Objective To observe the effect of different concentration Flos Buddlejae(Mi Meng Hua,密蒙花)granules on apoptosis factors of lacrimal gland cells of castrated male rabbits,and study the treatment effects of Flos Budd...Objective To observe the effect of different concentration Flos Buddlejae(Mi Meng Hua,密蒙花)granules on apoptosis factors of lacrimal gland cells of castrated male rabbits,and study the treatment effects of Flos Buddlejae(Mi Meng Hua,密蒙花)granules in the dry eye model of castrated male rabbits.Methods Flos Buddlejae(Mi Meng Hua,密蒙花)raw material was made into granules.Thirty healthy adult New Zealand rabbits were randomly divided into five groups,six rabbits in each group.Group A:blank group,Group B:model group,Group C:Flos Buddlejae(Mi Meng Hua,密蒙花)granule group,Group D:placebo group,Group E:testosterone group.Except for group A,all rabbits underwent removal of bilateral testis and epididymis.Rabbits in group C were administered Flos Buddlejae(Mi Meng Hua,密蒙花)granules(100 mg/kg),three times per day;rabbits in group D were administered normal saline,three times per day.Rabbits in group E were injected with testosterone propionate(0.5 mL/kg)in the thigh muscle,every 3 days.All rabbits were tested by Schirmer I test(SIT)and tear film break-up time(BUT)before operation and 4 weeks after operation.After 4 weeks,all rabbits were sacrificed by air embolism and clipping of the lacrimal gland.Apoptosis factors,including Bax,Bcl-2,Fas and FasL of lacrimal gland cells were characterized by immunohistochemistry,and resulting data were statistically analyzed.Results(1)Comparison of SIT and BUT before and after operation:There were statistically significant differences between groups B and D(P<0.01),but not among other groups(P>0.05).(2)Comparison of apoptosis factors Bax,Bcl-2 Fas and FasL:In a comparison of groups B and D,there was no statistically significant difference after operation(P>0.05).In a comparison of the other groups,there were statistically significant differences(P<0.01).In comparisons among A,C and E groups,there were no statistically significant differences(P>0.05).Conclusion Compared with androgen,Flos Buddlejae(Mi Meng Hua,密蒙花)granules caused similar but slightly weaker depression of Bax,Fas and FasL,and increased expression of Bcl-2.展开更多
Objective:To investigate the effect of Gubi Tongxiao granules on vascular formation and cell apoptosis in the process of glucocorticoid-induced femoral head necrosis.Methods:Thirty experimental New Zealand rabbits wer...Objective:To investigate the effect of Gubi Tongxiao granules on vascular formation and cell apoptosis in the process of glucocorticoid-induced femoral head necrosis.Methods:Thirty experimental New Zealand rabbits were randomly divided into groups.Except the control group,the animal model of steroid-induced femoral head necrosis was established by lipopolysaccharide combined with glucocorticoids.After successful modeling,the experimental group was given Gubi Tongxiao granules intragastric treatment,the model and control group were given the same amount of normal saline intragastric treatment,once a day,for 8 weeks,the experimental animals were sacrificed,the bilateral femoral head of each group was taken out for hematoxylin-eosin staining,and protein CD34,CYR61 and VEGF were immunohistochemical staining localization.TUNEL staining was used to observe the apoptosis of femoral head cells.The expression levels of apoptosis proteins Bax and Bcl-2 were detected by WB.The expression of VEGF eNOs Bax and Bcl-2 genes was analyzed by RT-qPCR.Results:Gubi Tongxiao granules reduced the number of hollow bone lacunae and apoptotic positive cells(P<0.01),and up-regulated the expressions of CD34,CYR61 and VEGF in femoral head tissue.Compared with control group,the expression of pro-apoptotic protein Bax increased and the expression of anti-apoptotic protein Bcl-2,VEGF and eNOs mRAN decreased significantly in the model group(P<0.01),the opposite results were obtained in the experimental group after Gubi Tongxiao granule intervention(P<0.05).Conclusion:Gubi Tongxiao granules can improve the expression of angiogenic genes and regulate apoptosis-related proteins in the bone tissue of rabbit model with steroid-induced necrosis of femoral head,promote angiogenic differentiation and inhibit apoptosis,and thus achieve the effect of treating steroid-induced necrosis of femoral head.展开更多
BACKGROUND： Some experiments have demonstrated that melatonin （N-aceyl-5-methoxytryptamine, Mel） has antioxidationHow. ever, whether it has neuroprotective effect in the ischemia/reperfusion injury of central nervo...BACKGROUND： Some experiments have demonstrated that melatonin （N-aceyl-5-methoxytryptamine, Mel） has antioxidationHow. ever, whether it has neuroprotective effect in the ischemia/reperfusion injury of central nervous system is unclear. OBJECTIVE： To observe the protective effect of Mel on ischemia/reperfusion-induced cerebellar neuronal apoptosis of rats, and the action mechanism. DESIGN： Controlled observation experiment. SETTING： Department of Biochemistry and Molecular Biology, Tongji Medical College, Huazhong University of Science and Technology. MATERIALS： Eight Sprague-Dawley rats aged 7 - 8 days and weighing 10 - 12 g were provided by Medical Experimental Animal Center, Tongji Medical College, Huazhong University of Science and Technology. Anti-cytochrome C monoclonal antibody was purchased from R ＆ D Company; 7-dichlorodihydrofluorescein diacetate（DCFH-DA）, rhodamine 123 and Mel were purchased from Sigma Company （USA）. Lactate dehydrogenase （LDH） kit was purchased from Nanjing Jiancheng Bioengineering Institute. METHODS： This experiment was carried out in the laboratory for Department of Biochemistry and Molecule Biology, Tongji Medical College between October 2002 and March 2004. Cerebellar neurons of rats were cultured in vitro. After oxygen-glucose deprivation （OGD） for 90 minutes, 1 × 10^-4, 1 × 10^-6, 1 × 10^-9 mol/L Mel was added, respectively, namely high-, middle-, and low-concentration Mel groups. Cells, which were cultured by OGD, served as model group, and control group, in which OGD intervention was omitted, was set. ①Cytochrome C level of mitochondrial cells in each group was detected by ELISA method. ②LDH activity in the cell culture fluid was measured, and cell membrane permeability change was analyzed. The cells in the Mel group with the lowest LDH activity served as Mel treatment group, i.e. cells were cultured with OGD, and then Mel was added; Meanwhile, Mel prevention group was set, i.e. Mel was added before OGD. Intervention was not changed in the model group and control group. ③ DNA level was analyzed and cell apoptosis was observed by agarose gel electrophoresis（AGE）. ④Mitochondrial transmembrane potential of cells, and apoptotic way in each group were analyzed by confocal laser scanning microscopy. MAIN OUTCOME MEASURES： ①Mitochondrial cytochrome C level of cerebellar nerve cells. ②LDH activity of cerebellar nerve cells. ③ DNA AGE results.④Mitochondrial transmembrane potential change. RESULTS： ①Mitochondrial cytochrome C level of cerebellar nerve cells： cytochrome C was obviously released at 6 hours of OGD-reperfusion. Mel inhibited the release of cytochrome C in dose-dependent manner. ②LDH activity of cerebellar nerve cells： LDH activity （A value） was significantly lower in the high- and middle-concentration Mel groups than in the model group （P 〈 0.05）. LDH activity （A value） in the low-concentration Mel group was 0.415 0±0.012 9, indicating that Mel could decrease LDH activity of OGD-treated cell supernatant and promote membrane stablization in dose-dependent manner.③AGE results of DNA： 1 × 10^-9 mol/L was considered as the best concentration of melatonin. Cell DNA was extracted for AGE. Results presented typical ladder shape, indicating apoptosis appeared, while apoptosis was lessened in the Mel treatment group and Mel prevention group.④Mitochondrial transmembrane potential change： Experimental results showed that green fluorescein was evenly distributed in cerebellar granule cells cultured normally, and the axons of neurons were very clear. The body of neurons was condensed and the axons disappeared after cerebellar granule cells undergoing OGD injury. Mel could completely reverse the effect of OGD. CONCLUSION： Mel can enhance cerehellar neuronal membrane stabilization of rats in dosedependent manner, and suppress OGD-induced apoptosis of cerehellar granule cells by preventing against mitochondrial apoptosis.展开更多
OBJECTIVE:To explore whether fat mass and obesity associated proteins(FTO) is an important target of Qiteng Xiaozhuo granules(QTXZG,芪藤消浊颗粒) medicated serum in regulating proliferation and apoptosis of glomerular...OBJECTIVE:To explore whether fat mass and obesity associated proteins(FTO) is an important target of Qiteng Xiaozhuo granules(QTXZG,芪藤消浊颗粒) medicated serum in regulating proliferation and apoptosis of glomerular mesangial cells.METHODS:Medicated serum was obtained from Sprague-Dawley(SD) rats administered intragastrically with QTXZG decoction.The optimal concentration and intervention time of medicated serum were selected with the cell counting kit 8 assay.Cell proliferation was assessed by 5-ethynyl-2’-deoxyuridine(Ed U) and cell apoptosis was investigated using flow cytometry.The expression of FTO,Proliferating cell nuclear antigen,Cyclin D1,B-cell lymphoma 2(Bcl2) and BCL2 assaciated X was detected by Western blot and Real-time quantitative polymerase chain reaction,respectively.Quantification of the m6A RNA methylation was utilized to determine the total level of m6A methylation modification.RESULTS:Ed U and flow cytometry assays revealed that QTXZG medicated serum can remarkably inhibit proliferation and promote apoptosis of lipopolysaccharide(LPS)-induced human glomerular mesangial cells(HGMCs).The FTO overexpression plasmid could inhibit proliferation and promote apoptosis of LPS-induced HGMCs.The FTO inhibitor(FB23-2) can significantly attenuate the effect of QTZXG medicated serum on inhibiting excessive proliferation and promoting apoptosis.QTXZG medicated serum can significantly increase FTO expression and decrease the level of m6A methylation modification.CONCLUSIONS:FTO is a key target for QTXZG medicated serum in inhibiting excessive proliferation and promoting apoptosis of human glomerular mesangial cells.展开更多
基金the National Natural Science Foundation of China (NSFC) (No.30772824 and No.81574031)225 Project of High-Level Medical Talents of Hunan Province+4 种基金Research Project of Science and Technology Department of Hunan Province (No.2015SF2016-6)Research Project of Hunan Provincial Development and Reform Commission (No.[2014]658)Major Project of Changsha Science and Technology Plan (K1501014-31)Construction Project of Key Discipline of Chinese Ophthalmology of State Administration of Traditional Chinese MedicineConstruction Project of Key Discipline of Ophthalmology and Otolaryngology of Chinese Medicine of Hunan Province
文摘Objective To observe the effect of different concentration Flos Buddlejae(Mi Meng Hua,密蒙花)granules on apoptosis factors of lacrimal gland cells of castrated male rabbits,and study the treatment effects of Flos Buddlejae(Mi Meng Hua,密蒙花)granules in the dry eye model of castrated male rabbits.Methods Flos Buddlejae(Mi Meng Hua,密蒙花)raw material was made into granules.Thirty healthy adult New Zealand rabbits were randomly divided into five groups,six rabbits in each group.Group A:blank group,Group B:model group,Group C:Flos Buddlejae(Mi Meng Hua,密蒙花)granule group,Group D:placebo group,Group E:testosterone group.Except for group A,all rabbits underwent removal of bilateral testis and epididymis.Rabbits in group C were administered Flos Buddlejae(Mi Meng Hua,密蒙花)granules(100 mg/kg),three times per day;rabbits in group D were administered normal saline,three times per day.Rabbits in group E were injected with testosterone propionate(0.5 mL/kg)in the thigh muscle,every 3 days.All rabbits were tested by Schirmer I test(SIT)and tear film break-up time(BUT)before operation and 4 weeks after operation.After 4 weeks,all rabbits were sacrificed by air embolism and clipping of the lacrimal gland.Apoptosis factors,including Bax,Bcl-2,Fas and FasL of lacrimal gland cells were characterized by immunohistochemistry,and resulting data were statistically analyzed.Results(1)Comparison of SIT and BUT before and after operation:There were statistically significant differences between groups B and D(P<0.01),but not among other groups(P>0.05).(2)Comparison of apoptosis factors Bax,Bcl-2 Fas and FasL:In a comparison of groups B and D,there was no statistically significant difference after operation(P>0.05).In a comparison of the other groups,there were statistically significant differences(P<0.01).In comparisons among A,C and E groups,there were no statistically significant differences(P>0.05).Conclusion Compared with androgen,Flos Buddlejae(Mi Meng Hua,密蒙花)granules caused similar but slightly weaker depression of Bax,Fas and FasL,and increased expression of Bcl-2.
基金Natural Science Research Project of Higher Education Department of Anhui Province(No.KJ2020A0403)Natural Science Foundation of Anhui Province(No.2008085MH281)。
文摘Objective:To investigate the effect of Gubi Tongxiao granules on vascular formation and cell apoptosis in the process of glucocorticoid-induced femoral head necrosis.Methods:Thirty experimental New Zealand rabbits were randomly divided into groups.Except the control group,the animal model of steroid-induced femoral head necrosis was established by lipopolysaccharide combined with glucocorticoids.After successful modeling,the experimental group was given Gubi Tongxiao granules intragastric treatment,the model and control group were given the same amount of normal saline intragastric treatment,once a day,for 8 weeks,the experimental animals were sacrificed,the bilateral femoral head of each group was taken out for hematoxylin-eosin staining,and protein CD34,CYR61 and VEGF were immunohistochemical staining localization.TUNEL staining was used to observe the apoptosis of femoral head cells.The expression levels of apoptosis proteins Bax and Bcl-2 were detected by WB.The expression of VEGF eNOs Bax and Bcl-2 genes was analyzed by RT-qPCR.Results:Gubi Tongxiao granules reduced the number of hollow bone lacunae and apoptotic positive cells(P<0.01),and up-regulated the expressions of CD34,CYR61 and VEGF in femoral head tissue.Compared with control group,the expression of pro-apoptotic protein Bax increased and the expression of anti-apoptotic protein Bcl-2,VEGF and eNOs mRAN decreased significantly in the model group(P<0.01),the opposite results were obtained in the experimental group after Gubi Tongxiao granule intervention(P<0.05).Conclusion:Gubi Tongxiao granules can improve the expression of angiogenic genes and regulate apoptosis-related proteins in the bone tissue of rabbit model with steroid-induced necrosis of femoral head,promote angiogenic differentiation and inhibit apoptosis,and thus achieve the effect of treating steroid-induced necrosis of femoral head.
基金the National Science Foundation of Hygienic Committee of Hubei Province, No.WJ01510
文摘BACKGROUND： Some experiments have demonstrated that melatonin （N-aceyl-5-methoxytryptamine, Mel） has antioxidationHow. ever, whether it has neuroprotective effect in the ischemia/reperfusion injury of central nervous system is unclear. OBJECTIVE： To observe the protective effect of Mel on ischemia/reperfusion-induced cerebellar neuronal apoptosis of rats, and the action mechanism. DESIGN： Controlled observation experiment. SETTING： Department of Biochemistry and Molecular Biology, Tongji Medical College, Huazhong University of Science and Technology. MATERIALS： Eight Sprague-Dawley rats aged 7 - 8 days and weighing 10 - 12 g were provided by Medical Experimental Animal Center, Tongji Medical College, Huazhong University of Science and Technology. Anti-cytochrome C monoclonal antibody was purchased from R ＆ D Company; 7-dichlorodihydrofluorescein diacetate（DCFH-DA）, rhodamine 123 and Mel were purchased from Sigma Company （USA）. Lactate dehydrogenase （LDH） kit was purchased from Nanjing Jiancheng Bioengineering Institute. METHODS： This experiment was carried out in the laboratory for Department of Biochemistry and Molecule Biology, Tongji Medical College between October 2002 and March 2004. Cerebellar neurons of rats were cultured in vitro. After oxygen-glucose deprivation （OGD） for 90 minutes, 1 × 10^-4, 1 × 10^-6, 1 × 10^-9 mol/L Mel was added, respectively, namely high-, middle-, and low-concentration Mel groups. Cells, which were cultured by OGD, served as model group, and control group, in which OGD intervention was omitted, was set. ①Cytochrome C level of mitochondrial cells in each group was detected by ELISA method. ②LDH activity in the cell culture fluid was measured, and cell membrane permeability change was analyzed. The cells in the Mel group with the lowest LDH activity served as Mel treatment group, i.e. cells were cultured with OGD, and then Mel was added; Meanwhile, Mel prevention group was set, i.e. Mel was added before OGD. Intervention was not changed in the model group and control group. ③ DNA level was analyzed and cell apoptosis was observed by agarose gel electrophoresis（AGE）. ④Mitochondrial transmembrane potential of cells, and apoptotic way in each group were analyzed by confocal laser scanning microscopy. MAIN OUTCOME MEASURES： ①Mitochondrial cytochrome C level of cerebellar nerve cells. ②LDH activity of cerebellar nerve cells. ③ DNA AGE results.④Mitochondrial transmembrane potential change. RESULTS： ①Mitochondrial cytochrome C level of cerebellar nerve cells： cytochrome C was obviously released at 6 hours of OGD-reperfusion. Mel inhibited the release of cytochrome C in dose-dependent manner. ②LDH activity of cerebellar nerve cells： LDH activity （A value） was significantly lower in the high- and middle-concentration Mel groups than in the model group （P 〈 0.05）. LDH activity （A value） in the low-concentration Mel group was 0.415 0±0.012 9, indicating that Mel could decrease LDH activity of OGD-treated cell supernatant and promote membrane stablization in dose-dependent manner.③AGE results of DNA： 1 × 10^-9 mol/L was considered as the best concentration of melatonin. Cell DNA was extracted for AGE. Results presented typical ladder shape, indicating apoptosis appeared, while apoptosis was lessened in the Mel treatment group and Mel prevention group.④Mitochondrial transmembrane potential change： Experimental results showed that green fluorescein was evenly distributed in cerebellar granule cells cultured normally, and the axons of neurons were very clear. The body of neurons was condensed and the axons disappeared after cerebellar granule cells undergoing OGD injury. Mel could completely reverse the effect of OGD. CONCLUSION： Mel can enhance cerehellar neuronal membrane stabilization of rats in dosedependent manner, and suppress OGD-induced apoptosis of cerehellar granule cells by preventing against mitochondrial apoptosis.
基金National Natural Science Foundation of China:Lnc NONRATG001910.2 Competitively Binds miR-339 to Regulate the Mechanism of Syk Involvement in the Pathogenesis of Chronic Nephritis and the Intervention of Qiteng Xiaozhuo Particles (No.81973546)the Key Scientific Research Projects of Natural Science in Colleges and Universities in Anhui Province:Study on the Mechanism of FTO-Mediated FOXO6 mRNA m6A Modification Regulating the Proliferation and Apoptosis of Glomerular Mesangial Cells (No.2022AH050747)Natural General Projects of Chaohu University:Expression and Target Analysis of miR-155-5p in Kidney Tissue of CGN Rats (No.XLY-201912)
文摘OBJECTIVE:To explore whether fat mass and obesity associated proteins(FTO) is an important target of Qiteng Xiaozhuo granules(QTXZG,芪藤消浊颗粒) medicated serum in regulating proliferation and apoptosis of glomerular mesangial cells.METHODS:Medicated serum was obtained from Sprague-Dawley(SD) rats administered intragastrically with QTXZG decoction.The optimal concentration and intervention time of medicated serum were selected with the cell counting kit 8 assay.Cell proliferation was assessed by 5-ethynyl-2’-deoxyuridine(Ed U) and cell apoptosis was investigated using flow cytometry.The expression of FTO,Proliferating cell nuclear antigen,Cyclin D1,B-cell lymphoma 2(Bcl2) and BCL2 assaciated X was detected by Western blot and Real-time quantitative polymerase chain reaction,respectively.Quantification of the m6A RNA methylation was utilized to determine the total level of m6A methylation modification.RESULTS:Ed U and flow cytometry assays revealed that QTXZG medicated serum can remarkably inhibit proliferation and promote apoptosis of lipopolysaccharide(LPS)-induced human glomerular mesangial cells(HGMCs).The FTO overexpression plasmid could inhibit proliferation and promote apoptosis of LPS-induced HGMCs.The FTO inhibitor(FB23-2) can significantly attenuate the effect of QTZXG medicated serum on inhibiting excessive proliferation and promoting apoptosis.QTXZG medicated serum can significantly increase FTO expression and decrease the level of m6A methylation modification.CONCLUSIONS:FTO is a key target for QTXZG medicated serum in inhibiting excessive proliferation and promoting apoptosis of human glomerular mesangial cells.
