1.Oxylipidomics Combined with Transcriptomics Reveals Mechanism of Jianpi Huogu Prescription in Treating Steroid-induced Osteonecrosis of Femoral Head in Rats
Lili WANG ; Qun LI ; Zhixing HU ; Qianqian YAN ; Liting XU ; Xiaoxiao WANG ; Chunyan ZHU ; Yanqiong ZHANG ; Weiheng CHEN ; Haijun HE ; Chunfang LIU ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):190-199
ObjectiveTo unveil the mechanism of Jianpi Huogu prescription (JPHGP) in ameliorating the dyslipidemia of steroid-induced osteonecrosis of the femur head (SONFH) by oxylipidomics combined with transcriptomics. MethodsSixty SD rats were assigned into normal, model, low-, medium-, and high-dose (2.5, 5, 10 g·kg-1, respectively) JPHGP, and Jiangushengwan (1.53 g·kg-1) groups. Lipopolysaccharide was injected into the tail vein at a dose of 20 μg·kg-1 on days 1 and 2, and methylprednisolone sodium succinate was injected at a dose of 40 mg·kg-1 into the buttock muscle on days 3 to 5. The normal group received an equal volume of normal saline. Drug administration by gavage began 4 weeks after the last injection, and samples were taken after administration for 8 weeks. Hematoxylin-eosin staining was conducted to reveal the histopathological changes of the femoral head, and the number of adipocytes, the rate of empty bone lacunae, and the trabecular area were calculated. Micro-computed tomography was used for revealing the histological and histomorphometrical changes of the femoral head. Enzyme-linked immunosorbent assay was employed to measure the serum levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), apolipoprotein A1 (ApoA1), and apolipoprotein B (ApoB). At the same time, the femoral head was collected for oxylipidomic and transcriptomic detection. The differential metabolites and differential genes were enriched and analyzed, and the target genes regulating lipid metabolism were predicted. The predicted target proteins were further verified by molecular docking, immunohistochemistry, and Western blot. ResultsCompared with the normal group, the model group showcased thinning of the femoral head, trabecular fracture, karyopyknosis, subchondral cystic degeneration, increases in the number of adipocytes and the rate of empty bone lacunae (P<0.01), a reduction in the trabecular area (P<0.01), decreases in BMD, Tb.Th, Tb.N, and BV/TV, and increases in Tb.Sp and BS/BV (P<0.01). Compared with the model group, the JPHGP groups showed no obvious thinning of the femoral head or subchondroidal cystic degeneration. The high- and medium-dose JPHGP groups presented declines in the number of adipocytes and the rate of empty bone lacunae, an increase in the trabecular area (P<0.05, P<0.01), rises in BMD, Tb.Th, Tb.N, and BV/TV, and decreases in Tb.Sp and BS/BV (P<0.05, P<0.01). Compared with the normal group, the model group showcased raised serum levels of TG, TC, LDL, and ApoB and lowered serum levels of HDL and ApoA1 (P<0.01). Compared with the model group, the JPHGP groups had lowered serum levels of TG, TC, LDL, and ApoB (P<0.05, P<0.01) and a risen serum level of ApoA1 (P<0.05, P<0.01). Moreover, the serum level of HDL in the high-dose JPHGP group increased (P<0.01). A total of 19 different metabolites of disease set and drug set were screened out by oxylipidomics of the femoral head, and 119 core genes with restored expression were detected by transcriptomics. The enriched pathways were mainly concentrated in inflammation, lipids, apoptosis, and osteoclast differentiation. Molecular docking, immunohistochemistry, and Western blot results showed that compared with the normal group, the model group displayed increased content of 5-lipoxygenase (5-LO) and peroxisome proliferator-activated receptor γ (PPARγ) in the femoral head (P<0.01). Compared with the model group, medium- and high-dose JPHGP reduced the content of 5-LO and PPARγ (P<0.05, P<0.01). ConclusionJPHGP can restore the levels of oxidized lipid metabolites by regulating the 5-LO-PPARγ axis to treat SONFH in rats. Relevant studies provide experimental evidence for the efficacy mechanism of JPHGP in the treatment of SONFH.
2.Oxylipidomics Combined with Transcriptomics Reveals Mechanism of Jianpi Huogu Prescription in Treating Steroid-induced Osteonecrosis of Femoral Head in Rats
Lili WANG ; Qun LI ; Zhixing HU ; Qianqian YAN ; Liting XU ; Xiaoxiao WANG ; Chunyan ZHU ; Yanqiong ZHANG ; Weiheng CHEN ; Haijun HE ; Chunfang LIU ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):190-199
ObjectiveTo unveil the mechanism of Jianpi Huogu prescription (JPHGP) in ameliorating the dyslipidemia of steroid-induced osteonecrosis of the femur head (SONFH) by oxylipidomics combined with transcriptomics. MethodsSixty SD rats were assigned into normal, model, low-, medium-, and high-dose (2.5, 5, 10 g·kg-1, respectively) JPHGP, and Jiangushengwan (1.53 g·kg-1) groups. Lipopolysaccharide was injected into the tail vein at a dose of 20 μg·kg-1 on days 1 and 2, and methylprednisolone sodium succinate was injected at a dose of 40 mg·kg-1 into the buttock muscle on days 3 to 5. The normal group received an equal volume of normal saline. Drug administration by gavage began 4 weeks after the last injection, and samples were taken after administration for 8 weeks. Hematoxylin-eosin staining was conducted to reveal the histopathological changes of the femoral head, and the number of adipocytes, the rate of empty bone lacunae, and the trabecular area were calculated. Micro-computed tomography was used for revealing the histological and histomorphometrical changes of the femoral head. Enzyme-linked immunosorbent assay was employed to measure the serum levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), apolipoprotein A1 (ApoA1), and apolipoprotein B (ApoB). At the same time, the femoral head was collected for oxylipidomic and transcriptomic detection. The differential metabolites and differential genes were enriched and analyzed, and the target genes regulating lipid metabolism were predicted. The predicted target proteins were further verified by molecular docking, immunohistochemistry, and Western blot. ResultsCompared with the normal group, the model group showcased thinning of the femoral head, trabecular fracture, karyopyknosis, subchondral cystic degeneration, increases in the number of adipocytes and the rate of empty bone lacunae (P<0.01), a reduction in the trabecular area (P<0.01), decreases in BMD, Tb.Th, Tb.N, and BV/TV, and increases in Tb.Sp and BS/BV (P<0.01). Compared with the model group, the JPHGP groups showed no obvious thinning of the femoral head or subchondroidal cystic degeneration. The high- and medium-dose JPHGP groups presented declines in the number of adipocytes and the rate of empty bone lacunae, an increase in the trabecular area (P<0.05, P<0.01), rises in BMD, Tb.Th, Tb.N, and BV/TV, and decreases in Tb.Sp and BS/BV (P<0.05, P<0.01). Compared with the normal group, the model group showcased raised serum levels of TG, TC, LDL, and ApoB and lowered serum levels of HDL and ApoA1 (P<0.01). Compared with the model group, the JPHGP groups had lowered serum levels of TG, TC, LDL, and ApoB (P<0.05, P<0.01) and a risen serum level of ApoA1 (P<0.05, P<0.01). Moreover, the serum level of HDL in the high-dose JPHGP group increased (P<0.01). A total of 19 different metabolites of disease set and drug set were screened out by oxylipidomics of the femoral head, and 119 core genes with restored expression were detected by transcriptomics. The enriched pathways were mainly concentrated in inflammation, lipids, apoptosis, and osteoclast differentiation. Molecular docking, immunohistochemistry, and Western blot results showed that compared with the normal group, the model group displayed increased content of 5-lipoxygenase (5-LO) and peroxisome proliferator-activated receptor γ (PPARγ) in the femoral head (P<0.01). Compared with the model group, medium- and high-dose JPHGP reduced the content of 5-LO and PPARγ (P<0.05, P<0.01). ConclusionJPHGP can restore the levels of oxidized lipid metabolites by regulating the 5-LO-PPARγ axis to treat SONFH in rats. Relevant studies provide experimental evidence for the efficacy mechanism of JPHGP in the treatment of SONFH.
3.Analysis of IL-2R β gene promoter region polymorphism in Han males from a high radiation background area in Yangjiang City
Ying TANG ; Jing WANG ; Liting YAN ; Wenting ZHONG ; Cuiju WEN ; Shibiao SU
China Occupational Medicine 2025;52(1):45-49
Objective To analyze the distribution characteristics of the single nucleotide polymorphism (SNP) site rs76206423 in the promoter region of the interleukin-2 receptor (IL-2R) β gene among Han males in a high radiation background area (HBRA) in Yangjiang City. Methods A total of 48 male participants from Tangkou Town, Yangxi County, Yangjiang City (HBRA group), and 51 male participants from Hengpo Town, Enping City (control group) were selected as the research subjects using the random number table method. Peripheral venous blood samples of participants from both groups were collected, and genomic DNA was extracted. The genotyping and allele frequency distribution of the rs76206423 (A/G) site in the IL-2R β promoter region was detected among the participants in both groups using the SNP detection method. The difference of allele frequencies between population in HBRA group and five area of East Asia, South Asia, Africa, Europe, and the Americas published in the Human Genome Project database from National Center for Biotechnology Information were analyzed. Results The allele frequencies of rs76206423 of population in both groups conformed to Hardy-Weinberg equilibrium (P>0.05). In the HBRA group, the AA genotype was predominant (64.6%), while the AG genotype was the most common in the control group (51.0%), with a significant difference (P<0.05). Population in both groups showed a predominance of the variant allele A (78.1% and 72.5%, respectively), with no significant difference (P>0.05). The frequency of the G allele of rs76206423 in the population in HBRA group was higher than those in South Asian, African, European, and American populations (all P<0.01), but showed no significant difference compared with East Asian populations (P>0.05). Conclusion In the Han male population from the HBRA in Yangjiang City, the rs76206423 site in the IL-2R β gene promoter region is predominantly composed of the wild-type A allele and AA genotype, indicating genetic stability and a relatively high degree of variation at this locus.
4.Expert consensus on digital intraoral scanning technology
Jie YOU ; Wenjuan YAN ; Liting LIN ; Wen-Zhen GU ; Yarong HOU ; Wei XIAO ; Hui YAO ; Yaner LI ; Lihui MA ; Ruini ZHAO ; Junqi QIU ; Jianzhang LIU ; Yi ZHOU
Journal of Prevention and Treatment for Stomatological Diseases 2024;32(8):569-577
Digital intraoral scanning is a hot topic in the field of oral digital technology.In recent years,digital intra-oral scanning has gradually become the mainstream technology in orthodontics,prosthodontics,and implant dentistry.The precision of digital intraoral scanning and the accuracy and stitching of data collection are the keys to the success of the impression.However,the operators are less familiar with the intraoral scanning characteristics,imaging process-ing,operator scanning method,oral tissue specificity of the scanned object,and restoration design.Thus far,no unified standard and consensus on digital intraoral scanning technology has been achieved at home or abroad.To deal with the problems encountered in oral scanning and improve the quality of digital scanning,we collected common expert opin-ions and sought to expound the causes of scanning errors and countermeasures by summarizing the existing evidence.We also describe the scanning strategies under different oral impression requirements.The expert consensus is that due to various factors affecting the accuracy of digital intraoral scanning and the reproducibility of scanned images,adopting the correct scanning trajectory can shorten clinical operation time and improve scanning accuracy.The scanning trajec-tories mainly include the E-shaped,segmented,and S-shaped methods.When performing fixed denture restoration,it is recommended to first scan the abutment and adjacent teeth.When performing fixed denture restoration,it is recommend-ed to scan the abutment and adjacent teeth first.Then the cavity in the abutment area is excavated.Lastly,the cavity gap was scanned after completing the abutment preparation.This method not only meets clinical needs but also achieves the most reliable accuracy.When performing full denture restoration in edentulous jaws,setting markers on the mucosal tissue at the bottom of the alveolar ridge,simultaneously capturing images of the vestibular area,using different types of scanning paths such as Z-shaped,S-shaped,buccal-palatal and palatal-buccal pathways,segmented scanning of dental arches,and other strategies can reduce scanning errors and improve image stitching and overlap.For implant restora-tion,when a single crown restoration is supported by implants and a small span upper structure restoration,it is recom-mended to first pre-scan the required dental arch.Then the cavity in the abutment area is excavated.Lastly,scanning the cavity gap after installing the implant scanning rod.When repairing a bone level implant crown,an improved indi-rect scanning method can be used.The scanning process includes three steps:First,the temporary restoration,adjacent teeth,and gingival tissue in the mouth are scanned;second,the entire dental arch is scanned after installing a standard scanning rod on the implant;and third,the temporary restoration outside the mouth is scanned to obtain the three-di-mensional shape of the gingival contour of the implant neck,thereby increasing the stability of soft tissue scanning around the implant and improving scanning restoration.For dental implant fixed bridge repair with missing teeth,the mobility of the mucosa increases the difficulty of scanning,making it difficult for scanners to distinguish scanning rods of the same shape and size,which can easily cause image stacking errors.Higher accuracy of digital implant impres-sions can be achieved by changing the geometric shape of the scanning rods to change the optical curvature radius.The consensus confirms that as the range of scanned dental arches and the number of data concatenations increases,the scanning accuracy decreases accordingly,especially when performing full mouth implant restoration impressions.The difficulty of image stitching processing can easily be increased by the presence of unstable and uneven mucosal mor-phology inside the mouth and the lack of relatively obvious and fixed reference objects,which results in insufficient ac-curacy.When designing restorations of this type,it is advisable to carefully choose digital intraoral scanning methods to obtain model data.It is not recommended to use digital impressions when there are more than five missing teeth.
5.Exploration on the Molecular Mechanism of Shenling Baizhu Powder in Improving Adipose Tissue Inflammation in Children with Obesity Based on Network Pharmacology and Animal Experimental Verification
Changmiao NIU ; Liting LIANG ; Yongkai YIN ; Xiaoning YAN ; Kaixi ZHANG ; Gengtian NIU
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(12):27-34
Objective To explore the effective components and possible targets and mechanism of Shenling Baizhu Powder in improving adipose tissue inflammation in children with obesity with the help of network pharmacology and molecular docking method;To conduct preliminary verification through animal experiments.Methods The active components and targets of Shenling Baizhu Powder were screened through the TCMSP database.GeneCards,OMIM,DisGeNET databases were used to obtain the disease targets.An active component-target network was constructed by taking the intersection of drug and disease targets.Protein-protein interaction networks were constructed and core targets were obtained for GO and KEGG pathway enrichment analysis.Molecular docking validation was performed to key components and core targets.Animal experiments were conducted by establishing a model of young obese rats induced by high fat diet,and Shenling Baizhu Powder were given for gavage.The contents of TG and TC in serum and mRNA expressions of TNF-α,IL-1β,INF-γ and Fas in rat adipose tissue of epididymis were detected.Results The key components of Shenling Baizhu Powder to improve the microinflammation of obesity in children were piperlonguminine,acacetin,luteolin,denudatin B,mandenol,inermine,etc.There were 515 common drug-disease targets,and the core targets were TP53,SRC,PIK3R1,HSP90AA1 and PIK3CA.The results of GO enrichment analysis included inflammatory response,positive regulation of MAPK cascade,membrane raft,protein serine/threonine/tyrosine kinase activity,etc.KEGG pathway enrichment analysis results included metabolic,immune,endocrine,cancer and related liver disease signaling pathways.The results of molecular docking showed that the key components could play a regulatory role on the core target.Animal experiments showed that Shenling Baizhu Powder could improve microinflammation and metabolic indexes of model rats.Conclusion Shenling Baizhu Powder can act on multiple targets through various active components and multiple signaling pathways,adjusting inflammatory response and immune process,alleviating insulin resistance,regulating glucose and lipid metabolism,thereby improving adipose tissue inflammatory in children with obesity.
6.Association between coronary artery stenosis and myocardial injury in patients with acute pulmonary embolism: A case-control study
Yinjian YANG ; Chao LIU ; Jieling MA ; Xijie ZHU ; Jingsi MA ; Dan LU ; Xinxin YAN ; Xuan GAO ; Jia WANG ; Liting WANG ; Sijin ZHANG ; Xianmei LI ; Bingxiang WU ; Kai SUN ; Yimin MAO ; Xiqi XU ; Tianyu LIAN ; Chunyan CHENG ; Zhicheng JING
Chinese Medical Journal 2024;137(16):1965-1972
Background::The potential impact of pre-existing coronary artery stenosis (CAS) on acute pulmonary embolism (PE) episodes remains underexplored. This study aimed to investigate the association between pre-existing CAS and the elevation of high-sensitivity cardiac troponin I (hs-cTnI) levels in patients with PE.Methods::In this multicenter, prospective case-control study, 88 cases and 163 controls matched for age, sex, and study center were enrolled. Cases were patients with PE with elevated hs-cTnI. Controls were patients with PE with normal hs-cTnI. Coronary artery assessment utilized coronary computed tomographic angiography or invasive coronary angiography. CAS was defined as ≥50% stenosis of the lumen diameter in any coronary vessel >2.0 mm in diameter. Conditional logistic regression was used to evaluate the association between CAS and hs-cTnI elevation.Results::The percentage of CAS was higher in the case group compared to the control group (44.3% [39/88] vs. 30.1% [49/163]; P = 0.024). In multivariable conditional logistic regression model 1, CAS (adjusted odds ratio [OR], 2.680; 95% confidence interval [CI], 1.243–5.779), heart rate >75 beats/min (OR, 2.306; 95% CI, 1.056–5.036) and N-terminal pro-B type natriuretic peptide (NT-proBNP) >420 pg/mL (OR, 12.169; 95% CI, 4.792–30.900) were independently associated with elevated hs-cTnI. In model 2, right CAS (OR, 3.615; 95% CI, 1.467–8.909) and NT-proBNP >420 pg/mL (OR, 13.890; 95% CI, 5.288–36.484) were independently associated with elevated hs-cTnI. Conclusions::CAS was independently associated with myocardial injury in patients with PE. Vigilance towards CAS is warranted in patients with PE with elevated cardiac troponin levels.
7.A case report of COVID-19-associated acute necrotizing encephalopathy
Li ZHAO ; Xianzhi LI ; Liting YAN ; Yi LI ; Xiaoqi WANG ; Xiao YANG
Chinese Journal of Nervous and Mental Diseases 2024;50(3):156-158
This article reports one case of adult COVID-19-associated acute necrotizing encephalopathy(ANEC).The patient developed disturbance of consciousness and seizure on the 12th day after SARS-CoV-2 infection.Imaging showed significant swelling and signal changes in the bilateral thalamus,brainstem,cerebral hemisphere and cerebellar hemisphere,which were consistent with the characteristic images of Acute necrotizing encephalitis(ANE).Although methylprednisolone shock therapy and high-dose human immunoglobulin therapy were given early,the patient died.ANEC often starts quickly and progresses rapidly,unconsciousness and seizure are the main manifestations.Imaging features of thalamic and subtentorial symmetry and multifocal lesions are specific for diagnosis,but the treatment and prognosis still face challenges and need further study.
8.Effect of tanshinone ⅡA on angiotensin Ⅱ-induced proliferation and migration of vascular smooth muscle cells
Liting LIU ; Zhiqian XIONG ; Yan JIANG ; Chaojiang SU ; Shuai ZHANG ; Zongyang LIU
Journal of China Medical University 2024;53(5):439-445
Objective To determine whether Tanshinone ⅡA(Tan ⅡA)has an effect on angiotensin Ⅱ(Ang Ⅱ)-induced prolifera-tion and migration of vascular smooth muscle cells(VSMCs),phenotypic switching,or autophagy.Methods In this study,murine aortic smooth muscle cell lines were treated with Ang Ⅱ to establish a model of cell proliferation.Different concentrations of Tan ⅡA were added and effects on cell proliferation and migration were determined by cell counting using a CCK-8 assay and a cell scratch assay,respectively.Alpha-smooth muscle actin(α-SMA),a marker of contractile VSMCs,was detected by Western blotting.Expression levels of osteopontin(OPN)and the autophagy-related proteins(p62,Beclin-1,LC3A/B)were assayed to determine the effect of Tan ⅡA on VSMC phenotypic transformation and autophagy.Cells were treated with the autophagy inhibitor 3-methyladenine(3-MA),combined with Tan ⅡA,and then cell proliferation,migration and expression levels of phenotypic markers and autophagy-related proteins were assessed.Results After Ang Ⅱ treatment,the proliferation and migratory capacity of VSMCs was significantly enhanced,and phenotypic transformation was sig-nificantly inhibited by Ang Ⅱ.Western blotting revealed that Ang Ⅱ reduced the expression of α-SMA,increased the expression of OPN,p62,Beclin-1,and LC3A/B,and that these effects increased with higher Tan ⅡA concentrations.After the addition of 3-MA to the treated cells,the proliferation and migration of VSMCs induced by Ang Ⅱ was inhibited,the expression of α-SMA was increased,and the expres-sion of OPN,p62,Beclin-1,and LC3A/B was decreased,which was consistent with the effect of Tan ⅡA.Conclusion Tan ⅡA may have inhibitory effects on phenotypic transformation,proliferation,migration,and autophagy of Ang Ⅱ-treated VSMC,suggesting that the inhi-bition of the proliferation and migration may be regulated by autophagy.
9.A comparative study of iodine content in serum, plasma and whole blood
Liting YANG ; Zijun YAO ; Zhen WANG ; Han ZHANG ; Yan WANG ; Huailan GUO
Chinese Journal of Endemiology 2023;42(6):502-506
Objective:To evaluate the consistency of individual iodine nutrition levels by serum iodine, plasma iodine and whole blood iodine, and to provide reference for iodine-related epidemiological investigation.Methods:Healthy adults aged 18 - 59 years were recruited from the Research Center of Environment and Health in Water Source Area of South-to-North Water Diversion of Hubei University of Medicine. Whole blood sample was collected and serum and plasma were separated. The content of iodine in serum, plasma and whole blood was determined by inductively coupled plasma mass spectrometry (ICP-MS), and the linear relationship, precision and accuracy of the standard curve of the detection method were evaluated. The difference of three kinds of blood iodine levels was analyzed by variance analysis of compatibility group design, and Passing-Bablok regression and Bland-Altman plot were used to evaluate the consistency between serum iodine and plasma iodine.Results:The linear range of iodine in serum, plasma and whole blood was 0.0 - 25.0 μg/L, and the correlation coefficients ( R2) were all > 0.999. The relative standard deviation of 8 mixed blood samples ranged from 1.9% to 4.3% ( n = 6), and the determination results of blood iodine certified standard substances were all within the reference range. The recovery rate of the added standard ranged from 99% to 106%. The iodine levels in serum, plasma and whole blood of 50 volunteers were (57.31 ± 8.06), (57.49 ± 8.50) and (33.89 ± 5.40) μg/L, respectively, and there was no statistically significant difference between serum iodine and plasma iodine ( P = 0.904). The results of Passing-Bablok regression showed that there was no statistically significant difference in bias between serum iodine and plasma iodine ( P = 0.538). The Bland-Altman plot indicated that the difference between serum iodine and plasma iodine was within the consistency limit. Conclusion:The results of plasma iodine and serum iodine are in good agreement, and plasma iodine can be used as an evaluation index of individual iodine nutrition level. But there is no consistency between whole blood iodine and serum iodine.
10.Alleviation of amyloid beta-protein induced neurotixicity and synaptic damage in PC12 cells by downregulation of serine racemase
Chunmei TAO ; Zheng WU ; Xuejing CHEN ; Liting FAN ; Hongting YAN ; Yusong GE
Chinese Journal of Neuromedicine 2022;21(2):109-118
Objective:To investigate the role of down-regulating serine racemase (SRR) in alleviating the β-amyloid peptide (Aβ) induced neurotoxicity and synaptic damage and possible mechanism in PC12 cells.Methods:(1) PC12 cells cultured in vitro were divided into 0, 20, 40 and 80 μmol/L Aβ 25-35 treatment groups; they were treated with 0, 20, 40 and 80 μmol/L Aβ 25-35 for 24 h, respectively; cell counting kit (CCK)-8 was used to detect the survival rate of cells in each group, and Western blotting was used to detect the SRR protein expression. PC12 cells were treated with 40 μmol/L Aβ 25-35 for 0, 12, 24 and 48 h, respectively; cell survival and SRR protein expression were detected by CCK-8 and Western blotting, respectively. (2) PC12 cells were divided into control group, nonsense sequence group, SRR small interfering RNA (siRNA) group 1, SRR siRNA group 2, and SRR siRNA group 3; cells in the later three groups were transfected with SRR nonsense sequence or different SRR siRNA sequences, respectively; 48 h after that, Western blotting was used to detect the SRR protein expression of cells in each group, and SRR siRNA with best effect was selected for subsequent experiments. (3) PC12 cells were divided into control group, AD group, AD+nonsense sequence group, and AD+SRR siRNA group; cells in the latter two groups were transfected with nonsense sequence or SRR siRNA for 48 h, respectively; cells in the latter three groups were added 40 μmol/L Aβ 25-35, and cells in the control group were added same amount of solvent; 24 h after treatment, the SRR protein expression was detected by Western blotting, cell survival was detected by CCK-8, cell apoptosis was detected by Hoechst 33258 fluorescent staining, Caspase 3 activity was detected by enzyme linked immunosorbent assay, and the expressions of activated Caspase 3, N-methyl- D aspartate (NMDA) receptor-associated proteins and postsynaptic dense protein 95 (PSD95) were detected by Western blotting. Results:(1) The survival rate of cells in 0, 20, 40 and 80 μmol/L Aβ 25-35 treatment groups was successively decreased and the SRR protein expression was successively increased, with significant differences ( P<0.05); PC12 cells treated with 40 μmol/L Aβ 25-35 for 0, 12, 24 and 48 h had successively decreased survival rate and successively increased SRR protein expression, with significant differences ( P<0.05). (2) The SRR protein expressions in the SRR siRNA group 1, SRR siRNA group 2 and SRR siRNA3 group 3 were significantly decreased as compared with those in the control group and nonsense sequence group ( P<0.05), and the decrease in the SRR siRNA group 2 was the most obvious. (3) As compared with the control group, the cells in the AD group had significantly increased SRR protein expression and apoptosis rate, statistically decreased cell survival rate, significantly increased Caspase 3 activity and activated Caspase 3 protein expression, significantly increased protein expressions of NMDA receptor 2A (NMDAR2A) and NMDA receptor 2B(NMDAR2B), and statistically decreased PSD95 protein expression ( P<0.05); as compared with cells in the AD group, cells in the AD+SRR siRNA group had significantly decreased SRR protein expression and apoptosis rate, statistically increased cell survival rate, significantly decreased Caspase 3 activity and activated Caspase 3 protein expression, significantly decreased NMDAR2A protein expression, and statistically increased PSD95 protein expression ( P<0.05). Conclusion:Down-regulation of SRR expression can reduce the NMDAR2A protein expression, alleviate the over-activation of NMDA receptor, reduce the cell apoptosis, improve cell survival rate, protect nerve cells, increase PSD95 protein expression, and alleviate synaptic damage in PC12 cells.


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