Low Selenium and Low Protein Exacerbate Myocardial Damage in Keshan Disease by Affecting the PINK1/Parkin-mediated Mitochondrial Autophagy Pathway

Objective Keshan disease(KD)is a myocardial mitochondrial disease closely related to insufficient selenium(Se)and protein intake.PTEN induced putative kinase 1(PINK1)/Parkin mediated mitochondrial autophagy regulates various physiological and pathological processes in the body.This study aimed to elucidate the relationship between PINK1/Parkin-regulated mitochondrial autophagy and KD-related myocardial injury.Methods A low Se and low protein animal model was established.One hundred Wistar rats were randomly divided into 5 groups(control group,low Se group,low protein group,low Se+low protein group,and corn from KD area group).The JC-1 method was used to detect the mitochondrial membrane potential(MMP).ELISA was used to detect serum creatine kinase MB(CK-MB),cardiac troponin I(cTnI),and mitochondrial-glutamicoxalacetic transaminase(M-GOT)levels.RT-PCR and Western blot analysis were used to detect the expression of PINK1,Parkin,sequestome 1(P62),and microtubule-associated proteins1A/1B light chain 3B(MAP1LC3B).Results The MMP was significantly decreased and the activity of CK-MB,cTnI,and M-GOT significantly increased in each experimental group(low Se group,low protein group,low Se+low protein group and corn from KD area group)compared with the control group(P<0.05 for all).The mRNA and protein expression levels of PINK1,Parkin and MAP1LC3B were profoundly increased,and those of P62 markedly decreased in the experimental groups compared with the control group(P<0.05 for all).Conclusion Low Se and low protein levels exacerbate myocardial damage in KD by affecting the PINK1/Parkin-mediated mitochondrial autophagy pathway.

Research progress in mitochondrial autophagy mediated by BNIP3

Mitochondrial autophagy is widely found in mammals,and plays an important role in maintaining mitochondrial balance and mitochondrial quality control in cells.In this review,we reviewed the research progress of BNIP3-mediated mitochondrial autophagy and diseases in recent 5 years,providing new ideas for clinical diagnosis and treatment.

The Fang’s hypoglycemic formula improves insulin resistance in type 2 diabetes mellitus by regulating mitochondrial autophagy

Background:To investigate the pharmacological effects of Fangshi Jiangtang decoction(FSJT)on type 2 diabetes mellitus(T2DM)model rats and explore its mechanism of action from the perspective of mitochondrial autophagy.Methods:Sixty Sprague Dawley rats were randomly divided into six groups after one week of adaptive feeding:Control group,T2DM model group,metformin group(0.2 g/kg by gavage),and FSJT low,medium,and high dose groups(9.5,19,38 g/kg by gavage).Except for the Control group,the other five groups were given a high-fat diet.The treatment lasted for 8 weeks,and blood glucose levels were measured weekly.Eight weeks later,blood samples were collected from the rats,and serum was separated for the determination of HbA1c,oral glucose tolerance test,and homeostatic model assessment for insulin resistance index.The pancreas of the rats was collected,weighed,and fixed.The same part of the pancreas was used for hematoxylin-eosin.Kits were used to detect triglycerides,total cholesterol,interleukin-1β,interleukin-6,tumor necrosis factor-α,malondialdehyde,glutathione peroxidase,and superoxide dismutase in pancreatic tissue to assess the effects of FSJT on inflammation and oxidative stress in T2DM rats.Western blot analysis was performed to detect the expression of VDAC1,TOM20,COXⅣ,PINK1,Parkin,beclin1,light chain 3,and selective autophagy adaptor protein P62 to evaluate the effects of FSJT on mitochondrial autophagy in T2DM model rats.Results:Compared with the T2DM model group,FSJT intervention significantly reduced blood glucose,HbA1c,oral glucose tolerance test,and homeostatic model assessment for insulin resistance index in T2DM model rats,alleviated pancreatic tissue lesions,reduced levels of total cholesterol,triglycerides,interleukin-1β,interleukin-6,tumor necrosis factor-α,and malondialdehyde,increased glutathione peroxidase and superoxide dismutase activities,downregulated the expression of VDAC1,TOM20,COXⅣ,and P62 proteins,and upregulated the expression of PINK1,Parkin,Beclin1,and light chain 3 proteins.Conclusion:FSJT can improve insulin resistance in T2DM by promoting the activation of mitochondrial autophagy.

Effects of Qingguang’an Granules on mitochondrial autophagy of retinal ganglion cells in rats with chronic ocular hypertension

Objective To investigate the effect and underlying mechanism of Qingguang’an Granules(青光安颗粒剂,QGAG)on mitochondrial autophagy(mitophagy)of retinal ganglion cells(RGCs)in rats with chronic ocular hypertension(COH).Methods Sixty Sprague Dawley(SD)rats,half males and half females,were randomly assigned to three groups:the control,model,and QGAG(2.5 g/kg)groups,with 20 rats in each group.Rats’model of COH was established by cauterizing episcleral veins in the model group and QGAG group.Three weeks after successful modeling,rats in the QGAG group were intra-gastrically administered with QGAG,while rats in the control group and the model group received an equal dose of normal saline.After three months of intragastric administration,intraocular pressure(IOP)of all rats was measured.The mitophagy was monitored by the immunofluorescence method,the mitochondrial membrane potential was measured using the JC-1 method,and the morphological changes of mitophagy in RGCs were observed by transmission electron microscopy.Meanwhile,rat RGCs were labeled using the fluorescent gold method,and RGCs density in each group was calculated.Moreover,RGCs apoptosis was observed by TdT-mediated dUTP Nick-End Labeling(TUNEL)assay.Finally,the expression levels of Parkin,optineurin,microtubule-associated protein 1 light chain 3-Ⅱ/microtubule-associated protein 1 light chain 3-Ⅰ(LC3-Ⅱ/LC3-Ⅰ),recombinant lysosomal associated membrane protein 1(LAMP1),and B-cell lymphoma-2(Bcl-2)in RGCs were determined by Western blot assay.The corresponding mRNAs were detected through quantitative real-time polymerase chain reaction(qRT-PCR).Results The QGAG reduced IOP in COH rats,and inhibited mitophagy and apoptosis of RGCs(P<0.05).Besides,the QGAG significantly increased the expression levels of Parkin and Bcl-2(P<0.05),and inhibited the expression levels of optineurin,LAMP1,and LC3-Ⅱ/LC3-Ⅰ(P<0.05)in RGCs of COH rats.Conclusion The QGAG can inhibit mitophagy in RGCs of COH rats and show a protective effect against optic nerve damage caused by glaucoma,which may be mediated through the mitophagy ubiquitination via the Parkin/PINK1-related pathway.

A theoretical study of miRNA155-DNA methylation mediated mitokATP regulating mitochondrial autophagy in the improvement of myocardial ischemia-reperfusion injury by Qishen Yiqi Dropping Pills

In the state of acute myocardial ischemia,miRNA expression can regulate related genes and proteins,reduce myocardial cell damage,and thus play a protective role in the myocardium.However,the specific mechanism still needs to be further explored.Recent studies have found that the opening of the mitoKATP channel can regulate mitochondrial autophagy,and the initiation of miRNA-DNA methylation plays a regulatory role in inducing cell autophagy.The applicant research team previously found that Qishen Yiqi Dropping Pills could significantly improve myocardial ischemia by mediating MitokATP channels to regulate mitochondrial autophagy.,and animal experiments have confirmed that miR-155 plays a significant role in the aspect of autophagy regulates,inflammatory reaction and Vascular smooth muscle cell migration.Therefore,the applicant innovatively proposed that Qishen Yiqi Dropping Pills can regulate miRNA155-DNA methylation to mediate the opening of mitoKATP,thereby regulating mitochondrial autophagy and improving myocardial ischemia.In this paper,the association between mitochondrial autophagy and oxidative stress injury after myocardial ischemia was described,and the possible mechanism of Qisen Yiqi dropping pills regulating mitochondrial autophagy by regulating miRNA155-DNA methylation to mediate MitokATP to improve myocardial ischemia reperfusion injury was discussed,so as to provide theoretical ideas for related research.

Study on the mechanism of action of Lizhong Tang in ameliorating non-alcoholic fatty liver disease by regulating mitochondrial autophagy

Objective:To investigate the therapeutic efficacy and underlying mechanisms of LiZhong Tang in the context of non-alcoholic fatty liver disease(NAFLD).Methods:High-fat feed was used to induce NAFLD in rats,Blood and liver samples were collected to facilitate a comparative analysis of rat body mass and liver wet weight and calculate the liver index.Liver pathology was observed,while serum transaminase and blood lipid levels were measured.The protein expression levels of PINK1,Parkin,and LC-3II in rat liver were detected using Western Blot analysis.Results:Compared with the control group,the NAFLD rats exhibited a significant increase in body weight,liver wet weight,liver index,transaminase levels,and blood lipid levels.The expression levels of PINK1,Parkin,and LC3-II protein were significantly decreased(P<0.01).Following intervention with Lizhong Tang,rats in each herbal treatment group displayed a decrease in body weight,liver wet weight,liver index,se-rum transaminase,and blood lipid levels.The expression levels of PINK1,Parkin,and LC-3II rebounded(P<0.05),with the high-dose group demonstrating the most pronounced effects(P<0.01).Histopathological examination of liver tissue revealed that rats in the model group displayed disrupted hepatic lobule structure,swollen hepatocytes,disordered arrangement,and a multi-tude of varying-sized lipid vacuoles within the cytoplasm.Conversely,rats treated with different doses of the herbal remedy exhibited improvements in liver tissue pathology,with the high-dose group showing the most notable enhancement.Conclusion:Lizhong Tang can improve NAFLD disease by regulating mitochondrial autophagy.

Discussion on the Relationship Between Senescence Caused by Pi(Spleen)Deficiency and Mitochondrial Autophagy

The Pi(Spleen)is responsible for the formation and transportation,and it is in the center.It is the source of qi and blood generation and transformation,so it has an essential position.If the qi and blood are biochemically passive,the functions of the human body will age prematurely.Mitochondria are for energy synthesis and have always been a hot topic in modern medical research.Many of their functions are similar to those of the Pi in Chinese medicine.In Chinese medicine,strengthening the Pi is to enhance the Pi’s ability to transport and transform,and to speed up the Pi,which is similar to the ability of mitochondria to maintain the ability to remove damaged cells.This can speed up the body’s ability to absorb nutrients and delay aging.

Effect of phosphatase and tensin homolog-induced putative kinase 1/E3 ubiquitin ligase Parkin mediated mitochondrial autophagy on chronic kidney disease myocardial injury and the intervention mechanism of Shenshuai recipe(肾衰方)

OBJECTIVE:To study whether Shenshuai recipe(肾衰方,SSR)can play a protective role on chronic kidney disease myocardial injury model through phosphatase and tensin homolog-induced putative kinase 1(PINK1)/E3 ubiquitin ligase Parkin(Parkin)mitochondrial autophagy pathway.METHODS:Forty-eight nephrectomized rats were randomly divided into six groups:sham-operated group,model group,Benazepril group,low,medium and highdose groups of SSR.The rats were given the corresponding intervention for six weeks,then were sacrificed.Serum was examined by enzyme linked immunosorbent assay(ELISA).Cardiac ultrasound was used to detect cardiac function in 5/6 nephrectomized rats.Myocardial tissue was examined by light and electron microscopy;PINK1,Parkin,microtubule-associated protein 1 light chain 3 II(LC3B),sequestosome 1(P62),BECN1(Beclin-1)and dynamin-related protein 1(Drp-1)were measured by real time polymerase chain reaction(RT-PCR),Western blot(WB)and immunohistochemistry(IHC).RESULTS:The expression levels of blood urea nitrogen(BUN)and creatinine(SCr)in the model group were significantly higher than those in the sham-operated group,indicating that modeling was successful.SSR can protect myocardium by reducing the relative expression of creatine kinase myocardial isoenzyme and hypersensitivity cardiac troponin I(P<0.05).SSR can improve cardiac function in rats after ultrasound testing.SSR can improve the pathological manifestations of myocardial tissue after Masson staining.SSR can increase the number of autophagosomes and autophagiclysosomes in 5/6 nephrectomized rats(P<0.05).Determined by RT-PCR,WB and IHC,SSR can increase the relative expression of PINK1,Parkin,and LC3B(P<0.05),and decrease the relative expression of P62,Beclin-1 and Drp-1(P<0.05).CONCLUSIONS:The PINK1/Parkin mitochondrial autophagy pathway in myocardial tissues in 5/6 nephrectomy CKD myocardial injury rats was inhibited.SSR can activate PINK1/Parkin mitochondrial autophagy to enhance mitochondrial autophagy,and play a protective role in myocardial tissues.

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

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 PINK1,PARKIN m RNA and the ratio of LC3BⅡ/LC3BⅠin H/Rinduced H9C2 cells,inhibit the interaction between PARKIN and Ubiquitin in H/R-induced H9C2 cells,and further reduce the expression of TOM20,TIM23,PARL,Mfn1 and Mfn2 protein in H/R-induced H9C2 cells.The effect of resveratrol is consistent with that of autophagy activator on H/R-induced H9C2 cells.CONCLUSIONS:Resveratrol can protect H9C2 cells from H/R injury,which may be related to resveratrol promoting mitochondrial autophagy by activating PINK1/PARKIN signaling pathway.

The role of mitochondrial autophagy in Doxorubicininduced cardiotoxicity

Background Doxorubicin is a widely used drug in all kinds of chemotherapy, but its application is limited by its cardiac toxicity to some extent. Most of scholars have studied doxorubicin induced myocardial injury and cardiomyocytes death, but the specific mechanism remains unclear. Autophagy is a metabolic pathway of degrad- ing longevity protein and organelles by lysosome, especially selective autophagy which called mitochondrial au- tophagy exists in various cells. The normal range of mitochondrial autophagy helps to maintain the physiological function, otherwise, could lead to the development of disease. Recent studies have suggested that mitochondrial autophagy is involved in the progress of doxorubicin-induced cardiotoxicity. This article reviews the role of mito- chondrial autophagy in doxorubicin-induced cardiotoxicity, and how mitochondrial autophagy is involved in the occurrence and development of the progress, in order to provide a theoretical basis of prevention and treatment.

Cardioprotective effect of Yiqi Huoxue granule through regulation of mitophagy after myocardial infarction in rats

Objective:To investigate the cardioprotective effect of Yiqi Huoxue granule(YQHXG)in the regulation of autophagy in rats induced with myocardial infarction(MI).Methods:An acute MI animal model was established by ligation of the left anterior descending branch of the coronary artery in Sprague-Dawley rats.Besides,20 rats received sham operation were classified into a control group.The remaining 59 rats were randomly divided into MI model group(n=19),YQHXG group(n=20),and perindopril group(n=20).Relevant indicators on days 7 and 28 were observed in each group.Left ventricular function was determined by echocardiography.The structure and morphology of mitochondria,and the number of autophagic vesicles,were observed by transmission electron microscopy.The mRNA and protein expression levels of LC3,FUNDC1,Beclin-1,and BNIP3 were examined in the tissue of the MI marginal area.Results:Compared with the MI model group,YQHXG showed obvious improvements in cardiac functions.Observing the microscopic morphology of the heart tissue,myocardial tissue damage attenuated,autophagic signs of autophagosomes and autolysosomes reduced,vacuolization in mitochondria mitigated,and mitochondria arranged in order.YQHXG could reduce the degree of tissue lesion after MI and regulate the expression of autophagy-related molecules at different stages.On Day 7,YQHXG significantly downregulated the expression of Fundc1,Becn1,Bnip3 mRNA and reduced the levels of FUNDC1,Beclin-1,BNIP3,and LC3 B proteins expression(all P<.001).On Day 28,YQHXG could upregulate the expression of Becn1,Fundc1 and Bnip3 mRNA and increased the levels of the corresponding proteins expression(all P<.001).Besides,it also increased LC3 B protein expression level(P=.0344).Conclusion:YQHXG regulated the expression of mitochondrial autophagy-related factors in myocardial tissue and mitochondrial autophagic activity at different stages to protect the heart following MI.

Pitavastatin attenuates AGEs-induced mitophagy via inhibition of ROS generation in the mitochondria of cardiomyocytes

This study aimed to investigate whether pitavastatin protected against injury induced by advanced glycation end products products（AGEs） in neonatal rat cardiomyocytes,and to examine the underlying mechanisms.Cardiomyocytes of neonatal rats were incubated for 48 hours with AGEs（100 μg/mL）,receptor for advanced glycation end products（RAGE）,antibody（1 μg/mL） and pitavastatin（600 ng/mL）.The levels of p62 and beclinl were determined by Western blotting.Mitochondrial membrane potential（△Ψm） and the generation of reactive oxygen species（ROS） were measured through the JC-1 and DCFH-DA.In the AGEs group,the expression of beclinl was remarkably increased compared to the control group,while the expression of p62 was significantly decreased.AGEs also markedly decreased △Ψm and significantly increased ROS compared with the control group.After treatment with RAGE antibody or pitavastatin,the level of beclinl was markedly decreased compared with the AGEs group,but the level of p62 was remarkably increased.In the AGEs ＋ RAGE antibody group and AGEs＋ pitavastatin group,△Ψm was significantly increased and ROS was remarkably decreased compared with the AGEs group.In conclusion,AGEs-RAGE may induce autophagy of cardiomyocytes by generation of ROS and pitavastatin could protect against AGEs-induced injury against cardiomyocytes.

Autophagy in ischemic aged livers

Ischemia/reperfusion(I/R)injury inevitably occurs during liver resection and transplantation.Elderly patients poorly recover from these surgeries.This reduced reparative capacity with aging is causatively associated with decreased mitochondrial function after reperfusion.Mitochondrial autophagy(mitophagy)is a vital cellular process that timely clears abnormal and dysfunctional mitochondria.Impaired or insufficient autophagy can contribute to hepatocyte death after I/R.This review describes our current understanding of I/R injury and highlights new mechanistic correlation between sirtuin 1(SIRT1),mitofusin 2(MFN2),and autophagy in the pathogenesis of age-dependent hypersensitivity to reperfusion injury.The deacetylation of MFN2 by SIRT1 plays a pivotal role in the recovery from reperfusion injury by modulating the onset of autophagy.Targeting the SIRT1-MFN2 axis could be a new therapeutic target to reduce I/R injury in aged livers.

Bloodletting Acupuncture at Jing-Well Points on Hand Induced Autophagy to Alleviate Brain Injury in Acute Altitude Hypoxic Rats by Activating PINK1/Parkin Pathway

Objective: To explore the protective effect of bloodletting acupuncture at twelve Jing-well points on hand(BAJP) on acute hypobaric hypoxia(AHH)-induced brain injury in rats and its possible mechanisms.Methods: Seventy-five Sprague Dawley rats were divided into 5 groups by a random number table(n=15),including control, model, BAJP, BAJP+3-methyladenine(3-MA), and bloodletting acupuncture at non-acupoint(BANA, tail tip blooding) groups. After 7-day pre-treatment, AHH models were established using hypobaric oxygen chambers. The levels of S100B, glial fibrillary acidic protein(GFAP), superoxide dismutase(SOD), and malondialdehyde(MDA) in serum were measured by enzyme-linked immunosorbent assay. Hematoxylin-eosin staining and the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling method were used to assess hippocampal histopathology and apoptosis. Transmission electron microscopy assay was used to observe mitochondrial damage and autophagosomes in hippocampal tissues. Flow cytometry was used to detect mitochondrial membrane potential(MMP). The mitochondrial respiratory chain complexes Ⅰ, Ⅲ and Ⅳ activities and ATPase in hippocampal tissue were evaluated, respectively. Western blot analysis was used to detect the protein expressions of Beclin1, autophagy protein 5(ATG5), microtubule-associated protein 1 light chain 3 beta(LC3B), phosphatase and tensin homolog induced kinase 1(PINK1), and Parkin in hippocampal tissues. The mRNA expressions of Beclin1, ATG5 and LC3-Ⅱ were analyzed by quantitative real-time polymerase chain reaction. Results: BAJP treatment reduced hippocampal tissue injury and inhibited hippocampal cell apoptosis in AHH rats. BAJP reduced oxidative stress by decreasing S100B, GFAP and MDA levels and increasing SOD level in the serum of AHH rats(P<0.05 or P<0.01). Then, BAJP increased MMP, the mitochondrial respiratory chain complexes Ⅰ, Ⅲ and Ⅳ activities, and the mitochondrial ATPase activity in AHH rats(all P<0.01). BAJP improved mitochondrial swelling and increased the autophagosome number in hippocampal tissue of AHH rats. Moreover,BAJP treatment increased the protein and mRNA expressions of Beclin1 and ATG5 and LC3-Ⅱ/LC3-Ⅰratio in AHH rats(all P<0.01) and activated the PINK1/Parkin pathway(P<0.01). Finally, 3-MA attenuated the therapeutic effect of BAJP on AHH rats(P<0.05 or P<0.01). Conclusion: BAJP was an effective treatment for AHH-induced brain injury, and the mechanism might be through reducing hippocampal tissue injury via increasing the PINK1/Parkin pathway and enhancement of mitochondrial autophagy.

Mitophagy deficiency activates stimulator of interferon genes activation and aggravates pathogenetic cardiac remodeling

Stimulator of interferon genes(STING)has recentlybeen found to play a crucial role in cardiac sterile inflammation and dysfunction.The role of stimulator of interferon genes(STING)in cardiac sterile inflammation and dysfunction has been recently discovered.This study aims to examine the involvement of STING in pathological cardiac remodeling and the mechanisms that govern the activation of the STING pathway.To investigate this,transverse aortic constriction(TAC)was performed on STING knockout mice to induce pressure over-load-induced cardiac remodeling.Subsequently,cardiac function,remodeling,and inflammation levels were evaluated.The STING pathway was found to be activated in the pressure overload-stressed heart and angiotensin II(Ang Il)-stimulated cardiac fibroblasts.Loss of STING expression led to a significant reduction in inflammatory responses,mitochondrial fragmenta-tion,and oxidative stress in the heart,resulting in attenuated cardiac remodeling and dysfunc-tion.Furthermore,the exacerbation of pressure overload-induced sTING-mediated inflammation and pathological cardiac remodeling was observed when mitophagy was sup-pressed through the silencing of Parkin,an E3 ubiquitin ligase.Taken together,these findings indicate that STING represents a newly identified and significant molecule implicated in the process of pathological cardiac remodeling and that mitophagy is an upstream mechanism that regulates STING activation.Targeting STING may therefore provide a novel therapeutic strategy for pathological cardiac remodeling and heart failure.

Protective effect of Tongdu Tiaoshen acupuncture combined with Xiaoxuming decoction(小续命汤)on dopaminergic neurons in Parkinson’s disease model

OBJECTIVE:To explore the possible mechanism of Tongdu Tiaoshen acupuncture combined with Xiaoxuming decoction(小续命汤,XXMD)in the treatment of Parkinson’s disease(PD).METHODS:C57BL/6 mice were randomly divided into eight groups(n=12),including blank group,model group,medication group,acupuncture group,high-dose XXMD group(XXMD-H),low-dose XXMD group(XXMD-L),acupuncture combined with high-dose XXMD group(A+H),and acupuncture combined with low-dose XXMD group(A+L).After treatment for 6 weeks,dopamine(DA)neurons and the pathological changes of tyrosine hydroxylase(TH)positive cells were observed.The enzyme-linked immunosorbent assay(ELISA)was used to measure the content of DA and the level of interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-10(IL-10)and tumor necrosis factor alpha(TNF-α).The m RNA level of PINK1 and Parkin and the protein expression of Nix,PINK1 and Parkin in the substantia nigra were also detected.RESULTS:Combination treatment effectively ameliorated the symptoms of PD.Compared with model group,combined treatment significantly up-regulated the protein expression of Nix,Parkin and PINK1 and the m RNA levels of PINK1 and Parkin in the substantia nigra(P<0.0001,P<0.001,P<0.01 or P<0.05).Furthermore,the levels of pro-inflammation cytokines were obviously decreased after combination therapy,while IL-10 content was increased remarkably(P<0.01).CONCLUSION:Compared with each treatment alone,combination therapy improved the pathological damage of DA neurons of PD mice more effectively.The possible mechanism may be attributed to the up-regulated level of mitochondrial autophagy and improved mitochondrial function.These results provide fresh insight into the mechanism of co-treatment with Tongdu Tiaoshen acupuncture and XXMD for PD.

Chemical Synthesis of Structurally Defined Phosphorylated Ubiquitins Suggests Impaired Parkin Activation by Phosphorylated Ubiquitins with a Non-Phosphorylated Distal Unit

Mutations in genes encoding PINK1(PTEN-induced kinase 1)and Parkin(E3 ubiquitin ligase)are identified in familial Parkinson’s disease.However,it remains unclear whether the phosphorylated Ub chains activate wild-type Parkin(w-Parkin)or phosphorylated Parkin(p-Parkin),with the consequent expulsion of the damaged mitochondria.