| Objective: Established PCOS rats model by subcutaneously injecting DHEA and INS ; To compare the expression of Heat Shock Protein 27 and Heat Shock Protein 90 in ovary of PCOS group and control group for investigating the possible role of Heat Shock Protein 27 and Heat Shock Protein 90 in the pathogenesy of PCOS.Method: Twenty-five female Sprague-Dawley(SD)rats were studied. Ten control rats were injected subcutaneously with 0.2 ml sesame oil for 28 days, while the rest 15 rats were injected subcutaneously with dehydroepiandrosterone (DHEA) and Insuline for 28 days. When all rats are 70 days old, we give them 20 days of continuous vaginal smear, observing the variation of their estrous cycle. Ten rats with irregular estrous cycle were used as PCOS models. At the end of the vaginal smear Examination, all rats were sacrificed and ovaries removed. Aorta blood was collected and the blood serum was separated by centrifugation. Concentrations of serum T, E2, P, LH, FSH, FPG, FINS were determined , LH/FSH were calculated. One side of the ovaries were stored in -80℃liquid nitrogen ,then HSP27 and HSP90 in PCOS group and control group were measured and evaluated by semi-quantitative RT-PCR(reverse transcription polymerase chain reaction) and del image analysis system. Another side of the ovaries were fixed in 4% PFA ,then were HSP27 and HSP90 in PCOS group and control group were detected and evaluated by immunohistochemistry and image analysis system.Results: (1) The ovary weight of rats in PCOS models(42.45±0.45) was heavier than that of control group(28.07±0.29)(t = 29.513 , P<0.05). Ovarian follicles in each developmental stage were observed in control rats. The ovaries of rats treated with DHEA+INS demonstrated a PCOS-like appearance, which contained large follicular cysts with poorly developed granulosa cells and many atretic follicles oocyte and corona radiatea were disappered and little soma were found. Compared with the control group, the serum T concentration in PCOS group were increased (P<0.05), the serum LH concentration were increased(t = 51.998 , P<0.05), LH/FSH were increased(t = 38.62 , P<0.05),the serum P concentration were decreased(t = 6.195 , P<0.05), the serum E2 concentration were unaltered(t = 0.547 , P>0.05), the serum FSH concentration were unaltered(t = 1.438 , P>0.05). The level of FINS were significantly higher in PCOS group than that in control group(t = 74.302 , P<0.05). The level of FPG were higher in PCOS group than that in control group(t = 19.474 , P<0.05).(2)The expression of ovary HSP27 mRNA in PCOS group(0.743±0.027) was significantly higher than the level in control group(0.639±0.041)(t =6.699, P< 0.05); The expression of ovary HSP90 mRNA in PCOS group(0.499±0.064) was significantly higher than the level in control group(0.391±0.060)(t =3.893, P< 0.05).(3) HSP27 expression was observed in the oocyte of both primordial follicles and vesicular graafianae. In control group(0.697±0.044),the oocyte of both primordial follicles and vesicular graafianae were intensely stained,which was significantly more intense compared to the PCOS group (0.277±0.023)(t =26.751,P< 0.05);.In control group(0.808±0.066),HSP90 expression was observed in oocytes , granulosa cells and interstitium cells , these cells were intensely stained,which was significantly more intense compared to the PCOS group(0.301±0.028)( t = 22.362, P<0.05).Conclusion: (1) The expression of HSP27 in the control group were higher than that in PCOS group.(2)The expression of HSP27 in the control group were higher than that in PCOS group. |
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