1.Effect of Yishen Jiangzhuo Granules on Mitochondrial Dynamic Protein and Apoptosis in Renal Tubule of Rats with Chronic Kidney Diseases
Minlin ZHENG ; Qianqian ZHAN ; Xiaoxia FANG ; Guang LIU ; Qin SAN ; Wenjiang FAN ; Yanan WANG ; Jianting WANG ; Shiwei RUAN
World Science and Technology-Modernization of Traditional Chinese Medicine 2025;27(9):2673-2686
Objective Exploring the therapeutic mechanism of Yishen Jiangzhuo Granules(YSJZG)on chronic kidney disease(CKD)based on mitochondrial dynamics and apoptosis of renal tubular epithelial cells.Methods The CKD model of rats with 5/6 nephrectomy was adopted and divided into 6 groups according to random number table:sham operation control group,model group,emodin group(500 mg/kg/d),Yishen Jiangzhuo granule low,middle and high dose groups.After 8 weeks of treatment with YSJZG,serum creatinine(SCR)and urea nitrogen(BUN),pathological changes of renal cortex,mitochondrial morphology and ultrastructure were detected,and mitochondrial kinetic protein in renal tubular epithelial cells was detected by immunohistochemistry(Drp1,Fis1)and fusion proteins(Opa1,Mfn1)were detected by Western blot,and apoptotic proteins(CytC,Bax)in cytoplasm and mitochondria were detected by real-time PCR.Results Renal injury:Compared with the model group,YSJZG groups significantly reduced the levels of SCR and BUN,renal tubular degeneration and necrosis,and mitochondrial structural damage in rats.Renal tubule mitochondrial dynamic protein:Compared with the model group,the expression of division proteins Drp1 and Fis1 was downregulated,the expression of fusion proteins Opa1 and Mfn1 was upregulated,and transmission electron microscopy observed that the mitochondrial fragmentation changes were relatively mild.Apoptosis related indicators and mtDNA copy number of renal tubular cells:Compared with the model group,the content of Bax protein in renal tubular epithelial cells of YSJZG groups increased significantly in cytoplasm(P<0.05)and decreased significantly in mitochondria(P<0.05).The content of CytC protein decreased significantly in cytoplasm(P<0.05)and increased significantly in mitochondria(P<0.05).The copy number of mtDNA increased significantly(P<0.05),and the total levels of SMAC,CytC and Bax mRNA decreased significantly(P<0.05).Correlation between mitochondrial dynamic protein and apoptosis in renal tubular cells:Pearson correlation analysis showed that the expression of Drp1 and Fis1 was negatively correlated with the expression of CytC in mitochondria,and positively correlated with the expression of CytC in cytoplasm.The expression levels of fusion proteins Opa1 and Mfn1 showed a significant positive correlation with CytC expression in mitochondria,and a significant negative correlation with CytC expression in cytoplasm.Conclusion YSJZG can significantly delay the progression of CKD,and its mechanism may be achieved by regulating mitochondrial dynamics of renal tubular epithelial cells,thereby inhibiting endogenous cell apoptosis pathway.
2.Effect of Yishen Jiangzhuo Granules on Mitochondrial Dynamic Protein and Apoptosis in Renal Tubule of Rats with Chronic Kidney Diseases
Minlin ZHENG ; Qianqian ZHAN ; Xiaoxia FANG ; Guang LIU ; Qin SAN ; Wenjiang FAN ; Yanan WANG ; Jianting WANG ; Shiwei RUAN
World Science and Technology-Modernization of Traditional Chinese Medicine 2025;27(9):2673-2686
Objective Exploring the therapeutic mechanism of Yishen Jiangzhuo Granules(YSJZG)on chronic kidney disease(CKD)based on mitochondrial dynamics and apoptosis of renal tubular epithelial cells.Methods The CKD model of rats with 5/6 nephrectomy was adopted and divided into 6 groups according to random number table:sham operation control group,model group,emodin group(500 mg/kg/d),Yishen Jiangzhuo granule low,middle and high dose groups.After 8 weeks of treatment with YSJZG,serum creatinine(SCR)and urea nitrogen(BUN),pathological changes of renal cortex,mitochondrial morphology and ultrastructure were detected,and mitochondrial kinetic protein in renal tubular epithelial cells was detected by immunohistochemistry(Drp1,Fis1)and fusion proteins(Opa1,Mfn1)were detected by Western blot,and apoptotic proteins(CytC,Bax)in cytoplasm and mitochondria were detected by real-time PCR.Results Renal injury:Compared with the model group,YSJZG groups significantly reduced the levels of SCR and BUN,renal tubular degeneration and necrosis,and mitochondrial structural damage in rats.Renal tubule mitochondrial dynamic protein:Compared with the model group,the expression of division proteins Drp1 and Fis1 was downregulated,the expression of fusion proteins Opa1 and Mfn1 was upregulated,and transmission electron microscopy observed that the mitochondrial fragmentation changes were relatively mild.Apoptosis related indicators and mtDNA copy number of renal tubular cells:Compared with the model group,the content of Bax protein in renal tubular epithelial cells of YSJZG groups increased significantly in cytoplasm(P<0.05)and decreased significantly in mitochondria(P<0.05).The content of CytC protein decreased significantly in cytoplasm(P<0.05)and increased significantly in mitochondria(P<0.05).The copy number of mtDNA increased significantly(P<0.05),and the total levels of SMAC,CytC and Bax mRNA decreased significantly(P<0.05).Correlation between mitochondrial dynamic protein and apoptosis in renal tubular cells:Pearson correlation analysis showed that the expression of Drp1 and Fis1 was negatively correlated with the expression of CytC in mitochondria,and positively correlated with the expression of CytC in cytoplasm.The expression levels of fusion proteins Opa1 and Mfn1 showed a significant positive correlation with CytC expression in mitochondria,and a significant negative correlation with CytC expression in cytoplasm.Conclusion YSJZG can significantly delay the progression of CKD,and its mechanism may be achieved by regulating mitochondrial dynamics of renal tubular epithelial cells,thereby inhibiting endogenous cell apoptosis pathway.
3.Study on the Prevention and Treatment of Postoperative Abdominal Adhesion by Inhibiting Epithelial-Mesenchymal Transition of Peritoneal Mesothelial Cells with Supplemented Buyang Huanwu Decoction
Minlin ZHENG ; Wenjiang FAN ; Yanan WANG ; Qianqian ZHAN
World Science and Technology-Modernization of Traditional Chinese Medicine 2024;26(6):1458-1470
Objective To evaluated the efficacy of Supplemented Buyang Huanwu Decoction in preventing and treating postoperative abdominal adhesions.Methods One hundred and eight male SD rats were randomly divided into ordinary group,mold group,Sodium hyaluronate group and Supplemented Buyang Huanwu Decoction group.The animals were sacrificed 7,14 and 28 days after the operation,Visual scoring was performed,and samples were taken from the injury site for hematoxylin-eosin staining and Masson staining,and scanning electron microscope to observe the morphology of peritoneal mesothelial cells in the modeling site.Immunohistochemical method was used to observe the expression of epithelial mesenchymal transition(EMT)related cell marker proteins E-cadherin and α-SMA in the adhesive site.Results ①Efficacy of Supplemented Buyang Huanwu Decoction in the prevention and treatment of postoperative abdominal adhesions:The Supplemented Buyang Huanwu Decoction group can reduce the Diamod visual adhesion score(P<0.05),as well as the adhesion levels of HE staining and Masson staining(P<0.05).②Degree of damage to peritoneal mesothelial cells:Scanning electron microscopy showed that in the field of view of the Supplemented Buyang Huanwu Decoction group,paving like peritoneal mesothelial cells could be seen,indicating that the Supplemented Buyang Huanwu Decoction can improve the damage of peritoneal mesothelial cells on the serosal side of the cecum in PAA.③Expression of Epithelial mesenchymal transition-related indicators:The expression of E-Cadherin protein in the Supplemented BuyangHuanwu Decoction group increased(P<0.05).The expression of α-SMA protein in the Supplemented Buyang Huanwu Decoction group decreased(P<0.05).Conclusion The effect of Supplemented Buyang Huanwu Decoction in the prevention and treatment of postoperative abdominal adhesions is exact,which mechanism may be by reducing Epithelial mesenchymal transition of peritoneal mesothelial cells,reducing the deposition of extracellular matrix,thereby reducing peritoneal adhesion.
4.Effect of pentraxin-3 on memory improvement and Aβ expression in 5×FAD mice
Qi ZHOU ; Zheng XIE ; Minlin LAI ; Pengpeng XING ; Zhao WANG ; Yaoyuan DONG ; Changlin LIAN ; Boyang LIU ; Hongbo GUO
Chinese Journal of Neuromedicine 2024;23(4):325-332
Objective:To explore the effect of pentraxin 3 (PTX3) on memory improvement and Aβ expression in Alzheimer's disease (AD) model mice.Methods:(1) Ten 5-month-old 5×FAD mice were randomly divided into PTX3 group and model group ( n=5); 5 C57BL/6 wild-type mice at the same age were selected as control group; mice in the PTX3 group and control group were stereotactically injected 4 μL 0.5 g/L PTX3 or same dose of phosphate buffered saline (PBS); Morris water maze test was used to detect the learning and memory abilities, Y maze test was used to detect the short-term memory, and ELISA was used to obsevre the contents of Aβ 40 and Aβ 42 in the brain hemisphere. (2) Twenty-five 3-month-old 5×FAD mice were randomly divided into model group, 2 μg/kg PTX3 group, 4 μg/kg PTX3 group, 8 μg/kg PTX3 group, and 16 μg/kg PTX3 group ( n=5); 5 C57BL/6 wild-type mice at the same age were selected as control group; mice in the PTX3 groups were intranasally injected 2, 4, 8, and 16 μg/kg PTX3, respectively; those in the model group and control group were intranasally injected same dose of PBS; injection was given once every 96 h for a total of 7 times. Morris water maze test was used to detect the learning and memory abilities, Y maze test was used to detect the short-term memory, and ELISA was used to obsevre the contents of Aβ 40 and Aβ 42 in the hippocampus. Results:(1) Compared with the model group, the PTX3 group had significantly shorter platform latency, higher percentage of exploration time and higher percentage of spontaneous alternations ( P<0.05). Compared with those in model group ([63.38±21.42] pg/mL, [29.77±6.11] pg/mL), the concentrations of Aβ 40 and Aβ 42 in the brain tissues of PTX3 group ([15.87±2.11] pg/mL, [16.55±1.95] pg/mL) were statistically lower ( P<0.05). (2) Compared with the model group, the 16 μg/kg PTX3 group had significantly shorter escape latency and higher percentage of exploration time ( P<0.05); compared with the model group, the 2 μg/kg PTX3 group and 16 μg/kg PTX3 group had significantly higher percentage of spontaneous alternations ( P<0.05). The contents of Aβ 40 and Aβ 42 in the hippocampus of 8 μg/kg PTX3 group and 16 μg/kg PTX3 group were statistically lower compared with those in the model group ( P<0.05). Conclusion:PTX3 may attenuate cognitive deficits and decrease Aβ expression in the brain or hippocampus tissues of 5×FAD mice with AD.

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