| Part Ⅰ Theoretical ResearchObjective To exploring "hepatorenal homology" literature,ancient and modern famous doctor of traditional Chinese medical theory,combing the "hepatorenal homology" theory origin,preliminary clearly related to male reproductive use of traditional Chinese medical theory of "liver and kidney homology" prescription,"therapy with kidney and liver"general law,medicine,medical theory,etc.to explore the theory essence of the liver and kidney homologous,preliminary interpretation of the liver and kidney as "liver controlling dispersion and kidney reserving essence" scientific connotation of reproductive regulation.Methods To analysis the Qing,Han and Tang dynasties in ancient books of traditional Chinese medicine,during the Song,Jin,Yuan,Ming and Qing dynasties that representative physicians,medical books,mining theory of "liver and kidney homology" origin and evolution of law;study theory of various famous doctor medical research on male reproductive related with"hepatorenal homology" theory of "therapy with kidney and liver" prescription general law,medicine,medical theory,etc.① Using zhejiang university of Chinese medicine library collection and library union catalog of zhejiang province,by computer information query,to explore "hepatorenal homology","blood and essence homology"," therapy with kidney and liver" as subjects in professional literature database,combining with the consulting experts and the screening of related literature.Collecting Chinese ancient books of TCM such as<Huang di’s Canon of Medicine>,<Qianjin Yao Fang>,<General Collection for Holy Relief>,during the song,Jin,Yuan,Ming and Qing dynasties that representative physicians,medical books,summed up the prescription,medicine,medical theory aspects such as the universal law of mining "hepatorenal homology" theory origin and evolution of law;②Collecting the modern research related to male reproductive "hepatorenal homology" theory,"therapy with kidney and liver" prescription,medication and medical aspects of theoretical research and experimental research literature,summarized the theory of liver and kidney homologous modern research characteristics and general law.Results ①Literature of traditional Chinese medicine on liver and kidney homologous linked to male reproductive was recorded in<Huang di’s Canon of Medicine>,after Han and Tang dynasties in all previous dynasties,the acknowledgement in writing is rich,"hepatorenal homology"thoughts formed in the Ming dynasty,Li Zhongzi’s<essential readings for medical professionals required>was to establish a sign of "hepatorenal homology",in Qing dynasty science of seasonal febrile diseases focus on liver and kidney homologous with protecting fluid,modern physicians applied this theory in the reproductive ②In recent years,the experimental studies on " hepatorenal homology" focuses on depression,diabetes,osteoporosis,liver disease,infertility,modern disease research,from the hypothalamuspituitary-target axis(thyroid,thymus gland,adrenal gland and gonad)disease research.③The doctors of traditional Chinese medicine in the modern had done literature and empirical researches about "hepatorenal homology",but liver and kidney homologous associated with reproduction was still lack of systematic and deeper experimental research of traditional Chinese medicine.Conclusion ①Male branch diseases treated from liver and kidney disease has rich connotation,"hepatorenal homology" theory can effectively guide the clinical and experimental research of male infertility;②The "hepatorenal homology" theory from TCM viscera-state study as the foundation,deep into the network coordination level of gene expression,signal,further interpret the scientific connotation of the theory;③Liver and kidney as closely associated with male reproductive,male "hepatorenal homology" theory can guide clinical practice and experimental research,the mechanism of liver and kidney homologous reproductive regulation may be realized through network approach to regulation which include the endocrine,immunity,cell apoptosis,oxidative stress,etc.Part Ⅱ Experimental ResearchObjective To investigate the apoptosis of spermatogenic cell and the protective effect of Tongjingling decoction on spermatogenic function in testis of rats with varicocele by observing apoptosis of spermatogenic cell through mitochondrial pathway induced by ROS and JNK/P38MAPK signaling.Methods Experimental left varicocele was induced by partial ligation of the left renal vein in 90 Wistar rats,which were randomly divided into 6 groups as following:shame operation group,model group,Tongjingling groups with different dose,control group,with 15 cases in each group.After 4 weeks models were made successfully,medicine was given for two months.Tongjingling groups were given Tongjingling decoction 1 millilitre per 100 gram avoirdupois.(low dose comprise crude drug 0.6 gram per milliliter;middle dose comprise crude drug 1.2 gram per milliliter;high dose comprise crude drug 2.4 gram per millilitre),control group given crude drug 1.2g/ml,false operation group and model group were given the same dosage of normal saline.8 weeks later the rats were killed and the testes were selected,histopathology changes were observed by hematoxylin and eosin stain,Function of spermiogenesis was evaluated by Johnsen’s method and the changes of structure and ultrastructure of testis tissue were observed under light microscope or transmission electron microscope,JNK/P38MAPK mRNA was observed by RT-PCR;meanwhile apoptosis cells and expression of p-JNK/p-P38MAPK were detected by FCM and Western-bolt respectively;reactive oxygen species(ROS),Ca2+,mitochondrial trans membrane potential change,apoptosis and DNA content in spermatogenic cells all were detected by FCM,to measure the expressions of Cyt-C,AIF and Bax by IHC.Results ①The degree of histopathology changes in Tongjingling groups was markedly lower than that in mode group,such as basel membrane of contorted seminiferous tubules thickened,and spermatogenesis stoppage were lower than that in model group.②Stoppage of sperm cell cycles and apoptosis index in spermatogenic cells of mode group were distinctly higher than that in the sham operation group and Tongjingling groups(p<0.01),the contrast between Tongjingling groups and model group was manifest(p<0.05).③ROS content sperm of cells in Model group and Ca2+content were increased,mitochondrial membrane potential(MMP)decreased significantly,compared with control group,compared with model group had obvious difference(p<0.05),and elves dose groups in sperm cells ROS,Ca2+content is reduced,MMP rise,compared with model group,there are significant differences(p<0.05).④Cyt C,AIF,Bax protein expression of model group testicular tissue of rats;Compared with control group,is significant difference(p<0.01,p<0.05).Tong elves dose group of testicular tissue Cyt C,AIF,Bax protein expression decreased,compared with model group,control group,with significant difference(p<0.05).⑤JNK/p38MAPK mRNA expression of Tongjingling group and model group testicular tissue were lower,compared with control group,with significant difference(p<0.05);Model group JNK protein increased,p-p38MAPK higher protein phosphorylation,and,compared with control group,with significant difference(p<0.01,p<0.05),p-JNK/p-p38MAPK protein expression can be decreased in model group which have significant difference among control group,model group(p<0.05).Conclusion ①Tongjingling have negative effect on the apoptosis of spermatogenic cell,can relive the pathological harm of testicular,protect sperm cells mitochondria structure,and improve sperm production function in testis of experimental rats with varicocele;②Tongjingling can regulate sperm cell cycle,reduce apoptosis of spermatogenic related with mitochondria oxidative stresss,which is likely to be realized by the way to regulate sperm cells ROS,Ca2+,MMPS and Cyt C,AIF,Bax expression of testicular tissue;③Tongjingling can improve sperm production and decrease apoptosis of spermatogenic cell by the way that Tongjingling modulate cell mitochondria oxidative stress and JNK/p38MAPK signaling pathway activated by ROS mediated,which be concerned with sperm cells apoptosis,it may be one key mechanism of Tongjingling in improving testicular function in rats with varicodcle... |
PDF Full Text Request