| Objective: To evaluate the anti-aging effects of water extracts and ethanol extracts from five root mongolian traditional medicinal herbs named Pentamon using an animal model of oxidative stressMethods: Healthy adult male Wistar rats were randomly divided into 8 groups,with 8 rats in each group.The groups are blank group,cisplatin(CP)model group,positive control DHEA group(5.166 mg/kg),positive control qinaskul group(12.4 mg/kg),low-dose(497 mg/kg)water extract group,high-dose(994 mg/kg)water extract group,low-dose(43.736 mg/kg)ethanol extract group and high-dose(262.416mg/kg)ethanol extract groups of Pentamon.After 15 days of administration,the animals were sacrificed,and the serum,testicular tissue and epididymal tail sperm were taken to detect relevant indexes.Except for the blank group,the other groups were intraperitoneally injected with CP at 7.5 mg/kg on the 10 th day for modeling.(1)Modeling: Pathological sections of testicular tissue were routinely prepared and pathological changes were observed under a microscope after HE staining.(2)Apoptosis: The m RNA expression levels of caspase-3 and beta cell leukemia-2 protease(BCL-2)in testis were detected by Q-PCR.(3)Testicular function: The sperm concentration and viability were determined by using a Nikon DS microscope and NIS-Elements software.And the testosterone content in the serum was measured by the Elisa method.(4)Oxidative stress: The catalase(CAT),glutathione(GSH)and malondialdehyde(MDA)in the serum were detected by biochemical methods.The m RNA expression level of superoxide disproportionation enzyme 2(SOD2)in testis was detected by Q-PCR.(5)Mitochondrial stability: The ROS content in the testis was detected by chemiluminescence assay and the m RNA expression level of uncoupling protein 2(UCP2)in the testis was detected by Q-PCR.(6)Telomere length: The telomerase content in the serum was measured by the Elisa method.Results(1)Compared with the blank group,the testicular weight of the CP model group was significantly lower(P<0.05).From the pathological section,the seminal tubule basement membrane was significantly thickened and the spermatogenic cells and sertoli cells were not arranged neatly and the layers were reduced and the number of mature sperm in the seminiferous tube was reduced and testicular tissue damage occurred.This prove that the CP animal model has been succeed.There was a recover in testicular weight after administration but the trend in the DHEA group and the high-dose water extracts group was not significant(P>0.05).The result of pathological section shows that the damage of testicular tissue in the all drug-administered group recovered and the structure,arrangement,density and hierarchical integrity of spermatogenic cells and sertoli cells were close to the blank group and a number of .mature sperm in the seminiferous tube was observed.(2)Compared with the blank group,the m RNA expression level of Casp-3 was significantly up-regulated and the BCL-2 was down-regulated in the CP model group(P<0.01),which proved that the testis tissue of the model group ocour apoptosis.After administration,the m RNA expression levels of Casp-3 in the qinaskul group,DHEA group,low-dose ethanol extract group and high-dose water extract group were significantly down-regulated(P<0.05).But there is no significant difference in other groups(P>0.05).The m RNA expression levels of BCL-2 in qinaskul group and DHEA group were significantly up-regulated(P<0.01)and in the low-dose ethanol extract group was significantly higher(P<0.05).There was no significant difference in other groups(P>0.05).(3)Compared with the blank group,the sperm motility and concentration of the model group decreased(P<0.01,P<0.05)and the content of serum Testosterone also decreased significantly(P<0.01),which proved that the function of testicular tissue in the model group has been weakened.After administration,the viability of sperm was significantly higher than that of the model group(P<0.01).The concentration of sperm in low-dose ethanol extract group and the qinaskul group was significantly higher than that of the model group(P<0.05).There was no significant difference between the other groups(P>0.05).The content of Testosterone was significantly increased in the low-dose ethanol extract group,the high-dose water extract group and the DHEA group(P<0.05).And there was no significant difference in other groups(P>0.05).(4)Compared with the blank group,the serum MDA and GSH contents in the model group decreased(P<0.05).The of CAT content and the m RNA expression level of SOD2 in the testis decreased significantly(P<0.01),indicating the oxidative stress effect in the model group has been enhanced.After administration,the MDA content of five low-dose ethanol extract group and the low-dose water extract group decreased significantly(P<0.01)and the qinaskul group decreased(P<0.05).There was no significant difference in other groups(P>0.05).The GSH content of the low-dose ethanol extract group and the high-dose water extract group increased significantly(P<0.01)and the low-dose water extract group and qinaskul group increased(P<0.05).There was no significant difference in other groups(P>0.05).The CAT content of the qinaskul group and the DHEA group increased significantly(P<0.01)and the low-dose and high-dose ethanol extract group and low-dose water extract group increased(P<0.05).There is no significant difference in other groups(P>0.05).The m RNA expression level of SOD2 of the low-dose and high-doseethanol extract group and qinaskul group increased(P<0.05).There was no significant difference in other groups(P>0.05).(5)Compared with the blank group,the ROS content and UCP2 m RNA expression level of the model group increased(P<0.01,P<0.05),indicating that the membrane of mitochondrial became unstable the model group.After administration,the ROS content of the low-dose ethanol extract group and the DHEA group was significantly decreased(P<0.01).The ROS content in the qinaskul group was increased(P<0.05).There was no significant difference in other groups(P>0.05).The UCP2 m RNA expression levels of low-dose,high-dose water extract group and qinaskul group were significantly decreased(P<0.01),and the DHEA group was decreased(P<0.05).There was no significant difference in other groups(P>0.05).(6)Compared with the blank group,the content of serum Telomerase of the model group decreased(P<0.05),indicating that the telomere of the model group was likely to be shortened.After administration,the content of Telomerase in the low-dose ethanol extract group and in the high-dose water extract group increased(P<0.05).The content of Telomerase in DHEA group increased significantly(P<0.01).There was no significant difference in other groups(P>0.05).Conclusion:(1)The CP-induced testicular injury model was successfully established.The water and ethanol extracts of Pentamon have protective effects on testicular damage.(2)Water extracts and ethanol extracts of Pentamon all inhibited testicular injury and restored testicular function by enhancing sperm motility,concentration and Testosterone content.The oxidative stress effect was attenuated by increasing the content of SOD,GSH and CAT and reducing the content of MDA.Water extracts and ethanol extracts of Pentamon improve mitochondrial function by down-regulating ROS and UCP2 content.They can a Iso prevent the telomere from shortening and anti-aging. |
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