Effect of Wenyang Zhenshuai Granules on autophagy and apoptosis of myocardial cells in septic rats via regulating miR-132-3p/UCP2 expression / 中国中药杂志

This study aimed to investigate the effect of Wenyang Zhenshuai Granules(WYZSG) on autophagy and apoptosis of myocardial cells in rats with sepsis via regulating the expression of microRNA-132-3p(miR-132-3p)/uncoupling protein 2(UCP2). Sixty SD rats were randomly divided into modeling group(n=50) and sham operation group(n=10). The sepsis rat model was constructed by cecal ligation and perforation in the modeling group. The successfully modeled rats were randomly divided into WYZSG low-, medium-and high-dose groups, model group and positive control group. Rats in the sham operation group underwent opening and cecum division but without perforation and ligation. Hematoxylin-eosin(HE) staining was used to observe the pathological changes of rat myocardial tissue. Myocardial cell apoptosis was detected by TdT-mediated dUTP nick end labeling(TUNEL) assay. Real-time quantitative polymerase chain reaction(RT-qPCR) was performed to detect the expression of miR-132-3p and the mRNA expressions of UCP2, microtubule-associated protein light chain 3(LC3-Ⅱ/LC3-Ⅰ), Beclin-1 and caspase-3 in rat myocardial tissue. The protein expressions of UCP2, LC3-Ⅱ/LC3-Ⅰ, Beclin-1 and caspase-3 in myocardial tissue were detected by Western blot. Dual luciferase reporter assay was used to verify the regulatory relationship between miR-132-3p and UCP2. The myocardial fibers of sepsis model rats were disordered, and there were obvious inflammatory cell infiltration as well as myocardial cell edema and necrosis. With the increase of the WYZSG dose, the histopathological changes of myocardium were improved to varying degrees. Compared with the conditions in the sham operation group, the survival rate and left ventricular ejection fraction(LVEF) of rats in the model group, positive control group and WYZSG low-, medium-and high-dose groups were decreased, and the myocardial injury score and apoptosis rate were increased. Compared with the model group, the positive control group and WYZSG low-, medium-and high-dose groups had elevated survival rate and LVEF, and lowered myocardial injury score and apoptosis rate. The expression of miR-132-3p and the mRNA and protein expressions of UCP2 in myocardial tissue in the model group, positive control group and WYZSG low-, medium-and high-dose groups were lower, while the mRNA and protein expressions of LC3-Ⅱ/LC3-Ⅰ, Beclin-1 and caspase-3 were higher than those in the sham operation group. Compared with model group, the positive control group and the WYZSG low-, medium-and high-dose groups had an up-regulation in the expression of miR-132-3p and the mRNA and protein expressions of UCP2, while a down-regulation in the mRNA and protein expressions of LC3-Ⅱ/LC3-Ⅰ, Beclin-1 and caspase-3. WYZSG inhibited excessive autophagy and apoptosis of myocardial cells in septic rats and improved myocardial injury, possibly by regulating the expression of miR-132-3p/UCP2.

Chinese medicine Jiangzhuo mixture regulates glucose and lipid metabolism in obese rats through TLR4/I κB α/NF- κB signaling pathway / 浙江大学学报·医学版

OBJECTIVES@#To explore the mechanism of Chinese medicine Jiangzhuo mixture regulating glucose and lipid metabolism in obese rats.@*METHODS@#Thirty healthy male SD rats were randomly divided into normal control group, model control group, and Jiangzhuo mixture treatment group, with 10 rats in each group. The rats in the normal control group were fed with normal diet, the obesity model was induced by feeding high-fat diet in the model control group and the Jiangzhuo mixture treatment group, the rats in the treatment group were given with Jiangzhuo mixture 50 g/kg by gavage. After 8 weeks of intervention, the blood glucose (GLU), total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) levels were measured in the three groups. Quantitative reverse transcription PCR were used to detect the expression levels of PR domain containing 16 (PRDM16) and uncoupling protein 1 (UCP1) in white and brown adipose tissues of the rats in each group; Western blotting was used to detect the expression of PRDM16 in the white and brown adipose tissue of rats, and Toll-like receptor 4 (TLR4), nuclear factor-κB (NF-κB) and inhibitor of NF-κB alpha (IκBα) in the white adipose tissue; immunohistochemistry was used to detect the expression of UCP1 protein in white and brown adipose tissues.@*RESULTS@#Compared with the normal control group, the white fat weight (P<0.01), white fat coefficient (P<0.05) and Lee's coefficient (P<0.01) were significantly increased in the model control group; the contents of GLU, TC, TG and LDL-C were all increased, and the content of TG was significantly increased (P<0.05) in the model control group. The mRNA and protein expression levels of PRDM16 and UCP1 in white fat and brown fat were significantly decreased (P<0.05) in the model control group. Compared with the model control group, the white fat weight and white fat coefficient and Lee's coefficient were significantly reduced in the Jiangzhuo mixture treatment group (all P<0.01), the levels of GLU, TC, TG, and LDL-C in the the treatment group were all reduced, and the content of TG was reduced more obviously (P<0.01); expression levels of PRDM16 and UCP1 mRNA and protein were increased in brown and white adipose tissue. Compared with the normal control group, the expression levels of TLR4, phospho-IκBα and NF-κB-p65 proteins in white adipose tissue of the model control group were significantly increased (all P<0.01), while the expression levels of these proteins in the treatment group were significantly lower than those in the model control group (all P<0.05).@*CONCLUSIONS@#Jiangzhuo mixture can alleviate high-fat diet-induced increase in body fat, abnormal expression of biochemical indexes and promote the expression of key proteins including UCP1 and PRDM16 in white and brown adipose tissues by regulating TLR4/IκBα/NF-κB signaling pathway.

The role of LncRNA00602 in Ad36-induced differentiation of adipocytes / 中华内分泌代谢杂志

Objective:To investigate the possible role of long non-coding RNA (LncRNA) 00602 in promoting browning in adipocytes induced by adenovirus type 36 (Ad36).Methods:According to Ad36 infection, adipose tissue samples of obese patients were divided into Ad36-negative group and Ad36-infected group. Realtime fluorescent quantitative PCR (qRT-PCR) was used to detect the changes in the expression of LncRNA00602 mRNA in omental adipose tissue of the two groups, and analyze the differences between the two groups. The correlation between waist-to-hip ratio, systolic blood pressure, diastolic blood pressure, fasting blood glucose, triacylglyceride and other indicators of the patients in the group with LncRNA00602 mRNA expression were analyzed. HE staining was used to detect the size of adipocytes in the omental adipose tissue of the Ad36 negative group and the Ad36 infection group. qRT-PCR and Western blotting were used to detect the mRNA and protein expression levels of uncoupling protein 1 (UCP1) and PR domain containing 16 (PRDM16) in omental adipose tissue of two groups of patients. Human adipose-derived stem cells (hADSC) were isolated and cultured, using Ad36 to induce differentiation, and divided into control group and LncRNA00602 knockdown group. On 0, 2, and 4 days after LncRNA00602 knockdown, fluoroboron dipyrrole (BODIPY) and mitochondrial red fluorescence (Mito-Tracker Red) were used to stain intracellular lipid droplets and mitochondria. At the same time, qRT-PCR and Western blotting were used to detect changes in the expression of UCP1 and PRDM16.Results:The expression of LncRNA00602 gene in the Ad36 infection group was higher than that in the Ad36 negative group (all P<0.05). The expression of LncRNA00602 in the Ad36 negative group was not significantly different from the above clinical indicators, while the expression of LncRNA00602 was negatively correlated with serum fasting blood glucose and triacylglyceride ( r=-0.522, -0.486, P<0.05) in the Ad36 infection group; HE staining showed that the average adipocyte area of the Ad36 infection group was smaller than that of the Ad36 negative group. At the same time, UCP1 and PRDM16 gene expression were higher than the negative group (all P<0.05). At the cellular level, on the 2nd and 4th days after knockdown of LncRNA00602, the lipid droplet area of adipocytes in the LncRNA00602 knockdown group was larger than that of the control group, the number of mitochondria decreased compared with the control group, and difference was statistically significant ( P<0.05 or P<0.01); Compared with the control group, there was significantly lower expression of the browning marker genes UCP1, PRDM16, and protein in the adipocytes in the LncRNA00602 knockdown group (all P<0.05). Conclusion:In Ad36-induced adipocyte differentiation, LncRNA00602 may positively regulate the expression of UCP1, PRDM16 and lipid droplet metabolism, and promote the browning of adipocytes.

Effects of N-Oleoylglycine and Oleate on Mitochondrial UCP1-independent Thermogenesis / 中国生物化学与分子生物学报

Besides UCP1-dependent thermogenesis pathways, UCP1-independent thermogenesis pathways also could increase heat production in adipose tissue to combat obesity. N-Acyl amino acids (NAAs) have been suggested as novel endogenous uncouplers to induce mitochondria UCP1-independent thermogenesis in adipose tissue. Here, we use mouse skeletal muscle C2C12 cells which lack of UCP1 as UCP1 negative cell models. Comparing with its corresponding common fatty acid—oleate, one of the NAAs—N-Oleoylglycine (NOGly), which is highly expressed in the plasma of HFD mice, is selected to study their effects and mechanisms on mitochondrial thermogenesis. We found that 60 μmol / L oleate could induce mitochondrial oxidative phosphorylation protein levels, as well as increase mitochondria thermogenesis-related genes (COX8b, DIO2, UCP3) expression (P < 0. 05) . However, 60 μmol / L NOGly damaged the production and oxidative phosphorylation of mitochondria, significantly down-regulated expression of thermogenic genes (PGC1a, COX8b, COX2, DIO2, UQCRFS1and UCP3) (P< 0. 01), induced the production of reactive oxygen species (ROS) in the mitochondria, and enhanced the oxidative stress in cells. Our study found that oleate can induce UCP1-independent thermogenesis under 60 μmol / L addition dose, whereas NOGly does not due to the induction of oxidative stress in cells.

Effect of uncoupling protein 2 gene on radiation sensitivity of Siha cells / 中华放射医学与防护杂志

Objective@#To investigate whether silencing UCP2 can sensitize cervical cancer cell line Siha to radiation.@*Methods@#Siha cells were transfected with UCP2 siRNA and then irradiated by X-ray. The radiosensitivity of Siha cells was verified by colony formation, CCK-8, apoptosis and immunofluorescence assays. The mitochondrial membrane potential and the production of reactive oxygen species (ROS) were detected to further explore the related mechanism.@*Results@#RT-PCR and Western blot assays showed that the expression of UCP2 in Siha cells was increased after irradiation and the UCP2 siRNA successfully silenced the expression of UCP in cells. According to the survival curves, the D0, Dq, N and SF2 were 1.54, 1.31, 2.31 Gy and 0.52 for siUCP2 group, 2.50, 3.64, 4.30 Gy and 0.83 for blank control group, and 3.34, 2.16, 1.91 Gy and 0.69, for siNC group, respectively. The radiosensitivity enhancement ratio of silent group was 0.62 and 0.46, compared with blank control group and negative control group, respectively. The proliferative activity of cells in the silent group was lower than that in the control group (t=13.2, P<0.05). Apoptosis levels in the silent group were significantly higher than those in the control group after irradiation(t=3.14, P<0.05). At 4 h after irradiation, the ROS production in the silent group was significantly higher than that in the control group (t=19.10, P<0.05). At 24 h after irradiation, the mitochondrial membrane potential of Siha cells in the silent group was significantly lower than that in the control group (t=4.18, P<0.05).@*Conclusions@#The radiosensitivity of Siha cells is enhanced after UCP2 silencing, and thus UCP2 may applicable as a new target for radiosensitization of cervical cancer cells.

Effect of Yinfupian and Yangfupian on Brown Adipose Tissue of Yang Deficiency Mice / 中国实验方剂学杂志

Objective:To observe the expression of brown adipose tissue (BAT), cells, proteins and corresponding genes in Yang deficiency model mice induced by Rhei Radix et Rhizoma suspension, and to explore the thermogenesis of processed products of Aconiti Lateralis Radix Praeparata with Jianchang faction characteristics. Method:Twenty mice, half male and half female, were randomly selected as the normal female and male groups. And the other 80 mice were administrated with Rhei Radix et Rhizoma suspension (the content of 0.25 g·mL-1) to establish Yang deficiency model, after the model was established, they were randomly divided into the model female and male groups, female and male groups of Shengfupian, female and male groups of Yinfupian, female and male groups of Yangfupian, 10 mice in each group. Mice were intragastric administrated with corresponding medical solution for two weeks (1.54 g·kg-1·d-1) according to groups. Normal group and model group were given equal volume distilled water. After administration, BAT of scapular region of mice was collected and the changes of BAT cells were observed by hematoxylin-eosin (HE) staining. The expression of uncoupling protein 1 (UCP1) and its mRNA were detected by Western blot and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). Result:Compared with the normal group of the same sex, the proportion of BAT in the model group decreased significantly (P<0.01). Compared with the model group of the same sex, the proportion of BAT in female mice from Shengfupian and Yinfupian groups increased significantly (P<0.01), while there was no significant difference between each administration group and model group in the male mice. Compared with normal mice of the same sex, there were many scattered vacuoles in BAT cells of the model group, and fewer cells could be observed due to larger vacuoles. Compared with the model group of the same sex, BAT cells in mice from the Shengfupian group showed fewer vacuoles, smaller cells and tight arrangement, the density of BAT cells in mice from the Yangfupian group also increased significantly, while the vacuoles in BAT cells of mice from the Yinfupian group decreased relatively and the cells did not increase significantly. Compared with the same sex mice, the expression level of UCP1 in the model group and the normal group was statistically significant (P<0.05, P<0.01). In the female mice, the expression level of UCP1 in Yangfupian group was significantly higher than that in the model group (P<0.05), each administration group of male mice was significantly different from that of the model group of the same sex (P<0.05), of which Yangfupian was the most significant. The relative expression of UCP1 mRNA in the model group was significantly lower than that in the normal group of the same sex (P<0.05, P<0.01). In the female mice, compared with the model group, the relative expression levels of UCP1 mRNA in Yangfupian group, Shengfupian group and Yinfupian group increased significantly (P<0.05, P<0.01), compared with Yangfupian group, the relative expression levels of UCP1 mRNA in Shengfupian and Yinfupian were also significantly different (P<0.05). In the male mice, compared with the model group, the relative expression of UCP1 mRNA in Yangfupian group was significantly increased (P<0.01), but there was no significant difference in Shengfupian group and Yinfupian group, in addition, compared with Yangfupian group, the relative expression of UCP1 mRNA in Shengfupian group and Yinfupian group had significant difference (P<0.05). Conclusion:Shengfupian, Yinfupian and Yangfupian all have obvious improvement on Yang deficiency syndrome induced by Rhei Radix et Rhizoma suspension. The mechanism may be to promote the expression of UCP1 protein and its mRNA and enhance the activity of BAT. And the effect of Yangfupian is the best.

Effect of uncoupling protein 2 gene on radiation sensitivity of Siha cells / 中华放射医学与防护杂志

Objective To investigate whether silencing UCP2 can sensitize cervical cancer cell line Siha to radiation.Methods Siha cells were transfected with UCP2 siRNA and then irradiated by Xray.The radiosensitivity of Siha cells was verified by colony formation,CCK-8,apoptosis and immunofluorescence assays.The mitochondrial membrane potential and the production of reactive oxygen species (ROS) were detected to further explore the related mechanism.Results RT-PCR and Western blot assays showed that the expression of UCP2 in Siha cells was increased after irradiation and the UCP2 siRNA successfully silenced the expression of UCP in cells.According to the survival curves,the D0,Dq,N and SF2 were 1.54,1.31,2.31 Gy and 0.52 for siUCP2 group,2.50,3.64,4.30 Gy and 0.83 for blank control group,and 3.34,2.16,1.91 Gy and 0.69,for siNC group,respectively.The radiosensitivity enhancement ratio of silent group was 0.62 and 0.46,compared with blank control group and negative control group,respectively.The proliferative activity of cells in the silent group was lower than that in the control group (t =13.2,P＜0.05).Apoptosis levels in the silent group were significantly higher than those in the control group after irradiation (t=3.14,P＜0.05).At 4 h after irradiation,the ROS production in the silent group was significantly higher than that in the control group (t=19.10,P＜0.05).At 24 h after irradiation,the mitochondrial membrane potential of Siha cells in the silent group was significantly lower than that in the control group (t =4.18,P ＜ 0.05) Conclusions The radiosensitivity of Siha cells is enhanced after UCP2 silencing,and thus UCP2 may applicable as a new target for radiosensitization of cervical cancer cells.

Effect of exercise-induced irisin on myocardial fibrosis / 中国组织工程研究

BACKGROUND: Myocardial fibrosis is an important topic in modern medical research, and its development is closely related to common heart diseases such as arrhythmia and chronic heart failure. Exercise intervention can significantly improve myocardial fibrosis, but there is no systematic and comprehensive understanding of the mechanism by which exercise improves myocardial fibrosis as well as the effects of different types of exercises on myocardial fibrosis. To date, it is still unclear about how exercise triggers the production of irisin against myocardial fibrosis. OBJECTIVE: To comprehensively review the exercise-induced production of irisin and its effect on myocardial fibrosis, and reveal its myocardial protection, so as to improve heart function and provide fundamental basis for preventing against common heart diseases, such as arrhythmia and chronic heart failure. METHODS: A search of ELSEVIER, Web of Science, CNKI, WanFanga, VIP and Taiwan Academic Literature Database was performed for articles regarding exercise, irisin, and myocardial fibrosis. The deadline for publication was August 2019. According to the inclusion and exclusion criteria, 58 articles were eligible for review. RESULTS AND CONCLUSION: Long-term and single exercise in human experiments has been shown to improve muscular and circulatory irisin levels, which has been better verified in animal experiments. A few experimental results indicate that long-term exercise has no significant effect on blood irisin levels, which may be due to different research subjects, exercise methods, exercise intensity, and exercise frequency. However, the specific mechanism is still unclear. Exercise can improve myocardial fibrosis by acting on myocardial mitochondrial stabilization, energy metabolism, oxidative stress and inflammatory response. The occurrence of myocardial fibrosis results from the regulation of neuroendocrine and oxidative stress and inflammatory responses. Irisin can influence the processes of oxidative stress and inflammation related to the mechanism of myocardial fibrosis, by inhibiting ROS/p38MAPK/NF-κB signaling pathway, endogenous reactive oxygen species and ROS-NLRP3 inflammation signaling pathway, and regulating the expression of uncoupling protein 2 and mitochondrial homeostasis. Therefore, exercise may improve myocardial fibrosis by upregulating the expression of irisin, thus providing myocardial protection.

Anti-Hypertensive Action of Fenofibrate via UCP2 Upregulation Mediated by PPAR Activation in Baroreflex Afferent Pathway / 神经科学通报·英文版

Fenofibrate, an agonist for peroxisome proliferator-activated receptor alpha (PPAR-α), lowers blood pressure, but whether this action is mediated via baroreflex afferents has not been elucidated. In this study, the distribution of PPAR-α and PPAR-γ was assessed in the nodose ganglion (NG) and the nucleus of the solitary tract (NTS). Hypertension induced by drinking high fructose (HFD) was reduced, along with complete restoration of impaired baroreceptor sensitivity, by chronic treatment with fenofibrate. The molecular data also showed that both PPAR-α and PPAR-γ were dramatically up-regulated in the NG and NTS of the HFD group. Expression of the downstream signaling molecule of PPAR-α, the mitochondrial uncoupling protein 2 (UCP2), was up-regulated in the baroreflex afferent pathway under similar experimental conditions, along with amelioration of reduced superoxide dismutase activity and increased superoxide in HFD rats. These results suggest that chronic treatment with fenofibrate plays a crucial role in the neural control of blood pressure by improving baroreflex afferent function due at least partially to PPAR-mediated up-regulation of UCP2 expression and reduction of oxidative stress.

Effects of uncoupling protein 2 overexpression on myocardial mitochondrial dynamics in sepsis rats / 中华危重病急救医学

Objective@#To investigate the effects of uncoupling protein 2 (UCP2) overexpression on mitochondrial dynamics (mitochondrial division and fusion) of sepsis myocardial injury in rats.@*Methods@#Forty male Sprague-Dawley (SD) rats were randomly divided into four groups (n = 10): sham operation group (Sham group) using normal saline instead of transfection and simulating cecal ligation and perforation (CLP); CLP group using normal saline instead of transfection, performing CLP to induce sepsis; adeno-associated virus (AAV) group using CLP after myocardial transfection with empty virus; UCP2 overexpression group (UCP2 group) CLP was performed 3 weeks after AAV-UCP2 (1×1015 vg/L, a total of 60 μL) myocardial transfection. The rats in each group were examined by echocardiography at 24 hours after the CLP, and then the rats were sacrificed immediately to harvest myocardial tissue. Myocardial ultrastructural changes were observed under the electron microscope, the expression of regulatory proteins related to myocardial mitochondrial dynamics [optic atrophy 1 (Opa1), dynamin-related protein 1 (Drp1) and fission 1 (Fis1)] were detected by Western Blot, and the level of mitochondrial adenosine triphosphate (ATP) production was detected by chemiluminescence.@*Results@#①The echocardiographic results showed that there was no significant difference in left ventricular mass (LVM) and stroke volume (SV). Compared with Sham group, left ventricular diastolic anterior wall thickness (LVAWd), left ventricular systolic anterior wall thickness (LVAWs), left ventricular diastolic posterior wall thickness (LVPWd), left ventricular systolic posterior wall thickness (LVPWs), left ventricular ejection fraction (LVEF) and left ventricular short axis shortening rate (LVFS) were significantly increased in CLP group and AAV group, while left ventricular systolic diameter (LVEDs), left ventricular diastolic diameter (LVEDd), left ventricular end-systolic volume (LVESV), and left ventricular end-diastolic volume (LVEDV) were significantly decreased. Compared with CLP group and AAV group, LVAWs, LVEF, LVFS were significantly decreased in UCP2 group, and LVEDs, LVEDV and LVESV were significantly increased [LVAWs (mm): 3.82±0.42 vs. 4.34±0.30, 4.44±0.12; LVEF: 0.921±0.038 vs. 0.979±0.019, 0.991±0.010; LVFS: (65.33±6.56)% vs. (80.11±8.23)%, (85.31±6.11)%; LVEDs (mm): 1.81±0.36 vs. 0.89±0.54, 0.60±0.17; LVEDV (μL): 137.09±50.05 vs. 89.72±53.04, 85.42±40.99; LVESV (μL): 10.48±4.59 vs. 2.48±3.52, 2.58±2.50, all P < 0.05]. ② Electron microscope showed that the structure of myocardial fibers in the Sham group was clear and aligned with complete intervertebral disc and mitochondrial structure, no damage to mitochondrial membranes, and tight arrangement of cristae. In CLP group and AAV group, muscle fiber breakage, sarcoplasmic reticulum expansion, severe mitochondrial swelling and even cristage structure disorder were observed. In the UCP2 group, only myocardial fiber edema was observed, and the muscle fiber structure was more complete than that of Sham group and AAV group. The mitochondria were slightly swollen and the cristae were intact.③ Western Blot showed that there was no significant difference in the expression of Opa1 and Fis1 in the four groups. The expression of Drp1 in CLP group and AAV group were significantly higher than that in Sham group. The expression of Drp1 in UCP2 group was significantly lower than that in CLP group and AAV group (Drp1/β-actin: 1.01±0.03 vs. 1.39±0.03, 1.49±0.03, both P < 0.05).④ The results of immunofluorescence showed that the ATP content of CLP group and AAV group were significantly lower than that of Sham group; the ATP content of UCP2 group was significantly higher than that of CLP group and AAV group (μmol/L: 1.99±0.15 vs. 1.10±0.17, 1.13±0.19, both P < 0.05).@*Conclusion@#UCP2 overexpression can significantly improve the systemic systolic function of myocardium in sepsis rats, protect myocardial mitochondrial ultrastructure, inhibit mitochondrial division, and improve mitochondrial ATP synthesis.

Effects of uncoupling protein 2 overexpression on myocardial mitochondrial dynamics in sepsis rats / 中华危重病急救医学

Objective To investigate the effects of uncoupling protein 2 (UCP2) overexpression on mitochondrial dynamics (mitochondrial division and fusion) of sepsis myocardial injury in rats. Methods Forty male Sprague-Dawley (SD) rats were randomly divided into four groups (n = 10): sham operation group (Sham group) using normal saline instead of transfection and simulating cecal ligation and perforation (CLP); CLP group using normal saline instead of transfection, performing CLP to induce sepsis; adeno-associated virus (AAV) group using CLP after myocardial transfection with empty virus; UCP2 overexpression group (UCP2 group) CLP was performed 3 weeks after AAV-UCP2 (1×1015 vg/L, a total of 60 μL) myocardial transfection. The rats in each group were examined by echocardiography at 24 hours after the CLP, and then the rats were sacrificed immediately to harvest myocardial tissue. Myocardial ultrastructural changes were observed under the electron microscope, the expression of regulatory proteins related to myocardial mitochondrial dynamics [optic atrophy 1 (Opa1), dynamin-related protein 1 (Drp1) and fission 1 (Fis1)] were detected by Western Blot, and the level of mitochondrial adenosine triphosphate (ATP) production was detected by chemiluminescence. Results ①The echocardiographic results showed that there was no significant difference in left ventricular mass (LVM) and stroke volume (SV). Compared with Sham group, left ventricular diastolic anterior wall thickness (LVAWd), left ventricular systolic anterior wall thickness (LVAWs), left ventricular diastolic posterior wall thickness (LVPWd), left ventricular systolic posterior wall thickness (LVPWs), left ventricular ejection fraction (LVEF) and left ventricular short axis shortening rate (LVFS) were significantly increased in CLP group and AAV group, while left ventricular systolic diameter (LVEDs), left ventricular diastolic diameter (LVEDd), left ventricular end-systolic volume (LVESV), and left ventricular end-diastolic volume (LVEDV) were significantly decreased. Compared with CLP group and AAV group, LVAWs, LVEF, LVFS were significantly decreased in UCP2 group, and LVEDs, LVEDV and LVESV were significantly increased [LVAWs (mm): 3.82±0.42 vs. 4.34±0.30, 4.44±0.12;LVEF: 0.921±0.038 vs. 0.979±0.019, 0.991±0.010; LVFS: (65.33±6.56)% vs. (80.11±8.23)%, (85.31±6.11)%;LVEDs (mm): 1.81±0.36 vs. 0.89±0.54, 0.60±0.17; LVEDV (μL): 137.09±50.05 vs. 89.72±53.04, 85.42±40.99;LVESV (μL): 10.48±4.59 vs. 2.48±3.52, 2.58±2.50, all P < 0.05]. ② Electron microscope showed that the structure of myocardial fibers in the Sham group was clear and aligned with complete intervertebral disc and mitochondrial structure, no damage to mitochondrial membranes, and tight arrangement of cristae. In CLP group and AAV group, muscle fiber breakage, sarcoplasmic reticulum expansion, severe mitochondrial swelling and even cristage structure disorder were observed. In the UCP2 group, only myocardial fiber edema was observed, and the muscle fiber structure was more complete than that of Sham group and AAV group. The mitochondria were slightly swollen and the cristae were intact.③ Western Blot showed that there was no significant difference in the expression of Opa1 and Fis1 in the four groups. The expression of Drp1 in CLP group and AAV group were significantly higher than that in Sham group. The expression of Drp1 in UCP2 group was significantly lower than that in CLP group and AAV group (Drp1/β-actin:1.01±0.03 vs. 1.39±0.03, 1.49±0.03, both P < 0.05).④ The results of immunofluorescence showed that the ATP content of CLP group and AAV group were significantly lower than that of Sham group; the ATP content of UCP2 group was significantly higher than that of CLP group and AAV group (μmol/L: 1.99±0.15 vs. 1.10±0.17, 1.13±0.19, both P < 0.05). Conclusion UCP2 overexpression can significantly improve the systemic systolic function of myocardium in sepsis rats, protect myocardial mitochondrial ultrastructure, inhibit mitochondrial division, and improve mitochondrial ATP synthesis.

Effects of uncoupling protein 2 overexpression on myocardial mitochondrial dynamics in sepsis rats / 中华危重病急救医学

Objective To investigate the effects of uncoupling protein 2 (UCP2) overexpression on mitochondrial dynamics (mitochondrial division and fusion) of sepsis myocardial injury in rats. Methods Forty male Sprague-Dawley (SD) rats were randomly divided into four groups (n = 10): sham operation group (Sham group) using normal saline instead of transfection and simulating cecal ligation and perforation (CLP); CLP group using normal saline instead of transfection, performing CLP to induce sepsis; adeno-associated virus (AAV) group using CLP after myocardial transfection with empty virus; UCP2 overexpression group (UCP2 group) CLP was performed 3 weeks after AAV-UCP2 (1×1015 vg/L, a total of 60 μL) myocardial transfection. The rats in each group were examined by echocardiography at 24 hours after the CLP, and then the rats were sacrificed immediately to harvest myocardial tissue. Myocardial ultrastructural changes were observed under the electron microscope, the expression of regulatory proteins related to myocardial mitochondrial dynamics [optic atrophy 1 (Opa1), dynamin-related protein 1 (Drp1) and fission 1 (Fis1)] were detected by Western Blot, and the level of mitochondrial adenosine triphosphate (ATP) production was detected by chemiluminescence. Results ①The echocardiographic results showed that there was no significant difference in left ventricular mass (LVM) and stroke volume (SV). Compared with Sham group, left ventricular diastolic anterior wall thickness (LVAWd), left ventricular systolic anterior wall thickness (LVAWs), left ventricular diastolic posterior wall thickness (LVPWd), left ventricular systolic posterior wall thickness (LVPWs), left ventricular ejection fraction (LVEF) and left ventricular short axis shortening rate (LVFS) were significantly increased in CLP group and AAV group, while left ventricular systolic diameter (LVEDs), left ventricular diastolic diameter (LVEDd), left ventricular end-systolic volume (LVESV), and left ventricular end-diastolic volume (LVEDV) were significantly decreased. Compared with CLP group and AAV group, LVAWs, LVEF, LVFS were significantly decreased in UCP2 group, and LVEDs, LVEDV and LVESV were significantly increased [LVAWs (mm): 3.82±0.42 vs. 4.34±0.30, 4.44±0.12;LVEF: 0.921±0.038 vs. 0.979±0.019, 0.991±0.010; LVFS: (65.33±6.56)% vs. (80.11±8.23)%, (85.31±6.11)%;LVEDs (mm): 1.81±0.36 vs. 0.89±0.54, 0.60±0.17; LVEDV (μL): 137.09±50.05 vs. 89.72±53.04, 85.42±40.99;LVESV (μL): 10.48±4.59 vs. 2.48±3.52, 2.58±2.50, all P < 0.05]. ② Electron microscope showed that the structure of myocardial fibers in the Sham group was clear and aligned with complete intervertebral disc and mitochondrial structure, no damage to mitochondrial membranes, and tight arrangement of cristae. In CLP group and AAV group, muscle fiber breakage, sarcoplasmic reticulum expansion, severe mitochondrial swelling and even cristage structure disorder were observed. In the UCP2 group, only myocardial fiber edema was observed, and the muscle fiber structure was more complete than that of Sham group and AAV group. The mitochondria were slightly swollen and the cristae were intact.③ Western Blot showed that there was no significant difference in the expression of Opa1 and Fis1 in the four groups. The expression of Drp1 in CLP group and AAV group were significantly higher than that in Sham group. The expression of Drp1 in UCP2 group was significantly lower than that in CLP group and AAV group (Drp1/β-actin:1.01±0.03 vs. 1.39±0.03, 1.49±0.03, both P < 0.05).④ The results of immunofluorescence showed that the ATP content of CLP group and AAV group were significantly lower than that of Sham group; the ATP content of UCP2 group was significantly higher than that of CLP group and AAV group (μmol/L: 1.99±0.15 vs. 1.10±0.17, 1.13±0.19, both P < 0.05). Conclusion UCP2 overexpression can significantly improve the systemic systolic function of myocardium in sepsis rats, protect myocardial mitochondrial ultrastructure, inhibit mitochondrial division, and improve mitochondrial ATP synthesis.

Mechanism of Baoyuan Jiedu Decoction in Alleviating Muscle Atrophy in Apcmin/+ Cachexia Mice / 中国实验方剂学杂志

Objective: To study the effect of Baoyuan Jiedu decoction on serum interleukin-6 (IL-6) content, expression of muscle atrophy F-box 1(Atrogin-1), muscle ring finger-1 (MuRF-1), uncoupling proteins-2 (UCP-2), uncoupling proteins-3 (UCP-3) in Apcmin/+ mice, in order to explore the mechanism in improving muscle atrophy in cancer cachexia model. Method: The 14-week-old Apcmin/+ cachexia mice model was randomly divided into model group, Baoyuan Jiedu decoction group (23 g·kg-1) and megestrol group (0.024 g·kg-1). C57BL/6J mice were normal group, with 10 mice in each group, and given continuous intragastric administration for 12 weeks. The quality of gastrocnemius muscle and the transverse diameter of muscle fibers were measured. The content of IL-6 in serum of Apcmin/+ cachexia mice was detected by enzyme-linked immunosorbent assay (ELISA). The expressions of Atrogin-1, MuRF-1, UCP-2, UCP-3 mRNA and protein in gastrocnemius muscle were detected by Western blot and quantitative real-time fluorescence polymerase chain reaction (Real-time PCR). Result: Compared with the normal group, the weight of gastrocnemius muscle and the transverse diameter of fibers in the model group decreased significantly (PPPPConclusion: Reduction of the concentration of IL-6 in serum and the down-regulation of the expressions of Atrogin-1, MuRF-1, UCP-2 and UCP-3 genes may be the possible mechanism of Baoyuan Jiedu decoction in alleviating muscle atrophy in Apcmin/+ cachexia mice model.

The effects of oxidative stress induced by occlusal interference and the regulatory mechanism of UCP3 on masseter in rats / 实用口腔医学杂志

The occlusal interference model in rats was established. In the masseter the level of MDA was increased, but both SOD and GPX were decreased in the presence of the occlusal interference, while the expression of UCP3 was higher than that in the control group (P ＜ 0. 05) .The findings indicate that the occlusal interference can induce the oxidative damage of masseter in rats, which has a correlation with UCP3.

Expression of peroxisome proliferator-activated receptor γ and uncoupling protein-1 after sleeve gastrectomy in obese diabetic rat / 中华内分泌外科杂志

Objective To investigate the expression of peroxisome proliferator-activated receptor γ (PPARγ) and uncoupling protein-1 (UCP-1) after sleeve gastrectomy in Zucker rats and to discuss the weight loss mechanisms.Metbods 30 male Zucker rats aged 10 weeks were randomly divided into 3 groups:the operation group (10 rats),the sham operation group(10 rats) and the diet-pairing group (10 rats).The rats were decapitated to retrieve the retroperitoneal adipose.mRNA and protein expressions of PPARγ and UCP-1 were detected by RT-PCR and Western blot.Results As for the operation group,the weight decreased significantly after the operation compared to the other two groups((250±5.8) g,(370±10.0) g,(310±9.6) g,respectively,P＜0.05).The expressions of PPARγ and UCP-1 gene of mRNA and protein were all significantly higher in the operation group (P＜0.05).Conclusions SG can up-regulate the expressions of thermogenic gene PPARγand UCP-1 in adipose in Zucker rats,browning the white adipose tissue,which was one of the important mechanisms of weight loss.

Down-regulated uncoupling protein 2 expression suppresses Fadu cell proliferation / 解剖学报

Objective The effect of uncoupling protein 2(UCP2) on the Fadu cells of the hypopharyngeal cancer was investigated in this study. The siRNA technology was used to study the effect of UCP2 on cell cycle and cell proliferation. iMeanwhile, cisplatin, a commonly chemotherapeutic drug, was researched on the effect of proliferation in Fadu cells after down-regulated UCP2 gene. Methods Flow cytometry/PI staining was used to observe the effect of UCP2 on Fadu cell cycle. Clone formation assay was used to detect the effect of down-regulated UCP2 on the proliferation of Fadu cells. MTT assay and Bnid assay were used to determine whether UCP2 enhanced the sensitivity of Fadu cells to cisplatin and to detect the effect of UCP2 on the proliferation of Fadu. Results The present result showed that down-regulated UCP2 expression inhibited Fadu cell cycle in G,/S phase and cell colony formation, but had no synergistic effect on inhibition of Fadu proliferation by cisplatin; The expression of UCP2 was significantly inhibited by genipin, and then inhibited cell proliferation in a time-concentration-dependent manner. Conclusion The proliferation of Fadu cells was weakened and the cell cycle was inhibited in G,/S phase after down-regulated UCP2, suggesting that UCP2 may play an important adjust role in the development in Fadu of hypopharyngeal cancer.

Change of gene expressions in perirenal adipose tissues of overweight and obese subjects / 中华内分泌代谢杂志

Objective@#To determine the change of gene expressions in human perirenal adipose tissue (PAT) and oblique abdomen subcutaneous adipose tissue (SAT) of overweight and obese subjects.@*Methods@#Ninety-seven patients, including 35 overweight/obese patients and 62 non-obese patients, who underwent renal surgery were included. The clinical data and various gene expressions in PAT and SAT of two groups were analyzed.@*Results@#Body mass index, waist circumference, systolic blood pressure, resting heart rate, fasting blood glucose, and serum creatinine were significantly higher in overweight/obese patients than those in non-obese patients(P<0.05 or P<0.01). Compared with non-obese patients, PAT adipocytes showed bigger, and the expressions of uncoupling protein-1 (UCP1) mRNA and protein were markedly lower in overweight/obese patients. The mRNA expressions of CIDEA and adiponectin in PAT of overweight/obese patients were significantly lower, while leptin, monocyte chemotactic protein 1, interleukin (IL)-6, and IL-10 mRNA expressions were significantly higher. There were no significant differences in the related gene expressions of SAT between the two groups.@*Conclusions@#Different parts of adipose tissue reveal various characteristics, and play different roles in the occurrence and development of obesity. UCP1 expression is decreased in PAT of overweight/obese patients, with the changes of adipocytokine expressions. (Chin J Endocrinol Metab, 2018, 34: 567-572)

Correlation between circulating uncoupling protein 2 level and severity of coronary artery lesions in patients with stable coronary artery disease / 实用医学杂志

Objective To investigate the correlation between circulating uncoupling protein 2(UCP2) level and severity of coronary artery disease(including Gensini score and criminal vessel counts)in patients with stable coronary artery disease(SCAD),and to analyze the predictive value of circulating UCP2 and urine acid (UA)for SCAD. Methods Three hundred and thirty patients from June 2015 to June 2017 were enrolled. Two hundred and forty patients with SCAD(SCAD group),90 patients without coronary artery disease(control group) were diagnosed. The circulating UCP2 level was detected by enzyme linked immunosorbent assay (ELISA) sandwich method. Results The levels of circulating UCP2 and UA in SCAD group were higher than those in the control group(UCP2[1.60(0.67,4.60)ng/mL]vs.[0.42(0.28,0.59)ng/mL](P<0.01),UA[(365.74 ± 66.06) μmol/L] vs. [(268.11 ± 45.81)μmol/L],P < 0.01). Multivariate logistic regression analysis showed that UCP2 (OR = 1.010 ,95% CI :1.001 ~ 1.020 ,P = 0.025)and UA(OR = 1.039 ,95% CI :1.007 ~ 1.072 ,P < 0.05)were independently associated with SCAD. Correlation analysis showed that the circulating UCP2 level was positively correlated with Gensini score(r=0.780,P<0.01)and criminal vessel counts(r=0.543,P<0.01). The receiver operating characteristic curve(ROC)showed that the optimal cutoff point of the circulating UCP2 level predicting SCAD was 0.64 ng/mL,and the sensitivity was 0.833 and the specificity was 0.944. No significant difference was observed in area under the curve between circulating UCP2 and UA(ΔAUC). Conclusion The high circulating UCP2 level indicates more severe coronary lesions in patients with SCAD. Circulating UCP2 level may be a new indicator of predicting SCAD,equal to the traditional oxidative stress related indicator of serum UA.

Effect and mechanism of uncoupling protein 2 on cardiomyocyte apoptosis induced by high glucose, high lipid and high uric acid / 解放军医学杂志

Objective To investigate the effects of uncoupling protein 2 (UCP2) on the myocardial cells of mice with type 2 diabetes mellitus combined with hyperuricemia (HUA), and clarify the mechanism thereof. Methods The mouse cardiac myocytes (MCM) cultured with 25mmol/L high glucose (HG) medium were divided into two groups: HG plus 300μmol/L sodium palmitate for 18 hours as high glucose and high fat (HG+HF) group, and HG+HF plus 1500μmol/L uric acid (UA) for 18 hours as HG+HF+HUA group. Then the myocardial cells in HG+HF+HUA group, by use or not use UCP2 inhibitor genipin, were further divided into two groups: vehicle group and genipin group. In order to verify the mechanism of UCP2 in myocardial cells injury caused by high glucose, high lipid and high uric acid, the myocardial cells were divided again into genipin group and genipin+N-acetylcysteine (NAC) group. Accordingly, the apoptosis of myocardial cells were measured by flow cytometry at specific time, the mRNA and protein expressions of UCP2 were determined by q-PCR and Western blotting, and the levels of reactive oxygen species (ROS) were detected by DHE staining and ELISA. Results The apoptosis rate of myocardial cells increased obviously, and the expression levels of UCP2 decreased and of ROS elevated significantly in HG+HF+HUA group than in HG+HF group (P<0.05). As the expression levels of UCP2 decreased by genipin intervention, the apoptosis rate of myocardial cells and ROS level in HG+HF+HUA group increased more obviously (P<0.05). In contrast, such an effect was reversed by the application of antioxidants NAC (P<0.05). Conclusion UCP2 can inhibit oxidative stress and alleviate the apoptosis of myocardial cells induced by high glucose, high fat and high uric acid.

Mast cells promote the beiging of white adipose tissue from cold shock / 中华内分泌代谢杂志

This article is the translation of Mast Cells Promote Seasonal White Adipose Beiging in Humans, published in the Diabetes in May, 2017(Diabetes 2017,66:1237-1246), with the consent of Diabetes.This study mainly investigated the mechanism of the beiging of white adipose tissue(WAT).Studies in rodents suggested that cold shock and a number of immune mediators were important for the beiging of subcutaneous white adipose tissue (SC-WAT).This study observed the seasonal beiging of SC-WAT from lean humans, measured the gene expression of various immune cell markers, and performed multivariate analysis of the gene expression data to identify genes that could predict uncoupling protein-1(UCP-1) expression.The mechanism of the induction of UCP-1 from adipocytes by mast cells was also explored by in vitro experiment.The results showed that TIB64 mast cells responded to cold shock by releasing histamine and interleukin 4 (IL-4) which promoted the expression of UCP-1 and the lipolysis of 3T3-L1 adipocytes.Pharmacological block of the degranulation of mast cells potently inhibited the release of histamine and adipocyte UCP-1 mRNA induction by conditioned medium.In summary, mast cells are an important immune cell type in the beiging of WAT which sense colder temperatures, and release factors that promote UCP-1 expression.