1.Investigation of Effect of Different Drying Conditions on Appearance Characteristics and Internal Indicators of Pinelliae Rhizoma Based on Standardization
Suqing LIU ; Xueli ZHANG ; Jing ZHANG ; Cong YANG ; Changfu YANG ; Jun YU ; Bingpeng ZHENG ; Huiwu LI ; Yanhua JIANG ; Chang LIN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):208-215
ObjectiveTo explore the effect of different drying conditions on the appearance and intrinsic quality indicators of Pinelliae Rhizoma for screening suitable drying conditions, so as to provide reference for its standardized production and quality evaluation. MethodsDifferent dried samples of Pinelliae Rhizoma were prepared by lime-assisted sweating method and intermittent drying method. Visual analysis was employed to measure the color brightness values(L*) of the surface, cross-section and powder of the samples, texture analyzer was used to determine the hardness of the samples under different drying conditions. The total starch content was calculated by measuring the contents of amylose and amylopectin in the samples with ultraviolet-visible spectroscopy. High performance liquid chromatography(HPLC) was used to determine the contents of seven nucleoside components(uracil, hypoxanthine, uridine, inosine, guanosine, β-thymidine and adenosine) in the samples. Pearson correlation analysis was conducted to explore the correlation between the external characteristics and intrinsic indicators of the different dried samples. Principal component analysis(PCA) was used to comprehensively rank the data of various indicators, and partial least squares-discriminant analysis(PLS-DA) was used to screen differential components with variable importance in the projection(VIP) value>1. Furthermore, the difference between the optimal drying condition for Pinelliae Rhizoma and the traditional sun-drying method was explored by independent samples t-test. ResultsWith the increase of temperature, the color of the intermittently dried samples gradually deepened, while their hardness gradually decreased. Concurrently, the contents of extract, total starch, uridine and adenosine exhibited an upward trend, whereas the contents of uracil, hypoxanthine and inosine displayed a downward trajectory. Compared with the intermittent drying group, the content of extract in the samples subjected to lime-assisted sweating increased. With the increase of lime dose, the hardness and the total content of nucleoside components in the samples showed a downward trend, while the total starch content showed an upward trend. Correlation analysis showed that the comprehensive score of L* was negatively correlated with the contents of uracil, hypoxanthine and inosine, and positively correlated with the contents of uridine, guanosine and adenosine. Hardness was negatively correlated with adenosine content, and positively correlated with the contents of inosine, uracil and hypoxanthine. Through comprehensive consideration and comprehensive score of principal components, the method of 5% lime-mixed sweating for 6 days emerged as the top-ranking approach. Except for the extract, the results of independent samples t-test showed that there was no significant difference between the 5% lime-mixed sweating for 6 days and the traditional sun-drying in terms of other content indicators. ConclusionThe whiteness and firmness of Pinelliae Rhizoma exhibit significant correlations with its chemical composition, while uridine, uracil, guanosine, adenosine and inosine are the key constituents responsible for the quality difference of Pinelliae Rhizoma under different drying conditions. The lime-assisted sweating method optimized in this study can be proposed as a viable alternative to the traditional sun-drying method. This method not only ensures the quality of the medicinal material but also effectively reduces the drying time and prevents mold contamination, which provides a valuable reference for the standardization of drying conditions and the establishment of quality evaluation criteria for Pinelliae Rhizoma.
2.Mechanism of astragalin in allevating ulcerative colitis in mice through modulation of the intestinal flora
Jing HUANG ; Yanhua LIAO ; Xinying MO ; Yuting YANG ; Weizhe JIANG
China Pharmacy 2025;36(14):1709-1716
OBJECTIVE To explore the potential mechanisms of astragalin (AG) in allevating ulcerative colitis (UC) in mice through modulation of the intestinal flora. METHODS Male C57BL/6 mice were randomly divided into normal group (CON group), model group [dextran sodium sulfate (DSS) group], 5-aminosalicylic acid group (5-ASA group), AG low-dose group and high-dose group (AGL and AGH groups), with 8 mice in each group. The mice UC model was established by drinking 3% DSS solution continuously for 7 days in all groups except the CON group. After that, 3% DSS solution was replaced by water, and the mice of each drug group were gavaged with the corresponding drug solution. Mice in the CON and DSS groups were gavaged with an equal volume of normal saline, once a day, for 7 days. After the last gavage, the body weight change index, disease activity index (DAI) score, colon length and spleen index, and levels of inflammatory factors (tumor necrosis factor-α, interleukin-1β, interleukin-6) were compared among the mice in each group; pathological changes in colonic tissues of the mice were observed in each group, and the pathological score and the percentage of goblet cells were compared; mRNA expressions of barrier-related factors [occludin and ZO-1] and inflammation-related factors [silencing information regulatory factor 1 (SIRT1), c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (p38 MAPK)] were detected in each group of mice; the changes in the intestinal flora of mice in each group were analyzed and the contents of intestinal metabolites short-chain fatty acids (SCFAs) was determined. Using DSS and AG-treated fecal bacterial liquid as an intervention, the mechanism of anti-UC effect of AG was further verified by a fecal microbiota transplant experiment. RESULTS Compared with the CON group, the intestinal mucosal structure of mice in the DSS group was severely damaged, with obvious infiltration of inflammatory cells collapsing the wall; their body weight change index, colon length, the percentage of goblet cells, mRNA expressions of occludin, ZO-1 and SIRT1, Chao1 and Shannon indexes, and contents of acetic acid and butyric acid were significantly reduced, shortened or down-regulated (P<0.05); however, DAI score, spleen index, levels of inflammatory factors, pathological score, as well as mRNA expressions of p38 MAPK and JNK, were all significantly increased or up-regulated (P<0.05). Compared with the DSS group, colon tissue lesions of AG mice in all dose groups showed different degrees of improvement, and the above quantitative indexes were generally regressed (P<0.05), and the intervention effect of AG-treated fecal bacterial fluid was basically the same as that of AG. CONCLUSIONS AG can improve relevant symptoms in UC mice and reduce their inflammatory response and colonic histopathological changes. The above effects may be related to regulating the diversity of intestinal flora in mice, increasing the contents of butyric acid and propionic acid, and promoting the repair of the colonic mucosal barrier, thus regulating the expressions of genes related to the SIRT1/p38 MAPK inflammatory pathway.
3.Changes of macular retinal structure and microcirculation in patients with pituitary adenoma observed by optical coherence tomography and optical coherence tomography angiography
Zhihui LIAO ; Xin YANG ; Kailun LU ; Tongtong DAI ; Yanhua PANG
International Eye Science 2024;24(2):203-209
AIM: To observe the changes of macular retinal structure and microcirculation in patients with pituitary adenoma(PA)by optical coherence tomography(OCT)and optical coherence tomography angiography(OCTA).METHODS: Cross-sectional study. A total of 40 PA patients treated at the department of neurosurgery, Affiliated Hospital of Guangdong Medical University from September 2021 to March 2023 were included as PA group, and 42 age- and gender-matched healthy volunteers were selected as normal control group. All patients underwent visual field, OCT and OCTA examinations, and the correlation of ocular parameters in PA patients was analyzed.RESULTS:The vessel density(VD)of each retinal layer in the macular area of the PA group was lower than that of the normal control group, and the superficial vascular complex(SVC)-VD in the macular area was positively correlated with the thickness of the macular ganglion cell complex(mGCC)(except the nasal side of the inner ring and the lower part of the outer ring; P<0.05). The thickness of mGCC in each quadrant of the macular area and the thickness of the circumpapillary retinal nerve fiber layer(CP-RNFL)in each quadrant were negatively correlated with the mean defect(MD)value of the visual field(P<0.05), and the area of the foveal avascular zone(FAZ)was positively correlated with the MD value(P<0.05).CONCLUSION:The combination of OCT and OCTA can fully understand the microscopic changes of retinal structure and microcirculation function in PA patients, which is of great value in evaluating the preoperative visual function of PA patients.
4.Mechanism of Yiqi Jiedu Formula Against Ischemic Stroke Based on Microbial-gut-brain Axis
Jialin YANG ; Bingjie CAI ; Yuhao DAI ; Shuting LI ; Keke ZHANG ; Yanhua GAO ; Qiman ZHANG ; Ying ZHANG ; Jianying SHEN ; Shaojing LI
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(6):91-100
ObjectiveTo explore the possible mechanism of the Yiqi Jiedu formula (YQ) in treating ischemic stroke (IS) from the perspective of the microbial-gut-brain axis (MGBA). MethodRats were randomly divided into five groups, with six in each group, including sham surgery group, model group, and low, medium, and high dose YQ groups (1, 5, and 25 mg·kg-1). Except for the sham surgery group, all other groups were established with a middle cerebral artery occlusion (MCAO) model using the thread occlusion method. The success of modeling was determined through neurobehavioral scoring, and the protective effect of YQ on IS was evaluated. Then, the changes in gut microbiota before and after MCAO modeling and YQ administration were compared using 16S rDNA sequencing technology, and the possible biological pathways related to the effect of this formula were analyzed. The expression of inflammatory factors such as interleukin-6 (IL-6), interleukin-17A (IL-17A), and interleukin-10 (IL-10) in serum was detected by enzyme-linked immunosorbent assay (ELISA). Western blot was used to detect the expression of tight junction proteins ZO-1 and Occludin in brain and intestinal tissue, and hematoxylin-eosin staining (HE) was used to observe pathological changes in the cerebral cortex and colon, so as to validate the possible mechanism of action. ResultYQ significantly improved the neurobehavioral score of MCAO rats (P<0.01) and played a good regulatory role in intestinal microbial disorders caused by enriched pathogens and opportunistic pathogens during the acute phase. Among them, significantly changed microorganisms include Morgentia, Escherichia Shigella, Adlercreutzia, and Androbacter. Bioinformatics analysis found that these bacteria may be related to the regulation of inflammation in the brain. Compared with the blank group, the detection of inflammatory factors in the serum of IS model rats showed an increase in inflammatory factors IL-6 and IL-17A (P<0.01) and a decrease in the content of anti-inflammatory factor IL-10 (P<0.01). Compared with the model group, the content of inflammatory factors IL-6 and IL-17A in the serum of the treatment group decreased (P<0.05), and that of anti-inflammatory factor IL-10 increased (P<0.01). The expression results of barrier proteins ZO-1 and Occludin in brain and intestinal tissue showed that the expression levels of both decreased in IS model rats (P<0.05), while the expression levels of both increased in the treatment group (P<0.05). ConclusionAcute cerebral ischemia can lead to an imbalance of intestinal microbiota and damage to the intestinal barrier, and it can increase intestinal permeability. YQ can regulate intestinal microbiota imbalance caused by ischemia, inhibit systemic inflammatory response, and improve the disruption of the gut-blood brain barrier, preventing secondary cascade damage to brain tissue caused by inflammation. The MGBA may be an important mechanism against the IS.
5.Preliminary study on the reduction of streak artifacts in thyroid CT examination with optimized position in prone position
Zhiyu JIA ; Yanhua ZHOU ; Yaqian LONG ; Hong ZHANG ; Xinguan YANG
Journal of Practical Radiology 2024;40(4):650-653,673
Objective To explore the factors influencing the image quality of thyroid CT and the application value of optimization schemes in reducing streak artifacts in thyroid CT examinations.Methods A retrospective review of the data of 141 patients who underwent thyroid enhanced CT examinations was conducted to analyze the factors affecting image quality.After determining the influencing factors,a self-controlled study was conducted on 48 patients who underwent thyroid CT examinations to analyze the feasibility of the optimization scheme.Results The patients'body mass index(BMI),neck circumference,pharynx-jaw angle,and pharynx-jaw angle-to-neck circumference ratio showed statistically significant differences in their impact on image quality(P<0.05).Compared to the conventional scheme,the optimization scheme showed significantly higher subjective and objective image evaluations.The optimization scheme also significantly improved the diagnostic performance of the images,with statistically significant differences(P<0.05).There was no statistically significant difference in radiation dose between the two positions(P>0.05).Conclusion The optimization scheme can effectively reduce streak artifacts in thyroid CT examinations.
6.Effect of patent foramen ovale on development of post-operative stroke in patients undergoing non-cardiac surgery: a meta-analysis
Chen CHEN ; Xiaoli YANG ; Xiaotian SUN ; Yunzhi SHEN ; Huiwen JIANG ; Yanhua YIN
Chinese Journal of Anesthesiology 2024;44(3):292-296
Objective:To evaluate the effect of patent foramen ovale on the development of post-operative stroke in the patients undergoing non-cardiac surgery using a meta-analysis approach.Methods:A comprehensive search was conducted across multiple databases including PubMed, Embase, Web of Science, CINAHL, Cochrane Library, China Biomedical Literature Database, Wanfang Data Knowledge Service Platform, and China Journal Full Text Database.The inclusion criteria encompassed studies assessing the correlation between patent foramen ovale and post-operative stroke.The primary outcome measure focused on the incidence of post-operative stroke, and secondary outcome measures comprised mortality, myocardial infarction rate, and readmission rate within 30 days after surgery. The quality of literature meeting the inclusion criteria was evaluated and data were extracted, and then meta-analysis was conducted using RevMan 5.4 software.Results:Eight retrospective cohort studies involving 21 142 237 patients were included.The results of meta-analysis showed that patent foramen ovale was associated with post-operative stroke and readmission within 30 days after surgery.There were no significant differences in all-cause mortality and myocardial infarction rates between patent foramen ovale group and mon-patent foramen ovale group ( P>0.05). Conclusions:Patent foramen ovale can increase the risk of post-operative stroke in the patients undergoing non-cardiac surgery.
7.Effects of Yiqi tongmai formula on atherosclerosis in ApoE-/- mice and its mechanism
Quanna REN ; Yuhan CHEN ; Kun WANG ; Ming MA ; Yanhua YANG ; Yu ZHANG ; Xu ZHAO
China Pharmacy 2024;35(18):2232-2237
OBJECTIVE To investigate the effects of Yiqi tongmai formula on atherosclerosis (AS) in ApoE-/- mice and its mechanism. METHODS Forty ApoE-/- mice were randomly divided into model group, positive control group [atorvastatin calcium, 2.6 mg/(kg·d)], and low-dose, medium-dose and high-dose groups of Yiqi tongmai formula [0.46, 0.91, 1.82 g/(kg·d), by raw material], with 8 mice in each group. Eight C57BL/6J mice were selected as the normal group. Except for the normal group, the other groups were given a high-lipid diet and relevant drug or normal saline intragastrically, once a day, for 12 consecutive weeks. After the last medication, the serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) as well as the contents of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and monocyte chemoattractant protein-1 (MCP-1) were measured in mice. The proportion of aortic plaque area in each group of mice was detected and calculated, and the pathological morphological changes of the aortic sinus were observed; the protein phosphorylation levels of aortic phosphoinositide 3-kinase (PI3K), protein kinase B (aka Akt) and mammalian target of rapamycin (mTOR) were examined. RESULTS Compared with the model group, the serum levels of TC, TG and LDL-C and the contents of TNF-α, IL-1β and MCP-1 (including low-dose group) were decreased significantly in medium-dose and high-dose groups of Yiqi tongmai formula, while the content of HDL-C in high-dose group was increased significantly (P<0.05 or P<0.01); aortic plaques of the mice were reduced in Yiqi tongmai formula groups to different extents, and pathological changes such as lipid deposition and inflammatory cell infiltration were relieved to different extents; the proportion of aortic plaque area, the protein phosphorylation levels of PI3K, Akt and mTOR in aortic tissue were significantly reduced in medium-dose and high-dose groups of Yiqi tongmai formula (P<0.05 or P<0.01). CONCLUSIONS Yiqi tongmai formula can improve lipid metabolism, reduce inflammatory response, and delay plaque development in AS mice. Its effect may be related to the inhibition of PI3K/Akt/mTOR signaling pathway activation.
8.Recognition of whole blood transfusion in civilian trauma resuscitation
Weijuan MA ; Shaomin REN ; Zhongsi YANG ; Yanhua ZHANG
Chinese Journal of Blood Transfusion 2024;37(5):602-606
There has been a growing amount of evidence that a balanced blood component transfusion with roughly equal ratio of units of packed red blood cells,fresh frozen plasma,and platelets leads to better outcomes in massive transfusion re-suscitation of trauma victims.Therefore,the great interest of massive transfusion protocol(MTP)with whole blood was a-roused.Low titer group O whole blood(LTOWB)is implemented in routine use for civilian prehospital ambulance services in large trauma centers of most European and American countries nowadays.There is a growing body of evidence to date to support that early use of LTOWB in patients with life-threatening bleeding improve their survival.In view of the current situ-ation of whole blood supply in our country,most trauma resuscitation guidelines still recommend balanced component trans-fusion for MTPs in the early stage of resuscitation.The research and application of LTOWB abroad will be introduced in this article.
9.Effect of Yiqi Tongmai Formula in Intervening in Liver Inflammation and Lipidosis in ApoE-∕- Mice Based on NF-κB/NLRP3 Signaling Pathway
Yuhan CHEN ; Quanna REN ; Kun WANG ; Tingting SU ; Yanhua YANG ; Ming MA ; Yu ZHANG ; Xu ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(15):64-72
ObjectiveTo investigate the effect of Yiqi Tongxin formula (YQTM) on liver inflammation in apolipoprotein E-∕- (ApoE-∕-) mice by regulating the nuclear transcription factor-κB (NF-κB)/NOD-like receptor protein 3 (NLRP3) signaling pathway. MethodForty ApoE-∕- mice were randomly divided into a model group, an atorvastatin group (positive drug group), and low-, medium-, and high-dose YQTM groups (0.39, 0.78, 1.56 g·kg-1). Each drug administration group was given the corresponding concentration of the drug by gavage on the basis of high-fat feeding for 12 consecutive weeks. Eight C57BL/6J mice were used as a blank group and fed with normal chow. After 12 weeks, oil red O staining and Masson staining were used to observe the aortic lesions in mice and to determine whether the modeling was successful. Oil red O staining was used to observe the lipidosis in the livers of mice. Hematoxylin-eosin (HE) staining was used to observe the tissue lesions in the livers of mice. Masson staining was used to observe the distribution of collagen fibers in the livers of mice. Enzyme markers were used to detect the total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL-C), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) in mouse serum, as well as total cholesterol (TC) and triglyceride (TG) in the liver. Interleukin-1β (IL-1β) and IL-18 were detected in mouse liver by enzyme-linked immunosorbent assay (ELISA). Immunohistochemistry (IHC) was utilized to observe the expression regions of NF-κB and NLRP3 in the livers of mice. Western blot was employed to detect the protein expression levels of NF-κB, NF-κB inhibitory protein (IκB), IκB kinase β (IKKβ), phosphorylated NF-κB (p-NF-κB), phosphorylated IκB (p-IκB), phosphorylated IKK β (p-IKKβ), NLRP3, and Caspase-1 in the livers of mice. ResultCompared with the blank group, the model group showed severe aortic lipidosis, and the intracellular fat droplets in the livers aggregated in large quantities. The cytoplasm was filled with fat vacuoles(P<0.01). The serum levels of TG, TC, LDL-C, AST, and ALT were significantly elevated in the mice(P<0.01). TG and TC levels were elevated in the liver(P<0.01). The levels of IL-1β and IL-18 in liver tissue, as well as the protein expression levels of NF-κB, IκB, IKKβ, p-NF-κB, p-IκB, p-IKKβ, NLRP3, and Caspase-1 in the liver were significantly elevated(P<0.01). Compared with the model group, the aortic arch plaques of mice in each YQTM group were attenuated, and the fat aggregation in the liver was reduced. The inflammatory cell infiltration was alleviated(P<0.05,P<0.01). The serum levels of TG, TC, LDL-C, AST, and ALT were significantly reduced(P<0.05,P<0.01). TG and TC levels in the liver were reduced. The IL-1β and IL-18 levels in liver tissue, as well as protein expression levels of NF-κB, IκB, IKKβ, p-NF-κB, p-IκB, p-IKKβ, NLRP3, and Caspase-1 in the liver were significantly reduced(P<0.05,P<0.01). ConclusionThe intervention mechanism of YQTM on liver inflammation in ApoE-∕- mice may be related to the down-regulation of the NF-κB/NLRP3 signaling pathway.
10.Exploring Mechanism of Neferine in Promoting Vascular Regeneration Against Cerebral Ischemia Based on Mitochondrial MCU Channel
Qiman ZHANG ; Yanhua GAO ; Wenjie WU ; Wei YANG ; Chen LIU ; Shuting LI ; Bingjie CAI ; Jialin YANG ; Ying ZHANG ; Jing MA ; Shaojing LI
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(24):103-113
ObjectiveTo investigate the mechanism of neferine(Nef) in promoting vascular regeneration against cerebral ischemia through modulation of mitochondrial calcium uniporter(MCU) ion channel. MethodTaking the area of subintestinal vessels in microvascular deficiency zebrafish as an index, the vascular regenerative efficacy of Nef was evaluated, and the median effective concentration(EC50) was calculated. Rats were randomly divided into a sham operation group, a model group, a positive drug group(butylphthalide, 6 mg·kg-1), and Nef low, medium, and high dose groups(0.125, 0.625, 3.125 μg·kg-1). Except for the sham operation group, the middle cerebral artery occlusion(MCAO) model was established in other groups. After modeling, the groups were administered the corresponding dose of drugs by gavage, while the sham operation and model groups received equal volumes of saline, once a day for 7 consecutive days. Neurobehavioral scores were assessed for each group of rats, and the infarct rate of ischemic brain tissue was calculated by 2,3,5-triphenyltetrazolium chloride(TTC) staining. The regional cerebral blood flow(rCBF) of each group was measured using a speckle contrast imaging. Immunofluorescence and Western blot were conducted to detect the expression of vascular endothelial growth factor(VEGF), platelet endothelial cell adhesion molecule-1(CD31), and hypoxia-inducible factor-1α(HIF-1α) proteins in each group. Human umbilical vein endothelial cells(HUVECs) were divided into the normal group, model group, positive drug group(astragaloside Ⅳ, 10 μmol·L-1), and Nef group (32 nmol·L-1). In the verification of mitochondrial protection of Nef and its mechanism in promoting vascular regeneration, the spermine(MCU agonist) and Nef+spermine group were added. HUVECs model of oxygen-glucose deprivation(OGD) was established in all groups except the normal group, the cell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay, and cell migration ability was evaluated through scratch and tube formation assays. Fluorescent probes(Rhod-2 AM, Fluo-3 AM, JC-1, Calcein AM) and a cellular energy metabolism analyzer were used to analyze the mitochondrial protective effects of Nef. Molecular docking was performed to predict the binding ability of Nef with MCU and HIF-1α, and Western blot was used to detect the effects of Nef on the protein expressions of MCU, B-cell lymphoma-2 associated X protein(Bax), Caspase-3 and HIF-1α in the OGD model HUVECs. ResultThe results of vascular regeneration in microvascular deficiency zebrafish showed that compared to the normal group, the area of subintestinal vessels in the model group significantly decreased(P<0.01). Compared to the model group, different concentrations of Nef could significantly increase the area of subintestinal vessels(P<0.01), with the maximum tolerated concentration of 10.24 μmol·L-1 and the EC50 of 0.23 μmol·L-1. Anti-cerebral ischemia results on MCAO rats showed that compared to the sham operation group, the model group had a significant decrease in rCBF and a significant increase in infarct rate, while CD31 expression significantly decreased(P<0.01), and VEGF and HIF-1α protein expressions significantly increased(P<0.05). Compared to the model group, the treated groups showed significant increases in rCBF, significant reductions in infarct volume, and significant increases in CD31, VEGF, and HIF-1α protein expression(P<0.01). Cell experiment results showed that compared to the normal group, the model group had decreased cell viability and migration ability, increased intracellular Ca2+ and mitochondrial Ca2+ levels, reduced mitochondrial permeability transition pore(MPTP) opening, and decreased mitochondrial energy metabolism capability, with increased expressions of MCU, Bax, Caspase-3 and HIF-1α proteins(P<0.05, P<0.01). Compared to the model group, the Nef group showed increased cell viability and migration ability, decreased intracellular Ca2+ and mitochondrial Ca2+ levels, increased MPTP opening, enhanced mitochondrial energy metabolism capability, decreased expressions of MCU, Bax and Caspase-3 proteins, and increased HIF-1α protein expression(P<0.05, P<0.01). ConclusionNef can stabilize mitochondrial membrane potential and inhibit mitochondrial apoptosis. By down-regulating the expression of MCU, it suppresses the activation of intracellular Bax and Caspase-3 while activating the HIF-1α signaling pathway, enhancing the expression of VEGF and CD31, thereby promoting vascular regeneration to treat ischemic brain injury.

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