique was used to detect the levels of NLRP3, ASC, Caspase-1, and OTULIN proteins in rat brain tissue. ResultsCompared with the normal group, the pain threshold of rats in the model group showed a continuous significant decrease (P<0.01). The pathological damage of brain tissue was significant (P<0.01), and the inflammatory levels of IL-1, IL-6, IL-8, and TNF-α in serum were significantly elevated (P<0.01). The level of the SP in the brain tissue was significantly elevated (P<0.01), and the level of β-EP was significantly reduced (P<0.01), while the level of OTULIN was significantly reduced, and NLRP3, ASC, and Caspase-1 protein levels were significantly elevated (P<0.01). After administration of the drug, compared with the model group, the pain threshold of each dose group of the Cranial Painkiller pills' extract powder and the Cranial Painkiller granules group significantly increased (P<0.01). The inflammatory levels of IL-1, IL-6, IL-8, and TNF-α and SP levels significantly decreased (P<0.01), and the β-EP levels were significantly elevated (P<0.01), while the levels of OTULIN protein were significantly elevated (P<0.05, P<0.01), and the levels of NLRP3, ASC proteins were decreased (P<0.01)in high dose Cranial Painkiller pills' extract powder. Meanwhile, compared with those in the model group, the trigeminal ganglion lesions of rats in the Cranial Painkiller pills' extract powder and Cranial Painkiller granules groups showed different degrees of improvement (P<0.05, P<0.01). ConclusionThe Cranial Painkiller pills' extract powder has significant therapeutic effects on the rat model of trigeminal neuralgia induced by infraorbital injection of Talci Pulvis, and its mechanism is related to the improvement of OTULIN-regulated neuroinflammation.
10.Research on Regulatory Mechanism of Verbenalin on HCoV-229E-infected Macrophage Injury Based on Mitophagy
Qiyue SUN ; Lei BAO ; Zihan GENG ; Ronghua ZHAO ; Shuran LI ; Xihe CUI ; Jingsheng ZHANG ; Xian LIU ; Rui XIE ; Xiaolan CUI ; Shanshan GUO ; Jing SUN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):29-37
ObjectiveTo investigate the protective effect and mechanism of verbenalin on mouse mononuclear macrophage leukemia cells (RAW264.7) damaged by human coronavirus (HCoV)-229E infection, thereby providing experimental evidence for its development and application. MethodsRAW264.7 macrophages were infected with different concentrations of HCoV-229E to establish a coronavirus-induced macrophage injury model using the cell counting kit-8 (CCK-8) assay for assessing cell proliferation and viability. Cells were randomly divided into four groups: normal control, verbenalin group (125 μmol·L-1), model group (HCoV-229E), and HCoV-229E + verbenalin group (HCoV-229E + 125 μmol·L-1 verbenalin). Cell viability was measured using the CCK-8 assay, and the maximum non-toxic concentration (CC0), half-maximal cytotoxic concentration (CC50), half-maximal effective concentration (EC50), and selectivity index (SI) of verbenalin were calculated. Calcein/PI double staining was used to assess cell viability and cytotoxicity, and JC-1 staining was applied to evaluate changes in mitochondrial membrane potential (MMP). mito-Keima adenovirus labeling was used to assess mitophagy levels in each group. ResultsA macrophage infection model was successfully established by infecting RAW264.7 cells with the original concentration of HCoV-229E for 36 h. The CC0 of verbenalin was 125 μmol·L-1. The CC50 was 448.25 μmol·L-1. The EC50 against HCoV-229E-infected cells was 46.28 μmol·L-1, and the SI was 9.68. Compared with the normal group, the model group showed significantly reduced cell survival rate (P<0.01), increased cell death rate (P<0.01), decreased MMP (P<0.01), and suppressed mitophagy (P<0.01). In contrast, verbenalin treatment significantly improved cell survival rate (P<0.01), reduced cell death rate (P<0.01), alleviated MMP loss (P<0.01), and enhanced mitophagy levels (P<0.01) compared with the model group. ConclusionVerbenalin can enhance the survival rate of macrophages following HCoV-229E infection. The underlying mechanism may be associated with the activation of mitophagy, maintenance of MMP stability, and alleviation of mitochondrial damage.


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