1.Study on quality control of Jinbei oral liquid based on fingerprint,chemical pattern recognition and multi-index content determination
Jing TIAN ; Weiliang CUI ; Yuanfang ZANG ; Bing WANG ; Huifen LI ; Aijun ZHANG ; Fei XUE ; Yingying XIE ; Yongqiang LIN
China Pharmacy 2026;37(13):1704-1709
OBJECTIVE To establish a quality control method for Jinbei oral liquid based on multi-wavelength switching high performance liquid chromatography (HPLC) fingerprint, chemical pattern recognition and multi-index content determination. METHODS A total of 15 batches of Jinbei oral liquid were used as test samples. The Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine ( Version 2012 ) was adopted to establish multi-wavelength switching HPLC fingerprints, followed by chromatographic peak identification and similarity evaluation. Cluster analysis, principal component analysis and orthogonal partial least squares-discriminant analysis were applied to conduct chemical pattern recognition on the 15 batches of samples. The multi-wavelength switching HPLC method was used to simultaneously determine the contents of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, forsythoside A, baicalin, salvianoli c acid B and wogonoside in samples. RESULTS A total of 25 common peaks were calibrated in the fingerprints of 15 batches of Jinbei oral liquid, among which 7 common peaks were unambiguously identified. The similarity of all samples was higher than 0.960. Chemical pattern recognition results showed that samples S1-S8 were clustered into group 1, and samples S9-S15 were clustered into group 2. Baicalin, wogonoside, neochlorogenic acid, chlorogenic acid and salvianolic acid B were identified as the differential quality markers. The average contents of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, forsythoside A, baicalin, salvianolic acid B and wogonoside in 15 batches of Jinbei oral liquid were 0.213 7, 0.085 3, 0.104 9, 0.287 3, 0.420 1, 0.062 1, 0.176 0 mg/mL, respectively. CONCLUSIONS The established quality control method for Jinbei oral liquid combining multi-wavelength switching HPLC fingerprint, chemical pattern recognition and multi-index content determination is stable and reliable, which can provide a reference for the formulation of quality standards of this preparation.
2.Study on quality control of Jinbei oral liquid based on fingerprint,chemical pattern recognition and multi-index content determination
Jing TIAN ; Weiliang CUI ; Yuanfang ZANG ; Bing WANG ; Huifen LI ; Aijun ZHANG ; Fei XUE ; Yingying XIE ; Yongqiang LIN
China Pharmacy 2026;37(13):1704-1709
OBJECTIVE To establish a quality control method for Jinbei oral liquid based on multi-wavelength switching high performance liquid chromatography (HPLC) fingerprint, chemical pattern recognition and multi-index content determination. METHODS A total of 15 batches of Jinbei oral liquid were used as test samples. The Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine ( Version 2012 ) was adopted to establish multi-wavelength switching HPLC fingerprints, followed by chromatographic peak identification and similarity evaluation. Cluster analysis, principal component analysis and orthogonal partial least squares-discriminant analysis were applied to conduct chemical pattern recognition on the 15 batches of samples. The multi-wavelength switching HPLC method was used to simultaneously determine the contents of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, forsythoside A, baicalin, salvianoli c acid B and wogonoside in samples. RESULTS A total of 25 common peaks were calibrated in the fingerprints of 15 batches of Jinbei oral liquid, among which 7 common peaks were unambiguously identified. The similarity of all samples was higher than 0.960. Chemical pattern recognition results showed that samples S1-S8 were clustered into group 1, and samples S9-S15 were clustered into group 2. Baicalin, wogonoside, neochlorogenic acid, chlorogenic acid and salvianolic acid B were identified as the differential quality markers. The average contents of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, forsythoside A, baicalin, salvianolic acid B and wogonoside in 15 batches of Jinbei oral liquid were 0.213 7, 0.085 3, 0.104 9, 0.287 3, 0.420 1, 0.062 1, 0.176 0 mg/mL, respectively. CONCLUSIONS The established quality control method for Jinbei oral liquid combining multi-wavelength switching HPLC fingerprint, chemical pattern recognition and multi-index content determination is stable and reliable, which can provide a reference for the formulation of quality standards of this preparation.
3.Scientific basis for acupuncture combined with neural stem cells for repairing spinal cord injury
Xiaomeng HUANG ; Zhilan ZHANG ; Wenya SHANG ; Jing HUANG ; Huilin WEI ; Bing LI ; Yafeng REN
Chinese Journal of Tissue Engineering Research 2025;29(19):4111-4121
BACKGROUND:Spinal cord injury is a neurological disorder caused by traumatic or non-traumatic events,often leading to severe functional impairment below the injured segment.In recent years,neural stem cell transplantation has been considered to have significant therapeutic potential in regulating the inflammatory response after spinal cord injury,inhibiting excessive proliferation of glial scars,and promoting nerve regeneration. OBJECTIVE:To review and discuss the potential mechanism of action of acupuncture and neural stem cell transplantation therapy in inhibiting spinal cord injury-induced secondary injury,and to delve into the scientific basis for its treatment of spinal cord injury. METHODS:PubMed,Elsevier,WanFang,and CNKI databases were searched using"spinal cord injury,acupuncture,neural stem cells,SDF-1α/CXCR4 axis"as Chinese and English search terms.Totally 96 articles were finally included.The research findings of acupuncture combined with neural stem cells in the treatment of spinal cord injury were summarized and analyzed,and the mechanism of this combination therapy in the treatment of secondary injury after spinal cord injury was summarized. RESULTS AND CONCLUSION:(1)The stromal-derived factor 1α(SDF-1α)/chemokine receptor 4(CXCR4)axis plays a crucial role in neural stem cell transplantation for spinal cord injury.This signaling mechanism not only affects neural stem cell migration,proliferation,and differentiation,but is also a key factor in determining the efficiency of stem cell homing to the injury site.Therefore,the regulation of targeting this axis is of great significance in enhancing the therapeutic effect of spinal cord injury.(2)Acupuncture,as a traditional Chinese medicine therapy,shows unique advantages in the regulation of secondary injury in spinal cord injury.It can effectively reduce secondary injury after spinal cord injury by regulating inflammatory response,inhibiting apoptosis,improving microcirculation,reducing glial scar formation,and counteracting oxidative stress.(3)Acupuncture was also able to influence the expression and function of the SDF-1α/CXCR4 axis,thereby enhancing the homing and survival ability of neural stem cells and promoting nerve regeneration and functional recovery.(4)The therapy combining acupuncture and stem cell transplantation is an innovative treatment strategy for spinal cord injury and suitable for repairing neural circuits.It combines the wisdom of traditional Chinese medicine with the advantages of modern biotechnology,providing a new treatment option for spinal cord injury patients.However,this combination therapy is still in the research and exploration stage,and its long-term efficacy and safety need to be further verified.(5)Taken together,acupuncture and neural stem cell transplantation for the treatment of spinal cord injury has great potential for clinical application,but in-depth research and optimization of treatment options are still needed.In the future,we look forward to further revealing the efficacy mechanism and optimal indications of this therapy through more clinical trials and mechanism studies,so as to bring better hope of recovery and more efficient therapeutic effects to spinal cord injury patients.
4.Xiaoyao Shukun Decoction Treats Sequelae of Pelvic Inflammatory Disease by Regulating Neutrophil Extracellular Traps via PI3K/Akt/mTOR Pathway
Jing PAN ; Bing ZHANG ; Chunxiao DANG ; Jinxiao LI ; Pengfei LIU ; Xiao YU ; Yuchao WANG ; Jinxing LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(15):69-78
ObjectiveTo investigate how Xiaoyao Shukun decoction (XYSKD) regulates the formation and release of neutrophil extracellular traps (NETs) via the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway, thereby reducing inflammation, inhibiting the excessive proliferation of fibroblasts in pelvic adhesion tissue, decreasing adhesion and fibrosis, and repairing the tissue damage in sequelae of pelvic inflammatory disease (SPID). MethodsA total of 84 Wistar rats were randomly allocated into seven groups: blank, model, XYSKD (8 mg·kg-1), mTOR agonist (10 mg·kg-1), mTOR agonist + XYSKD (10 mg·kg-1+8 mg·kg-1), mTOR inhibitor (2 mg·kg-1), and mTOR inhibitor + XYSKD (2 mg·kg-1+8 mg·kg-1). The rat model of SPID was constructed by starvation, fatigue, and ascending Escherichia coli infection. After 14 days of drug intervention, the ultrastructure of fibroblasts in the pelvic adhesion tissue was observed by transmission electron microscopy. The general morphology of the uterus, fallopian tube, and ovary was observed by laparotomy. The levels of interleukin-1β (IL-1β), interleukin-17 (IL-17), and tumor necrosis factor-α (TNF-α) in the peritoneal flushing fluid were determined by enzyme-linked immunosorbent assay (ELISA). The expression of myeloperoxidase (MPO) and citrullinated histone 3 (H3) in the fallopian tube was detected by immunofluorescence. Western blot and Real-time quantitative polymerase chain reaction (Real-time PCR) were employed to determine the relative protein and mRNA levels, respectively, of neutrophil elastase (NE), intercellular adhesion molecule-1 (CD54), α-smooth muscle actin (α-SMA), H3, PI3K, and Akt. ResultsCompared with the blank group, the model group presented a large number of collagen fibers in bundles, numerous cytoplasmic folds of fibroblasts, reduced or absent mitochondrial cristae, and disordered and expanded endoplasmic reticulum. By laparotomy, extensive pelvic congestion, connective tissue hyperplasia, thickening and hardening of the tubal end near the uterus, and tubal and ovarian adhesion or cyst were observed in the model group. In addition, the model group showed raised levels of IL-1β, IL-17, and TNF-α in the peritoneal flushing fluid (P<0.01), increased average fluorescence intensities of MPO and H3 (P<0.01), and up-regulated protein and mRNA levels of NE, H3, CD54, PI3K, and Akt (P<0.01). Compared with the model group, the mTOR agonist group showed increased fibroblasts and cytoplasmic folds, absence of mitochondrial cristae, endoplasmic reticulum dilation, and evident collagen fiber hyperplasia. Pelvic adhesions were observed to cause aggravated damage to the uterine, fallopian tube, and ovarian tissues. The levels of IL-1β, IL-17, and TNF-α in the peritoneal lavage fluid elevated (P<0.01) and the average fluorescence intensities of MPO and H3 enhanced (P<0.01) in the mTOR agonist group. In contrast, the XYSKD group and the mTOR inhibitor group showcased decreased fibroblasts and collagen fibers, alleviated mitochondrial crista loss and endoplasmic reticulum dilation, improved morphology and appearance of the uterine, fallopian tube, and ovarian tissues, lowered levels of IL-1β, IL-17, and TNF-α in the peritoneal lavage fluid (P<0.01), decreased average fluorescence intensities of MPO and H3 (P<0.01), and down-regulated protein and mRNA levels of NE, H3, CD54, PI3K, and Akt (P<0.05). Compared with the mTOR agonist group, the mTOR agonist + XYSKD group showed alleviated pathological changes in the pelvic tissue, declined levels of IL-1β, IL-17, and TNF-α (P<0.01), decreased average fluorescence intensities of MPO and H3 (P<0.01), and down-regulated protein levels of NE, H3, CD54, α-SMA, p-PI3K/PI3K, and p-Akt/Akt (P<0.01) and mRNA levels of NE, H3, CD54, α-SMA, PI3K, and Akt (P<0.01). Compared with the mTOR inhibitor group, the mTOR inhibitor + XYSKD group demonstrated reduced pathological severity of the pelvic tissue, reduced levels of IL-1β, IL-17, and TNF-α (P<0.01), decreased average fluorescence intensities of MPO and H3 (P<0.01), and down-regulated protein and mRNA levels of NE and CD54 (P<0.05). ConclusionXYSKD can inhibit the excessive formation and release of NETs via PI3K/Akt/mTOR to ameliorate the inflammatory environment and reduce fibrosis and adhesion of the pelvic tissue, thereby playing a role in the treatment of SPID. It may exert the effects by lowering the levels of IL-1β, IL-17, and TNF-α and down-regulating the expression of NE, H3, CD54, α-SMA, PI3K, and Akt in the pelvic adhesion tissue.
5.Exploration of evaluation criteria based on the biological variation in the external quality assessment for basic semen analysis in China.
Xi-Yan WU ; Jin-Chun LU ; Xin-Hua PENG ; Jing-Liang HE ; Dao WANG ; Cong-Ling DAI ; Wen-Bing ZHU ; Gang LIU ; Wei-Na LI
Asian Journal of Andrology 2025;27(5):621-626
This study explores whether the current external quality assessment (EQA) level and acceptable bias for basic semen analysis in China are clinically useful. We collected data of semen EQA from Andrology laboratories in the Hunan Province (China) in 2022 and searched for data in the published literature from January 2000 to December 2023 in China. On the basis of these data, we analyzed the coefficients of variation and acceptable biases of different quality control materials for basic semen analysis through robust statistics. We compared these findings with quality specifications based on biological variation from optimal, desirable, and minimum levels of bias to seek a unified and more suitable semen EQA bias evaluation standard for China's national conditions. Different sources of semen quality control material exhibited considerable variation in acceptable biases among laboratories, ranging from 8.2% to 56.9%. A total of 50.0% of the laboratories met the minimum quality specifications for progressive motility (PR), whereas 100.0% and 75.0% of laboratories met only the minimum quality specifications for sperm concentration and total motility (nonprogressive [NP] + PR), respectively. The Z value for sperm concentration and PR+NP was equivalent to the desirable performance specification, whereas the Z value for PR was equivalent only to the minimum performance specification. This study highlights the feasibility of operating external quality assessment schemes for basic semen analysis using quality specifications based on biological variation. These specifications should be unified among external quality control (EQC) centers based on biological variation.
Semen Analysis/standards*
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Humans
;
China
;
Male
;
Quality Control
;
Sperm Motility
;
Sperm Count/standards*
6.Genetic profiling and intervention strategies for phenylketonuria in Gansu, China: an analysis of 1 159 cases.
Chuan ZHANG ; Pei ZHANG ; Bing-Bo ZHOU ; Xing WANG ; Lei ZHENG ; Xiu-Jing LI ; Jin-Xian GUO ; Pi-Liang CHEN ; Ling HUI ; Zhen-Qiang DA ; You-Sheng YAN
Chinese Journal of Contemporary Pediatrics 2025;27(7):808-814
OBJECTIVES:
To investigate the molecular epidemiology of children with phenylketonuria (PKU) in Gansu, China, providing foundational data for intervention strategies.
METHODS:
A retrospective analysis was conducted on 1 159 PKU families who attended Gansu Provincial Maternity and Child Care Hospital from January 2012 to December 2024. Sanger sequencing, multiplex ligation-dependent probe amplification, whole exome sequencing, and deep intronic variant analysis were used to analyze the PAH gene.
RESULTS:
For the 1 159 children with PKU, 2 295 variants were identified in 2 318 alleles, resulting in a detection rate of 99.01%. The detection rates were 100% (914/914) in 457 classic PKU families, 99.45% (907/912) in 456 mild PKU families, and 96.34% (474/492) in 246 mild hyperphenylalaninemia families. The 2 295 variants detected comprised 208 distinct mutation types, among which c.728G>A (14.95%, 343/2 295) had the highest frequency, followed by c.611A>G (4.88%, 112/2 295) and c.721C>T (4.79%, 110/2 295). The cumulative frequency of the top 23 hotspot variants reached 70.28% (1 613/2 295), and most variant alleles were detected in exon 7 (29.19%, 670/2 295).
CONCLUSIONS
Deep intronic variant analysis of the PAH gene can improve the genetic diagnostic rate of PKU. The development of targeted detection kits for PAH hotspot variants may enable precision screening programs and enhance preventive strategies for PKU.
Humans
;
Phenylketonurias/epidemiology*
;
Female
;
Male
;
Retrospective Studies
;
Phenylalanine Hydroxylase/genetics*
;
Mutation
;
Child, Preschool
;
China/epidemiology*
;
Child
;
Infant
7.Clinical characteristics of trimethoprim-sulfamethoxazole-induced rash during treatment of pertussis in children.
Bing-Song WANG ; Kai-Hu YAO ; Xian-Yi ZHANG ; Jing WU ; Fei YING ; Li-Min DONG
Chinese Journal of Contemporary Pediatrics 2025;27(10):1227-1232
OBJECTIVES:
To study the clinical characteristics of rashes induced by trimethoprim-sulfamethoxazole (TMP-SMZ) in children treated for pertussis and to inform safe medication practices.
METHODS:
A retrospective analysis was conducted on 238 children diagnosed with pertussis and treated with TMP-SMZ at Wuhu First People's Hospital from January to August 2024. The incidence and clinical features of rashes were summarized.
RESULTS:
Of 238 children, 34 (14.3%) developed rashes; 19 (55.9%) were boys, and the 5 to <10-year age group accounted for the highest proportion (70.6%, 24/34). A history of allergic disease was present in 50.0% (17/34). Rashes typically appeared on or after day 7 of therapy (82%, 28/34) and were predominantly erythematous or maculopapular eruptions (97%, 33/34); 71% (24/34) were pruritic. Fever occurred in 56% (19/34); among those who were tested for respiratory viruses, 77% (10/13) were positive for viruses such as rhinovirus and adenovirus. After discontinuation of TMP-SMZ, rashes resolved within 3 days in 97% (33/34) of patients (41% within 1 day; 56% within more than 1 but within 3 days). There was no significant difference in rash incidence between photoprotection and non-photoprotection groups (P>0.05).
CONCLUSIONS
TMP-SMZ for pertussis can induce rashes, particularly in children aged 5 to <10 years. The eruption is usually a pruritic erythematous or maculopapular rash, with over half of cases accompanied by fever and frequent concomitant viral infections. Most rashes resolve within 3 days after drug withdrawal. The potential association between the rash and sun exposure warrants further investigation.
Humans
;
Male
;
Child, Preschool
;
Trimethoprim, Sulfamethoxazole Drug Combination/therapeutic use*
;
Child
;
Female
;
Exanthema/chemically induced*
;
Retrospective Studies
;
Infant
;
Whooping Cough/drug therapy*
;
Adolescent
8.PI-RADS v2.1 score combined with PSA density for diagnosis of clinically significant prostate cancer in the PSA grey zone by MRI-TRUS cognitivefusion-guided transperineal targeted prostate biopsy.
Yue LI ; Shan ZHOU ; Jing CHEN ; Fei MAO ; Xiao-Bing NIU ; Li SUN ; Ming XU ; Jin-Tao LIU
National Journal of Andrology 2025;31(1):50-54
OBJECTIVE:
To assess the value of the Prostate Imaging Reporting and Data System version 2.1 (PI-RADS v2.1) score combined with PSA density (PSAD) in the diagnosis of clinically significant prostate cancer (CSPCa) in the PSA grey zone by MRI-TRUS cognitive fusion-guided transperineal targeted prostate biopsy.
METHODS:
This retrospective study included 327 male patients with total PSA (tPSA) levels of 4-10 μg/L undergoing MRI-TRUS cognitive fusion-guided transperineal targeted prostate biopsy in our hospital between January 2021 and December 2023. According to the pathological results, we divided the patients into a CSPCa (n = 44) and a non-CSPCa group (n = 283), collected their clinical and imaging data, and subjected them to statistical analysis.
RESULTS:
The age, tPSA level, PSAD and PI-RADS score were significantly higher, while the free PSA (fPSA) level, f/tPSA ratio and prostate volume remarkably lower in the CSPCa than in the non-CSPCa group (P<0.05). The areas under the curve (AUCs) of PSAD, PI-RADS score and their combination were 0.772, 0.730 and 0.801, with sensitivities of 63.63%, 70.45% and 72.73%, and specificities of 84.10%, 75.62% and 83.75%, respectively (P<0.01). With PSAD 0.2 μg/(ml·cm3) as the best cut-off value and based on the PI-RADS scores, the patients were divided into two groups for analysis. In the patients with PI-RADS scores 2 and 5, the AUCs were 0.534 and 0.643, with sensitivities of 16.67% and 63.64%, and specificities of 85.14% and 64.29%, with no statistically significant differences (P= 0.784, P= 0.228), and in those with PI-RADS scores 3 and 4, the AUCs were 0.794 and 0.843, with sensitivities of 57.14% and 80.00%, and specificities of 87.14% and 81.82%, with statistically significant differences (P= 0.009, P<0.001).
CONCLUSION
PI-RADS v2.1 score combined with PSAD can effectively improve the diagnostic efficiency of CSPCa in the PSA grey zone by MRI-TRUS cognitive fusion-guided transperineal targeted prostate biopsy and serve as a guide for selection of prostate biopsy.
Humans
;
Male
;
Prostatic Neoplasms/diagnostic imaging*
;
Retrospective Studies
;
Prostate-Specific Antigen
;
Magnetic Resonance Imaging
;
Image-Guided Biopsy
;
Prostate/pathology*
;
Aged
;
Middle Aged
9.Observation on the Therapeutic Effect of Bushen Jianpi Formula in Treating Type 2 Diabetes Mellitus Complicated with Non-alcoholic Fatty Liver Disease
Yan LI ; Feng LIU ; Jiewen ZHANG ; Jing LIN ; Bing JI
Journal of Guangzhou University of Traditional Chinese Medicine 2025;42(11):2705-2713
Objective To evaluate the clinical efficacy of Bushen Jianpi Formula(BSJPF)in treating type 2 diabetes mellitus(T2DM)complicated with non-alcoholic fatty liver disease(NAFLD)of spleen-kidney deficiency type.Methods Seventy-two patients with T2DM complicated with NAFLD of spleen-kidney deficiency type admitted to Clifford Hospital from June 2023 to September 2024 were randomized into treatment group(n=36,receiving lifestyle intervention+metformin+BSJPF)and control group(n=36,lifestyle intervention+metformin alone)for 12 weeks.TCM syndrome scores,liver function markers,ultrasonographic grading of fatty liver,glycolipid metabolic parameters were observed,and the clinical efficacy and safety were assessed.Results(1)Regarding dropouts,during the study,2 patients in the treatment group dropped out,while none occurred in the control group.Ultimately,34 patients in the treatment group and 36 in the control group completed efficacy evaluation.(2)In terms of clinical efficacy,after 12 weeks of treatment,the total effective rate was 88.24%(30/34)in the treatment group versus 66.67%(24/36)in the control group.Intergroup comparison(by chi-square test)showed significantly superior efficacy in the treatment group(P<0.05).(3)For liver function indicators,after treatment,serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),and γ-glutamyl transpeptidase(GGT)levels decreased significantly in both groups compared to those before treatment(P<0.05),with substantially greater reduction in the treatment group(P<0.05).(4)Regarding fatty liver ultrasound grading,both groups showed improvement after treatment(P<0.05),with significantly greater enhancement in the treatment group(P<0.05).(5)For glucose-lipid metabolism markers,both groups exhibited decreased fasting blood glucose(FBG),2-hour postprandial glucose(2hPG),glycated hemoglobin(HbA1c),fasting insulin(FINS),insulin resistance index(HOMA-IR),total cholesterol(TC),triglycerides(TG),and low-density lipoprotein cholesterol(LDL-C)levels(P<0.05),with treatment group showing markedly greater reductions(P<0.05);both groups demonstrated increased high-density lipoprotein cholesterol(HDL-C)levels(P<0.05),with treatment group showing significantly greater elevation(P<0.05).(6)In TCM syndrome score assessment,both groups showed reduced scores after treatment(P<0.05),with treatment group demonstrating significantly greater improvement(P<0.05).(7)Regarding safety,routine blood/urine/stool tests,renal function indicators,and electrocardiograms remained normal in both groups.The adverse reaction rate was 2.94%(1/34)in treatment group versus 5.56%(2/36)in control group,with no statistically significant difference between groups(P>0.05).Conclusion BSJPF combined with metformin demonstrates superior efficacy to metformin alone for patients with T2DM complicated with NAFLD of spleen-kidney deficiency type.It is effective in relieving clinical symptoms,enhancing TCM syndrome efficacy,improving liver enzymes,fatty liver grading and insulin resistance,and regulating glycolipid metabolism.
10.Mechanism of senegenin in improving lipopolysacchride-induced inflammatory response of BV2 microglial cell
Bing-Tao MU ; Min-Fang GUO ; Jing-Wen YU ; Jia-Lei CAO ; Feng-Jun YANG ; Si-Wei JIA ; Qing SU ; Tao MENG ; Cun-Gen MA ; Jie-Zhong YU ; Li-Juan SONG
Medical Journal of Chinese People's Liberation Army 2025;50(2):188-196
Objective To investigate the mechanism by which Senegenin(SEN)alleviates microglial inflammatory response through the nuclear factor erythroid 2-related factor 2(Nrf2)/NOD-like receptor protein 3(NLRP3)pathway.Methods BV2 mouse microglia cells were randomly divided into control group,model group,SEN group and MCC950 group.Cells in control group were not treated,and cells in model group were added with 1 μg/ml lipopolysaccharide(LPS);Cells in SEN group were added with 1 μg/ml LPS+4 μmol/L SEN,and cells in MCC950 group were added with 1 μg/ml LPS+10 μmol/L MCC950 for 24 hours.CCK-8 method was used to detect the effect of different concentrations of SEN on the viability of BV2 cells.Griess method was used to determine the release amount of nitric oxide(NO)in the supernatant.Real-time fluorescent quantitative PCR was used to determine the mRNA expression levels of NLRP3,lymphocyte apoptosis-associated spect-like protein containing a CARD(ASC),caspase-1,interleukin(IL)-1β and IL-18 mRNA.Immunofluorescence staining was used to detect the expression levels of ASC,IL-1β,Nrf2 and heme oxygenase-1(HO-1).Western blotting was used to detect the expression levels of NLRP3,caspase-1,ASC,IL-1β,IL-18,Nrf2,HO-1,nuclear factor kappa B(NF-κB)and inducible nitric oxide synthase(iNOS).Results The results of CCK-8 method showed that there was no significant difference in the viability of BV2 cells treated with 2~20 μmol/L SEN compared with control group(P>0.05).Compared with control group,the viability of BV2 cells in model group decreased significantly(P<0.05).Compared with model group,the viability of BV2 cells in 4 μmol/L SEN group was significantly restored(P<0.05).Compared with control group,the results of Griess method showed that the release amount of NO in cells of model group increased significantly(P<0.05);the results of real-time PCR showed that the expression levels of NLRP3,ASC,caspase-1,IL-1β and IL-18 mRNA in cells of model group increased significantly(P<0.05);the results of Western blotting showed that the protein expression levels of NLRP3,ASC,caspase-1,IL-1β and IL-18 proteins in cells of model group increased significantly(P<0.05),and the immunofluorescence staining results showed that the expression levels of iNOS and NF-κB protein in cells of model group increased,and the expression levels of Nrf2 and HO-1 decreased,with statistically significant differences(P<0.05).Compared with model group,the release amount of NO in cells of SEN group and MCC950 group decreased,and the expression levels of NLRP3,ASC,caspase-1,IL-1β and IL-18 mRNA and proteins decreased,with statistically significant differences(P<0.05);in the SEN group,the expression levels of iNOS and NF-κB decreased,and immunofluorescence staining showed that Nrf2 was translocated into the nucleus,and the expression levels of Nrf2 and HO-1 proteins increased significantly,with statistically significant differences(P<0.05).Conclusions SEN could alleviate the inflammatory response of mouse microglia cells induced by LPS and inhibit the activation and expression of NLRP3 inflammasome,with an effect comparable to that of the inflammasome inhibitor MCC950.The mechanism may be related to the regulation of the expression of upstream factors Nrf2 and HO-1.

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