Drug-induced Ovarian Depletion In Rat Model Induced By Tripterygium Wilfordii Construction And Intervention Study Of Tonifying Kidney Method

ObjectiveThe purpose of this study is to construct model based on a modified RA(rheumatoid arthritis) model, LRAM (Like rheumatoid arthritis model), of tripterygium wilfordii on drug-induced rat ovarian function loss model system platform; Preliminary exploration of tonifying kidney method and modified Shoutai pills on the ovarian function in rat models of protection and restoration of the role, for the study of drug-induced ovarian function loss model series and expand modified Shoutai pills clinical research lay the foundation.Methods and results1. LRAM female SD rats induced by tripterygium wilfordii drug-induced ovarian depletion model construction and estimation1.1LRAM rat inflammation model construction and estimationMethods11～12months age SPF level female SD rat40, are randomly divided into A、B、C and D four groups, each group of10, D is the blank control group,A,B and C three groups is the LRAM, like rheumatoid arthritis model, inflammation model group, each group of10.Takes Freund’s complete adjuvant0.2mL each time,injects in model group each rat left full pad,fully various0.1mL, the control group rat injects the0.1mL physiological saline, measures the rat full volume and rat quality, the rat vagina cast-off cell inspection, observes the rat ordinary circumstances, observes continuously for21days. Main outcome measures:Rat ordinary circumstances, rat full volume, rat oestral cycle. ResultsThree model group rat joints successively present the joint to be red and swelling, the joint motion barrier becoming impatient chronic inflammatory response, had no significant effects on the rat oestral cycle in rat model with stability and repeatability.1.2LRAM female SD rats induced by tripterygium wilfordii drug-induced ovarian depletion model construction and estimationMethods6months age SPF level female SD rat40, are randomly divided into A, B, C and D four groups, each group of10, A, B and C three groups carry on LRAM to produce the mold, and then, A, B two sets of tripterygium glycosides tablet (Glucosidorum tripterygll totorum, GTT) model with different doses, C as a model in the control group, does not participate in the GTT model, D for the controlgroup.Group A(LRAM+GTT75mg):LRAM:pick up0.2mL Freund’s complete adjuvant, injected within the group each rat foot pad on the left side, front and back foot0.1mL, measuring volume of rats, for21days in a row. Re-GTT75mg model:75mg/kg/dGTT after intragastric and intragastric administration of lmL/100g daily scheduled testing quality, vaginal exfoliated cells in experimental rat behavior observation and, for30consecutive days.Group B(LRAM+GTT50mg):LRAM:(as same as Group A). Re-GTT50mg model:50mg/kg/d GTT intragastric, intragastric administration of1mL/100g daily scheduled testing quality, vaginal exfoliated cells in experimental rat behavior observation and, for30consecutive days.Group C(LRAM control group):LRAM:(as same as Group A). Then,as a model in the control group, with3mL/d gastric saline irrigation, timing detection quality of laboratory rats, daily behavioral observation and exfoliated cells of the vagina, for30days.Group D(Blank control group):with3mL/d gastric saline irrigation, timing detection quality of laboratory rats, daily behavioral observation and exfoliated cells of the vagina, for30days.ResultsThere are rat ovarian structure and features real injury and ovarian reserves features declined on Group A&Group B, rat uterine injury degree more light. LRAM female SD rats induced by tripterygium wilfordii drug-induced ovarian depletion model has stability and can be repeatd.2. Healthy female SD rats induced by tripterygium wilfordii drug-induced ovarian depletion model construction and estimation Research purposes:as a parallel control model for experiment1, provides controlled parameters for experiment1.Methods6months age SPF level healthy female SD rat30, are randomly divided into A, B, and C three groups, each group of10, where A, B set to GTT modules with different doses, C is set for the control group.Group A (GTT75mg):75mg/kg/dGTT intragastric, intragastric administration of lmL/100g daily scheduled testing quality, vaginal exfoliated cells in experimental rat behavior observation and, for30consecutive days.Group B(GTT50mg):50mg/kg/d GTT intragastric, intragastric administration of lmL/100g daily scheduled testing quality, vaginal exfoliated cells in experimental rat behavior observation and, for30consecutive days.Group C(Control Group):with3mL gastric saline irrigation, for30consecutive days.ResultsThere are rat ovarian structure and features real injury and ovarian reserves features declined on Group A&Group B, rat uterine injury degree more light. Healthy female SD rats induced by tripterygium wilfordii drug-induced ovarian depletion model has stability and can be repeatd.3. Designing drug-induced ovarian depletion in rat model induced by tripterygium wilfordii based on age-related principle Methods3, and6, and11-12months age SPF level female SD the40only, the age paragraph rat respectively random is divided into Ai, and Bi, and Ci, and Di four group, each group10only, which, Ai, and Bi two group for LRAM made model, then, Ai, group again for GTT75mg dose made model, Bi group as model control group, Di as blank control group, not participation GTT made model, Ci directly for GTT75mg dose made model, i=3,6and12.Group Ai(LRAM+GTT75mg):LRAM:pick up0.2mL Freund’s complete adjuvant, injected within the group each rat foot pad on the left side, front and back foot0.1mL, measuring volume of rats, for21days in a row. Re-GTT75mg model:75mg/kg/dGTT after intragastric and intragastric administration of1mL/100g daily scheduled testing quality, vaginal exfoliated cells in experimental rat behavior observation and, for30consecutive days.Group Bi (LRAM control group):LRAM:(as same as Group A). Then, as a model in the control group, with3mL/d gastric saline irrigation, timing detection quality of laboratory rats, daily behavioral observation and exfoliated cells of the vagina, for30days.Group Ci(GTT75mg):75mg/kg/dGTT after intragastric and intragastric administration of1mL/100g daily scheduled testing quality, vaginal exfoliated cells in experimental rat behavior observation and, for30consecutive days.Group Di (Control group):with3mL/d gastric saline irrigation, timing detection quality of laboratory rats, daily behavioral observation and exfoliated cells of the vagina, for30days.ResultsThere are rat ovarian structure and features real injury and ovarian reserves features declined on Group Ai&Group Ci, i=3,6, and12and there are clear difference in injury between different age groups and different designing style of drug-induced ovarian depletion in rat model induced by tripterygium wilfordii.4. Effect of Modified ShouTai pills antagonist of tripterygium wilfordii on ovarian function in healthy female SD ratMethods6months age SPF level healthy female SD rat50, are randomly divided into A, B, C, D and E five groups, each group of10, where A, B and C three groups respectively with modified shoutai pill high dose, the low dose and estradiol valerate piece conventionaldosage fill the stomach, carries on GTT75mg to produce the mold again synchronously, D group directly carry on the GTT75mg dosage to produce the mold, E is the blank control group, dose not participate to produce the mold.ResultsModified Shoutai pills early the antagonism intervention Tripterygium wilfordii reproductive toxicity, can maintain the healthy female SD rat ovary structure is basically complete, maintains or restores some model rat oestral cycles, obviously increases the model rat primordial follicle counting average value level, obviously declines model rat blood serum FSH and FSH/LH ratio average value level, obviously declines the GDF-9mRNA relative expression quantity average value level.5. Ovarian protective effect of Modified Shoutai pills on LRAM and healthy female rat induced by Tr ipterygium Wilfordii Hook source ovarian function loss modelMethods6months age SPF level female SD rat100, are randomly divided into A1, A2, B1, B2, C1, C2, D1, D2, E and F groups,10groups, each group of10.1suppose based on the healthy female SD rat,2suppose based on LRAM female SD rat. A1and A2suppose for Modified Shoutai pills high dose group, B1and B2suppose for Modified Shoutai pills low dose group, C1and C2suppose for estradiol valerate tablets regular dose group, D1and D2suppose for GTT75mg model and LRAM+GTT75mg model, E suppose is the LRAM model control group,F suppose to share the blank control group.A1,B1,C1and D, group daily morning9when timing detection experimental rat quality, and then to75mg/kg/dGTT gavage, gastric perfusion of1mL/100g, and vaginal cytology and behavior observation, for30consecutive days, respectively modified with perfusion of high dose of Modified Shoutai pills solution, low dose of Modified Shoutai pills solution, estradiol valerate tablets solution and3mL physiological saline,30consecutive days.A2, B2, C2, and D2group were first made LRAM:apply0.2mL Freund’s complete adjuvant was injected into the left side, hin the group of each rat foot pad, and the measurement of0.1mL, rat paw volume, continuous observation for21days. Then, the daily morning9when timing detection in experimental rats with gastric perfusion of quality,75mg/kg/dGTT, gastric perfusion of1mL/100g, and vaginal cytology and behavior observation, for30consecutive days, then change to the perfusion of high dose of Modified Shoutai pills solution, low dose of Modified Shoutai pills solution, estradiol valerate tablets solution and3mL physiological saline,30consecutive days. E group:LRAM:(with A2), then, the daily morning9when timing detection experimental rats fed with3mL quality, physiological saline, and vaginal cytology and behavior observation, for60consecutive days. Group F:daily morning9when timing detection experimental rats fed with3mL quality, physiological saline, and vaginal cytology and behavior observation, for60consecutive days.Results Modified Shoutai pills late rehabilitation intervention of Tripterygium wilfordii reproductive toxicity have certain effect, it can repair model rat ovarian structure and reproductive function, the onset point Modified Shoutai pills cumulative dosage and the model itself is closely related to the degree of influence of Tripterygium wilfordii reproductive toxicity. Modified Shoutai pills late repair intervention on two models made repair effect induced by Tripterygium Wilfordii Hook ovarian ovarian function in rat model of drug-induced loss difference.Conclusion1. This is the first time in the like RA model (LRAM) based on the established model of ovarian function loss in SD rats. This composite model can reflect the reproductive toxicity of therapeutic drugs, simulate the clinical phenomenon, is a method of molding a convenient and stable. It is proved that this drug-induced ovarian damage model based on POF model of disease with Tripterygium healthy female rats based on differences. The main effect of Tripterygium wilfordii reproductive toxicity target organ for ovarian, uterus had no significant effect on the;2. On the basis of Tiangui time limit theory, age-related principle, rat source ovarian function loss model induced by the establishment of three ages. The theory of traditional Chinese medicine thinking in experimental study, functional ovarian drug-induced loss characteristics simulation of different age groups, provide an ideal platform to study the pathogenesis of further study in different age groups of ovarian dysfunction and intervention program.3Construction Based on Tripterygium wilfordii drug-induced ovarian function loss model of LRAM based on the day and Tripterygium wi Ifordii ovarian function loss model and Decyl time limit (age-related principle), and establish the evaluation system of ovarian function in a rat model of platform, formed more complete, for further research on drug source provides the technical specification of ovarian function loss disease;4Tonifying kidney can effectively protect the ovarian function in rats from or reduce drug reproductive toxicity damage, Modified Shoutai pill is an effective prescription for tonifying kidney, ovarian function in rats model of maintenance, defense and repair of exogenous drugs reproductive toxicity damage on ovarian function.