1.Quality Evaluation of Lycii Cortex and Roasted Lycii Cortex Based on Fingerprint and Content Determination
Yihuan WU ; Wenli ZENG ; Xuemei QIN ; Zongxin SHI ; Chengcheng HUANG ; Yuntao DAI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):165-172
ObjectiveTo establish fingerprint profiles and a quantitative determination method for Lycii Cortex, providing a scientific basis for the formulation of quality standards for Lycii Cortex and its roasted products. MethodsHigh performance liquid chromatography(HPLC) was developed for the quantitative method for determining kukoamine B in Lycii Cortex and its roasted products on an Alphasil XD-C18 CH column(4.6 mm×250 mm, 5 μm). HPLC fingerprint profiles were established for 10 batches of Lycii Cortex and its roasted products, and ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used to identify the common peaks based on reference standards, literature and MS information. Quality evaluation indicators included yield of decoction pieces, appearance properties, content of kukoamine B, and fingerprint profiles. The temperature and time of the roasting process were investigated to select the optimal preparation process, which was then verified. Additionally, chemical pattern recognition was combined to assess the differences in the chemical composition of Lycii Cortex before and after roasting, as well as among samples from different origins. ResultsQuantitative analysis indicated that the contents of kukoamine B in Lycii Cortex and its roasted products were 0.35%-5.51% and 0.24%-4.15%, respectively. The transfer rate of kukoamine B was 58.6%-78.9% after roasting. The fingerprint profile analysis demonstrated that the method established in this study effectively separated kukoamine B from other components in the samples and distinctly differentiated it from its impurity peak, cis-N-caffeoylputrescine. The HPLC fingerprint profiles of Lycii Cortex and its roasted products showed high similarity(all above 0.95), with 7 common peaks identified and five common components, including kukoamine B, cis-N-caffeoylputrescine, N-coumaroyl tyramine, feruloyltyramine, and glucosyringic acid, confirmed. Process optimization confirmed that baking at 110 ℃ for 20 min was a stable and feasible method for roasting Lycii Cortex. Principal component analysis and cluster analysis showed that there was little difference in the chemical composition between raw and roasted Lycii Cortex, but the quality of Lycii Cortex from different origins differed greatly. ConclusionThis study successfully established the fingerprint profiles and a quantitative method for the effective component kukoamine B in Lycii Cortex and roasted Lycii Cortex. The qualitative and quantitative analyses clarified that the impact of the roasting process on the chemical composition of Lycii Cortex was less significant than the variations due to its geographical origin. The findings of this study offer a reference for the development of quality evaluation methods and the establishment of quality standards for Lycii Cortex and its processed products.
2.The distribution pattern of traditional Chinese medicine syndromes and influencing factors for primary liver cancer: An analysis of 415 cases
Zhiyao SHI ; Xiaofei FAN ; Yu GAO ; Shaojian REN ; Shiyu WU ; Xixing WANG
Journal of Clinical Hepatology 2025;41(1):84-91
ObjectiveTo investigate the influencing factors for traditional Chinese medicine (TCM) syndromes of primary liver cancer, and to provide a theoretical basis for the TCM syndrome differentiation and standardized treatment of liver cancer. MethodsTCM syndrome differentiation was performed for 415 patients who were admitted to Shanxi Institute of Traditional Chinese Medicine and were diagnosed with primary liver cancer based on pathological or clinical examinations from January 2019 to December 2023. The chi-square test was used for comparison of categorical data between groups, and the unordered polytomous logistic regression model was used to investigate the influencing factors for TCM syndromes of liver cancer. ResultsThe common initial symptoms of the 415 patients with primary liver cancer included pain in the liver area (31.81%), abdominal distension (25.30%), abdominal pain (15.18%), and weakness (13.98%), and the main clinical symptoms included poor appetite (70.84%), fatigue (69.16%), pain in the liver area (67.47%), poor sleep (59.04%), abdominal distension (53.01%), and constipation (52.53%). There were significant differences in TCM syndromes between patients with different sexes, courses of the disease, clinical stages, Child-Pugh classes, presence or absence of intrahepatic and extrahepatic metastasis, and presence or absence of transcatheter arterial chemoembolization (TACE) and radiofrequency ablation (all P<0.05). The logistic regression analysis showed that male sex was a risk factor for damp-heat accumulation (odds ratio [OR]=2.036, P=0.048) and the syndrome of spleen-kidney Yang deficiency (OR=5.240, P<0.001); a course of disease of<1 year was a risk factor for damp-heat accumulation (OR=2.837, P=0.004) and syndrome of Qi stagnation and blood stasis (OR=2.317, P=0.021), but it was a protective factor against syndrome of spleen-kidney Yang deficiency (OR=0.385, P=0.005); Child-Pugh class A/B was a protective factor against liver-kidney Yin deficiency (OR=0.079, P<0.001); intrahepatic metastasis was a risk factor for liver-kidney Yin deficiency (OR=5.117, P=0.003) and syndrome of spleen-kidney Yang deficiency (OR=3.303, P=0.010); TACE was a protective factor against liver-kidney Yin deficiency (OR=0.171, P<0.001) and syndrome of spleen-kidney Yang deficiency (OR=0.138, P<0.001); radiofrequency ablation was a risk factor for damp-heat accumulation (OR=4.408, P<0.001) and liver-kidney Yin deficiency (OR=32.036, P<0.001). ConclusionSex, course of disease, Child-Pugh class, intrahepatic metastasis, TACE, and radiofrequency ablation are the main influencing factors for TCM syndromes of liver cancer.
3.Modified Shaofu Zhuyutang Mediates VEGF/PI3K/Akt/eNOS Signaling Pathway to Inhibit Angiogenesis in Endometriosis
Jiaxing WANG ; Qi SHI ; Quansheng WU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):81-90
ObjectiveTo explore the mechanism by which modified Shaofu Zhuyutang inhibits angiogenesis in endometriosis via the vascular endothelial growth factor (VEGF)/phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/endothelial nitric oxide synthase (eNOS)-nitric oxide (NO) signaling pathway. MethodsEighty-four female SD rats were randomly assigned into blank, sham operation, model, positive control (gestrinone, 0.25 mg·kg-1), high-, medium-, and low-dose (30, 15, 7.5 g·kg-1, respectively) traditional Chinese medicine (TCM, modified Shaofu Zhuyutang) groups. A rat model of endometriosis was established by the autotransplantation method. After successful modeling, rats in the drug intervention groups were administrated with corresponding agents by gavage, and those in the blank, sham operation, and model groups received an equal volume of distilled water. After 28 days of gavage, rats were administrated with oxytocin, and the number and latency period of writhing responses were observed. Serum samples from each group, ectopic lesions from modeling groups, and uteri from blank and sham operation groups were collected. Hematoxylin-eosin staining was used to observe the pathological morphology of endometriotic lesions. Immunohistochemistry was employed to observe the expression of angiogenesis-specific markers cluster of differentiation 34 antigen (CD34) and friend leukemia virus integration-1 (FLI-1). Enzyme-linked immunosorbent assay and the nitrate reductase method were employed to determine the serum levels of VEGF and NO, respectively. Western blot was employed to measure the protein levels of VEGF, PI3K, phosphorylated PI3K (p-PI3K), Akt, phosphorylated Akt (p-Akt), eNOS, and phosphorylated eNOS (p-eNOS). Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was conducted to determine the mRNA levels of VEGF, PI3K, Akt, and eNOS. ResultsThe blank group and the sham operation group had no significant changes in the number and latency period of writhing responses, serum VEGF and NO levels, protein levels of VEGF, p-PI3K/PI3K, p-Akt/Akt, and p-eNOS/eNOS, and mRNA levels of VEGF, PI3K, Akt, and eNOS. The model group showed an increase in the number and a reduction in the latency period of writhing responses, enlargement of ectopic endometrial tissue in the abdominal wall, with stromal hyperplasia, glandular dilation, and increased vasculature. In addition, the modeling led to increased positive expression of CD34 and FLI-1, elevated serum VEGF and NO levels, and up-regulated protein levels of VEGF, p-PI3K/PI3K, p-Akt/Akt, and p-eNOS/eNOS and mRNA levels of VEGF, PI3K, Akt, and eNOS (P<0.01). Compared with the model group, the gestrinone and high-, medium-, and low-dose TCM groups showed a significant reduction in the number of writhing responses, a significant prolongation of the latency period, reduced ectopic endometrial tissue in the abdominal wall, alleviated pathological damage, and reduced positive expression of CD34 and FLI-1. The gestrinone group and the high- and medium-dose TCM groups showed lowered serum VEGF and NO levels as well as down-regulated protein levels of VEGF, p-PI3K/PI3K, p-Akt/Akt, and p-eNOS/eNOS and mRNA levels of VEGF, PI3K, Akt, and eNOS. Moreover, the low-dose TCM group showed reductions in the serum VEGF level, the protein levels of VEGF, p-PI3K/PI3K, p-Akt/Akt, and p-eNOS/eNOS, and the mRNA levels of VEGF and eNOS (P<0.05, P<0.01). ConclusionModified SShaofu Zhuyutang can inhibit angiogenesis in endometriosis by antagonizing the abnormal activation of the VEGF/PI3K/Akt/eNOS-NO signaling pathway, thereby preventing the occurrence, development, and deterioration of endometriosis.
4.Effects of loneliness, social support and social media use on post-traumatic stress symptoms among post-earthquake adolescents
Zhaoran GUO ; Qicheng CHE ; Zhihong WU ; Wei SHI
Sichuan Mental Health 2025;38(1):53-58
BackgroundPost-traumatic stress symptoms (PTSS) is highly prevalent in adolescents who have experienced earthquake, which seriously threatens their physical and mental health, yet there is currently a lack of research on the effects of loneliness, social support and social media use on PTSS among post-earthquake adolescents. ObjectiveTo assess the PTSS among adolescents experiencing MS6.0 Luxian, Sichuan, earthquake on 16 September 2021, and to investigate the effects of loneliness, social support and social media use on PTSS, so as to provide references for the intervention of PTSS among post-earthquake adolescents. MethodsOn November 12, 2021, simple random sampling technique was used to select 2 522 post-earthquake adolescents in Luxian county of Luzhou city in Sichuan province. All subjects were assessed using Post-Traumatic Stress Disorder Checklist for DSM-5 (PCL-5), Multidimensional Scale of Perceived Social Support (MSPSS), Short-form UCLA Loneliness Scale (ULS-3) and Social Media Use Scale (SM-10). Binary Logistic regression was used to determine the factors influencing PTSS among post-earthquake adolescents. ResultsPTSS was detected in 91 (3.61%) adolescents. Binary Logistic regression revealed that perceived social support from family members (OR=0.926, 95% CI: 0.879~0.976) was a protective factor for PTSS among post-earthquake adolescents. Lack of companionship (OR=1.764, 95% CI: 1.141~2.727), feeling isolated (OR=2.037, 95% CI: 1.282~3.236), and viewing negative emotional response of disaster victims through social media (OR=1.615, 95% CI: 1.291~2.020) were risk factors for PTSS among post-earthquake adolescents. ConclusionLack of companionship, feeling isolated, and viewing negative emotional response of disaster victims through social media pose a negative impact on PTSS among post-earthquake adolescents, while perceived social support from family members exert a positive impact on PTSS among post-earthquake adolescents. [Funded by Humanity and Social Science Youth foundation of Ministry of Education of China (number, 22YJC190019); Natural Science Foundation of Sichuan Province (number, 2023NSFSC1486)]
5.Modified Shaofu Zhuyutang Mediates VEGF/PI3K/Akt/eNOS Signaling Pathway to Inhibit Angiogenesis in Endometriosis
Jiaxing WANG ; Qi SHI ; Quansheng WU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):81-90
ObjectiveTo explore the mechanism by which modified Shaofu Zhuyutang inhibits angiogenesis in endometriosis via the vascular endothelial growth factor (VEGF)/phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/endothelial nitric oxide synthase (eNOS)-nitric oxide (NO) signaling pathway. MethodsEighty-four female SD rats were randomly assigned into blank, sham operation, model, positive control (gestrinone, 0.25 mg·kg-1), high-, medium-, and low-dose (30, 15, 7.5 g·kg-1, respectively) traditional Chinese medicine (TCM, modified Shaofu Zhuyutang) groups. A rat model of endometriosis was established by the autotransplantation method. After successful modeling, rats in the drug intervention groups were administrated with corresponding agents by gavage, and those in the blank, sham operation, and model groups received an equal volume of distilled water. After 28 days of gavage, rats were administrated with oxytocin, and the number and latency period of writhing responses were observed. Serum samples from each group, ectopic lesions from modeling groups, and uteri from blank and sham operation groups were collected. Hematoxylin-eosin staining was used to observe the pathological morphology of endometriotic lesions. Immunohistochemistry was employed to observe the expression of angiogenesis-specific markers cluster of differentiation 34 antigen (CD34) and friend leukemia virus integration-1 (FLI-1). Enzyme-linked immunosorbent assay and the nitrate reductase method were employed to determine the serum levels of VEGF and NO, respectively. Western blot was employed to measure the protein levels of VEGF, PI3K, phosphorylated PI3K (p-PI3K), Akt, phosphorylated Akt (p-Akt), eNOS, and phosphorylated eNOS (p-eNOS). Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was conducted to determine the mRNA levels of VEGF, PI3K, Akt, and eNOS. ResultsThe blank group and the sham operation group had no significant changes in the number and latency period of writhing responses, serum VEGF and NO levels, protein levels of VEGF, p-PI3K/PI3K, p-Akt/Akt, and p-eNOS/eNOS, and mRNA levels of VEGF, PI3K, Akt, and eNOS. The model group showed an increase in the number and a reduction in the latency period of writhing responses, enlargement of ectopic endometrial tissue in the abdominal wall, with stromal hyperplasia, glandular dilation, and increased vasculature. In addition, the modeling led to increased positive expression of CD34 and FLI-1, elevated serum VEGF and NO levels, and up-regulated protein levels of VEGF, p-PI3K/PI3K, p-Akt/Akt, and p-eNOS/eNOS and mRNA levels of VEGF, PI3K, Akt, and eNOS (P<0.01). Compared with the model group, the gestrinone and high-, medium-, and low-dose TCM groups showed a significant reduction in the number of writhing responses, a significant prolongation of the latency period, reduced ectopic endometrial tissue in the abdominal wall, alleviated pathological damage, and reduced positive expression of CD34 and FLI-1. The gestrinone group and the high- and medium-dose TCM groups showed lowered serum VEGF and NO levels as well as down-regulated protein levels of VEGF, p-PI3K/PI3K, p-Akt/Akt, and p-eNOS/eNOS and mRNA levels of VEGF, PI3K, Akt, and eNOS. Moreover, the low-dose TCM group showed reductions in the serum VEGF level, the protein levels of VEGF, p-PI3K/PI3K, p-Akt/Akt, and p-eNOS/eNOS, and the mRNA levels of VEGF and eNOS (P<0.05, P<0.01). ConclusionModified SShaofu Zhuyutang can inhibit angiogenesis in endometriosis by antagonizing the abnormal activation of the VEGF/PI3K/Akt/eNOS-NO signaling pathway, thereby preventing the occurrence, development, and deterioration of endometriosis.
6.Expert Consensus on Clinical Application of Qinbaohong Zhike Oral Liquid in Treatment of Acute Bronchitis and Acute Attack of Chronic Bronchitis
Jian LIU ; Hongchun ZHANG ; Chengxiang WANG ; Hongsheng CUI ; Xia CUI ; Shunan ZHANG ; Daowen YANG ; Cuiling FENG ; Yubo GUO ; Zengtao SUN ; Huiyong ZHANG ; Guangxi LI ; Qing MIAO ; Sumei WANG ; Liqing SHI ; Hongjun YANG ; Ting LIU ; Fangbo ZHANG ; Sheng CHEN ; Wei CHEN ; Hai WANG ; Lin LIN ; Nini QU ; Lei WU ; Dengshan WU ; Yafeng LIU ; Wenyan ZHANG ; Yueying ZHANG ; Yongfen FAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):182-188
The Expert Consensus on Clinical Application of Qinbaohong Zhike Oral Liquid in Treatment of Acute Bronchitis and Acute Attack of Chronic Bronchitis (GS/CACM 337-2023) was released by the China Association of Chinese Medicine on December 13th, 2023. This expert consensus was developed by experts in methodology, pharmacy, and Chinese medicine in strict accordance with the development requirements of the China Association of Chinese Medicine (CACM) and based on the latest medical evidence and the clinical medication experience of well-known experts in the fields of respiratory medicine (pulmonary diseases) and pediatrics. This expert consensus defines the application of Qinbaohong Zhike oral liquid in the treatment of cough and excessive sputum caused by phlegm-heat obstructing lung, acute bronchitis, and acute attack of chronic bronchitis from the aspects of applicable populations, efficacy evaluation, usage, dosage, drug combination, and safety. It is expected to guide the rational drug use in medical and health institutions, give full play to the unique value of Qinbaohong Zhike oral liquid, and vigorously promote the inheritance and innovation of Chinese patent medicines.
7.Effects and mechanism of Qiangxin decoction on mitochondrion of rats with chronic heart failure
Meiling MAO ; Jianqi LU ; Zhide ZHU ; Yan PANG ; Liyu XIE ; Jiayong CHEN ; Xinyu WU ; Xiang XIAO ; Junshen LU ; Weiqi SHI
China Pharmacy 2025;36(2):160-165
OBJECTIVE To investigate the effects and potential mechanism of Qiangxin decoction on mitochondrion of rats with chronic heart failure (CHF). METHODS The CHF model was established by ligating the left anterior descending branch of the coronary artery. Modeled rats were divided into model group, Qiangxin decoction low-dose and high-dose groups (12.25, 24.50 g/kg, calculated by crude drug), and chemical medicine group (Sacubitril valsartan sodium tablets, 10.42 mg/kg), with 10 rats in each group; control group was set up without treatment. Each group of rats was orally administered with the corresponding medication or normal saline twice a day for 28 consecutive days. After the last medication, the contents of N-terminal pro-brain natriuretic peptide (NT-proBNP) and adenosine triphosphate (ATP) in serum and phosphatidic acid (PA) and cardiolipin (CL) in myocardial tissue were all detected; the pathological damage and collagen fibrosis of rat myocardial tissue were observed; the apoptosis of myocardial cells was determined; the ultrastructure of myocardial tissue was observed; the protein expressions of mitofusin 1 (Mfn1), Mfn2, optic atrophy protein 1 (OPA1) and dynamin-related protein 1 (Drp1) were all detected in myocardial tissue. RESULTS Compared with control group,the serum content of NT-proBNP, apoptotic rate of myocardial cells, and relative expressions of S-OPA1 and Drp1 proteins were all increased significantly; serum content of ATP,contents of PA and CL, and relative expressions of Mfn1, Mfn2 and L-OPA1 proteins were all significantly reduced (P<0.05). There were abnormal membrane tissue structure in various layers of myocardial tissue, degeneration and necrosis of myocardial cells, and severe fibrosis; the mitochondria were swollen, with reduced or absent cristae, and uneven matrix density. After intervention with Qiangxin decoction, the levels of the aforementioned quantitative indicators in serum and myocardial tissue of rats (excluding CL content in the Qiangxin decoction low- dose group) were significantly reversed (P<0.05); the pathological damage of myocardial tissue had significantly improved, fibrosis had significantly reduced, mitochondrial morphology tended to be normal, cristae had increased, and matrix density was uniform. CONCLUSIONS Qiangxin decoction can regulate myocardial mitochondrial function and structural integrity of CHF rats, thereby improving myocardial energy metabolism and antagonizing myocardial fibrosis, the mechanism of which may be associated with activating PA/Mfn/CL signaling pathway.
8.Effects and mechanism of Qiangxin decoction on mitochondrion of rats with chronic heart failure
Meiling MAO ; Jianqi LU ; Zhide ZHU ; Yan PANG ; Liyu XIE ; Jiayong CHEN ; Xinyu WU ; Xiang XIAO ; Junshen LU ; Weiqi SHI
China Pharmacy 2025;36(2):160-165
OBJECTIVE To investigate the effects and potential mechanism of Qiangxin decoction on mitochondrion of rats with chronic heart failure (CHF). METHODS The CHF model was established by ligating the left anterior descending branch of the coronary artery. Modeled rats were divided into model group, Qiangxin decoction low-dose and high-dose groups (12.25, 24.50 g/kg, calculated by crude drug), and chemical medicine group (Sacubitril valsartan sodium tablets, 10.42 mg/kg), with 10 rats in each group; control group was set up without treatment. Each group of rats was orally administered with the corresponding medication or normal saline twice a day for 28 consecutive days. After the last medication, the contents of N-terminal pro-brain natriuretic peptide (NT-proBNP) and adenosine triphosphate (ATP) in serum and phosphatidic acid (PA) and cardiolipin (CL) in myocardial tissue were all detected; the pathological damage and collagen fibrosis of rat myocardial tissue were observed; the apoptosis of myocardial cells was determined; the ultrastructure of myocardial tissue was observed; the protein expressions of mitofusin 1 (Mfn1), Mfn2, optic atrophy protein 1 (OPA1) and dynamin-related protein 1 (Drp1) were all detected in myocardial tissue. RESULTS Compared with control group,the serum content of NT-proBNP, apoptotic rate of myocardial cells, and relative expressions of S-OPA1 and Drp1 proteins were all increased significantly; serum content of ATP,contents of PA and CL, and relative expressions of Mfn1, Mfn2 and L-OPA1 proteins were all significantly reduced (P<0.05). There were abnormal membrane tissue structure in various layers of myocardial tissue, degeneration and necrosis of myocardial cells, and severe fibrosis; the mitochondria were swollen, with reduced or absent cristae, and uneven matrix density. After intervention with Qiangxin decoction, the levels of the aforementioned quantitative indicators in serum and myocardial tissue of rats (excluding CL content in the Qiangxin decoction low- dose group) were significantly reversed (P<0.05); the pathological damage of myocardial tissue had significantly improved, fibrosis had significantly reduced, mitochondrial morphology tended to be normal, cristae had increased, and matrix density was uniform. CONCLUSIONS Qiangxin decoction can regulate myocardial mitochondrial function and structural integrity of CHF rats, thereby improving myocardial energy metabolism and antagonizing myocardial fibrosis, the mechanism of which may be associated with activating PA/Mfn/CL signaling pathway.
9.Wen-Shen-Tong-Du Decoction promoting spinal cord injury repair in mice
Ruihua ZHAO ; Sixian CHEN ; Yang GUO ; Lei SHI ; Chengjie WU ; Mao WU ; Guanglu YANG ; Haoheng ZHANG ; Yong MA
Chinese Journal of Tissue Engineering Research 2025;29(6):1118-1126
BACKGROUND:Previous studies have confirmed that Wen-Shen-Tong-Du Decoction can promote the recovery of spinal cord injury by inhibiting pyroptosis of splenic B cells,promoting the phagocytosis of myelin debris by microvascular endothelial cells,affecting the migration and infiltration of microglia,promoting the recovery of damaged neurons,and decreasing neuronal apoptosis after spinal cord injury,but the mechanism of this is still not clear. OBJECTIVE:To investigate the effect of Wen-Shen-Tong-Du Decoction on the triggering receptor expressed on myeloid cells 2(TREM2)and PI3K/Akt signaling pathways in mice following spinal cord injury. METHODS:Thirty-six C57BL/6 mice were selected and randomly divided into a sham-operation group,a model group and a Wen-Shen-Tong-Du Decoction group,with 12 mice in each group.In the model and Wen-Shen-Tong-Du Decoction groups,mouse models of T10 spinal cord injury were prepared by the modified Allen's method.On the 1st day after modeling,the Wen-Shen-Tong-Du Decoction group was given Wen-Shen-Tong-Du Decoction by gavage,and the sham-operation group and the model group were given saline by gavage once a day for 28 days.During the drug administration period,mouse motor function was evaluated by Basso Mouse Scale score and inclined plane test.On the 7th and 28th days after modeling,hematoxylin-eosin staining was used to observe the histopathological changes in the spinal cord tissue of the mice;immunofluorescence double staining was used to detect the protein expression of ionized calcium binding adaptor molecule 1(IBA1)and TREM2;and western blot assay was used to detect the expression of TREM2,PI3K,p-PI3K,Akt,p-Akt,Bcl2,Bax and Caspase3 in spinal cord tissue. RESULTS AND CONCLUSION:Basso Mouse Scale scores and inclined plane test results indicated that the motor function of the mouse hindlimbs was declined after spinal cord injury,and Wen-Shen-Tong-Du Decoction significantly improved motor function in mice with spinal cord injury.Hematoxylin-eosin staining results revealed that Wen-Shen-Tong-Du Decoction significantly ameliorated the pathological structure of spinal cord tissue compared with the model group,manifesting as reduced degrees of dorsal white matter and neuronal atrophy,decreased cytoplasmic vacuolization,and reduced inflammatory cell infiltration.Immunofluorescence double staining results showed that on the 7th day after modeling,the protein expression of IBA1 and TREM2 in the model group was lower than that in the sham-operation group(P<0.05),and the protein expression of IBA1 and TREM2 in the Wen-Shen-Tong-Du Decoction group was higher than that in the model group(P<0.05);on the 28th day after modeling,the protein expression of TREM2 in the model group was lower than that in the sham-operation group(P<0.05),and the protein expression of TREM2 in the spinal cord tissue of the mice in the Wen-Shen-Tong-Du Decoction group was higher than that in the model group(P<0.05).Western blot results analysis demonstrated that on the 7th day after modeling,compared with the sham-operation group,the model group exhibited a significant reduction in TREM2,PI3K,and Bcl2/Bax(P<0.05),as well as a significant increase in p-Akt,Bax and p-Akt/Aktp-PI3K(P<0.05);compared with the model group,the Wen-Shen-Tong-Du Decoction group showed a significant increase in TREM2,PI3K,p-PI3K,Akt,p-Akt,Bcl2,p-PI3K/PI3K,p-Akt/Ak,and Bcl2/Bax(P<0.05),as well as a significant decrease in Bax and Caspase3 protein expression(P<0.05).On the 28th day after modeling,compared with the sham-operation group,the model group exhibited a significant reduction in TREM2,PI3K,p-PI3K,Akt,p-Akt,Bcl2 and Bcl2/Bax(P<0.05),as well as a significant increase in Bax protein expression(P<0.05);compared with the model group,the Wen-Shen-Tong-Du Decoction group showed a significant increase in TREM2,PI3K,Akt,p-Akt,Bcl2,and Bcl2/Bax(P<0.05),as well as a significant decrease in Bax protein expression(P<0.05).To conclude,Wen-Shen-Tong-Du Decoction may activate the PI3K/Akt signaling pathway by up-regulating the expression of TREM2 protein in microglia,and then inhibit neuronal apoptosis,thus exerting neuroprotective effects and promoting the repair of spinal cord injury.
10.Mechanism of Piezo-type mechanosensitive ion channel component 1 in rat pressure injury
Jiaqi SUN ; Lu BIAN ; Wentao SHI ; Xuechao WU ; Xiaojie LU
Chinese Journal of Tissue Engineering Research 2025;29(8):1578-1584
BACKGROUND:The mechanisms underlying the occurrence of pressure injuries are complex,and it is not entirely clear which factors play a central role in the development of pressure injuries and how these factors operate. OBJECTIVE:To investigate the relationship between Piezo-type mechanosensitive ion channel component 1(Piezo1)and the occurrence of pressure injuries. METHODS:(1)Cellular experiment:Human immortalized keratinocytes(HaCaT)were treated with Yoda1,a Piezo1 agonist,at different concentrations.Cell viability,calcium ion influx,Piezo1,and apoptosis-related protein expression were detected.(2)Animal experiment:Twelve Sprague-Dawley rats were randomly divided into a control group and three experimental groups,with three rats in each group.The control group was not subjected to pressure,while in the three experimental groups,magnets with a thickness of 1,2,and 3 mm were used to press on both sides of the rats'back for 1 hour,respectively,to establish the animal models of pressure injuries.After modeling,all traumatic tissues were excised and subjected to hematoxylin-eosin,Masson,immunofluorescence staining and western blot assay. RESULTS AND CONCLUSION:Cellular experiments:The results of live/dead cell staining showed that HaCaT cell apoptosis increased with the increase of Yoda1 concentration(0,2.5,5,and 10 μmol/L),and calcium ion influx increased with the increase of Yoda1 concentration(0,5,and 10 μmol/L),as well as with the prolongation of treatment time.Western blot assay results showed an increase in the expression of BAX,TG2,and PIEZO1 and a decrease in the expression of the expression of Bcl-2 protein in HaCaT cells in 5 and 10 μmol/L Yoda1 groups compared with the control group(0 μmol/L Yoda1).Animal experiments:The results of hematoxylin-eosin and Masson staining showed that the skin structure of the three experimental groups was damaged at the compression site,there was subcutaneous fat liquefaction and necrosis,and collagen was sparse and disorganized,and damage to the skin structure at the compression site was aggravated with the increase of magnet thickness.Immunofluorescence staining and western blot results showed that compared with the control group,the expression of BAX,TG2,Yap1 and PIEZO1 proteins was elevated,and the expression of Bcl-2 proteins was lowered in the three experimental groups.Moreover,the expression of related proteins showed more significant changes with the increase of magnet thickness(pressure).To conclude,skin compression activates PIEZO1,leading to a significant influx of calcium ions.As the pressure increases,this ultimately results in cell apoptosis due to calcium overload.

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