1.Efficacy Connotation and Mechanisms of Shudi Qiangjin Pills Against Steroid-induced Osteonecrosis of Femoral Head Based on "Disease-Syndrome-Formula" Association Network
Zhijian CHEN ; Suya ZHANG ; Longlong DING ; Guixin ZHANG ; Bo LIU ; Baohong MI ; Yanqiong ZHANG ; Na LIN ; Weiheng CHEN ; Chunzhu GONG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):88-99
ObjectiveTo elucidate the efficacy connotation of Shudi Qiangjin pills (SQP) against liver and kidney deficiency in steroid-induced osteonecrosis of femoral head (SONFH) from the perspective of the "disease-syndrome-formula" association and to clarify the underlying mechanisms based on in vivo and in vitro experiment validation. MethodsThe chemical components and the corresponding putative targets of SQP were collected from the Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine (TCMIP) v2.0, the Encyclopedia of Traditional Chinese Medicine (ETCM) v2.0, and HERB databases. The SONFH-related genes were identified based on the differential expression profiles of peripheral blood of patients with SONFH compared to the healthy volunteers, and the disease phenotype-related targets were collected from the TCMIP v2.0 database. Then, the interaction network of "SONFH-related genes and candidate targets of SQP" was constructed based on "gene-gene interaction information", and the major network targets were screened by calculating the topological characteristic values of the network followed by the functional mining according to the Kyoto Encyclopedia of Genes and Genomes (KEGG) database and the SoFDA database. After that, the SONFH rat model was prepared by lipopolysaccharide combined with methylprednisolone injection, and 2.5, 5, 7.5 g·kg-1 SQP (once per day, equivalent to 1, 2, and 3 times the clinical equivalent dose, respectively) or 7.3×10-3 g·kg-1 of alendronate sodium (ALS, once per week, equivalent to the clinical equivalent dose) was given for 8 weeks. The effect characteristics of SQP and ALS in the treatment of SONFH were evaluated by micro-computed tomography scanning, hematoxylin and eosin staining, alkaline phosphatase (ALP) staining, immunohistochemical staining, enzyme-linked immunosorbent assay, and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)staining, and a comparative efficacy analysis was conducted with ALS. In addition, SONFH cell models were prepared by dexamethasone stimulation of osteoblasts, and the intervention was carried out with the medicated serum of SQP at the aforementioned three doses. Cell counting kit-8, ALP staining, ALP activity assay, alizarin red staining, and flow cytometry were employed to investigate the regulatory effect of SQP on osteoblasts. The expression levels of osteogenesis-related proteins and key factors of the target signaling axis were detected by quantitative real-time polymerase chain reaction and Western blot. ResultsThe network analysis results demonstrated that the candidate targets of SQP primarily exerted their therapeutic effects through key signaling pathways, including phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt), lipid metabolism and atherosclerosis, prolactin, chemokines, and neurotrophic factors pathways. These pathways were significantly involved in critical biological processes such as muscle and bone metabolism and the regulation of the "neuro-endocrine-immune" network, thereby addressing both modern medical symptoms (e.g., delayed skeletal maturation and recurrent fractures) and traditional Chinese medicine (TCM) symptoms (e.g., fatigue, aversion to cold, cold limbs, and pain in the limbs and joints in patients with SONFH characterized by liver and kidney deficiency syndrome. Among these pathways, the PI3K/Akt signaling pathway exhibited the highest degree of enrichment. The in vivo experimental results demonstrated that starting from the 4th week after modeling, the modeling group exhibited a significant reduction in body weight compared to the control group (P<0.05). After six weeks of treatment, all dosage groups of SQP showed significantly higher body weights compared to the model group (P<0.01). Compared with the normal group, the model group exhibited significant decreases in bone mineral density (BMD), bone volume fraction (BV/TV), trabecular number (Tb.N), osteocalcin (OCN), alkaline phosphatase (ALP) levels in femoral head tissue, and serum bone-specific alkaline phosphatase (BALP) (P<0.01), along with significant increases in trabecular separation (Tb.Sp), empty lacunae rate in tissue, and apoptosis rate (P<0.01). In comparison to the model group, the SQP intervention groups showed significant improvements in BMD, BV/TV and Tb.N (P<0.01), significant reductions in Tb.Sp, empty lacunae rate and apoptosis rate (P<0.05), and significant increases in protein levels of OCN and ALP as well as BALP content (P<0.05). The in vitro experimental results revealed that all dosage groups of SQP medicated serum showed no toxic effects on osteoblast. Compared with the normal group, the model group displayed significant suppression of osteoblast proliferation activity, ALP activity, and calcified nodule formation rate (P<0.01), significant decreases in mRNA transcription levels of OCN and Runt-related transcription factor 2 (RUNX2) (P<0.01), significant reductions in protein content of osteopontin (OPN), typeⅠ collagen (ColⅠ)A1, B-cell lymphoma-2 (Bcl-2), PI3K, and phosphorylated (p)-Akt (P<0.01), and a significant increase in apoptosis rate (P<0.01). Compared with the model group, the SQP medicated serum intervention groups exhibited significant increases in proliferation activity, ALP activity, calcified nodule formation rate, mRNA transcription levels of OCN and RUNX2, and protein content of OPN, ColⅠA1, Bcl-2, PI3K, and p-Akt (P<0.05), along with a significant decrease in apoptosis rate (P<0.01). ConclusionSQP can effectively reduce the disease severity of SONFH with liver and kidney deficiency syndrome and improve bone microstructure, with the therapeutic effects exhibiting a dose-dependent manner. The mechanism may be related to its regulation of key processes such as muscle and bone metabolism and the correction of imbalances in the "neuro-endocrine-immune" network, thereby promoting osteoblast differentiation and inhibiting osteoblast apoptosis. The PI3K/Akt signaling axis is likely one of the key pathways through which this formula exerts its effects.
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.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.
4.Boosting with Omicron-specific mRNA vaccine or historical SARS-CoV-2 vaccines elicits discriminating immune responses against Omicron variants.
Yi WU ; Xiaoying JIA ; Namei WU ; Xinghai ZHANG ; Yan WU ; Yang LIU ; Minmin ZHOU ; Yanqiong SHEN ; Entao LI ; Wei WANG ; Jiaming LAN ; Yucai WANG ; Sandra CHIU
Acta Pharmaceutica Sinica B 2025;15(2):947-962
Booster vaccinations are highly recommended in combating the SARS-CoV-2 Omicron variant and its subvariants. However, the optimal booster vaccination strategies and related immune mechanisms with different prior vaccinations are under-revealed. In this study, we systematically evaluated the immune responses in mice and hamsters with different prime-boost regimens before their protective efficacies against Omicron were detected. We found that boosting with Ad5-nCoV, SWT-2P or SOmicron-6P induced significantly higher levels of neutralization activities against Omicron variants than CoronaVac and ZF2001 by eliciting stronger germinal center (GC) responses. Specifically, SOmicron-6P induced even stronger antibody responses against Omicron variants in CoronaVac and Ad5-nCoV-primed animals than non-Omicron-specific vaccines but with limited differences as compared to Ad5-nCoV and SWT-2P. In addition, boosting with a specific vaccine has the potential to remodel the existing immune profiles. These findings indicated that adenovirus-vectored vaccines and mRNA vaccines would be more effective than other types of vaccines as booster shots in combating Omicron infections. Moreover, the protective efficacies of the vaccines in booster vaccinations are highly related to GC reactions in secondary lymphatic organs. In summary, these findings provide timely important information on prime-boost regimens and future vaccine design.
5.pLM4ACP: a model for predicting anticancer peptides based on machine learning and protein language models.
Yitong LIU ; Wenxin CHEN ; Juanjuan LI ; Xue CHI ; Xiang MA ; Yanqiong TANG ; Hong LI
Chinese Journal of Biotechnology 2025;41(8):3252-3261
Cancer is a serious global health problem and a major cause of human death. Conventional cancer treatments often run the risk of impairing vital organ functions. Anticancer peptides (ACPs) are considered to be one of the most promising therapeutic agents against common human cancers due to their small sizes, high specificity, and low toxicity. Since ACP recognition is highly limited to the laboratory, expensive, and time-consuming, we proposed pLM4ACP, a model for predicting ACPs based on machine learning and protein language models. In this model, the protein language model ProtT5 was used to extract the features of ACPs, and the extracted features were input into the support vector machine (SVM) classification algorithm for optimization and performance evaluation. The model showcased significantly higher accuracy than other methods, with the overall accuracy of 0.763, F1-score of 0.767, Matthews correlation coefficient of 0.527, and area under the curve of 0.827 on the independent test set. This study constructs an efficient anticancer peptide prediction model based on protein language models, further advancing the application of artificial intelligence in the biomedical field and promoting the development of precision medicine and computational biology.
Machine Learning
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Antineoplastic Agents/chemistry*
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Humans
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Peptides/chemistry*
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Support Vector Machine
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Algorithms
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Computational Biology/methods*
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Neoplasms/drug therapy*
6.Application of quantitative grip strength training in postoperative patients with autogenous arteriovenous internal fistula
Siyi LI ; Yao LU ; Jing LIU ; Meibin ZHANG ; Huijin GONG ; Mingyan LI ; Yanqiong OUYANG
Chongqing Medicine 2024;53(11):1675-1678
Objective To formulate the quantitative grip strength training program for application in the postoperative patients with autogenous arteriovenous internal fistula,and to evaluate its effect on the mat-uration and initial use of autogenous arteriovenous internal fistula.Methods A total of 98 patients with ce-phalic venous radial arterial anastomosis internal fistula formation surgery in Shenzhen Hospital of Southern Medical University from September 2021 to November 2022 were selected as the study subjects by the conven-ience sampling method.According to the follow-up time,they were divided into the observation group (n=41) and control group (n=42).The observation group adopted the quantitative grip strength training for function-al exercise of the limb on the side of internal fistula,while the control group adopted the conventional grip training for functional exercise of the limb on the side of internal fistula.In postoperative 8 weeks,the matura-tion rate of internal fistula,natural blood flow amount of internal fistula,internal diameter of cephalic vein,pre-pump pressure used in the initial stage of internal fistula and the incidence rate of internal fistula complica-tions were evaluated in the two groups.Results Compared with the control group,the maturation rate of in-ternal fistula in the observation group was higher (97.6% vs. 83.3%).The inner diameter of cephalic vein and natural blood flow amount of internal fistula in the observation group were larger than those in the control group[(5.24±0.66)mm vs. (4.63±0.59)mm;(1215.38±562.99)mL/min vs. (955.75±341.94)mL/min],the pre-pump pressure used at the initial stage of internal fistula in the observation group was lower than that in the control group[(119.20±19.83)mmHg vs. (135.74±17.07)mmHg],and the differences were statistically significant (P<0.05).Conclusion The quantitative grip strength training could increase the postoperative maturity rate of patients's internal fistula,and is beneficial to the use in the initial stage of internal fistula.
7.Analysis of the current situation of psychological crisis vulnerability and influencing factors of the elderly multimorbidity
Dou FU ; Xinmei WANG ; Lili ZHANG ; Yanqiong OUYANG ; Shanshan LIU ; Jiaojiao WU ; Xiaojie MA ; Yandi TANG
Chinese Journal of Geriatrics 2024;43(9):1174-1179
Objective:To understand the current situation of psychological crisis vulnerability among elderly individuals with multimorbidity and analyze the factors that influence it, to provide insights for improving their coping abilities.Methods:A cross-sectional survey design was conducted at Renmin Hospital of Wuhan University from May 1 to November 30, 2022.The attitudes toward aging, sense of meaning of life, and vulnerability to psychological crisis were analysed among outpatients and inpatients.Statistical analysis was performed on the questionnaire results, and the influencing factors of vulnerability to psychological crisis in elderly patients with co-morbidities were analyzed using one-way linear regression and multivariate linear regression.Additionally, the correlation between aging attitudes, sense of meaning of life, and vulnerability to psychological crisis was examined using Pearson correlation analysis.Results:A total of 685 questionnaires were distributed, and 602 valid questionnaires were collected, resulting in a valid recovery rate of 87.9%.The total score for the sense of meaning of life in elderly co-morbid patients was(39.2±8.3), while the total score for aging attitudes was(80.2±13.5).The total score for psychological crisis vulnerability was(69.4±12.8), indicating a medium-high level of vulnerability.The results of multiple linear regression analysis revealed that the factors influencing psychological crisis vulnerability in elderly multimorbidity, in descending order, were residence status, economic situation, marital status, age, type of chronic disease, and hospitalization history in the past six months.Psychological crisis vulnerability in elderly multimorbidity showed a negative correlation with the sense of meaning of life and the attitude of aging( r=-0.315, -0.264, both P<0.01), while the attitude of aging exhibited a positive correlation with the sense of meaning of life( r=0.515, P<0.01). Conclusions:The vulnerability of elderly individuals with multimorbidity to psychological crises is influenced by several factors.Healthcare professionals should prioritize individuals who are elderly, residing in nursing institutions, widowed, financially disadvantaged, experiencing multiple illnesses, and not currently hospitalized.
8.Meta-analysis of Clinical Efficacy and"Disease-Symptom-Formula"Associated Mechanism Investigation of Shangke Jiegu Tablet Against Fractures
Xueting LIU ; Lin CHEN ; Suya ZHANG ; Haoying XI ; Ning ZHANG ; Na LIN ; Yanqiong ZHANG
Herald of Medicine 2024;43(12):1919-1929
Objective This study systematically evaluated the clinical efficacy of Shangke Jiegu tablet in the treatment of fracture,and explored the mechanism of action of Shangke Jiegu tablet and the compatibility of each efficacy group from the"Disease-Symptom-Formula"perspective.Methods Clinical research literatures on the use of Shangke Jiegu tablet for fracture intervention were retrieved from Chinese databases(CNKI,Wanfang Database,VIP database)and English databases(PubMed,Cochrane Library,EMbase),covering the period from the inception of the databases to January 2024.Risk assessment tools were used to evaluate the literature's quality,and the data were extracted and analyzed using Stata 16.0 software.Gene sets associated with fracture symptoms were identified through the TCMIP platform(version 2.0).Differential gene expression related to fractures was obtained from the GEO database.Chemical composition and candidate target profiles of the 12 herbs in Shangke Jiegu tablets were collected from TCMIP v 2.0.An interaction network between fracture-related genes and drug candidate targets was established,and core network targets were screened based on topological features,with functional enrichment analysis performed.Results A total of 14 articles were incorporated into the Meta-analysis,encompassing a total sample size of 1 293 cases,indicating an overall response rate of Shangke Jiegu tablets in fracture therapy(RR=1.24,95%CI:1.18-1.31,P<0.001).The"Disease-Symptom-Formula"association network analysis indicated that the pathways related to the putative targets of Shangke Jiegu tablet were primarily involved in bone healing,nerve and blood system regulation,and immune-inflammation regulation.Different efficacy groups within the prescriptions showed varying emphases on these roles.Conclusions Shangke Jiegu tablet may facilitate fracture healing by regulating blood and nervous systems,correcting immune-inflammatory imbalances,and maintaining bone and energy metabolism.The comprehensive effects include the dissipation of blood stasis,the promotion of blood circulation,the alleviation of swelling and pain,the regeneration of muscles and bones,and the clearance of heat and detoxification.These findings support the clinical advantages and positioning of Shangke Jiegu tablet.
9.Discussion on the Targeting Mechanism of Qingre Huoxue Formula (清热活血方) for Treatment of Rheumatoid Arthritis by Regulating “Inflammation-Bone Destruction”:Based on LRG1/TGF- β Signalling Axis
Weixiang LIU ; Yu SHI ; Yanqiong ZHANG ; Xun GONG ; Quan JIANG
Journal of Traditional Chinese Medicine 2024;65(7):691-696
Based on the previous publications, it is believed that damp-heat syndrome is the core syndrome of rheumatoid arthritis (RA), and Qingre Huoxue Formula (清热活血方) is an effective formula for the treatment of damp-heat syndrome of RA. “Inflammation-bone destruction” is a key pathological link of RA, and it is also the advantage of the effectiveness of Qingre Huoxue Formula. Leucine rich α-2-glycoprotein 1 (LRG1) can mediate the transforming growth factor-β (TGF-β) signalling pathway to participate in the pathogenic process of “inflammation-bone destruction” of RA, and it can be used as a target protein in the treatment of damp-heat syndrome of RA by Qingre Huoxue Formula. Accordingly, a scientific hypothesis was proposed that Qingre Huoxue Formula may regulate TGF-β signalling pathway mediated by LRG1 to improve “inflammation-bone destruction” of RA, and it was envisioned that the clinical effect of Qingre Huoxue Formula on LRG1 could be confirmed through clinical studies, and the mechanism of action of Qingre Huoxue Formula on the LRG1/TGF-β signalling axis as well as the influence of the expression or non-expression of the LRG1/TGF-β signalling axis on the therapeutic effectiveness of Qingre Huoxue Formula could be clarified through animal experiments.
10.Establishment and evaluation of the comorbidity animal model of knee osteoarthritis and hypertension with pattern of liver and kidney deficiency
Zhaochen MA ; Chu ZHANG ; Mingzhu XU ; Yudong LIU ; Weiheng CHEN ; Yanqiong ZHANG ; Na LIN
Journal of Beijing University of Traditional Chinese Medicine 2024;47(5):652-661
Objective To establish the animal model of comorbidity of knee osteoarthritis(KOA)and hypertension with pattern of liver and kidney deficiency and evaluate its characteristics of comorbidity and pattern.Methods Wistar-Kyoto(WKY)rats and spontaneously hypertensive rats(SHR)were assigned to the WKY control group(control group),hypertension combined with KOA sham-operation group(sham-operation group),hypertension combined with KOA group(model group),hypertension combined with KOA and liver-kidney deficiency pattern group(LKD group).The animal model of KOA combined with hypertension was prepared by anterior cruciate ligament transection(ACLT)in spontaneously hypertensive rats.ACLT combined with intramuscular injection of hydrocortisone was performed to prepare an animal model of KOA with liver-kidney deficiency(LKD)pattern type,combined with hypertension.Then,the related indexes of LKD syndrome were detected in turn,including the contents of thyroid stimulating hormone(TSH),testosterone(T),corticosterone(CORT),adrenocorticotropic hormone(ACTH)in serum,and enzyme activities of alanine transaminase(ALT),aspartate transaminase(AST),and alkaline phosphatase(ALP)in serum,the mass ratio of liver,kidney,spleen,thymus to the brain,body weight,anal temperature,activity situation,and emotion.Systolic blood pressure,diastolic blood pressure,and other blood pressure-related indices were also detected.The levels of serum tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β),plantar mechanical pain sensitivity threshold,weight difference score of both hind limbs,hind limb joint swelling,and quadruped gait parameters were also measured.Furthermore,hematoxylin-eosin,safranine-fast green,and Masson staining were performed to observe pathological changes,cartilage degeneration,and bone destruction of the knee joint,and the microstructure parameters of the tibia were detected by Micro-CT imaging.Results Compared to the model group,the contents of serum TSH,ACTH,T,CORT and the mass ratio of the kidney,spleen,and thymus to the brain in the LKD group decreased(P<0.05).Compared to the control group,the systolic blood pressure and diastolic blood pressure of the other three groups increased significantly(P<0.05).Compared to the sham-operation group,serum TNF-α and IL-1β levels increased,plantar mechanical pain threshold decreased,weight difference score of both hind limbs and joint swelling of the affected limb increased(P<0.05),and gait parameters(e.g.,gait length and standing time of the affected limbs)became abnormal in the model and LKD groups.Simultaneously,the cartilage surface defect of the rat knee joint was severe,the arrangement of the surface chondrocytes was altered,the cartilage layer became thinner,the muscle fibers increased,and the cartilage ossification was severe.Furthermore,the relative volume,thickness,and number of trabeculae of the knee joint decreased significantly(P<0.05).Conclusion The rat model established in this study is consistent with the clinical characteristics of integrated traditional Chinese and Western medicine in patients with comorbidities of hypertension and KOA with liver and kidney deficiency pattern.This rat model can characterize the typical symptoms of liver and kidney deficiency pattern.It has typical pathological changes in knee cartilage and subchondral bone tissues and can maintain a stable range of high systolic and diastolic blood pressure.It also explore the scientific connotation of simultaneous treatment of different diseases in traditional Chinese medicine,revealing the therapeutic mechanism and developing new drugs.

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