雌激素对大鼠咬肌ER和MHC mRNA表达的影响
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摘要
颞下颌关节紊乱病是口腔临床中的常见病和多发病，临床上常表现为颌面部的疼痛不适，病程迁延不愈，反复发作。TMD中青年女性发病率显著高于男性，并且多表现为肌功能紊乱。雌激素是一种作用广泛的类固醇类激素，也是女性最重要的激素之一。近来的研究发现雌激素对四肢骨骼肌的纤维构成、代谢等也有不同程度的影响，但对咀嚼肌研究的报道甚少。
本研究通过雌性未孕12周龄SD大鼠60只，随机分成3组：假手术组(Sham)、去卵巢组(Ovx)，雌激素替代组(去卵巢后加戊酸雌二醇替代组，Ovx／EV)，各20只。分别在实验开始4周及8周后，处死每组10只。将大鼠咬肌雌激素受体ERα和ERβ的mRNA，以及肌球蛋白重链MHC-Ⅰ、Ⅱa、Ⅱx和Ⅱb型的mRNA作为研究对象，利用SYBR GreenⅠ荧光实时定量PCR技术，来研究雌激素对咀嚼肌功能的影响，对颞下颌关节紊乱病的雌激素病因及治疗新途径进行初步探索。
根据实验观察得出以下结果：
1．大鼠咬肌组织有ERα和ERβ的表达，其中ERα表达主要在肌细胞核，表达强度为阳性。ERβ主要表达在肌细胞核和毛细血管内皮细胞核，表达强度为强阳性。但是大鼠咬肌ERαmRNA的表达明显强于ERβmRNA的表达。
2．大鼠咬肌组织MHC-Ⅱb mRNA的表达较丰富。
3．用大鼠去卵巢，结合雌激素替代法，可以建立雌激素对大鼠咬肌功能影响的实验动物模型。
4．4周去卵巢组ERαmRNA表达上调，与4周组的假手术组及雌激素替代组相比有统计学意义。其余组间没有统计学差异。
5．4周去卵巢组ERβmRNA表达上调，与8周去卵巢组，以及4、8周组的假手术组和雌激素替代组相比有统计学意义。其余组间没有统计学差异。
6．MHC-ⅠmRNA各组之间相比无统计学差异。
7．8周去卵巢组MHC-Ⅱa mRNA表达上调，与4、8周假手术组和去卵巢组，以及与4周雌激素替代组相比有统计学意义。其余组间没有统计学差异。
8．8周去卵巢组MHC-Ⅱx mRNA表达上调，与4、8周假手术组和去卵巢组，以及与4周雌激素替代组相比有统计学意义。其余组间没有统计学差异。
9．MHC-Ⅱb mRNA各组之间相比无统计学差异。
综上所述，本研究认为大鼠咬肌是雌激素的靶组织，雌激素水平下降，ERα和ERβmRNA的表达有上调后又下降恢复的趋势，而MHC-Ⅱa和Ⅱx mRNA表达上调。研究提示雌激素对大鼠咀嚼肌的功能有影响。
Temporomandibular disorder (TMD), characterised by pain and recurrence, is one of the most commonly seen diseases in dentistry clinic. The prevalence of TMD, mostly identified as functional disturbances in the masticatory system, has been well documented to be higher in young females than in males. Estrogen, a steroid compound with multifunctions, is one of the most important hormones in women. The effects of estrogen on the composition and metabolism of myofiber in limb skeletal muscles have been investigated extensively in recent years. However, the effect of estrogen on masticatory muscles has been rarely studied. This study is therefore undertaken to examine this aspect and to provide new data regarding the possible role of estrogen in the development of TMD.
Sixty twelve-week old female Sprague-Dawley rats were randomly divided into 3 groups: (1) control (Sham surgery), (2) ovariectomy without estrogen replacement (Ovx) treatment, (3) ovariectomy with estradiol valerate replacement (Ovx/EV) treatment. Half of the animals were sacrificed at 4 and 8 weeks, respectively, after surgery and the masseter muscles were removed for analysis. Real time PCR quantification with SYBR Green I was employed to study the mRNA expression of estrogen receptor alpha (ERα) and beta (ERβ), myosin heavy chain (MHC) I、IIa、IIx and IIb .
The results showed:
1. Expression of ERαand ERβwas detected in normal rat masseter muscle with ERαmainly in myonuclei while ERβin both myonuclei and endothelial cell nuclei of blood capillaries. ERαmRNA expression was stronger than ERβmRNA.
2. Strong expression of MHC- IIb mRNA in masseter of rat was found.
3. Ovx/EV is a good animal model for studying the effect of estrogen on rat masticatory function.
4. At week 4, the expression of ERαmRNA was significantly higher in the Ovx group than in the Ovx/EV group.
5. The expression of ERβmRNA in the Ovx group at week 4 was significantly higher than at week 8. The expression of ERβmRNA in the Ovx group at week 4 was also significantly higher than the sham and Ovx/EV groups at both week 4 and week 8 time points.
6. MHC-I mRNA expression did not differ among groups.
7. MHC-IIa mRNA expression was higher in the Ovx group at week 8 than its expression in the Sham, Ovx, Ovx/EV groups at week 4 and the sham group at week 8.
8. MHC-IIx mRNA expression was higher in the Ovx group at week 8 than its expression in the Sham, Ovx, Ovx/EV groups at week 4 and the sham group at week 8.
9. MHC-IIb mRNA expression was not significantly different among groups.
The results suggest that the rat masseter muscle is one of the targets of estrogen.In response to estrogen level decrease, the expression of ERαand ERβmRNA tend to be upregulated initially and then be restoreded, while the expression of MHC- IIa and IIx mRNA tend to be upregulated constantly. It is concluded that estrogen does have effect on the masticatory function of female rats.
本研究通过雌性未孕12周龄SD大鼠60只，随机分成3组：假手术组(Sham)、去卵巢组(Ovx)，雌激素替代组(去卵巢后加戊酸雌二醇替代组，Ovx／EV)，各20只。分别在实验开始4周及8周后，处死每组10只。将大鼠咬肌雌激素受体ERα和ERβ的mRNA，以及肌球蛋白重链MHC-Ⅰ、Ⅱa、Ⅱx和Ⅱb型的mRNA作为研究对象，利用SYBR GreenⅠ荧光实时定量PCR技术，来研究雌激素对咀嚼肌功能的影响，对颞下颌关节紊乱病的雌激素病因及治疗新途径进行初步探索。
根据实验观察得出以下结果：
1．大鼠咬肌组织有ERα和ERβ的表达，其中ERα表达主要在肌细胞核，表达强度为阳性。ERβ主要表达在肌细胞核和毛细血管内皮细胞核，表达强度为强阳性。但是大鼠咬肌ERαmRNA的表达明显强于ERβmRNA的表达。
2．大鼠咬肌组织MHC-Ⅱb mRNA的表达较丰富。
3．用大鼠去卵巢，结合雌激素替代法，可以建立雌激素对大鼠咬肌功能影响的实验动物模型。
4．4周去卵巢组ERαmRNA表达上调，与4周组的假手术组及雌激素替代组相比有统计学意义。其余组间没有统计学差异。
5．4周去卵巢组ERβmRNA表达上调，与8周去卵巢组，以及4、8周组的假手术组和雌激素替代组相比有统计学意义。其余组间没有统计学差异。
6．MHC-ⅠmRNA各组之间相比无统计学差异。
7．8周去卵巢组MHC-Ⅱa mRNA表达上调，与4、8周假手术组和去卵巢组，以及与4周雌激素替代组相比有统计学意义。其余组间没有统计学差异。
8．8周去卵巢组MHC-Ⅱx mRNA表达上调，与4、8周假手术组和去卵巢组，以及与4周雌激素替代组相比有统计学意义。其余组间没有统计学差异。
9．MHC-Ⅱb mRNA各组之间相比无统计学差异。
综上所述，本研究认为大鼠咬肌是雌激素的靶组织，雌激素水平下降，ERα和ERβmRNA的表达有上调后又下降恢复的趋势，而MHC-Ⅱa和Ⅱx mRNA表达上调。研究提示雌激素对大鼠咀嚼肌的功能有影响。
Temporomandibular disorder (TMD), characterised by pain and recurrence, is one of the most commonly seen diseases in dentistry clinic. The prevalence of TMD, mostly identified as functional disturbances in the masticatory system, has been well documented to be higher in young females than in males. Estrogen, a steroid compound with multifunctions, is one of the most important hormones in women. The effects of estrogen on the composition and metabolism of myofiber in limb skeletal muscles have been investigated extensively in recent years. However, the effect of estrogen on masticatory muscles has been rarely studied. This study is therefore undertaken to examine this aspect and to provide new data regarding the possible role of estrogen in the development of TMD.
Sixty twelve-week old female Sprague-Dawley rats were randomly divided into 3 groups: (1) control (Sham surgery), (2) ovariectomy without estrogen replacement (Ovx) treatment, (3) ovariectomy with estradiol valerate replacement (Ovx/EV) treatment. Half of the animals were sacrificed at 4 and 8 weeks, respectively, after surgery and the masseter muscles were removed for analysis. Real time PCR quantification with SYBR Green I was employed to study the mRNA expression of estrogen receptor alpha (ERα) and beta (ERβ), myosin heavy chain (MHC) I、IIa、IIx and IIb .
The results showed:
1. Expression of ERαand ERβwas detected in normal rat masseter muscle with ERαmainly in myonuclei while ERβin both myonuclei and endothelial cell nuclei of blood capillaries. ERαmRNA expression was stronger than ERβmRNA.
2. Strong expression of MHC- IIb mRNA in masseter of rat was found.
3. Ovx/EV is a good animal model for studying the effect of estrogen on rat masticatory function.
4. At week 4, the expression of ERαmRNA was significantly higher in the Ovx group than in the Ovx/EV group.
5. The expression of ERβmRNA in the Ovx group at week 4 was significantly higher than at week 8. The expression of ERβmRNA in the Ovx group at week 4 was also significantly higher than the sham and Ovx/EV groups at both week 4 and week 8 time points.
6. MHC-I mRNA expression did not differ among groups.
7. MHC-IIa mRNA expression was higher in the Ovx group at week 8 than its expression in the Sham, Ovx, Ovx/EV groups at week 4 and the sham group at week 8.
8. MHC-IIx mRNA expression was higher in the Ovx group at week 8 than its expression in the Sham, Ovx, Ovx/EV groups at week 4 and the sham group at week 8.
9. MHC-IIb mRNA expression was not significantly different among groups.
The results suggest that the rat masseter muscle is one of the targets of estrogen.In response to estrogen level decrease, the expression of ERαand ERβmRNA tend to be upregulated initially and then be restoreded, while the expression of MHC- IIa and IIx mRNA tend to be upregulated constantly. It is concluded that estrogen does have effect on the masticatory function of female rats.
引文
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1．易新竹，牙合学，人民卫生出版社，2003．12，89．
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