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Acupotomy alleviates knee osteoarthritis in rabbit by regulating chondrocyte mitophagy via Pink1-Parkin pathway

Journal of Traditional Chinese Medicine 2024年第3期44卷 468-477页

作者： ZHU Wenting GUO Changqing DU Mei MA Yunxuan CUI Yongqi CHEN Xilin GUO Changqing the Third Affiliated Hospital of Beijing University of Chinese Medicine School of Acupuncture-Moxibustion and Tuina Beijing University of Chinese Medicine Department of Medical Technology Shijiazhuang Medical College

OBJECTIVE: To investigate the effect of acupotomy, on mitophagy and the Pink1-Parkin pathway in chondrocytes from rabbits with knee osteoarthritis(KOA).METHODS: A KOA model was established via the modified Videman method. Rabbits were randomly divided into a control group(CON), KOA group and KOA + acupotomy group(Acu). Rabbits in the acupotomy group were subjected to acupotomy for 4 weeks after model establishment. The behavior of the rabbits before and after intervention was recorded. Cartilage degeneration was evaluated by optical microscopy and fluorescence microscopy. The level of mitophagy was evaluated by transmission electron microscopy,immunofluorescence and enzyme-linked immunosorbent assay(ELISA). The expression of phosphatase and tensin homolog(pten)-induced kinase 1(Pink1)-Parkin mitophagy pathway components was evaluated by immunofluorescence, Western blotting and real-time polymerase chain reaction.RESULTS: In rabbits with KOA, joint pain, mobility disorders and cartilage degeneration were observed, the Mankin score was increased, collagen type Ⅱ(Col-Ⅱ)expression was significantly decreased, mitophagy was inhibited, mitochondrial function was impaired, and factors associated with the Pink1-Parkin pathway were inhibited. Acupotomy regulated the expression of Pink1-Parkin pathway-related proteins, the mitophagy-related protein microtubule-associated protein-1 light chain-3,the translocase of the outer membrane, and the inner mitochondrial membrane 23; increased the colocalization of mitochondria and autophagosomes; promoted the removal of damaged mitochondria;restored mitochondrial adenosine-triphosphate(ATP) production;and alleviated cartilage degeneration in rabbits with KOA.CONCLUSIONS: Acupotomy played a role in alleviating KOA in rabbits by activating mitophagy in chondrocytes via the regulation of proteins that are related to the Pink1-Parkin pathway.

关键词： pten phosphohydrolase acupuncture therapy cartilage mitophagy osteoarthritis, knee

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Protective effect of resveratrol on rat cardiomyocyte H9C2 cells injured by hypoxia/reoxygenation by regulating mitochondrial autophagy via pten-induced putative kinase protein 1/Parkinson disease protein 2 signaling pathway

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Journal of Traditional Chinese Medicine 2022年第2期42卷 176-186页

作者： ZHAO Lixia SUN Wei BAI Decheng Institute of Integrated Traditional Chinese and Western Medicine School of Basic Medical SciencesLanzhou UniversityLanzhou 730000China School of Nursing Gansu University of Chinese MedicineLanzhou 730000China Key Laboratory of Dunhuang Medicine Ministry of EducationGansu University of Chinese MedicineLanzhou 730000China Cardiac Surgery the First Hospital of Lanzhou UniversityLanzhouGansu 730000China

OBJECTIVE:To investigate the protective effect of resveratrol on cardiomyocytes after hypoxia/reoxygenation intervention based on pten-induced putative kinase protein 1/Parkinson disease protein 2(PINK1/PARKIN)signaling pathway.METHODS:3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide was used to detect the effect of resveratrol on the viability of H9C2 cells;the hypoxia/reoxygenation(H/R)model was established in tri-gas incubator;2’,7’-Dichlorofluorescin diacetate staining was used to measure the content of reactive oxygen species(ROS);the changes of mitochondrial membrane potential was determined by 5,5’,6,6’-Tetrachloro-1,1’,3,3’-tetraethyl-imidacarbocyanine iodide staining;the changes of mitochondrial respiratory chain complex activity was evaluated by enzyme activity kits;flow cytometry was used to detect the ratio of apoptotic cells;transmission electron microscope was used to observe the ultrastructure of H9C2 cells;Western blot was used to detect the protein changes of mitochondrial 20 k Da outer membrane protein(TOM20),translocase of inner mitochondrial membrane 23(TIM23),presenilins associated rhomboid-like protein(PARL),PINK1,PARKIN and mitofusin 1(Mfn1),mitofusin 2(Mfn2),phosphotyrosine independent ligand for the Lck SH2 domain of 62 k Da(P62),microtubule-associated protein 1 light chain 3 beta(LC3B);the m RNA levels of PINK1 and PARKIN was detected by quantitative polymerase chain reaction;immunoprecipitation assay was used to detect the interaction between PARKIN and Ubiquitin.RESULTS:Resveratrol could inhibit the proliferation of H9C2 cells in a time-and concentration-dependent manner;however,pretreatment with low cytotoxic resveratrol could reduce the H/R-induced increase in cellular ROS levels,alleviate the loss of mitochondrial membrane potential induced by H/R,inhibit H/R-induced apoptosis of H9C2 cells,and protect the mitochondrial structure and respiratory chain of H9C2 cells from H/R damage.Resveratrol could further increase the levels of p62,PINK1,PARKIN protein,the expression of

关键词： resveratrol myocytes cardiac hypoxia pten phosphohydrolase Parkinson disease associated proteins mitochondrial autophagy

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Fuzheng Kang'ai decoction(扶正抗癌方) inhibits cell proliferation,migration and invasion by modulating mir-21-5p/human phosphatase and tensin homology deleted on chromosome ten in lung cancer cells

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Journal of Traditional Chinese Medicine 2022年第3期42卷 344-352页

作者： LI Jinhua LIU Chunping ZHAO Yueyang WU Wanyin SUN Pengtao LI Longmei YANG Xiaobing ZHOU Yushu Department of Ultrasound Guangdong Provincial Hospital of Chinese Medicinethe Second Affiliated Hospital of Guangzhou University of Chinese MedicineGuangzhou 510120China Department of Ultrasound Institute of Ultrasound in Musculoskeletal Sports MedicineGuangdong Second Provincial General HospitalGuangzhou 510317China State Key Laboratory of Dampness Syndrome of Chinese Medicine Guangdong Provincial Hospital of Chinese MedicineThe Second Affiliated Hospital of Guangzhou University of Chinese MedicineGuangzhou 510120China Department of Hematology Guangdong Provincial Hospital of Chinese Medicinethe Second Affiliated Hospital of Guangzhou University of Chinese MedicineGuangzhou 510120China Department of Oncology Clinical and Basic Research Team of TCM Prevention and Treatment of NSCLCGuangdong Provincial Hospital of Chinese Medicinethe Second Affiliated Hospital of Guangzhou University of Chinese MedicineGuangzhou 510120China

OBJECTIVE:To elucidate the potential molecular mechanism by which Fuzheng Kang’ai decoction(扶正抗癌方,FZKA)inhibits proliferation,migration,and invasion of lung cancer cells.METHODS:Varying FZKA concentrations were used to manage lung cancer cells(A549 and PC9).We employed:cell counting kit-8(CCK-8)and plate clone formation assays to examine the cell viability;flow cytometry(FCM)to analyze the cycle arrest;transwell and woundhealing assays to assess the cell invasion and migration,respectively.Further,a quantitative real-time polymerase chain reaction(q RT-PCR)assay was adopted to evaluate the miR-21-5p expression.For protein expression analysis,we employed the Western blot technique.Recombinant miR-21-5p overexpression adenovirus vector harboring GFP was constructed and transfected into A549 and PC9,after which we explored the effect of FZKA on miR-21-5p overexpression.RESULTS:Notably,treatment with FZKA inhibited viability,clone-formation ability,invasion,and migration of lung cancer cells.Mechanistically,FZKA markedly suppressed miR-21-5p expression but elevated the human phosphatase and tensin homology deleted on chromosome ten(pten)protein level in both A549 and PC9 cells.Over-expression of miR-21-5p lowered pten protein expression.Besides,overexpressed miR-21-5p levels with adenovirus antagonized FZKA-upregulated pten protein expression.CONCLUSION:The present study demonstrates how FZKA modulates cell biological behaviors,for instance,it impedes the proliferation by upregulating pten expression with miR-21-5p as the target.These findings unveil the potential novel molecular mechanisms from the micro RNA aspect by which FZKA suppresses the growth of human lung cancer cells.

关键词： lung neoplasms miR-21-5p pten phosphohydrolase Fuzheng Kang’ai decoction

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Mechanism of total glucosides from Chishao(Radix Paeoniae Rubra)on proliferation and apoptosis of hepatocellular carcinoma cells via phosphatase and tensin homolog deleted on chromosome ten/phosphatidylinositol 3-kinase/protein kinase B signaling pathway

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Journal of Traditional Chinese Medicine 2021年第5期41卷 677-683页

作者： FAN Bingbing LI Tianjiao MENG Xiansheng WANG Shuai BAO Yongrui WANG Fei College of Pharmacy Liaoning University of Traditional Chinese MedicineDalian 116600China College of Graduate Studies Liaoning University of Traditional Chinese MedicineShenyang 110847China

OBJECTIVE:To investigate the possible molecular mechanism of total glycosides of Chishao(Radix Paeoniae Rubra)(TG-RPR)on proliferation and apoptosis of hepatocellular carcinoma cells.METHODS:The proliferation of TG-RPR on Hep G2 cells was detected using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay.The apoptosis of Hep G2 cells was measured by annexin V-FITC/double staining.The phosphatase and tensin homolog deleted on chromosome ten(pten)/phosphatidylinositol 3-kinase(PI3 K)/protein kinase B(Akt)signaling pathway was evaluated by Western Blot and reverse transcription-polymerase chain reaction(RT-PCR).RESULTS:TG-RPR can up-regulation the expression of pro-apoptotic factors such as pten and BCL2-Associated X(Bax),down-regulation the expression of anti-apoptotic factors including B-cell lymphoma-2(Bcl-2),PI3 K,and Akt.CONCLUSION:TG-RPR significantly inhibits the proliferation of Hep G2 cells in a dose-dependent manner and promotes apoptosis.These results demonstrated TG-RPR has significant inhibitory effect on Hep G2 cells.These results identify a critical role of TG-RPR in proliferation and apoptosis of Hep G2 cells via modulating pten/PI3 K/Akt signaling pathway.TG-RPR may offer a promise as a potential pharmaceutical therapy for hepatocellular carcinoma.

关键词： carcinoma,hepatocellular glycosides pten phosphohydrolase phosphatidylinositol 3-kinase proto-oncogene proteins c-akt Chishao(Radix Paeoniae Rubra)

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Effect of compound Guizhu capsule on phosphate and tension homology deleted on chromsome ten and murine double mimute 2 gene expression in lung cancer of mice

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Journal of Traditional Chinese Medicine 2017年第4期37卷 461-465页

作者： Cao Yong Zhang Shuyong Zhang Ge Zhang Na Zhang Lixia Huang Yuanliang Yuan Naijun Tang Xiaojuan Du Jiajin Fang Jiuyun College of Traditional Chinese Medicine Jinan University Guangzhou 510632 China

OBJECTIVE: To observe the inhibitory effect of the Chinese herbal compound Guizhu capsule(CGZC)on lung cancer and explore the possible mechanism underlying its actions by evaluating its effects on the expression of phosphate and tension homology deleted on chromsome ten(pten) and murine double mimute 2(MDM2) in lung cancer model mice.METHODS: A mouse model of transplanted lung cancer was established by subcutaneous inoculation of cancer cells into the axillae of mice. Twenty-four hours later, the mice were weighed and randomly divided into the model control group, cisplatin group(DDP group), and high, moderate and low dosage CGZC groups, with ten mice in each group. The control mice received an equal volume of distilled water. DDP was intraperitoneally injected at 1 mg/kg in the DDP group, once a day for 3days. CGZC diluted with distilled water was admin-istered at 20 g/kg(10 times the clinical adult dosage) in the high-dose group, 10 g/kg(5 times the clinical adult dosage) in the moderate-dose group and 5 g/kg(2.5 times the clinical adult dosage) in the low-dose group once a day for 10 days. On the11 th day, the mice were weighed and killed. The tumor tissues were weighed and the tumor inhibition rate was calculated. pten and MDM2 protein expression were detected by immunohistochemical analysis of tumor tissues.RESULTS: The CGZC high-and moderate-dose groups showed a significant inhibitory effect on tumor growth(P 0.05). The CGZC high-dose group also showed enhanced expression of pten protein(P pten and decreasing the expression of MDM2 in the lung cancer tissues of the mice.

关键词：肺瘤 Transplantion Immunohistochemistry pten phosphohydrolase Proto-oncogene 蛋白质 c-mdm2 Ubiquitin 蛋白质 ligases 加重 Guizhu 囊

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