Quantum-dot-based Fluorescent Labeling For Detection Of Heat Shock Protein70and90in Ovary Tissues In Rats Of PCOS And PCOS With Type2Diabetes Mellitus

Aim; Polycystic ovary syndrome(PCOS)is clinical syndrome which has a kind of endocrine disorder and a variety of heterogeneous metabolic abnormalities, affecting4%～8%women. The etiology and pathogenesis of PCOS is still unclear. The metabolic abnormalities should not be ignored and be treated as a significant part to prevent and treat for a long time. Now the mechanism of PCOS developing into type2diabetes mellitus(2-DM) and its pathological physiology characteristics are still not clear. Heat shock protein(HSP)is a group of highly conserved protein molecule family which all prokaryotic cells and eukaryotic cells can produce under physiological, pathological and environmental factors (such as high temperature, lack of oxygen or virus infection, stress). It has various biological functions. And several studies have showed that HSP has close relationship with the incidence and development of2-DM. However the expression of HSP have not been reported in PCOS with2-DM. Therefore the studies for the expression of HSP in rat’s ovary tissues with PCOS and PCOS with2-DM can further define HSP’s pathogenesis in PCOS and its long-term complications of possible mechanism. Quantum dot is a new type of fluorescent probe, which has broad application prospects in many biomedical fields. This article used QDs-SA605fluorescene elabeling and the traditional method immunohistochemical(IHC)to detect the HSPs in the ovaries of rats of PCOS and PCOS with2-DM, to explore its application characteristics when compared with IHC and to the possible mechanism of how HSPs worked in the mechanism of PCOS and PCOS with2-DM. And it also was a primary study on quantum dot fluorescent to provide the basis for further expanding its application field.Methods:Female rats aged23-day were divided into three groups:control group, PCOS group and PCOS with2-DM group. The rats in PCOS group and PCOS with2-DM group were injected daily S. C. with sodium prasterone sulfate90μg/(g· d) for20days, then PCOS with2-DM group were intraperitoneally injected streptozotocin once in a dose of45mg/kg. All rats were feed with high fat and sugar feed. The ovarian weight and pathological morphology was observed. The sera levels of sex hormones, blood glucose(BG)and insulin(INS)were measured. QDs-SA605fluorescene elabeling and IHC staining analysis of heat shock protein (HSP)70and HSP90expression was performed on ovary tissues respectively, and analysis the similarities and differences of QDs-SA605and the traditional marking method of IHC for detection of HSP70and HSP90.Results:1.The rats induced by sodium prasterone sulfate had polycystic ovaries and hyperinsulinemia, showed the morphologic and hormonal changes which were similar to patients with PCOS. The rats induced by the sodium prasterone sulfate and streptozotocin showed the morphologic and hormonal changes which were similar to patients with PCOS with2-DM.2.HSP70and HSP90were detected in the ovary tissues of rats, which were expressed in granular cells and in the cytoplasm majorly. The tendency of intensity of HSP70was control group less than PCOS group, and both less than PCOS with2-DM group, while the tendency of intensity of HSP90was control group approximate equal to PCOS with2-DM group, and both more than PCOS group.3. QDs-SA605labeling can detect the expressions of HSP70and HSP90, and showed the same location and tendency of intensity of these two proteins. Because of the strong comparison of red and green flsorescence in QDs-SA605labeling, the expressions of HSPs were observed easily. Although intensity of the fluorescence was attenuated, the fluorescence did not disappear almost, and the specimens were still visible clearly for30days.Conclusions:This animal model induced by the sodium prasterone sulfate and streptozotocin is a good model of PCOS with2-DM, but the stability and perdurability of this animal model are still needed to be verified. There are HSP70and HSP90expressed in ovary tissues both for PCOS and PCOS with2-DM. QDs-SA605labeling takes advantage of the fluorescence intensity to accurate display the expressions of HSPs, and the QDs fluorescence can maintain visible long enough to be potentially valuable for biomedical practical application.