| Objective:In this experiment, the polycystic ovary syndrome(PCOS) model of rat were made.After successfully established the model, the Ganshao Decoction were used on the rat model.In order to explore the effect of Ganshao Decoction on PCOS forn the animal experimental method.,the sex hormone level, blood glucose, liver transaminase and ovarian tissue morphology of the PCOS rat were observed.Methods:The first part: Seventy SD female non pregnant rats,aged 6 weeks, in regular estrous cycle,were divided into two groups randomly,20 rats in the normal group(1%CMC1ml/100g/d intragastric administration, 21d) and 50 rats in the model group(letrozole 1mg/kg/d, intragastric administration,21d).Polycystic ovary syndrome(PCOS) rat models were established by letrozole(1mg/kg/d, 21d).Each group of rats were made stain smears daily to observe the changes of estrous cycle.After letrozole intragastric administration 3 weeks,the blood was collected from tail, centrifugal settling, the serum was taken, then we use automatic biochemical analyzer to detect the fasting blood glucose, serumalanine aminotransferase(ALT),aspartate aminotransferase(AST) of all rats; and use chemiluminescence method to measure the sex hormone levels in all rats;lastly we choose two rats form each group at random;the ovaries of which were resected and were put into 10% formalin for 24 hours. And then,tissue sectioned and HE dying. Mor-phologic changes were observed by ophthalmoscopy.The second part: The successful established model rats,were randomly divided into three groups:the PCOS model group(1%CMC1ml/100g/d, 21d),the Diane-35group(0.2mg/kg/d,21d) and Ganshao Decoction group(20mg/kg/d, 21d), 15 rats in each group,the normal group(1%CMC1ml/100g/d 21 d in total), after 3 weeks’ drug intervention,The blood was collected from tail, centrifugal settling, the serum was taken. Thenthe serum were detected the sex hormone levels, ALT, AST levels and fasting glucose.Finally,double ovaries of all the rats were resected and were put into 10%formalin for 24 hours. And then,tissue sectioned and HE dying.Mor-phologic changes were observed by ophthalmoscopy.Results:1.PCOS rat models were established by letrozole. Twelve days later,those rats were out of regular estrous cycle. Though the light microsocpe,leucocyte and a few keratinized cells were seen in the vagina smear,which pointed out that rats were at diestrus. Twenty-one days later, the level of serum sex hormone was tested. The level of LH was 6.28±0.64 m IU/ml, FSHwas 2.09±0.44 m IU/ml and T was 2.26±0.29ng/dl,the level were all higher than the normal control group, but the level of E2 was 55.21±5.83pg/ml, P was 18.56 ±2.95ng/ml,which were all lower than the normal control group,those pointed out the PCOS rat models were successfully established.2. After all kinds of drug intervention, in Chinese medicine group(Ganshao Decoction group) the level of LH was 3.91±0.47 m IU/ml,the level of T was 1.32 ± 0.27ng/dl, and the level of FSH was 1.15 ± 0.22 m IU/ml.Compared with the PCOS model group,the level were decreased significantly(P<0.05);Compared with the PCOS model group,in the western medicine group(Diane-35group) the level of LH was 4.52± 0.37 m IU/ml, the level of T was 1.64 ±0.34ng/dl, the level of FSH was 1.15± 0.22 m IU/ml, all levels were decreased significantly(P<0.05);Compared with the PCOS model group: in Ganshao Decoction group the level of E2 was 71.06 ± 4.15pg/ml, the level of P was 28.46 ±1.91ng/ml,both were increased significantly(P<0.05); inthe Diane-35 group, the level of E2 was 67.01± 2.87pg/ml, the level of P was 23.77±2.25ng/ml,both were increased significantly(P<0.05);Compared with the Diane-35 group,in Ganshao Decoction group the level of LH and T were decreased in significant difference(P<0.05), the level of E2 and P were increased significantly(P<0.05),the level of FSH was decreased with no significant difference(P >0.05).3.Morphological changes of ovarian tissue:Compared with the normal control group: the ovarian tissue of the model group rats’ looked pale,there were many vesicular structure on the surface; under the microscope, The majority of the follicle sinus increased significantly and became cystic dilatation, at the same time,the follicle oocytes and corona radiata disappeared,the layer of the granule cell layers decreased to 2 ~ 3.There was a fewvisible corpus luteum and most undevelopedmature small follicles, but the mature follicle and ovulation can’t be seen.Compared with the model group, Ganshao Decoction group rats were with ruddy ovarian surface and thinner white coating. Under the light microscope, there were a large number of mature follicles, with gracile follicular thecacell layer and the granule cell layer thickening to 6 ~ 8.There was rarely cystic immature follicle can be seen.Compared with the PCOS model group,the Diane-35 group rats were with ruddy ovarian appearance.Under the light microscope,there are many mature follicle,with clear structure and multilayer granular cell layer of morphologically intact, neatly arranged. There was no cystic follicular, but the number of the luteal and follicular were more than the PCOS model group。4.Changes of blood glucose and glutamic pyruvic transaminase levels :Compared with the normal control group, the level of the blood glucose of the PCOS model group was 5.91±0.68mmol/L,the level of ALT was 61.2 ± 9.89U/L, the level of AST was 219.91 ±31.27U/L level, which were increased significantly(P<0.05);Compared with the PCOS model group: the level of the blood glucose of the Ganshao Decoction group was 4.90±1.06mmol/L, the level of ALT was 65.93±5.48U/L, the level of AST was 195.091±18.11U/L,which were increased significantly(P<0.05);Compared with the PCOS model group: the level of the blood glucose of the Diane-35 group was 5.08±0.63mmol/L,the level of ALT was 71.47± 3.11U/L, which were increased significantly(P<0.05);But the level of AST was 239 ± 14.23U/L,which was no significant difference between the two group.(P>0.05);Compared with the Diane-35 group,the level of blood glucose of the Ganshao Decoction group changing had no significant difference(P>0.05), but the level of ALT and AST were decreased significantly(P<0.05).5. Changes of body weight in rats The average body weight of rats was 160±2.5g, after 21 days the weight of the normal control group was 180.33±2.58 g, increased 17.87±3.81 g,meanwhile,the weight of the PCOS model group was 195.4±4.64 g, increased 32.91±5.19 g. Compared withthe normal control group, there was significant difference of body weight increased in the rats of the PCOS model group(P<0.05)Also,the body weight growth increased greatly.(P<0.05).After all kinds of medicine intervention; the weight of the normal control group was208.67±3.04 g, the weight of the PCOS model group was 213.47±3.02 g, the weight of the Ganshao Decoction group was 208.53±3.18 g,the weight of the Diane-35 group was 206.6 ±3.74g; compared with the normal control group, the weight of the PCOS model group increased significantly(P<0.05), the weight between the remaining three groups had no significant difference(P>0.05).Conclusion:1.The experimental results indicate that it is a reliable,fairly ideal and high successful rate method to establish the polycystic ovary rat models by letrozole.2 Gan Shao Tang can reduce the level of FSH, LH, T and bloodas to stimulate follicular development and ovulation. This is maybe the glucose of the PCOS model group.In Gan Shao Tang trestment,the ovay issue are changed, so mechanism of Compound Ganshao patch to stimulate ovulation.3 Compared with the PCOS model group and Diane-35 group, the level of AST and ALT of the Ganshao Decoction group droped obviously(P<0.05), suggesting that this decoction has protective effect on the liver function, reflects the role of the advantages of traditional Chinese medicine. |
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