论文部分内容阅读
目的:研究多囊卵巢综合征(PCOS)患者脂肪组织瘦素受体后信号转导分子——JAK2和STAT3蛋白的表达及酪氨酸磷酸化程度以及JAK2激酶活性变化,探讨PCOS瘦素抵抗的分子机制。方法:采用酶联免疫吸附法(ELISA)检测PCOS合并胰岛素抵抗(IR)患者23例(PCOSIR组)、PCOS非IR患者18例(PCOS非IR组)及单纯子宫肌瘤患者23例(对照组)的血清瘦素水平;采用放射免疫法检测各组空腹血清胰岛素(FIN)水平,利用稳态模型(HOMA)计算IR指数(HO-MA-IR);采用免疫印迹法检测脂肪组织JAK2和STAT3蛋白表达及磷酸化程度;应用免疫沉淀、薄层层析及γ液闪计数方法检测脂肪组织JAK2激酶活性,并进行分析比较。结果:①PCOSIR组血清瘦素水平为(19.63±4.16)μg/L,明显高于PCOS非IR组的(15.54±4.26)μg/L和对照组的(15.29±2.56)μg/L,差异均有统计学意义(P<0.01);PCOS非IR组与对照组比较差异无统计学意义(P>0.05);②PCOSIR组JAK2和STAT3的蛋白表达分别为1.42±0.26和2.33±0.47,PCOS非IR组分别为1.36±0.18和2.10±0.24,对照组分别为1.29±0.11和2.45±0.36,3组比较差异均无统计学意义(P>0.05);③PCOSIR组JAK2和STAT3蛋白酪氨酸磷酸化程度分别为1.87±0.16和3.32±0.17,均明显低于PCOS非IR组的4.12±0.68和5.96±0.44及对照组的4.27±0.34和6.15±0.64,差异均有统计学意义(P<0.01);PCOS非IR组与对照组比较差异无统计学意义(P>0.05);④PCOSIR组JAK2激酶活性较PCOS非IR组和对照组均明显下降,差异均有统计学意义(P<0.01),PCOS非IR组与对照组比较差异无统计学意义(P>0.05)。结论:PCOSIR患者同时存在瘦素抵抗,其原因可能为瘦素受体后信号转导分子JAK2酪氨酸磷酸化受损,使JAK2活性降低,导致STAT3酪氨酸磷酸化异常和瘦素受体后信号转导障碍。
Objective: To study the expression of leptin receptor signal transducers JAK2 and STAT3, tyrosine phosphorylation and JAK2 kinase activity in adipose tissue of patients with polycystic ovary syndrome (PCOS) Molecular mechanism. Methods: Twenty three patients (PCOSIR group) with PCOS combined with insulin resistance (IR), 18 PCOS non-IR patients (PCOS non-IR group) and 23 patients with simple uterine fibroids were detected by enzyme linked immunosorbent assay (ELISA) ). The fasting serum insulin (FIN) was measured by radioimmunoassay. HO-MA-IR was calculated by steady-state model (HOMA). The expressions of JAK2 and STAT3 Protein expression and phosphorylation were measured. The activity of JAK2 kinase in adipose tissue was detected by immunoprecipitation, TLC and γ-liquid scintillation counting and analyzed. Results (1) Serum leptin level in PCOSIR group was (19.63 ± 4.16) μg / L, which was significantly higher than that in PCOS non-IR group (15.54 ± 4.26) μg / L and control group (15.29 ± 2.56) μg / L (P <0.05); ② The protein expression of JAK2 and STAT3 in PCOSIR group were 1.42 ± 0.26 and 2.33 ± 0.47, respectively, and PCOS non-IR group (1.36 ± 0.18 and 2.10 ± 0.24 respectively), while the control group was 1.29 ± 0.11 and 2.45 ± 0.36 respectively (P> 0.05). ③ The levels of tyrosine phosphorylation of JAK2 and STAT3 protein in PCOSIR group were respectively 1.87 ± 0.16 and 3.32 ± 0.17, respectively, which were significantly lower than those in PCOS non-IR group (4.12 ± 0.68 and 5.96 ± 0.44) and in control group (4.27 ± 0.34 and 6.15 ± 0.64) (P <0.01) There was no significant difference between non-IR group and control group (P> 0.05) .④The activity of JAK2 kinase in PCOSIR group was significantly lower than that in PCOS non-IR group and control group, the differences were statistically significant (P <0.01) There was no significant difference between the two groups (P> 0.05). Conclusions: The simultaneous presence of leptin resistance in PCOSIR patients may be due to impaired tyrosine phosphorylation of signal transducer JAK2 after leptin receptor, which leads to the decrease of JAK2 activity, resulting in abnormal tyrosine phosphorylation of STAT3 and leptin receptor Post-signal transduction disorders.