Apoptosis manifests in two major execution programs downstream of the death signal: the caspase pathway and organelle dysfunction. An important antiapoptosis factor, Bcl-2 protein, contributes in caspase pathway of ap...Apoptosis manifests in two major execution programs downstream of the death signal: the caspase pathway and organelle dysfunction. An important antiapoptosis factor, Bcl-2 protein, contributes in caspase pathway of apoptosis. Calcium, an important intracellular signal element in cells, is also observed to have changes during apoptosis, which maybe affected by Bcl-2 protein. We have previously reported that in Harringtonine (HT) induced apoptosis of HL-60 cells, there’s a change of intracellular calcium distribution, moving from cytoplast especially Golgi’s apparatus to nucleus and accumulating there with the highest concentration. We report here that caspase-3 becomes activated in HT-induced apoptosis of HL-60 cells, which can be inhibited by overexpression of Bcl-2 protein. No sign of apoptosis or intracellular calcium movement from Golgi’s apparatus to nucleus in HL-60 cells overexpressing Bcl-2 or treated with Ac-DEVD-CHO, a specific inhibitor of caspase-3. The results indicate that activated caspase-3 can promote the movement of intracellular calcium from Golgi’s apparatus to nucleus, and the process is inhibited by Ac-DEVD-CHO (inhibitor of caspase-3), and that Bcl-2 can inhibit the movement and accumulation of intracellular calcium in nucleus through its inhibition on caspase3. Calcium relocalization in apoptosis seems to be irreversible, which is different from the intracellular calcium changes caused by growth factor.展开更多
Geometry optimization at the B3LYP/6-31＋＋G＊ level of theory has been undertaken on clusters containing L-Met （L-methionine） or L-Cys （L-cysteine） surrounded by eight water molecules. The comparison of the struc...Geometry optimization at the B3LYP/6-31＋＋G＊ level of theory has been undertaken on clusters containing L-Met （L-methionine） or L-Cys （L-cysteine） surrounded by eight water molecules. The comparison of the structural parameters of L-Met and L-Cys with X-ray experimental values is in good agreement within 4.8%. This result shows that the privileged positions of water molecules and the possible hydrogen bonding network formed around the backbone of both AAs （amino acids） are adequate. Subsequent calculations of the harmonic vibrational modes followed by a post-processing treatment enable us to assign the vibrational modes of L-Met and L-Cys surrounded explicitly by eight water molecules. The frequencies of the assigned modes are in good agreement with available IR （infra red） and Raman values within 5%.展开更多
We evaluated the cardioprotective effects of ginsenoside Rg1 in a diabetic rat model induced with high-fat diet and intraperitoneal injection of streptozotocin. Ginsenoside Rg1 was injected intraperitoneal y for 12 we...We evaluated the cardioprotective effects of ginsenoside Rg1 in a diabetic rat model induced with high-fat diet and intraperitoneal injection of streptozotocin. Ginsenoside Rg1 was injected intraperitoneal y for 12 weeks. Myocardial injury indices and oxidative stress markers were determined. Changes in cardiac ultrastructure were evaluated with transmission electron microscopy. Myocardial apoptosis was assessed via terminal deoxynucleotidyl transferase （TDT）-mediated DNA nick-end labeling （TUNEL） and immunohistochemistry. Ginsenoside Rg1 was as-sociated with a significant dose-dependent reduction in serum levels of creatinine kinase MB and cardiac troponin I, and lessened ultrastructural disorders in diabetic myocardium, relative to the untreated diabetic model rats. Also, compared with the untreated diabetic rats, significant reductions in serum and myocardial levels of malondialdehyde were noted in the ginsenoside Rg1-treated groups, and increased levels of the antioxidants （superoxide dismutase, catalase, and glutathione peroxidase） were detected. TUNEL staining indicated reduced myocardial apoptosis in ginsenoside Rg1-treated rats, which may be associated with reduced levels of caspase-3 （CASP3） and increased levels of B-cell lymphoma-extra-large （Bcl-xL） in the diabetic myocardium. Ginsenoside Rg1 treatment of diabetic rats was associated with reduced oxidative stress and attenuated myocardial apoptosis, suggesting that ginsenoside Rg1 may be of potential preventative and therapeutic value for cardiovascular injury in diabetic patients.展开更多
Objective To observe the effect of electroacupuncture on the morphological change of the bladder tissue and the protein expression levels of cytochrome C, Caspase-3 and Caspase-9 in the bladder tissue of rats with neu...Objective To observe the effect of electroacupuncture on the morphological change of the bladder tissue and the protein expression levels of cytochrome C, Caspase-3 and Caspase-9 in the bladder tissue of rats with neurogenic bladder after suprasacral spinal cord injury, and to preliminarily explore its partial mechanism of action. Methods Forty-eight female SD rats were randomly divided into blank group（group B）, sham operation group（group S）, model group（group M） and electroacupuncture group（group EA） according to random number table method with 12 rats in each group. Neurogenic bladder models after suprasacral spinal cord injury were established by adopting modified spinal cord transection method. Electroacupuncture intervention was conducted at ＂Cìliáo（次髎 BL 32）＂＂Zhōngjí（中极 CV 3）＂ ＂Sānyīnjiāo（三阴交 SP 6）＂ and ＂Dàzhuī（大椎 GV 14）＂ on the 19 th day after modeling. After treatment for 7 consecutive days, the rats were sacrificed, and the bladder tissues were removed rapidly for microscopic observation of morphological change after HE stain and for determination of the protein expression levels of cytochrome C, Caspase-3 and Caspase-9 via Western blot method. Results① Under light microscope after HE stain, injury in varying degrees in muscle fiber arrangement and the shape and size of cells, etc. of bladder detrusor of rats in group M and group EA was found, and the degree of injury of detrusor in group M was severer than that in group EA;② comparison of the protein expression levels of cytochrome C, Pro-Caspase-9 and activated Caspase-9 between group M as well as group EA and group B as well as group S： the protein expression levels of cytochrome C, Pro-Caspase-9 and activated Caspase-9 in bladder tissue of rats in group M and group EA obviously increased（ P〈0.01, P〈0.05）; comparison between group EA and group M： the protein expression levels of cytochrome C, Pro-Caspase-9 and activated Caspase-9 in bladder tissue of rats in group EA obviously reduced（ P〈0.05）;③ comparison of the protein expression level of Caspase-3 between group M as well as group EA and group B as well as group S： the protein expression level of Pro-Caspase-3（35 KD） in group M and group EA reduced, and the protein expression level of activated Caspase-3（17 KD/19 KD） obviously increased（ P〈0.01, P〈0.05）; comparison between group EA andgroup M： the protein expression level of Pro-Caspase-3（35 KD） in group EA obviously increased, and the protein expression level of activated Caspase-3（17 KD/19 KD） obviously reduced（ P〈0.05）. Conclusion suprasacral spinal cord injury can increase the protein expression levels of cytochrome C, activated Caspase-3 and Caspase-9 in bladder tissues of rats, and electroacupuncture at BL 32, CV 3, SP 6 and GV 14 can down-regulate the protein expression levels of cytochrome C, activated Caspase-3 and Caspase-9 in bladder tissues of rats after suprasacral spinal cord injury and protect bladder tissue, which may be one of the mechanisms that electroacupuncture protects the involved bladder detrusor after suprasacral spinal cord injury.展开更多
Objective:To observe the effects of electroacupuncture(EA)on the protein and gene expressions of Bax,Caspase-3 and Bcl-2 in cerebral cortex of type 2 diabetic rats with cognitive impairment(CI),and to explore the mech...Objective:To observe the effects of electroacupuncture(EA)on the protein and gene expressions of Bax,Caspase-3 and Bcl-2 in cerebral cortex of type 2 diabetic rats with cognitive impairment(CI),and to explore the mechanism of EA in improving the learning and memory abilities.Methods:A total of 100 Sprague-Dawley(SD)rats were divided into a normal group(n=10)and a model group(n=90)by random number table method.Rats in the model group were intraperitoneally injected with a small dose of streptozotocin(STZ)to establish the type 2 diabetic models,after being fed with high-fat and high-sugar diet for 1 month.Twenty CI rats were selected from the 50 successful model rats by the Morris water maze(MWM)test and randomly divided into a model group and an EA group according to the blood glucose level and MWM data(n=10).Rats in the EA group received acupuncture at Zusanli(ST 36),Neiting(ST 44)and Yishu(Extra),of which Zusanli(ST 36)and Neiting(ST 44)were stimulated by EA apparatus,20 min/time,once a day for 6 d a week and 4 consecutive weeks.The rats in the model and the normal groups were fixed without treatment.After 4-week treatment,the random blood glucose level of the rats was measured;the learning and memory abilities of rats were measured by MWM;terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)assay was used to detect apoptotic cells;Western blot(WB)and real-time quantitative polymerase chain reaction(RT-qPCR)were used to detect the protein and gene expressions of Bax,Caspase-3 and Bcl-2 in cerebral cortex.Results:After modeling,the random blood glucose level and the escape latency tested by MWM were significantly increased,and the number of crossing the platform tested by the MWM was decreased in the EA and model groups,and were significantly different from those in the normal group(P<0.05 or P<0.01),while the differences between the model group and the EA group were not statistically significant(all P>0.05).After 4-week treatment,the random glucose level and the escape latency tested by MWM were significantly increased(both P<0.05),and the number of crossing the original platform tested by the MWM was significantly reduced(P<0.01),the protein and gene expressions of Bax and Caspase-3 were significantly increased(all P<0.001),the protein and gene expressions of Bcl-2 were significantly reduced(both P<0.001),and the number of neuron apoptosis was significantly increased(P<0.001)in the model group than in the normal group;the random blood glucose level was significantly reduced(P<0.05),the escape latency tested by MWM was significantly shortened(P<0.05),and the number of crossing the original platform tested by MWM was significantly increased(P<0.05),the protein and gene expressions of Bax and Caspase-3 were significantly reduced(all P<0.001),the protein and gene expressions of Bcl-2 were significantly increased(both P<0.001),and the number of neuron apoptosis was significantly reduced(P<0.001)in the EA group than in the model group.Conclusion:EA can improve the learning and memory damages induced by type 2 diabetic model rats with CI;the action mechanism may be achieved via anti-apoptosis.展开更多
Objective:By observing the effects of electroacupuncture(EA)on the apoptosis of conjunctival cells of rabbits with dry eye syndrome(DES)and the expressions of apoptosis-related proteins Caspase-3,Fas and Bcl-2,to disc...Objective:By observing the effects of electroacupuncture(EA)on the apoptosis of conjunctival cells of rabbits with dry eye syndrome(DES)and the expressions of apoptosis-related proteins Caspase-3,Fas and Bcl-2,to discuss the mechanism of EA in the treatment of DES from the perspective of cell apoptosis.Methods:Male New Zealand rabbits were randomly divided into a normal group(NG),a model group(MG),an EA group(EAG)and a sham EA group(SEAG).DES rabbit model was developed by eye drop of 0.1%benzalkonium chloride.The rabbit tear secretion and tear film break-up time(BUT)were measured;terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)assay was used to detect the apoptosis of conjunctival cells;the expressions of Caspase-3,Fas and Bcl-2 proteins in conjunctival cells were detected by immunohistochemistry.Results:Compared with the NG,the rabbit tear secretion decreased and the BUT was shortened in the MG(both P<0.01);compared with the MG and the SEAG,the rabbit tear secretion increased and the BUT was prolonged in the EAG(all P<0.05).Compared with the NG,the apoptosis of rabbit conjunctival cells increased(P<0.01),the expressions of Caspase-3 and Fas proteins increased(both P<0.05),and the expression of Bcl-2 protein decreased(P<0.01)in the MG;compared with the MG and the SEAG,the apoptosis of rabbit conjunctival cells decreased(both P<0.01),the expressions of Caspase-3 and Fas proteins decreased(all P<0.05),and the expression of Bcl-2 protein increased(both P<0.01)in the EAG.Conclusion:EA can inhibit the apoptosis of rabbit conjunctival cells,down-regulate the expressions of apoptosis-related proteins Caspase-3 and Fas,and up-regulate the expression of Bcl-2 protein,which may be one of the mechanisms of EA in treatment of DES.展开更多
基金the National Natural Science Foundation of China(Grant No. 39730160).
文摘Apoptosis manifests in two major execution programs downstream of the death signal: the caspase pathway and organelle dysfunction. An important antiapoptosis factor, Bcl-2 protein, contributes in caspase pathway of apoptosis. Calcium, an important intracellular signal element in cells, is also observed to have changes during apoptosis, which maybe affected by Bcl-2 protein. We have previously reported that in Harringtonine (HT) induced apoptosis of HL-60 cells, there’s a change of intracellular calcium distribution, moving from cytoplast especially Golgi’s apparatus to nucleus and accumulating there with the highest concentration. We report here that caspase-3 becomes activated in HT-induced apoptosis of HL-60 cells, which can be inhibited by overexpression of Bcl-2 protein. No sign of apoptosis or intracellular calcium movement from Golgi’s apparatus to nucleus in HL-60 cells overexpressing Bcl-2 or treated with Ac-DEVD-CHO, a specific inhibitor of caspase-3. The results indicate that activated caspase-3 can promote the movement of intracellular calcium from Golgi’s apparatus to nucleus, and the process is inhibited by Ac-DEVD-CHO (inhibitor of caspase-3), and that Bcl-2 can inhibit the movement and accumulation of intracellular calcium in nucleus through its inhibition on caspase3. Calcium relocalization in apoptosis seems to be irreversible, which is different from the intracellular calcium changes caused by growth factor.
文摘Geometry optimization at the B3LYP/6-31＋＋G＊ level of theory has been undertaken on clusters containing L-Met （L-methionine） or L-Cys （L-cysteine） surrounded by eight water molecules. The comparison of the structural parameters of L-Met and L-Cys with X-ray experimental values is in good agreement within 4.8%. This result shows that the privileged positions of water molecules and the possible hydrogen bonding network formed around the backbone of both AAs （amino acids） are adequate. Subsequent calculations of the harmonic vibrational modes followed by a post-processing treatment enable us to assign the vibrational modes of L-Met and L-Cys surrounded explicitly by eight water molecules. The frequencies of the assigned modes are in good agreement with available IR （infra red） and Raman values within 5%.
文摘We evaluated the cardioprotective effects of ginsenoside Rg1 in a diabetic rat model induced with high-fat diet and intraperitoneal injection of streptozotocin. Ginsenoside Rg1 was injected intraperitoneal y for 12 weeks. Myocardial injury indices and oxidative stress markers were determined. Changes in cardiac ultrastructure were evaluated with transmission electron microscopy. Myocardial apoptosis was assessed via terminal deoxynucleotidyl transferase （TDT）-mediated DNA nick-end labeling （TUNEL） and immunohistochemistry. Ginsenoside Rg1 was as-sociated with a significant dose-dependent reduction in serum levels of creatinine kinase MB and cardiac troponin I, and lessened ultrastructural disorders in diabetic myocardium, relative to the untreated diabetic model rats. Also, compared with the untreated diabetic rats, significant reductions in serum and myocardial levels of malondialdehyde were noted in the ginsenoside Rg1-treated groups, and increased levels of the antioxidants （superoxide dismutase, catalase, and glutathione peroxidase） were detected. TUNEL staining indicated reduced myocardial apoptosis in ginsenoside Rg1-treated rats, which may be associated with reduced levels of caspase-3 （CASP3） and increased levels of B-cell lymphoma-extra-large （Bcl-xL） in the diabetic myocardium. Ginsenoside Rg1 treatment of diabetic rats was associated with reduced oxidative stress and attenuated myocardial apoptosis, suggesting that ginsenoside Rg1 may be of potential preventative and therapeutic value for cardiovascular injury in diabetic patients.
基金Supported by National Natural Science Foundation of China:81473753
文摘Objective To observe the effect of electroacupuncture on the morphological change of the bladder tissue and the protein expression levels of cytochrome C, Caspase-3 and Caspase-9 in the bladder tissue of rats with neurogenic bladder after suprasacral spinal cord injury, and to preliminarily explore its partial mechanism of action. Methods Forty-eight female SD rats were randomly divided into blank group（group B）, sham operation group（group S）, model group（group M） and electroacupuncture group（group EA） according to random number table method with 12 rats in each group. Neurogenic bladder models after suprasacral spinal cord injury were established by adopting modified spinal cord transection method. Electroacupuncture intervention was conducted at ＂Cìliáo（次髎 BL 32）＂＂Zhōngjí（中极 CV 3）＂ ＂Sānyīnjiāo（三阴交 SP 6）＂ and ＂Dàzhuī（大椎 GV 14）＂ on the 19 th day after modeling. After treatment for 7 consecutive days, the rats were sacrificed, and the bladder tissues were removed rapidly for microscopic observation of morphological change after HE stain and for determination of the protein expression levels of cytochrome C, Caspase-3 and Caspase-9 via Western blot method. Results① Under light microscope after HE stain, injury in varying degrees in muscle fiber arrangement and the shape and size of cells, etc. of bladder detrusor of rats in group M and group EA was found, and the degree of injury of detrusor in group M was severer than that in group EA;② comparison of the protein expression levels of cytochrome C, Pro-Caspase-9 and activated Caspase-9 between group M as well as group EA and group B as well as group S： the protein expression levels of cytochrome C, Pro-Caspase-9 and activated Caspase-9 in bladder tissue of rats in group M and group EA obviously increased（ P〈0.01, P〈0.05）; comparison between group EA and group M： the protein expression levels of cytochrome C, Pro-Caspase-9 and activated Caspase-9 in bladder tissue of rats in group EA obviously reduced（ P〈0.05）;③ comparison of the protein expression level of Caspase-3 between group M as well as group EA and group B as well as group S： the protein expression level of Pro-Caspase-3（35 KD） in group M and group EA reduced, and the protein expression level of activated Caspase-3（17 KD/19 KD） obviously increased（ P〈0.01, P〈0.05）; comparison between group EA andgroup M： the protein expression level of Pro-Caspase-3（35 KD） in group EA obviously increased, and the protein expression level of activated Caspase-3（17 KD/19 KD） obviously reduced（ P〈0.05）. Conclusion suprasacral spinal cord injury can increase the protein expression levels of cytochrome C, activated Caspase-3 and Caspase-9 in bladder tissues of rats, and electroacupuncture at BL 32, CV 3, SP 6 and GV 14 can down-regulate the protein expression levels of cytochrome C, activated Caspase-3 and Caspase-9 in bladder tissues of rats after suprasacral spinal cord injury and protect bladder tissue, which may be one of the mechanisms that electroacupuncture protects the involved bladder detrusor after suprasacral spinal cord injury.
文摘Objective:To observe the effects of electroacupuncture(EA)on the protein and gene expressions of Bax,Caspase-3 and Bcl-2 in cerebral cortex of type 2 diabetic rats with cognitive impairment(CI),and to explore the mechanism of EA in improving the learning and memory abilities.Methods:A total of 100 Sprague-Dawley(SD)rats were divided into a normal group(n=10)and a model group(n=90)by random number table method.Rats in the model group were intraperitoneally injected with a small dose of streptozotocin(STZ)to establish the type 2 diabetic models,after being fed with high-fat and high-sugar diet for 1 month.Twenty CI rats were selected from the 50 successful model rats by the Morris water maze(MWM)test and randomly divided into a model group and an EA group according to the blood glucose level and MWM data(n=10).Rats in the EA group received acupuncture at Zusanli(ST 36),Neiting(ST 44)and Yishu(Extra),of which Zusanli(ST 36)and Neiting(ST 44)were stimulated by EA apparatus,20 min/time,once a day for 6 d a week and 4 consecutive weeks.The rats in the model and the normal groups were fixed without treatment.After 4-week treatment,the random blood glucose level of the rats was measured;the learning and memory abilities of rats were measured by MWM;terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)assay was used to detect apoptotic cells;Western blot(WB)and real-time quantitative polymerase chain reaction(RT-qPCR)were used to detect the protein and gene expressions of Bax,Caspase-3 and Bcl-2 in cerebral cortex.Results:After modeling,the random blood glucose level and the escape latency tested by MWM were significantly increased,and the number of crossing the platform tested by the MWM was decreased in the EA and model groups,and were significantly different from those in the normal group(P<0.05 or P<0.01),while the differences between the model group and the EA group were not statistically significant(all P>0.05).After 4-week treatment,the random glucose level and the escape latency tested by MWM were significantly increased(both P<0.05),and the number of crossing the original platform tested by the MWM was significantly reduced(P<0.01),the protein and gene expressions of Bax and Caspase-3 were significantly increased(all P<0.001),the protein and gene expressions of Bcl-2 were significantly reduced(both P<0.001),and the number of neuron apoptosis was significantly increased(P<0.001)in the model group than in the normal group;the random blood glucose level was significantly reduced(P<0.05),the escape latency tested by MWM was significantly shortened(P<0.05),and the number of crossing the original platform tested by MWM was significantly increased(P<0.05),the protein and gene expressions of Bax and Caspase-3 were significantly reduced(all P<0.001),the protein and gene expressions of Bcl-2 were significantly increased(both P<0.001),and the number of neuron apoptosis was significantly reduced(P<0.001)in the EA group than in the model group.Conclusion:EA can improve the learning and memory damages induced by type 2 diabetic model rats with CI;the action mechanism may be achieved via anti-apoptosis.
文摘Objective:By observing the effects of electroacupuncture(EA)on the apoptosis of conjunctival cells of rabbits with dry eye syndrome(DES)and the expressions of apoptosis-related proteins Caspase-3,Fas and Bcl-2,to discuss the mechanism of EA in the treatment of DES from the perspective of cell apoptosis.Methods:Male New Zealand rabbits were randomly divided into a normal group(NG),a model group(MG),an EA group(EAG)and a sham EA group(SEAG).DES rabbit model was developed by eye drop of 0.1%benzalkonium chloride.The rabbit tear secretion and tear film break-up time(BUT)were measured;terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)assay was used to detect the apoptosis of conjunctival cells;the expressions of Caspase-3,Fas and Bcl-2 proteins in conjunctival cells were detected by immunohistochemistry.Results:Compared with the NG,the rabbit tear secretion decreased and the BUT was shortened in the MG(both P<0.01);compared with the MG and the SEAG,the rabbit tear secretion increased and the BUT was prolonged in the EAG(all P<0.05).Compared with the NG,the apoptosis of rabbit conjunctival cells increased(P<0.01),the expressions of Caspase-3 and Fas proteins increased(both P<0.05),and the expression of Bcl-2 protein decreased(P<0.01)in the MG;compared with the MG and the SEAG,the apoptosis of rabbit conjunctival cells decreased(both P<0.01),the expressions of Caspase-3 and Fas proteins decreased(all P<0.05),and the expression of Bcl-2 protein increased(both P<0.01)in the EAG.Conclusion:EA can inhibit the apoptosis of rabbit conjunctival cells,down-regulate the expressions of apoptosis-related proteins Caspase-3 and Fas,and up-regulate the expression of Bcl-2 protein,which may be one of the mechanisms of EA in treatment of DES.
