荧光素酶报告基因检测系统原理 ppt

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Background
Mouse models
The development is similar to the process of type 2 diabetes
Hyperinsulinemia is one of common features among all the animal models of obesity, and also one of the earlis- t metabolic alterations observed in 4 models.
高胰岛素血症 血液胰岛素增加
血糖下降
暂时性血糖平衡
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Objective
1. Central melanocortin receptor signaling may play a role in the control of glucose homeostasis.
2.A hypothesis: the melanocortin agonist lowers serum insulin by stimulating the sympathetic drive to the pancreas known to inhibit insulin release.
At 60 min after 1nmol MTⅡ, the blood glucose test
The effect of melanocortin adminstration on serum insulin
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Results
Fig 1. 在ob/ob老鼠中，注射 黑皮质素激动剂对血清胰岛
素的影响。
2.芬妥胺对MTII在胰岛素分泌上的抑制作 用具有完全阻断效果。
3.芬妥胺对瘦素在胰岛素分泌上的抑制作 用具有部分阻断效果。
Background
Energy intake
Energy expenditure
Glucose homeostasis
The Central Melanocortin
system
Hypothesis: the central melanocortin system may be involved in the regulation of glucose homeostasis
The Central Melanocortin System Can Directly Regulate Serum Insulin Levels
From: Endocrinology (2000)
Author: Roger D. Cone et al.
Reporter：Ziqi Zhang
2015/6/22
to test the hypothesis
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Conclusion
Fig 2. MTII注射后降低血清胰岛素水平是通过刺激交 感神经而起作用的。
S: saline P: phentolamine A: ACSF M: MTⅡ L: Leptin
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Conclusion
1.芬妥胺对胰岛素分泌有促进作用，与文 献报道效果相同——芬妥胺能阻断交感 神经对胰岛素分泌的抑制作用。
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Process 2
Ob/ob mouse
Ip injection with phentolamine or saline
Icv injection of MTⅡ 、ACSF、Leptin
RIA assay for serum insulin
Phentolamine :芬妥胺 ，非特异性α-肾上腺素 受体的反向激动剂，对 交感神经对胰岛素分泌 的抑制有阻断作用。
3. Whether the hyperinsulinemia in the melanocortin obesity syndromes is primary to MC4R blockade, a development consequence of MC4R blockade, or secondary to te hyperphagia and obesity.
MC4R
MC3R
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BacBiblioteka Baiduground
Glucose homeostasis
外周组织对胰岛 素的敏感性
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Background
中央黑皮质素信号传 导缺陷（未知）
瘦素缺陷（有报道 ） 肥胖组织增生干扰胰 岛素作用（有报道）
其他未知
胰岛素敏感性下降
血糖升高
胰岛素抗性 胰岛补偿性分泌 胰岛素增加
血糖失衡 糖尿病
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Process 1
Ob/ob mouse
Icv injection of MTⅡ (0.3 、1、3nmol) 、ACSF、
Leptin
Ob/ob 老鼠是瘦素缺陷 老鼠，具有高胰岛素血 症。
MTII 是MC4R/MC3R 的 激动剂，是α-MSH的类 似物。 ACSF—人工脑脊液
After 30-60min, RIA assay for serum insulin
72.8%
30-60min
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Conclusion
1.A中与两个对照相比，MTII 降低血清中的胰岛素水平，并具 有剂量反应 2.B中注射MTII后一小时检测到血糖浓度有明显增加。
a.中央黑皮质素受体可能参与在血糖调节过程。 b.MTII对胰岛素分泌的抑制不是由于降低血糖浓度而间 接起作用的。
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Contents
Background Method process
Conclusion
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Background
Type 2 diabetes
Insulin resistance
The mechanistic relationship between obesity, hyperinsulinemia, insulin resistance, and diabetes is not entirely clear.
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Method
Surgical procedure
RIA assay for serum or plasma
insulin
Blood glucose and glucose tolerance test
Nonesterified fatty acid assay
Insulin tolerance test