| 1.Clinical partObjectiveThe research is to observe the influence of JJCFDTT combining with aerobic exercise on the body fat percentage and serum lipids blood sugar of patients with PCOS-IR based on previous studies on traditional Chinese and western medicine diagnosis and treatment scheme for PCOS combining with IR,and to further elaborate the important roles of traditional Chinese medicine on the treatment of PCOS-IR thus to provide a basis for clinical medication selection.MethodsThe patients accepted treatment at the Gynecology Outpatient Clinic of the First Affiliated Hospital of Guangzhou University of TCM from January 2016 to Oct.2016 was selected as the research object.The patients with phlegm-dampness syndrome PCOS-IR belonging to syndrome differentiation of traditional Chinese medicine were randomly divided into TCM treatment group and TCM+ aerobic exercise group.The therapeutic effects of TCM combining aerobic exercise on treatment of PCOS combining with IR were discussed through comparing the body weight,BMI values,body fat percentage,fasting glucose,fasting insulin value and IR values of the two groups.ResultsCases inclusion conditionsAccording to the diagnostic criteria,inclusion criteria and exclusion criteria,56 cases of patients complying with PCOS combining with IR from Jan.2016 to Oct.2016 were randomly divided into TCM treatment group and TCM+aerobic exercise group.Among them,there were 2 cases of patients in pregnancy and 1 case of patient fall-off in TCM treatment group;and there were 1 case of patient in pregnancy and 2 cases of patients fall-off in TCM+aerobic exercise group.The above cases were excluded.Therefore there were 25 cases of patients in both groups.(2)Basic conditions before treatmentAmong the 50 cases,a majority of them are aged ranging from 25-29 years old,approximately accounting for 38.8%.And the unmarried patients account for 35%.Two individual samples were selected for t test.The comparison of the age,stature,body weight,BMI value,fasting glucose,body fat percentage,fasting insulin value and IR of the TCM and TCM+ aerobic exercise group before the treatment shows that the difference was not statistically significant(P>0.05).(3)Research results of the therapeutic effects after treatment① The body weight(kg)of TCM treatment group is 63.24±5.932;and that(kg)of TCM+aerobic exercise group is 59.16±7.526;② The BMI value(kg/m2)of TCM treatment group is 24.77±2.08;and that(kg)of TCM+ aerobic exercise group is 23.16±2.66;③ The body fat percentage(%)of TCM treatment group is 27.12±3.04;and that(%)of TCM+ aerobic exercise group is 23.71±3.85;④ FBG of TCM treatment group(mmol/1)is 5.22±0.25;and that of TCM+aerobic exercise group is 5.07±0.22;⑤ Fasting insulin of TCM treatment group(mIU/ml)is 15.90±4.00;and that(mIU/ml)of TCM+aerobic exercise group is 13.48±4.43;⑥ The BMI value(kg/m2)of TCM treatment group is 3.68±0.88;and that(kg)of TCM+aerobic exercise group is 3.04±1.04;The differences on body weight,BMI values,body fat percentage,fasting insulin and IR value of both groups are of statistical significance(P<0.05).And the difference on fasting blood-glucose of the two groups is of no statistical significance(P>0.05).ConclusionJJCFDTT combining with aerobic exercise can significantly reduce the body fat percentage of phlegm-dampness type PCOS-IR patients,improve IR status,enhance the sensibility of insulin and reduce BMI.It is an effective scheme for prevention and treatment of PCOS-IR patients.The effect is superior to that of single TCM therapy.2.Experimental partObjectiveThe animal experiment is aimed to establish a PCOS-IR animal model thus to further discuss the impact of JJCFDTT combining with aerobic exercise on PI3K/PKB/mTOR signaling pathways of PCOS rats on the aspects of morphology,molecular biology and endocrinology thus to discuss the effective action mechanism.Methods(1)Establishment of rat PCOS combining with IR model(2)The cases were randomly divided into 8 groups(group 1:blank control group;group 2:model control group;group 3:aerobic exercise group;group 4:TCM dosage group;group 5:TCM dosage+aerobic exercise group;group 6:TCM high dosage group;group 7:TCM low dosage group;group 8:melbine group);different intervention measures were applied to different groups respectively.(3)Index inspection GOD method(Roche glucometer)is adopted to check the FBG.Radioimmunoassay is used to determine the fasting insulin,and calculate Homa IR;euzymelinked immunosorbent assay is to detect the LH,FSH and T in serum of rats;light microscope is used to observe the changes of ovarian tissue pathomorphism;Western Blot method is to detect the PI3K/PKB/mTOR protein level in rat skeletal muscle and ovarian tissue.Statistical analysisSPSS 22.0 software is adopted for statistical analysis.In case of equal variance,two independent samples were selected for t test of two groups of average values;ANOVA is adopted on multiple groups of average values;perform further pairwise comparison between the two groups by adopting Least Significant Difference t test.In case of heterogeneity of variance,rank sum test is adopted.ResultsRat PCOS combining with IR model was successfully established.The body weight,fasting insulin,fasting plasma glucose,testosterone value,follicle stimulating hormone and luteinizing hormone are higher than those of blank control group.And the difference is of statistical significance(P<0.05).It can be observed under the light microscope that the local form of ovary of control group is basically normal,and that corpus luteum and cumulus oospores coexist.Cystic expansion of follicles and immature small follicle are observed at the control group of model.The number of corpus luteum and cumulus oospores is obviously less than those of the blank control group which indicates that the control group of the model is free of ovulation or occasional ovulation.28 days after success modeling,the comparison of various indicators of blank control group and model control group is still of statistical significance(P<0.05)which indicates that COS-IR rat model is stable in the whole experimental process.(P<0.05)(一)After intervention treatment,the rat serum insulin resistance,internal secretion and body weight were compared.1.Body weightCompared with the blank control group,the body weight of the model group was significantly higher(P<0.05).Compared with the model group,body weights of the aerobic exercise group,the middle dose TCM group and the middle dose TCM+aerobic exercise group significantly decreased(P<0.05).Compared with the middle dose TCM+aerobic exercise group,the body weight of the metformin group was significantly higher(P<0.05).2.Fasting blood-glucoseCompared with the blank control group,fasting blood-glucose of the model group was significantly higher(P<0.05).Compared with the model group,fasting blood-glucose of each treatment group was significantly lower(P<0.05).3.Fasting insulinCompared with the blank control group,fasting insulin of the model group was significantly higher(P<0.05).Compared with model control group,fasting insulin of the the middle dose TCM + aerobic exercise group,the metformin group,the high dose TCM group,the middle dose TCM group and the low dose TCM group decreased significantly(P<0.05),and fasting insulin of the middle dose TCM group,the middle dose TCM+aerobic exercise group and the metformin group was significantly lower than that of the aerobic exercise group(P<0.05).4.Insulin resistance indexCompared with the blank control group,insulin resistance index of the model group was significantly higher(P<0.05).Compared with the model group,insulin resistance index of each intervention group was significantly lower(P<0.05),and insulin resistance index of the middle dose TCM group,the middle dose TCM+aerobic exercise group and the metformin group was significantly lower than that of the aerobic exercise group(P<0.05).5.Luteinizing hormoneCompared with the blank control group,luteinizing hormone of the model group was significantly higher(P<0.05).Compared with model control group,luteinizing hormone of the middle dose TCM+ aerobic exercise group decreased significantly(P<0.05).6.Follicle stimulating hormoneCompared with the blank control group,follicle stimulating hormone of the model group was significantly higher(P<0.05).Compared with the model group,the follicle stimulating hormone of the middle dose TCM+aerobic exercise group and the aerobic exercise group was significantly lower(P<0.05).Compared with the middle dose TCM+aerobic exercise group,follicle stimulating hormone of the high dose TCM group was significantly higher(P<0.05).7.TestosteroneCompared with the blank control group,testosterone of the model group was significantly higher(P<0.05).Compared with the model group,testosterone of the middle dose TCM group,the middle dose TCM+aerobic exercise group,the high dose TCM group and the low dose TCM group was significantly lower(P<0.05).Compared with the the middle dose TCM+aerobic exercise group,the testosterone of the aerobic exercise group was significantly higher(P<0.05).8.Changes of ovarian tissue pathomorphismThe local shape of ovarian tissue of the blank control group is normal.A large amount of corpus luteum and cumulus oophorus coexistence is observed.Cystic expansion of follicles and immature small follicle are observed at the control group of model.Theca cells thickening,corpus luteum,the number of corpus luteum,and granule cells are obviously less than those of control group.The local shape of TCM median dose+aerobic exercise group,and melbine group and blank control group are approximately close.However,the big follicles number of cystic dilatation of TCM median dose + aerobic exercise group and the immature small follicle are more than those of blank control group.The theca cells of melbine group and mesenchymal cell show obvious hyperplasia comparing to TCM median dose+ aerobic exercise group.The aerobic exercise group and TCM treatment groups with three concentration gradients present big follicles with cystic dilatation,mature follicle number and regular corpus luteum are significantly less than those of the control group.(三)Comparison of the relative transcript levels of rat skeletal muscle cells and ovary cells PI3K/PKB/mTOR proteinAfter 28-day treatment intervention,compare the model control group and blank control group.The protein expression level of PI3K/PKB has been significantly decreased(P<0.05).The mTOR protein expression level has been significantly increased(P<0.05).The PI3K,PKB and mTOR protein expression level of at skeletal muscle cells show basically consistent horizontal variation trend with PI3K,PKB and mTOR protein expression level of ovary cell.1.Expression level of PI3K/PKB protein in skeletal muscle tissuesCompared with the blank control group,the expression level of PI3K/PKB protein in skeletal muscle tissues of the model group significantly decreased(P<0.05).Compared with the model group,the expression levels of PI3K/PKB protein in skeletal muscle tissues of the low dose TCM group,the middle dose TCM group,the high dose TCM group,the aerobic exercise group,the middle dose TCM+aerobic exercise group and the metformin group were significantly higher(P<0.05).Compared with the the middle dose TCM+aerobic exercise group,expression levels of PI3K/PKB protein in skeletal muscle tissues of the low dose TCM group,the middle dose TCM group,the high dose TCM group and the aerobic exercise group were significantly lower(P<0.05).The expression levels of PI3K/PKB protein in skeletal muscle tissues of the high dose TCM group and the middle dose TCM group were significantly higher than that of the low dose TCM group(P<0.05).2.Expression level of PI3K/PKB protein in ovarian tissuesCompared with the blank control group,the expression level of PI3K/PKB protein in ovarian tissues of the model group significantly decreased(P<0.05).Compared with the model group,the expression levels of PI3K/PKB protein in ovarian tissues of the low dose TCM group,the middle dose TCM group,the high dose TCM group,the aerobic exercise group,the middle dose TCM+aerobic exercise group and the metformin group were significantly higher(P<0.05).Compared with the the middle dose TCM+aerobic exercise group,the expression levels of PI3K/PKB protein in ovarian tissues of the low dose TCM group,the middle dose TCM group,the high dose TCM group and the aerobic exercise group were significantly lower(P<0.05).The expression levels of PI3K/PKB protein in ovarian tissues of the high dose TCM group was significantly higher than those of the middle dose TCM group and the low dose TCM group(P<0.05).3.Expression level of mTOR protein in skeletal muscle tissueCompared with the blank control group,the expression level of mTOR protein in skeletal muscle tissues of the model group significantly improved(P<0.05).Compared with the model group,the expression levels of mTOR protein in skeletal muscle tissues of the low dose TCM group,the middle dose TCM group,the high dose TCM group,the aerobic exercise group,the middle dose TCM+aerobic exercise group and the metformin group were significantly lower(P<0.05).Compared with the the middle dose TCM+aerobic exercise group,the expression levels of mTOR protein in skeletal muscle tissues of the low dose TCM group,the middle dose TCM group,the high dose TCM group and the aerobic exercise group were significantly higher(P<0.05).The expression levels of mTOR protein in skeletal muscle tissues of the high dose TCM group was significantly lower than those of the middle dose TCM group and the low dose TCM group(P<0.05).4.Expression level of mTOR protein in ovarian tissuesCompared with the blank control group,the expression level of mTOR protein in ovarian tissues of the model group significantly improved(P<0.05).Compared with the model group,the expression levels of mTOR protein in ovarian tissues of the low dose TCM group,the middle dose TCM group,the high dose TCM group,the aerobic exercise group,the middle dose TCM+aerobic exercise group and the metformin group were significantly lower(P<0.05).Compared with the the middle dose TCM+aerobic exercise group,the expression levels of mTOR protein in ovarian tissues of the low dose TCM group,the middle dose TCM group,the high dose TCM group and the aerobic exercise group were significantly higher(P<0.05).Conclusions(一)Classical Poresky modeling method combining with high-fat diet can successfully induce stable obesity PCOS combining with IR rat model.(二)On the aspect of lose weight and serum laboratory tests,TCM median dose+aerobic exercise group and single aerobic exercise are superior to melbine group.On the aspect of improving the endocrine situation and IR,TCM median dose can play the same effects with that of melbine group.The effect is superior to that of all other treatment groups.The TCM treatment groups of three concentration gradients present no significant difference on the aspect of lose weight,improving internal secretion and IR.(三)On the aspect of ovarian morphologic changes,TCM median dose+aerobic exercise shows the optimal improvement status which indicates that TCM median dose+aerobic exercise is conducive to promoting the recovery of cystic follicles function,ovulation induction,and the improvement and maintaining of corpus luteum function.It mainly reflects on the recovery of promoting follicles cystic expansion,namely granular cell layers increasing,recurrence of cumulus oophorus and transparent tape,and increasing of corpus luteum number etc.The TCM treatment groups with three concentration gradients present no obvious difference on improving the ovary morphology of PCOS rat.(四)After the traditional Chinese medicine JJCFDTT combining with aerobic exercise working on skeletal muscle cell and ovary cell of PCOS-IR rat,the relative expression of the insulin signaling pathway protein PI3K/PKB can be up-regulated,and the relative expression of protein mTOR can be down-regulated.The action effects are equivalent to those of western medicine melbine.With the increasing of concentrations of different doses among TCM treatment groups,PI3K/PKB protein expression level has been gradually increased.mTOR protein expression level has been gradually decreased. |
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