| Objective:to control the quality of Cistanche deserticola from different producing areas,to study the therapeutic effect of Cistanche deserticola on rat testicular injury model and to explore the authenticity of Cistanche deserticola.Methods:1.Quality control of Cistanche deserticola from different producing areas:according to the method of Chinese Pharmacopoeia(2015 edition),Cistanche deserticola from Alashan,Kashgar,Aksu,Gansu and other producing areas of Inner Mongolia were identified by emblem and TLC.The content of echinacein in the medicinal materials was determined by high performance liquid chromatography(HPLC).two。.Pharmacological experiment(1)SPF male Wistar rats were randomly divided into 6 groups:blank group,cisplatin(CP)model group,positive control DHEA group(5.166mg/kg),high,middle and low dose groups of Cistanche deserticola,with 10 rats in each group.After 15 days of administration,the animals were killed,and the related indexes such as serum,testicular tissue and epididymal tail sperm were detected.Except the blank group,the other groups were injected intraperitoneally with CP of 6mg/kg on the 10th day.(1)Indexes of modeling:the pathological sections of testicular tissue were prepared routinely,and the pathological changes were observed under microscope after HE staining.(2)testicular function:sperm concentration and motility were measured by Nikon DS microscope and NIS-Elements software.(3)oxidative stress index:the index of peroxidative stress was detected by Q-PCR method and CAT,enzyme labeling method was used to detect the index of peroxidative stress reaction by SOD,visible spectrophotometry and CAT,enzyme labeling method.(4)using biochemical method to detect malondialdehyde(MDA)and superoxide dismutase(SOD)in testicular tissue.(5)Mitochondrial stability:the content of ROS and the expression level of mRNA in testicular tissue were detected by chemical fluorescence assay.3.Pharmacological experiment(2)SPF male Wistar rats were randomly divided into 7 groups with 10 rats in each group:blank group,cisplatin(CP)model group,positive control DHEA group(5.166mg/kg),Inner Mongolia Alashan,Xinjiang Kashgar,Xinjiang Aksu,Gansu and other producing areas.The experimental method is the same as the step pharmacological experiment(1).Results:1.Quality control of Cistanche deserticola from different areas:the medicinal materials all meet the identification standard of Chinese Pharmacopoeia(2015 edition);the microscopic identification results show that according to the microscopic characteristics of wood powder,there are reticulated ducts,threaded ducts,wood fibers and starch grains and other specific characteristics and easy to be detected;thin layer identification method was used to identify the same color spots in the chromatogram of Cistanche deserticola samples from different areas.The average content of echinacein in Cistanche deserticola from different producing areas was 0.5307mg/g per cent,that of Cistanche deserticola in Aksu was 0.3274 mg/g,that of Cistanche deserticola in Gansu was 0.2986mg/g per cent,and that of Cistanche deserticola in Kashi was 0.3268mg/g per cent.two。.In pharmacological experiment(1):(1),the testicular weight of CP model group was significantly lower than that of blank group(P<0 05).From the pathological section,the basal membrane of seminiferous tubules of the model testis was significantly thickened,the arrangement of spermatogenic cells and Sertoli cells were irregular and the levels decreased,the number of mature sperm in the lumen decreased,and testicular tissue injury occurred.It is proved that the CP modeling is successful.(2)the weight of testis increased after administration,but the recovery degree of DHEA group and Cistanche deserticola low and high dose group was not obvious(P>0.05),(3).From the results of pathological sections,the testicular tissue injury of the middle dose group recovered,the structure,arrangement,density and hierarchical integrity of spermatogenic cells and Sertoli cells were close to those of the blank group,and the number of mature spermatozoa in the lumen could be observed.(4)compared with the blank group,the sperm motility and concentration of the model group decreased(P<0.05),which proved that the testicular tissue function of the model group was weakened.After administration,the sperm motility in the treatment group was significantly higher than that in the model group(P<0.01).However,the concentration of sperm in the middle dose group was significantly higher than that in the model group(P<0.05),but there was no significant difference in other groups(P>0.05).(5)oxidative stress index:compared with the blank group,the contents of MDA and GSH in serum and testicular tissue in the model group increased,and the expression level of SOD GSH CAT T-AOC decreased significantly in the model group,indicating that the oxidative stress in the model group was enhanced.After administration,compared with the model group,the contents of MDA and ROS in the middle dose decreased,and the expression level of SOD "GSH"CAT "T-AOC increased significantly(P<0.05).3.In pharmacological experiment(2):(1),the testicular weight of Inner Mongolia Cistanche group increased after administration,but the recovery degree of DHEA group and other producing area Cistanche group was not obvious(P>0.005),(2).From the results of pathological sections,the testicular tissue damage of Inner Mongolia Cistanche group recovered,and the structure,arrangement,density and hierarchical integrity of spermatogenic cells and Sertoli cells were close to those of the blank group.A significant increase in the number of mature spermatozoa in the lumen was observed.(3)after administration,the sperm motility of Cistanche deserticola group in Inner Mongolia was significantly higher than that of Cistanche deserticola group from other producing areas(P<0.01).There was no significant difference in other groups(P>0.05).(4)compared with the model group,the content of MDA,ROS in serum and testicular tissue in Inner Mongolia Cistanche group decreased and the expression level of SOD "GSH" CAT "T-AOC increased significantly in Inner Mongolia Cistanche group.Conclusion:1.the model of testicular injury induced by CP was established successfully.2.in the quality control of Cistanche deserticola from Alxa,Inner Mongolia,Kashgar,Aksu,Gansu and other producing areas,the quality of Cistanche deserticola in Inner Mongolia is good,and the content of echinoside is higher than that of other groups.3.the model of testicular injury induced by CP is a typical model of oxidative stress.Medium dose of Cistanche deserticola could weaken the effect of oxidative stress.4.Inner Mongolia Cistanche deserticola has excellent therapeutic effect on testicular injury through pharmacological experiments.5.this experiment provides an effective basis for the authentic character of Cistanche deserticola in Alashan,Inner Mongolia. |
PDF Full Text Request