1.Protective Effects and Mechanism of Shenqi Qiangjing Granules on H2O2 Induced Injury of Mouse Spermatogonia by Inhibiting Oxidative Stress and Ferroptosis
Dianhui GAN ; Bingyu XIA ; Xin LI ; Kedao LAI ; Bin BIN ; Aicun TANG
Herald of Medicine 2025;44(3):371-376
Objective To explore the protective effects and mechanism of Shenqi Qiangjing granules containing serum on hydrogen peroxide(H2O2)induced oxidative damage and ferroptosis in mouse spermatogonia(GC-1 spg).Methods Thir-ty SPF grade healthy male SD rats were selected and randomly divided into blank group,levocarnitine group,and Shenqi Qiangjing granules group.After 7 days of intragastric administration,drug-containing serum was collected from each group.Using mouse sper-matogonia as a cell model,they were randomly divided into the normal control group,the model control group,blank serum group,levocarnitine containing serum group,and Shenqi Qiangjing granules containing serum group.Except for the normal control group,the other groups used hydrogen peroxide at a concentration of 600 μmol·L-1 to induce injury to mouse spermatogonia for 4 hours,and then established oxidative stress injury models,after 24 hours of medication intervention in each group.The survival rate of cells was detected using CCK-8 method;The levels of intracellular reactive oxygen species(ROS),malondialdehyde(MDA),catalase(CAT),glutathione(GSH),and superoxide dismutase(SOD)were detected by ELISA;The intracellular iron level was detected by iron ion colorimetry;The activities of Caspase-3 and Caspase-9 were detected by colorimetry;The mRNA levels of glu-tathione peroxidase 4(GPX4)and ACSL4 were determined by qRT-PCR.Results Compared with the normal control group,the cell proliferation activity of the model control group decreased significantly,the levels of ROS,MDA and Fe3+were significantly increased,while the activities of CAT,GSH and SOD were significantly decreased in the model control group,however,Caspase-3 and Caspase-9 activities were significantly increased,the results showed significant difference(P<0.05).Compare with the model control group,Shenqi Qiangjing granules containing serum could significantly increase the cell proliferation activity,decrease the levels of ROS,MDA and Fe3+,increase the activities of CAT,GSH and SOD,and decrease the activities of Caspase-3 and Caspase-9,the results showed significant difference(P<0.05).The qRT PCR results showed that compared with the model control group,the expression of GPX4 mRNA was upregulated and ACSL4 mRNA was downregulated in blank serum group containing Shenqi Qiangjing granules,and the differences were statistically significant(P<0.05).Conclusions Shenqi Qiangjing granules have significant protective effects on hydrogen peroxide-induced oxidative stress injury and ferroptosis of spermatogonia in mice.The mechanism may be related to the decrease of Caspase-3 and Caspase-9 activities and the inhibition of oxidative stress injury and ferroptosis.
2.Protective Effects and Mechanism of Shenqi Qiangjing Granules on H2O2 Induced Injury of Mouse Spermatogonia by Inhibiting Oxidative Stress and Ferroptosis
Dianhui GAN ; Bingyu XIA ; Xin LI ; Kedao LAI ; Bin BIN ; Aicun TANG
Herald of Medicine 2025;44(3):371-376
Objective To explore the protective effects and mechanism of Shenqi Qiangjing granules containing serum on hydrogen peroxide(H2O2)induced oxidative damage and ferroptosis in mouse spermatogonia(GC-1 spg).Methods Thir-ty SPF grade healthy male SD rats were selected and randomly divided into blank group,levocarnitine group,and Shenqi Qiangjing granules group.After 7 days of intragastric administration,drug-containing serum was collected from each group.Using mouse sper-matogonia as a cell model,they were randomly divided into the normal control group,the model control group,blank serum group,levocarnitine containing serum group,and Shenqi Qiangjing granules containing serum group.Except for the normal control group,the other groups used hydrogen peroxide at a concentration of 600 μmol·L-1 to induce injury to mouse spermatogonia for 4 hours,and then established oxidative stress injury models,after 24 hours of medication intervention in each group.The survival rate of cells was detected using CCK-8 method;The levels of intracellular reactive oxygen species(ROS),malondialdehyde(MDA),catalase(CAT),glutathione(GSH),and superoxide dismutase(SOD)were detected by ELISA;The intracellular iron level was detected by iron ion colorimetry;The activities of Caspase-3 and Caspase-9 were detected by colorimetry;The mRNA levels of glu-tathione peroxidase 4(GPX4)and ACSL4 were determined by qRT-PCR.Results Compared with the normal control group,the cell proliferation activity of the model control group decreased significantly,the levels of ROS,MDA and Fe3+were significantly increased,while the activities of CAT,GSH and SOD were significantly decreased in the model control group,however,Caspase-3 and Caspase-9 activities were significantly increased,the results showed significant difference(P<0.05).Compare with the model control group,Shenqi Qiangjing granules containing serum could significantly increase the cell proliferation activity,decrease the levels of ROS,MDA and Fe3+,increase the activities of CAT,GSH and SOD,and decrease the activities of Caspase-3 and Caspase-9,the results showed significant difference(P<0.05).The qRT PCR results showed that compared with the model control group,the expression of GPX4 mRNA was upregulated and ACSL4 mRNA was downregulated in blank serum group containing Shenqi Qiangjing granules,and the differences were statistically significant(P<0.05).Conclusions Shenqi Qiangjing granules have significant protective effects on hydrogen peroxide-induced oxidative stress injury and ferroptosis of spermatogonia in mice.The mechanism may be related to the decrease of Caspase-3 and Caspase-9 activities and the inhibition of oxidative stress injury and ferroptosis.
3.Protective Effect and Mechanism of Tadehaginoside on Hepatic Fibrosis Model Mice Induced by Carbon Tetra- chloride
Aicun TANG ; Yanfei WEI ; Xihua LIU ; Minggang WANG ; Qiuyu LU
China Pharmacy 2020;31(2):190-195
OBJECTIVE:To study the protective effect of tadehaginoside(TA)on liver fibrosis model mice induced by carbon tetrachloride(CCl4),and to investigate its mechanism. METHODS :Kunming mice were randomly divided into normal group , model group ,colchicines group (positive control ,0.2 mg/kg),TA low-dose ,medium-dose and high-dose groups (3,6,12 mg/kg),with 10 mice in each group. Those groups were intraperitoneally injected with 10% CCl4-olive oil solution (5 mL/kg)to induce liver fibrosis model twice a week ,for consecutive 8 weeks;except that ,the normal group was intraperitoneally injected with olive oil. From 3rd week ,the mice in each administration groups were given relevant medicine intragastrically ,normal group and model group were given constant volume 2% sodium carboxymethylcellulose solution intragastrically ,once a day ,for consecutive 6 weeks. The contents of ALT ,AST,HA and IL- 6 in serum of mice were test ed by ELISA. The contents of Hyp , SOD,MDA and GSH-Px in liver tissues were detected by spectrophotometry. mRNA expression of Col- Ⅰ,TIMP-1 and TIMP-2 in liver tissue was detected by RT-PCR. The protein expression of MMP- 2 and TGF-β1 in liver tissue was detected by Western blotting. RESULTS : Compared with normal group,the contents of ALT,AST,HA,IL-6,MDA and Hyp,the mRNA expression of Col- Ⅰ,TIMP-1 and TIMP- 2,as well as the protein exp ression of MMP- 2 and TGF-β1 were increased significantly ,while the contents of SOD and GSH-Px were decreased significantly (P<0.01). Compared with model group,the contents of ALT ,AST,HA,IL-6,MDA and Hyp ,the mRNA expression of Col- Ⅰ,TIMP-1 and TIMP- 2,as well as the protein expression of MMP- 2 and TGF-β1were decreased significantly ,while the contents of SOD and GSH-Px were increased significantly(P<0.05 or P<0.01). CONCLUSIONS :TA has a significant protective effect on liver tissue and anti-fibrosis effects in liver fibrosis model mice ,the mechanism of which may be associated with inhibiting lipid peroxidation and collagen synthesis , down-regulating the mRNA expression of Col- Ⅰ,TIMP-1 and TIMP- 2 as well as the protein expression of MMP- 2 and TGF-β1.
4.Protective Effects of Ethanol Extracts of Tadehagi triquetrum on Acute Liver Injury in Mice
Aicun TANG ; Zhaoni CHEN ; Tao LIANG ; Qiuyu LU
Herald of Medicine 2016;(3):242-245
Objective To investigate the protective effects of ethanol extracts of Tadehagi triquetrum ( TTOE) on car-bon tetrachloride ( CCl4 )-induced acute liver injury in mice. Methods Kunming mice were randomly divided into six groups:normal control group ( NC group) ,model control group,bifendate dropping pill group,low-,medium-and high-dose TTOE groups. The liver injury model was established by administration of CCl4 in all the groups except the NC group.The indexes of the liver, spleen and thymus were obtained.The activities of serum ALT,AST,ALP,LDH, albumin and T-AOC were measured.The activi-ties of SOD and GSH-PX and the contents of MDA,NO and GSH and Cyt P450 were also detected in hepatic tissues. Results TTOE at different doses could obviously reduce the indexes of the liver,thymus and spleen,which were (57.13±0.71),(32.44± 0.24),and (27.78±0.16),respectively,in high-dose TTOE group,and there were significant differences between the TTOE groups and model control group (P<0.01).The activities of ALT,AST,ALP and LDH were obviously decreased in high-dose TTOE groups,which were (65.59±8.23),(141.38±15.52),(2 462.4±253.6),(172.51±20.64),respectively,in the TTOE high-dose group (P<0.01).The serum levels of Alb and T-AOC were obviously increased,the contents of NO and MDA significantly decreased and the activities of SOD and GSH-PX and the contents of GSH Cyt P450 in liver tissues profoundly increased in TTOE groups when compared with those in model control group ( P<0.05 or P<0.01) . Conclusion TTOE could protect against acute hepatic injury induced by CCl4 in mice,which may be associated with the decrease in the activities of liver enzymes,anti-oxide free radical effect,decreased NO content and inhibited lipid peroxidation.

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