Dehydroepiandroesteron Accompanied Retinoic Acid
Enhances Differentiation of P19 Embryonal
Stem Cells into Neural Cells
Hossein Azizi, M.Sc.
1, Narges Zare Mehrjerdi, M.Sc.
1, Mirza Khalil Bahmani, Ph.D.
2,
Hossein Baharvand, Ph.D.
1, 3*
1. Stem Cells and Developmental Biology Department, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran
2. HIV & Hepatitis Research Center, Shiraz University of Medical Science, Shiraz, Iran 3. Developmental Biology Department, University of Science and Culture, Tehran, Iran
* Corresponding Address: P.O.Box: 19395-4644, Stem Cells and Developmental Biology Department, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran
Email: baharvand@royaninstitute.org Received: 6/Dec/2007, Accepted: 5/Jun/2008
Abstract
Objective: Dehydroepiandroesteron (DHEA) is a neurosteroid with potential effect on neu-rogenesis, neuronal survival and proliferation of neural progenitor cells. However there is no direct evidence for its biological effect during the differentiation of stem cell-derived neurons. The p19 line of embryonal carcinoma cells develops into neurons, astroglia and fibroblasts after exposure to retinoic acid (RA). This study was initiated to assess the effect of DHEA on neural cells derived from p19 embryonal carcinoma stem cells.
Materials and Methods: P19 cells were suspended in dulbecco’s modified eagle’s medium (DMED) containing fetal bovine serum (FBS) in bacterial-grade petri dishes in the presence of RA, DHEA and RA+DHEA for 6 days. Then cells were trypsinized for dispersion and re-placed in poly L- lysine (10μg/ml) coated tissue culture dishes without RA and DHEA for 4 days. The expression of neural markers Map-2, Tau, beta-tubulin III- clone Juj (Tuj1), astro-cyte marker GFAP and the percent of neurotransmitters tyrosin hydroxylase, glutamate, se-rotonin and actyl cholin transferase were evaluated by flowcytometry, immunocytochemistry and RT-PCR analysis.
Results: Flowcytometry analysis showed that about 63 ± 3% of the cells express neuronal marker Tuj1 and about 5 ± 1% of the cells express tyrosine hydroxylase neurotransmitters in RA treated groups. However when RA and DHEA were added to the culture medium, Tuj1 expression increased to about 74 ± 1% and tyrosine hydroxylase expression increased to 23 ± 2%.
Conclusion: Results showed that DHEA accompanied RA increased the number of Tuj1 and dopaminergic neurons that were derived from p19 embryonal carcinoma stem cells.
Keywords: Retinoic Acid, Neurogenesis, Dehydroepiandroesteron
Yakhteh Medical Journal, Vol 11, No 2, Summer 2009, Pages: 228-235
ﺰﻳﺎﻤﺗ
ﺮﺑ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﺎﺑ
ﻩﺍﺮﻤﻫ
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19395-4644 :
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:
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ﻱﺩﺎﻴﻨﺑ
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Email: baharvand@royaninstitute.org
:
ﻚﻴﻧﻭﺮﺘﻜﻟﺍ
ﺖﺴﭘ
87/3/16 :ﻪﻟﺎﻘﻣﺵﺮﻳﺬﭘ،86/10/15 :ﻪﻟﺎﻘﻣﺖﻓﺎﻳﺭﺩ
ﻩﺪﻴﻜﭼ
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ﺮﺛﺍ
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*
ﺎﻬﻧﺁ
ﺖﻴﻫﺎﻣ
ﻦﻴﻴﻌﺗ
ﻭ
P19ﻯﺎﻫ
ﻝﻮﻠﺳ
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ﻰﺒﺼﻋ
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ﻝﻮﻠﺳ
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ﺹﻮﺼﺨﻣ
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6
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P19ﻯﺎﻫ
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ﺵﻭﺭ ﻭ ﺩﺍﻮﻣ *
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
،ﻚﻴﺋﻮﻨﻴﺗﺭ
ﺪﻴﺳﺍ
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ﻝﻮﻠﺳ
ﻥﺎﻣﺯ
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ﺭﺩ
.
ﺪﺷ
ﺖﺸﻛ
ﻡﺮﺳ
ﻯﻭﺎﺣ
DMEMﻂﻴﺤﻣ
ﺎﺑ
ﻩﺍﺮﻤﻫ
ﻯﺮﺘﻛﺎﺑ
ﺮﺘﺴﺑ
ﻯﻭﺭ
ﺯﻭﺭ
4
ﺕﺪﻣ
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ﻭ
ﻩﺪﺷ
ﻪﻨﻴﺴﭙﻳﺮﺗ
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ﻝﻮﻠﺳ
.
ﺖﻓﺮﮔ
ﺭﺍﺮﻗ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
+
ﻚﻴﺋﻮﻨﻴﺗﺭ
ﺪﻴﺳﺍ
ﻭ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﺘﻴﺳﻭﺮﺘﺳﺁﺮﻛﺭﺎﻣ
،
Tuj1, MAP-2, Tauﻰﺒﺼﻋ
ﻯﺎﻫﺮﻛﺭﺎﻣ
ﻥﺎﻴﺑ
ﺪﺻﺭﺩ
ﺲﭙﺳ
.
ﺪﺷ
ﺖﺸﻛ
ﻦﻳﺰﻴﻟ
-
L-
ﻰﻠﭘ
ﺯﺍ
ﻩﺪﺷ
ﻩﺪﻴﺷﻮﭘ
ﺎﺑ
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻦﻴﻟﻮﻛ
ﻞﻴﺘﺳﺍﻭ
ﻦﻴﻧﻮﺗﻭﺮﺳ
،ﺕﺎﻣﺎﺗﻮﻠﮔ
،ﺯﻼﻴﺴﻛﻭﺭﺪﻴﻫ
ﻦﻳﺯﻭﺮﻴﺗ
ﻯﺎﻫﺮﺘﻴﻤﺴﻧﺍﺮﺗﻭﺭﻮﻧ
ﺪﺻﺭﺩ
،
(
GFAP)
.
ﺖﻓﺮﮔ
ﺭﺍﺮﻗ
ﻰﺳﺭﺮﺑ
ﺩﺭﻮﻣ
RT-PCRﻭ
ﻰﻤﻴﺷﻮﺘﻴﺳﻮﻧﻮﻤﻳﺍ
،ﻯﺮﺘﻣﻮﺘﻳﺎﺳﻮﻠﻓ
ﻦﻳﺯﻭﺮﻴﺗ
ﺎﻫ
ﻝﻮﻠﺳ
ﺪﺻﺭﺩ
5
±
1
ﺩﻭﺪﺣ
ﻭ
Tuj1ﺎﻫ
ﻝﻮﻠﺳ
ﺪﺻﺭﺩ
63
±
3
ﺩﻭﺪﺣ
ﻪﻛ
ﺩﺍﺩ
ﻥﺎﺸﻧ
ﻯﺮﺘﻣﻮﺘﻳﺎﺳﻮﻠﻓ
ﻯﺎﻫﺰﻴﻟﺎﻧﺁ
:ﺎﻫ
ﻪﺘﻓﺎﻳ *
ﻥﺎﻴﺑ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
+
ﻚﻴﺋﻮﻨﻴﺗﺭ
ﺪﻴﺳﺍ
ﺭﺎﻤﻴﺗ
ﺖﺤﺗ
ﻩﻭﺮﮔﺭﺩ
ﺎﻣﺍ
.
ﺪﻨﻛ
ﻰﻣ
ﻥﺎﻴﺑ
ﻚﻴﺋﻮﻨﻴﺗﺭ
ﺪﻴﺳﺍ
ﻩﻭﺮﮔ
ﺭﺩ
ﺍﺭ
ﺯﻼﻴﺴﻛﻭﺭﺪﻴﻫ
.
ﺖﻓﺎﻳ
ﺶﻳﺍﺰﻓﺍ
ﺪﺻﺭﺩ
23
±
2
ﺩﻭﺪﺣ
ﻪﺑ
ﺯﻼﻴﺴﻛﻭﺭﺪﻴﻫ
ﻦﻳﺯﻭﺮﻴﺗ
ﺮﺘﻴﻤﺴﻧﺍﺮﺗﻭﺭﻮﻧ
ﻭ
ﺪﺻﺭﺩ
74
±
1
ﺩﻭﺪﺣ
ﻪﺑ
Tuj1ﻭ
Tuj1ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺩﺍﺪﻌﺗ
ﻚﻴﺋﻮﻨﻴﺗﺭ
ﺪﻴﺳﺍ
ﻩﺍﺮﻤﻫ
ﻪﺑ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﻪﻛ
ﺪﻫﺩ
ﻰﻣ
ﻥﺎﺸﻧ
ﺞﻳﺎﺘﻧ
:ﻱﺮﻴﮔ
ﻪﺠﻴﺘﻧ *
.
ﺪﻫﺩ
ﻰﻣ
ﺶﻳﺍﺰﻓﺍ
ﺍﺭ
P19ﻝﻮﻠﺳ
ﺯﺍ
ﻩﺪﺷ
ﻖﺘﺸﻣ
ﻚﻳﺯﺮﻨﻴﻣﺎﭘﻭﺩ
ﻯﺎﻫ
ﺐﺼﻋ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
،ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻪﺑ
ﺰﻳﺎﻤﺗ
،ﻚﻴﺋﻮﻨﻴﺗﺭ
ﺪﻴﺳﺍ
:ﻥﺎﮔژﺍﻭﺪﻴﻠﻛ *
228-235 :ﺕﺎﺤﻔﺻ،88 ﻥﺎﺘﺴﺑﺎﺗ،2 ﻩﺭﺎﻤﺷ،ﻢﻫﺩﺯﺎﻳﻝﺎﺳ،ﻪﺘﺧﺎﻳﻰﻜﺷﺰﭘﻪﻣﺎﻨﻠﺼﻓ
ﻪﻣﺪﻘﻣ
ﺢﻄﺳ
.
ﺪﻨﻨﻛ
ﻰﻣ
ﺎﻔﻳﺍ
ﻰﻳﺍﺯ
ﺐﺼﻋ
ﻝﺮﺘﻨﻛ
ﺭﺩ
ﻰﻤﻬﻣ
ﺶﻘﻧ
ﺎﻫﺪﻴﺋﻭﺮﺘﺳﺍ
ﻯﺎﻫﺪﻨﻳﺍﺮﻓ
ﺎﻳ
ﺎﻫ
ﺱﺮﺘﺳﺍ
ﺪﻨﻧﺎﻣ
ﻰﺟﺭﺎﺧ
ﻊﻳﺎﻗﻭ
ﺯﺍ
ﻰﺧﺮﺑ
ﻪﺑ
ﺰﻐﻣ
ﻭ
ﻥﻮﺧ
ﺭﺩ
ﺎﻫﺪﻴﺋﻭﺮﺘﺳﺍ
ﺲﻜﺗﺭﻮﻛ
ﻯﺩﺎﻴﻨﺑ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺭﺩ
.(2
،
1)
ﺖﺳﺍ
ﻪﺘﺴﺑﺍﻭ
ﻦﺳ
ﻝﻮﻃ
ﺪﻨﻧﺎﻣ
ﻰﺘﺴﻳﺯ
ﻝﺎﻴﻠﮔﺮﻛﺭﺎﻣ
ﻥﺎﻴﺑ
ﺶﻳﺍﺰﻓﺍ
ﺐﺒﺳ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﻥﺎﺴﻧﺍ
ﻥﺎﻳﻭﺭ
Beta-)ﻰﺒﺼﻋ
ﺮﻛﺭﺎﻣ
ﻭ
(Glial Fibrillar Acidic Protein; GFAP)nﻪﻛ
ﺪﻨﺘﻓﺎﻳﺭﺩ
ﻥﺎﻘﻘﺤﻣ
.(3)
ﺩﻮﺷ
ﻰﻣ
(Tubulin III- Clone Tuj; Tuj1ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﺽﺮﻌﻣ
ﺭﺩ
ﻯﺩﺍﺯﻮﻧ
ﻥﺍﺭﻭﺩ
ﺭﺩ
ﻪﻛ
ﻰﻐﻟﺎﺑ
ﻯﺎﻫ
ﺕﺭ
Microtubule-Associated)ﻰﺒﺼﻋ
ﻦﻴﺌﺗﻭﺮﭘ
ﻥﺎﻴﺑ
،ﺪﻧﺮﻴﮔ
ﻲﻣ
ﺭﺍﺮﻗ
.(4)
ﺪﺑﺎﻳ
ﻰﻣ
ﺶﻳﺍﺰﻓﺍ
ﺎﻬﻧﺁ
ﭗﻣﺎﻛﻮﭙﻴﻫ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺭﺩ
(Protein; MAP-2
ﻰﺒﺼﻋ
ﺮﻛﺭﺎﻣ
ﻥﺎﻴﺑ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﻪﻛ
ﻩﺪﺷ
ﺺﺨﺸﻣ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺭﺩ
ﺍﺭ
(Neuronal Specific Nuclear Protein; NeuN).(5)
ﺩﻮﺷ
ﻰﻣ
ﻰﻳﺍﺯ
ﺐﺼﻋ
ﺶﻳﺍﺰﻓﺍ
ﺐﺒﺳ
ﻭ
ﻩﺩﺮﻛ
ﻚﻳﺮﺤﺗ
ﻎﻟﺎﺑ
ﺵﻮﻣ
ﭗﻣﺎﻛﻮﭙﻴﻫ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺮﺑ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﻭ
ﻚﻴﺋﻮﻨﻴﺗﺭ
ﺪﻴﺳﺍ
ﻦﻴﺑ
ﻰﻳﺍﺰﻓﺍ
ﻢﻫ
ﺮﺛﺍ
ﻚﻳ
ﻰﺒﺼﻋ
ﺮﻛﺭﺎﻣ
ﻥﺎﻴﺑ
ﺶﻳﺍﺰﻓﺍ
ﺚﻋﺎﺑ
ﺪﻧﺍﻮﺗ
ﻰﻣ
ﻪﻛ
ﻩﺪﺷ
ﺵﺭﺍﺰﮔ
ﻰﻧﺎﺴﻧﺍ
ﻯﺎﻣﻮﺘﺳﻼﺑﻭﺭﻮﻧ
ﺪﻴﺳﺍ
.(6)
ﺩﻮﺷ
(Growth-Associated Protein 43; GAP-43)ﻰﻌﻴﺒﻃ
ﺖﻓﺮﺸﻴﭘ
ﻭ
ﺪﺷﺭ
ﻆﻔﺣ
ﻯﺍﺮﺑ
،
Aﻦﻴﻣﺎﺘﻳﻭ
ﺯﺍ
ﻰﻘﺘﺸﻣ
ﻥﺍﻮﻨﻋ
ﻪﺑ
ﻚﻴﺋﻮﻨﻴﺗﺭ
ﺭﺩ
ﻪﻛ
ﻩﺩﻮﺑ
ﻰﻣﻮﻤﻋ
ﻩﺪﻨﻫﺩﺰﻳﺎﻤﺗ
ﻚﻳ
ﻩﺩﺎﻣ
ﻦﻳﺍ
.
ﺖﺳﺍ
ﻯﺭﻭﺮﺿ
ﺎﻫ
ﻝﻮﻠﺳ
ﻰﺒﺼﻋ
ﻢﺘﺴﻴﺳ
ﺭﺩ
ﺹﻮﺼﺧ
ﻪﺑ
ﻍﻮﻠﺑ
ﻭ
ﻰﻧﺎﻳﻭﺭ
ﻥﺍﺭﻭﺩ
ﺭﺩ
ﻒﻠﺘﺨﻣ
ﻯﺎﻫ
ﺖﻓﺎﺑ
ﻰﺒﺼﻋ
ﻪﻟﻮﻟ
ﺪﺷﺭ
ﻯﻮﮕﻟﺍ
.
ﺩﻮﺷ
ﻰﻣ
ﻰﺒﺼﻋ
ﺰﻳﺎﻤﺗ
ﺖﻓﺮﺸﻴﭘ
ﺚﻋﺎﺑ
ﻭ
ﺩﺭﺍﺩ
ﺩﻮﺟﻭ
ﻥﻮﭼ
ﻢﻫ
ﻰﻟﻮﻠﺳ
ﺝﺭﺎﺧ
ﻯﺎﻫﺍﺯ
ﺖﺨﻳﺭ
ﺮﻴﺛﺄﺗ
ﺖﺤﺗ
ﻰﻤﻜﺷ
-
ﻰﺘﺸﭘ
ﺭﻮﺤﻣ
ﺭﺩ
(
FGF) Fibroblast Growth Factors, Retinoic Acid, (BMP)Bone Morphogenetic Protein, Shh (Sonic Hedgehog)h
ﻯﺎﻫ
ﺶﺨﺑ
ﺯﺍ
ﻰﻜﻳ
ﻥﺍﻮﻨﻋ
ﻪﺑ
ﺪﺷﺭ
ﻝﺎﺣ
ﺭﺩ
ﻰﺒﺼﻋ
ﻪﻟﻮﻟ
.(8
،
7)
ﺩﺮﻴﮔ
ﻰﻣ
ﺭﺍﺮﻗ
ﺕﺎﻘﻴﻘﺤﺗ
.(9)
ﺖﺳﺍ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﺯﺍ
ﻯﺩﺎﻳﺯ
ﺮﻳﺩﺎﻘﻣ
ﻯﻭﺎﺣ
،ﻰﻧﺎﻳﻭﺭ
ﻰﻠﺻﺍ
ﺭﺩ
ﻰﺗﻻﻼﺘﺧﺍ
ﺩﺎﺠﻳﺍ
ﺚﻋﺎﺑ
ﻚﻴﺋﻮﻨﻴﺗﺭ
ﺪﻴﺳﺍ
ﺭﻮﻀﺣ
ﻡﺪﻋ
ﻪﻛ
ﻩﺩﺍﺩ
ﻥﺎﺸﻧ
ﺭﺩ
ﻰﻟﻮﻠﺳ
ﻯﺎﻫﺭﺎﺘﻓﺭ
ﻦﻴﻨﭽﻤﻫ
ﻭ
ﻉﺎﺨﻧ
ﻰﻜﻳژﻮﻟﻮﻓﺭﻮﻣ
ﻯﺎﻫ
ﻩﺪﻨﻫﺩ
ﻥﺎﻣﺯﺎﺳ
ﻪﻛ
ﺖﺳﺍ
ﻦﻳﺍ
ﺭﺩ
p19ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻯﺎﻫ
ﻰﮔﮋﻳﻭ
ﺯﺍ
.
ﺩﺩﺮﮔ
ﻰﻣ
ﻉﺎﺨﻧ
ﺪﺷﺭ
ﻰﻃ
،ﺪﻧﻮﺷ
ﻩﺩﺍﺩ
ﺖﺸﻛ
ﻯﺍ
ﻩﺩﻮﺗ
ﺕﺭﻮﺻ
ﻪﺑ
ﻰﻟﻮﻤﻌﻣ
ﺖﺸﻛ
ﻯﺎﻫ
ﻂﻴﺤﻣﺭﺩ
ﺮﮔﺍ
(10)
ﺪﻨﺑﺎﻳ
ﻰﻣ
ﺰﻳﺎﻤﺗ
ﻰﻧﺎﻳﻭﺭ
ﺝﺭﺎﺧ
ﻡﺭﺩﻭﺪﻧﺍ
ﻪﻴﺒﺷ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻪﺑ
ﺎﻫ
ﻝﻮﻠﺳ
ﺯﺍ
ﻯﺮﻴﮔ
ﻢﺸﭼ
ﺭﻮﻃ
ﻪﺑ
ﺪﻧﺮﻴﮕﺑ
ﺭﺍﺮﻗ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﺽﺮﻌﻣ
ﺭﺩ
ﻪﻛ
ﻰﻧﺎﻣﺯ
ﻭ
ﺽﺮﻌﻣ
ﺭﺩ
ﻰﺘﻗﻭ
ﺎﻣﺍ
.(11)
ﺪﻧﻮﺷ
ﻰﻣ
ﺖﻳﺍﺪﻫ
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺖﻤﺳ
ﻪﺑ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺖﻤﺳ
ﻪﺑ
ﺪﻧﺮﻴﮕﺑ
ﺭﺍﺮﻗ
Dimethyl Sulfoxide (DMSO)ﻰﺒﺳﺎﻨﻣ
ﻰﻟﻮﻠﺳ
ﻝﺪﻣ
ﺎﻫ
ﻝﻮﻠﺳ
ﻦﻳﺍ
ﻦﻳﺍﺮﺑﺎﻨﺑ
.(6)
ﺪﻧﻮﺷ
ﻰﻣ
ﺖﻳﺍﺪﻫ
ﻯﺍ
ﻪﭽﻴﻫﺎﻣ
ﺖﻴﻤﻫﺍ
.
ﺪﺷﺎﺑ
ﻰﻣ
ﻯﺍ
ﻪﭽﻴﻫﺎﻣ
ﻭ
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻦﻳﻮﻜﺗ
ﻰﺳﺭﺮﺑ
ﻯﺍﺮﺑ
ﻯﺎﻫ
ﻝﻮﻠﺳ
،ﻪﺘﻓﺎﻳ
ﺰﻳﺎﻤﺗﺮﻴﻏ
ﺖﻟﺎﺣ
ﺭﺩ
ﻪﻛ
ﺖﺳﺍ
ﻦﻳﺍ
ﺭﺩ
،p19ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺽﺮﻌﻣ
ﺭﺩ
ﻪﻛ
ﻰﻧﺎﻣﺯ
ﻦﻴﻨﭽﻤﻫ
ﻭ
ﺪﻨﺘﺴﻫ
ﻰﻜﻴﺘﻧژ
ﻯﺭﺎﻛ
ﺖﺳﺩ
ﻯﺍﺮﺑ
ﻰﺒﺳﺎﻨﻣ
ﻰﺒﺼﻋ
ﻢﺘﺴﻴﺳ
ﻪﻴﺒﺷ
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻪﺑ
ﺩﺮﻴﮕﺑ
ﺭﺍﺮﻗ
ﻚﻴﺋﻮﻨﻴﺗﺭ
ﺪﻴﺳﺍ
ﻖﺘﺸﻣ
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻪﻌﻟﺎﻄﻣ
ﺎﺑ
ﻦﻳﺍﺮﺑﺎﻨﺑ
.(12)
ﺪﺑﺎﻳ
ﻰﻣ
ﺰﻳﺎﻤﺗ
ﻯﺰﻛﺮﻣ
ﻪﻴﺣﺎﻧ
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻪﺑ
ﺍﺭ
ﺞﻳﺎﺘﻧ
ﻥﺍﻮﺗ
ﻰﻣ
،p19ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺯﺍ
ﻩﺪﺷ
ﺪﻴﺳﺍ
ﺮﺛﺍ
ﺩﺭﻮﻣ
ﺭﺩ
ﻯﺍ
ﻪﻌﻟﺎﻄﻣ
ﻥﻮﻨﻛ
ﺎﺗ
.
ﺩﺍﺩ
ﻢﻴﻤﻌﺗ
ﻯﺰﻛﺮﻣ
ﻰﺒﺼﻋ
ﻢﺘﺴﻴﺳ
ﻰﻧﺎﻳﻭﺭ
ﻯﺩﺎﻴﻨﺑ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺰﻳﺎﻤﺗ
ﺮﺑ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﻭ
ﻚﻴﺋﻮﻨﻴﺗﺭ
ﻪﻛ
ﺩﺍﺩ
ﻥﺎﺸﻧ
ﺎﻣ
ﻰﻠﺒﻗ
ﻪﻌﻟﺎﻄﻣ
.
ﺖﺳﺍ
ﻪﺘﻓﺮﮕﻧ
ﺕﺭﻮﺻ
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻪﺑ
ﺶﻳﺍﺰﻓﺍ
ﺐﺒﺳ
ﺪﻧﺍﻮﺗ
ﻰﻣ
ﻚﻴﺋﻮﻨﻴﺗﺭ
ﺪﻴﺳﺍ
ﻩﺍﺮﻤﻫ
ﻪﺑ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﻪﻌﻟﺎﻄﻣ
ﻦﻳﺍ
ﺭﺩ
ﺍﺬﻟ
.(13)
ﺩﻮﺷ
p19ﺯﺍ
ﻖﺘﺸﻣ
ﻰﺒﺼﻋ
ﻯﺎﻫﺯﺎﺳ
ﺶﻴﭘ
ﺮﻴﺜﻜﺗ
ﺍﺰﺠﻣ
ﺕﺭﻮﺻ
ﻪﺑ
ﺍﺭ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﻭ
ﻚﻴﺋﻮﻨﻴﺗﺭ
ﺪﻴﺳﺍ
ﺮﺛﺍ
ﺍﺪﺘﺑﺍ
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻪﺑ
p19ﻰﻧﺎﻳﻭﺭ
ﻰﻳﺎﻣﻮﻨﻴﺳﺭﺎﻛ
ﻯﺩﺎﻴﻨﺑ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺰﻳﺎﻤﺗ
ﺮﺑ
ﺰﻳﺎﻤﺗ
ﺮﺑ
ﺰﻴﻧ
ﺎﻬﻧﺁ
ﻥﺎﻣﺯ
ﻢﻫ
ﺮﺛﺍ
ﻪﺑ
ﻪﻣﺍﺩﺍ
ﺭﺩ
ﺲﭙﺳ
ﻩﺩﺍﺩ
ﺭﺍﺮﻗ
ﻰﺳﺭﺮﺑ
ﺩﺭﻮﻣ
ﻎﻟﺎﺑ
.
ﺖﺧﺍﺩﺮﭘ
ﻢﻴﻫﺍﻮﺧ
ﻰﺒﺼﻋ
ﺕﺎﻘﺘﺸﻣ
ﻪﺑ
ﺎﻫ
ﻝﻮﻠﺳ
ﻦﻳﺍ
ﺎﻫ
ﺵﻭﺭ
ﻭ
ﺩﺍﻮﻣ
p19
ﻩﺩﺭ
ﻰﻨﻴﻨﺟ
ﻰﻳﺎﻣﻮﻨﻴﺳﺭﺎﻛ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺯﺍ
ﻪﻌﻟﺎﻄﻣ
ﻦﻳﺍ
ﺭﺩ
ﻂﻴﺤﻣﺭﺩ
ﺎﻫ
ﻝﻮﻠﺳ
ﻦﻳﺍ
.
ﺪﺷ
ﻩﺩﺎﻔﺘﺳﺍ
(
ﺭﻮﺘﺳﺎﭘ
ﻪﺴﺳﻮﻣ
،ﻥﺍﺮﻬﺗ
)
15
ﻯﻭﺎﺣ
Dulbecco's Modified Eagle's Medium (DMEM)(
ﻦﻴﺴﻳﺎﻣﻮﺘﭘﺮﺘﺳﺍ
-
ﻦﻴﻠﻴﺳ
ﻰﻨﭘ
ﻭ
Fetal Bovine Serum (FBS)ﺪﺻﺭﺩ
.
ﺪﻧﺍ
ﻩﺪﺷ
ﻩﺩﺍﺩ
ﺖﺸﻛ
(
ﺮﺘﻴﻟ
ﻲﻠﻴﻣ
ﺮﺑ
ﺪﺣﺍﻭ
100)
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻪﺑ
ﺰﻳﺎﻤﺗ
ﺱﺎﺳﺍ
ﺮﺑ
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻪﺑ
p19ﻯﺩﺎﻴﻨﺑ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺰﻳﺎﻤﺗ
ﻩﻮﺤﻧ
ﺖﻈﻠﻏ
ﺎﺑ
ﺎﻫ
ﻝﻮﻠﺳ
ﻪﻛ
ﺐﻴﺗﺮﺗ
ﻦﻳﺪﺑ
.
ﺪﺷ
ﻡﺎﺠﻧﺍ
ﻰﻧﺎﻳﻭﺭ
ﻪﺒﺷ
ﻡﺎﺴﺟﺍ
ﻞﻴﻜﺸﺗ
ﺭﺩ
ﻥﻮﻴﺴﻧﺎﭙﺳﻮﺳ
ﺕﺭﻮﺻ
ﻪﺑ
ﺯﻭﺭ
6
ﺕﺪﻣ
ﻪﺑ
ﺮﺘﻴﻟ
ﻰﻠﻴﻣ
ﺮﺑ
ﻝﻮﻠﺳ
1/5 ×10
5ﻂﻴﺤﻣ
.
ﺪﺷ
ﺖﺸﻛ
ﻯﺮﺘﻛﺎﺑ
ﺖﺸﻛ
ﺹﻮﺼﺨﻣ
ﻰﻜﻴﺘﺳﻼﭘ
ﺖﺸﻛ
ﻑﻭﺮﻇ
ﻭ
ﻩﺩﻮﺑ
FBSﺪﺻﺭﺩ
5
ﺎﺑ
DMEMﻯﻭﺎﺣ
1-4
ﻯﺎﻫﺯﻭﺭ
ﺭﺩ
ﺎﻫ
ﻝﻮﻠﺳ
ﺕﺪﻣ
ﻦﻳﺍ
ﺭﺩ
.
ﺖﻓﺎﻳ
ﺶﻫﺎﻛ
ﺪﺻﺭﺩ
3
ﻪﺑ
5-6
ﻯﺎﻫﺯﻭﺭ
ﺭﺩ
ﻥﺁ
ﺭﺍﺪﻘﻣ
ﺕﺪﻣ
ﺭﺩ
ﺎﻫ
ﻝﻮﻠﺳ
ﻩﻭﺮﮔ
ﻦﻳﺍ
ﺭﺩ
)
ﻝﺮﺘﻨﻛ
ﻯﺎﻫ
ﻩﻭﺮﮔ
ﺭﺩ
ﺎﻫ
ﻝﻮﻠﺳ
ﻥﺎﻣﺯ
ﺭﺍﺮﻗ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﻭ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﺽﺮﻌﻣ
ﺭﺩ
ﺯﻭﺭ
6
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
،
(
ﺭﻻﻮﻣﻭﺮﻜﻴﻣ
1)
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
،
(
ﺖﻓﺮﮕﻧ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
+ (
ﺭﻻﻮﻣﻭﺮﻜﻴﻣ
4)
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﻭ
(
ﺭﻻﻮﻣﻭﺮﻜﻴﻣ
4)
ﻡﺎﺴﺟﺍ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺲﭙﺳ
.
ﺪﺷ
ﺭﺎﻤﻴﺗ
ﻢﻫ
ﺎﺑ
(
ﺭﻻﻮﻣﻭﺮﻜﻴﻣ
1)
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻦﻳﺰﻴﻟ
-Lﻰﻠﭘ
ﺯﺍ
ﻩﺪﺷ
ﻩﺪﻴﺷﻮﭘ
ﺮﺘﺴﺑ
ﻯﻭﺭ
ﻭ
ﻩﺪﺷ
ﺍﺪﺟ
ﻢﻫ
ﺯﺍ
ﻰﻧﺎﻳﻭﺭ
ﻪﺒﺷ
ﻭ
ﻪﺘﻓﺮﮔﺭﺍﺮﻗ
ﻰﺒﺼﻋ
ﺖﺸﻛ
ﻂﻴﺤﻣ
ﻯﻭﺎﺣ
(
ﺮﺘﻴﻟ
ﻲﻠﻴﻣ
ﺮﺑ
ﺭﻻﻮﻣﻭﺮﻜﻴﻣ
10)
.
ﺪﺷ
ﻰﺳﺭﺮﺑ
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻯﺎﻫ
ﺖﺴﺗ
ﺯﻭﺭ
4
ﺯﺍ
ﺪﻌﺑ
ﻯﺮﺘﻣﻮﺘﻳﺎﺳﻮﻠﻓ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻰﺻﺎﺼﺘﺧﺍ
ﻥژ
ﻰﺘﻧﺁ
ﻯﺍﺮﺑ
ﻯﺮﺘﻣﻮﺘﻳﺎﺳﻮﻠﻓ
ﻞﺣﺍﺮﻣ
10
6ﺩﻭﺪﺣ
ﻩﺪﺷ
ﺏﺎﺨﺘﻧﺍ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺩﺍﺪﻌﺗ
:
ﺩﻮﺑ
ﺮﻳﺯ
ﺡﺮﺷ
ﻪﺑ
ﻰﺒﺼﻋ
ﻝﻮﻠﺤﻣ
ﺎﺑ
ﻮﺷﻭ
ﺖﺴﺷ
ﻭ
ﻪﺟﺭﺩ
4
ﺎﻣﺩ
ﺶﻳﺎﻣﺯﺁ
ﻞﺣﺍﺮﻣ
ﻡﺎﻤﺗ
ﺭﺩ
ﻭ
ﺪﺻﺭﺩ
ﻚﻳ
FBSﻯﻭﺎﺣ
Phosphate Buffered Saline (PBS)eﻪﻓﺎﺿﺍ
ﺯﺍ
ﺪﻌﺑ
ﻪﻛ
ﻮﺤﻧ
ﻦﻳﺪﺑ
.
ﺪﺷ
ﻡﺎﺠﻧﺍ
ﺪﺻﺭﺩ
0/058
EDTAﻭ
5
ﺕﺪﻣ
ﻪﺑ
rpm2000
ﺭﻭﺩ
ﺎﺑ
ﺎﻫ
ﻝﻮﻠﺳ
،ﻮﺷﻭ
ﺖﺴﺷ
ﻝﻮﻠﺤﻣ
ﻥﺩﺮﻛ
ﺍﺪﺘﺑﺍ
ﺭﺩ
.
ﺪﺷ
ﻑﺬﺣ
ﻰﻳﻭﺭ
ﻝﻮﻠﺤﻣ
ﺖﻳﺎﻬﻧ
ﺭﺩ
ﻭ
ﻩﺪﺷ
ژﻮﻴﻔﻳﺮﺘﻧﺎﺳ
ﻪﻘﻴﻗﺩ
ﻪﺑﻮﻜﻧﺍ
ﻪﻘﻴﻗﺩ
30
ﺕﺪﻣ
ﻪﺑ
ﺪﺻﺭﺩ
4
ﺪﻴﺋﺪﻟﺎﻣﺮﻓﺍﺭﺎﭘ
ﺎﺑ
ﺮﻈﻧ
ﺩﺭﻮﻣ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻚﻳ
X-100
ﻥﻮﺘﻳﺮﺗ
ﺎﺑ
ﻭ
ﻩﺪﺷ
ﻩﺩﺍﺩ
ﻮﺷﻭ
ﺖﺴﺷ
ﺎﻫ
ﻝﻮﻠﺳ
ﺲﭙﺳ
.
ﺪﺷ
ﺎﻫ
ﻝﻮﻠﺳ
،ﻮﺷﻭ
ﺖﺴﺷ
ﺯﺍ
ﺪﻌﺑ
.
ﺪﺷ
ﺮﻳﺬﭘﺫﻮﻔﻧ
ﻪﻘﻴﻗﺩ
15
ﺕﺪﻣ
ﻪﺑ
ﺪﺻﺭﺩ
ﻯﺩﺎﺑ
ﻰﺘﻧﺁ
ﺎﻫ
ﻝﻮﻠﺳ
ﻪﺑ
.
ﺪﺷ
ﻪﺑﻮﻜﻧﺍ
ﻪﻘﻴﻗﺩ
15
ﺕﺪﻣ
ﻪﺑ
ﺪﺻﺭﺩ
10
ﻡﺮﺳ
ﺎﺑ
Tuj1(Sigma T5293) ،MAP-2 (Sigma M1406)
ﻪﻴﻟﻭﺍ
ﻝﺎﻴﻠﮔ
ﻯﺍ
ﻪﺘﺷﺭ
ﻯﺪﻴﺳﺍ
ﻦﻴﺌﺗﻭﺮﭘ
Tau (RD MAB 361)ﻦﻳﺯﻭﺮﻴﺗ
ﻯﺎﻫﺮﺘﻴﻤﺴﻧﺍﺮﺗﻭﺭﻮﻧ
،
(GFAP) (Chemicon MAB3402)ﻚﻴﻣﺎﺗﻮﻠﮔ
)
ﺕﺎﻣﺎﺗﻮﻠﮔ
،
(TH) (Sigma T1299)ﺯﻼﻴﺴﻛﻭﺭﺪﻴﻫ
ﻰﺴﻛﻭﺭﺪﻴﻫ
-5)
ﻦﻴﻧﻮﺗﻭﺮﺳ
،
(Sigma G5038)(
ﺯﻼﻴﺴﻛﻮﺑﺮﻛﺩ
ﺪﻴﺳﺍ
ﺯﺍﺮﻓ
ﺲﻧﺍﺮﺗ
ﻦﻴﻟﻮﻛ
ﻞﻴﺘﺳﺍ
ﻭ
(Sigma S5545) (5HTﻦﻴﻣﺎﺘﭙﻳﺮﺗ
ﺯﺍ
ﺪﻌﺑ
ﻭ
ﻩﺪﺷ
ﻪﻓﺎﺿﺍ
200/1
ﺖﻈﻠﻏ
ﺎﺑ
(Chemicon MAB5270)ﻪﻳﻮﻧﺎﺛ
ﻯﺎﻫ
ﻯﺩﺎﺑ
ﻰﺘﻧﺁ
،ﻮﺷﻭ
ﺖﺴﺷ
ﻭ
ﻯﺭﺍﺪﻬﮕﻧ
ﻪﻘﻴﻗﺩ
60
ﻭ
Goat Anti-Mouse IgG FITC (Chemicon AP308F)ﻪﻓﺎﺿﺍ
100/1
ﺖﻈﻠﻏ
(Sigma,F1262) Anti Rabbit IgG FITCﻮﺷﻭ
ﺖﺴﺷ
ﺎﻫ
ﻝﻮﻠﺳ
ﺲﭙﺳ
.
ﺪﺷ
ﻯﺭﺍﺪﻬﮕﻧ
ﻪﻘﻴﻗﺩ
30
ﺕﺪﻣ
ﻪﺑ
ﻭ
ﻩﺩﺮﻛ
ﻩﺎﮕﺘﺳﺩ
ﻂﺳﻮﺗ
،ﺪﺻﺭﺩ
ﻚﻳ
ﺪﻴﺋﺪﻟﺎﻣﺮﻓﺍﺭﺎﭘ
ﺎﺑ
ﺖﻴﺒﺜﺗ
ﺯﺍ
ﺪﻌﺑ
ﻭ
ﻩﺪﺷ
ﻩﺩﺍﺩ
Win MDI2/8
ﺭﺍﺰﻓﺍ
ﻡﺮﻧ
ﻭ
Bectin Dekensonﻝﺪﻣ
ﻯﺮﺘﻣﻮﺘﻳﺎﺳﻮﻠﻓ
.
ﺪﺷ
ﻩﺪﻧﺍﻮﺧ
8/2
ﺲﻧﺎﺳﺭﻮﻠﻓ
ﻮﻧﻮﻤﻳﺍ
ﻯﺰﻴﻣﺁ
ﮓﻧﺭ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻰﺻﺎﺼﺘﺧﺍ
ﻯﺎﻫ
ﻥژ
ﻰﺘﻧﺁ
ﻯﺍﺮﺑ
ﻰﻤﻴﺷﻮﺘﻴﺳ
ﻮﻧﻮﻤﻳﺍ
ﻞﺣﺍﺮﻣ
ﺪﻴﺋﺪﻟﺍ
ﺎﻣﺭﻮﻓﺍﺭﺎﭘ
ﻝﻮﻠﺤﻣ
ﺎﺑ
ﺎﻫ
ﻝﻮﻠﺳ
:
ﺪﺷﺎﺑ
ﻰﻣ
ﺮﻳﺯ
ﺡﺮﺷ
ﻪﺑ
ﻰﺒﺼﻋ
ﺭﺩ
ﺩﺍﺮﮔ
ﻰﺘﻧﺎﺳ
ﻪﺟﺭﺩ
ﺭﺎﻬﭼ
ﻯﺎﻣﺩ
ﺭﺩ
ﻪﻘﻴﻗﺩ
10
ﺕﺪﻣ
ﻪﺑ
ﺪﺻﺭﺩ
4
ﺪﺻﺭﺩ
ﻢﻫﺩﻭﺩ
Triton x-100ﻝﻮﻠﺤﻣ
ﺎﺑ
ﻭ
(
Fixed)
ﺖﻴﺒﺜﺗ
ﻝﺎﭽﺨﻳ
ﻩﺪﺷ
ﺮﻳﺬﭘﺫﻮﻔﻧ
ﻕﺎﺗﺍ
ﻯﺎﻣﺩ
ﺭﺩ
ﻪﻘﻴﻗﺩ
ﺞﻨﭘ
ﺕﺪﻣ
ﻪﺑ
PBSﺭﺩ
ﻩﺪﺷ
ﻪﻴﻬﺗ
ﻭ
ﻩﺪﺷ
ﻖﻴﻗﺭ
β-TubulinIII ،Tau ،GFAPﻪﻴﻟﻭﺍ
ﻯﺩﺎﺑ
ﻰﺘﻧﺁ
ﺎﺑ
ﺲﭙﺳ
ﻭ
4
ﺭﺩ
ﺯﻭﺭ
ﻪﻧﺎﺒﺷ
ﻚﻳ
ﺎﻳ
ﺩﺍﺮﮔ
ﻰﺘﻧﺎﺳ
ﻪﺟﺭﺩ
37
ﻯﺎﻣﺩ
ﺭﺩ
ﻪﻘﻴﻗﺩ
60
ﺕﺪﻣ
ﻪﺑ
ﻪﻳﻮﻧﺎﺛ
ﻯﺎﻫ
ﻯﺩﺎﺑ
ﻰﺘﻧﺁ
ﻮﺷﻭ
ﺖﺴﺷ
ﺯﺍ
ﺪﻌﺑ
.
ﺪﺷ
ﻪﺑﻮﻜﻧﺍ
ﺩﺍﺮﮔ
ﻰﺘﻧﺎﺳ
ﻪﺟﺭﺩ
ﺖﻈﻠﻏ
ﺎﺑ
Anti Rabbit IgG FITCﻭ
Goat anti-mouse IgG FITCﺭﺩ
ﺖﻋﺎﺳ
ﻚﻳ
ﺕﺪﻣ
ﻪﺑ
ﻭ
ﻩﺪﺷ
ﻪﻓﺎﺿﺍ
ﻪﻴﻟﻭﺍ
ﻯﺎﻫ
ﻯﺩﺎﺑ
ﻰﺘﻧﺁ
ﻪﻴﻠﻋ
200/1
.
ﺪﺷ
ﻯﺭﺍﺪﻬﮕﻧ
ﺩﺍﺮﮔ
ﻰﺘﻧﺎﺳ
ﻪﺟﺭﺩ
37
ﻯﺎﻣﺩ
RT-PCR
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻥژ
ﻥﺎﻴﺑ
ﻅﺎﺤﻟ
ﺯﺍ
ﺰﻳﺎﻤﺗ
10
ﺯﻭﺭ
ﺭﺩ
ﻝﻮﻠﺳ
10
6ﺩﻭﺪﺣ
RNA
،ﺭﻮﻈﻨﻣ
ﻦﻳﺍ
ﻯﺍﺮﺑ
.
ﺪﺷ
ﻰﺳﺭﺮﺑ
HB9, β-TubulinIII, THﻖﺑﺎﻄﻣ
ﻭ
RNX-plus solutionﻝﻮﻠﺤﻣ
ﺯﺍ
ﻩﺩﺎﻔﺘﺳﺍ
ﺎﺑ
ﺮﻈﻧ
ﺪﻣ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻝﻮﻠﺤﻣ
ﺯﺍ
ﻩﺩﺎﻔﺘﺳﺍ
ﺎﺑ
ﻪﻣﺍﺩﺍ
ﺭﺩ
.
ﺪﺷ
ﻯﺯﺎﺳﺍﺪﺟ
ﺖﻛﺮﺷ
ﻦﻳﺍ
ﻞﻛﻮﺗﻭﺮﭘ
ﺎﺑ
ﻩﺍﺮﻤﻫ
ﻰﻟﺎﻤﺘﺣﺍ
Reaction buffer ،DNAﻭ
Mgcl2ﻩﺍﺮﻤﻫ
ﻪﺑ
Dnase Iﻥﺩﻭﺰﻓﺍ
ﺎﺑ
Dnase Iﺖﻴﻟﺎﻌﻓ
ﺲﭙﺳ
.
ﺪﻳﺩﺮﮔ
ﻑﺬﺣ
،ﻩﺪﺷ
ﺝﺍﺮﺨﺘﺳﺍ
RNAﺎﺑ
ﻰﺴﻳﻮﻧﻭﺭ
ﺶﻨﻛﺍﻭ
ﺭﺩ
cDNAﺖﺧﺎﺳ
.
ﺪﺷ
ﻒﻗﻮﺘﻣ
EDTAﻡﺮﮔ
ﻰﻠﻴﻣ
25
ﺭﺩ
.
ﺪﺷ
ﻡﺎﺠﻧﺍ
Revert acid H minusﺖﻴﻛ
ﺯﺍ
ﻩﺩﺎﻔﺘﺳﺍ
ﺎﺑ
(
RT)
ﺱﻮﻜﻌﻣ
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻥژ
ﻰﺻﺎﺼﺘﺧﺍ
ﻯﺎﻫﺮﻤﻳﺍﺮﭘ
ﺯﺍ
ﻩﺩﺎﻔﺘﺳﺍ
ﺎﺑ
PCRﺶﻨﻛﺍﻭ
ﻪﻣﺍﺩﺍ
ﻪﺟﺭﺩ
93)
ﻪﻘﻴﻗﺩ
5 :
ﻪﻴﻟﻭﺍ
ﻰﮕﺘﺷﺮﺳﺍﻭ
:
ﺕﺭﻮﺻ
ﻪﺑ
PCRﻂﻳﺍﺮﺷ
.
ﺪﺷ
ﻡﺎﺠﻧﺍ
(
ﺩﺍﺮﮔ
ﻰﺘﻧﺎﺳ
ﻪﺟﺭﺩ
93)
ﻪﻴﻧﺎﺛ
45 :
ﻞﻜﻴﺳ
ﺮﻫ
ﻰﮕﺘﺷﺮﺳﺍﻭ
(
ﺩﺍﺮﮔ
ﻰﺘﻧﺎﺳ
ﺮﻛﺫ
ﻚﻳ
ﻝﻭﺪﺟ
ﺭﺩ
ﻪﻛ
ﻥژ
ﺮﻫ
ﻯﺎﻫﺮﻤﻳﺍﺮﭘ
Tmﻪﺑ
ﻪﺟﻮﺗ
ﺎﺑ
:
Annealing.
Extentionﻪﻴﻧﺎﺛ
45 :
ﻞﻜﻴﺳ
ﺮﻫ
Extention.
ﺖﺳﺍ
ﻩﺪﺷ
ﻯﺭﺎﻣﺁ
ﻥﻮﻣﺯﺁ
Parallel Experimental Design
ﺕﺭﻮﺻ
ﻪﺑ
ﺮﺿﺎﺣ
ﻪﻌﻟﺎﻄﻣ
ﻰﻳﺎﻣﻮﻨﻴﺳﺭﺎﻛ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻒﻠﺘﺨﻣ
ﻯﺎﻫ
ﻩﻭﺮﮔ
ﻥﺁ
ﺭﺩ
ﻪﻛ
ﺪﺷ
ﻰﺣﺍﺮﻃ
ﺯﺍ
ﻪﻌﻟﺎﻄﻣ
ﻦﻳﺍ
ﺭﺩ
.
ﺪﺷ
ﻰﺳﺭﺮﺑ
ﻭﺩ
ﻪﺑ
ﻭﺩ
ﺕﺭﻮﺻ
ﻪﺑ
P19ﻰﻨﻴﻨﺟ
ﺖﺴﺗ
ﺯﺍ
ﻭ
ﺎﻫ
ﻩﻭﺮﮔ
ﻪﻤﻫ
ﻦﻴﺑ
P-valueﻰﺳﺭﺮﺑ
ﻯﺍﺮﺑ
ANOVAﺖﺴﺗ
.
ﺪﺷ
ﻩﺩﺎﻔﺘﺳﺍ
ﻩﻭﺮﮔ
ﻭﺩ
ﻯﺎﻫ
ﻦﻴﮕﻧﺎﻴﻣ
ﻑﻼﺘﺧﺍ
ﻰﺳﺭﺮﺑ
ﻯﺍﺮﺑ
Tukeyﻪﺑ
p<
0/05
ﻭ
ﻩﺩﻮﺑ
ﺭﺎﺑ
ﻪﺳ
ﺎﻫ
ﻩﻭﺮﮔ
ﻯﺍﺮﺑ
ﺶﻳﺎﻣﺯﺁ
ﻯﺎﻫﺭﺍﺮﻜﺗ
ﻞﻗﺍﺪﺣ
.
ﺪﺷ
ﻪﺘﻓﺮﮔ
ﺮﻈﻧ
ﺭﺩ
ﻯﺭﺍﺩ
ﻰﻨﻌﻣ
ﻥﺍﻮﻨﻋ
ﺎﻫ
ﻪﺘﻓﺎﻳ
ﻪﺑ
(Feeder layer)ﻩﺪﻨﻨﻛ
ﻪﻳﺬﻐﺗ
ﻪﻳﻻ
ﻪﺑ
ﺯﺎﻴﻧ
ﻥﻭﺪﺑ
P19ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺪﻨﻨﻛ
ﻰﻣ
ﺪﺷﺭ
FBSﺪﺻﺭﺩ
15
ﻯﻭﺎﺣ
DMEMﻂﻴﺤﻣ
ﺭﺩ
ﻰﮔﺩﺎﺳ
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻪﺑ
P19ﻯﺩﺎﻴﻨﺑ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺰﻳﺎﻤﺗ
ﻩﻮﺤﻧ
.(1
Aﻞﻜﺷ
)
ﻑﻭﺮﻇ
ﺭﺩ
ﺎﻫ
ﻝﻮﻠﺳ
ﻦﻳﺍ
.
ﺪﺷ
ﻡﺎﺠﻧﺍ
ﻰﻧﺎﻳﻭﺭ
ﻪﺒﺷ
ﻡﺎﺴﺟﺍ
ﻞﻴﻜﺸﺗ
ﺱﺎﺳﺍ
ﺮﺑ
ﺪﻨﻫﺩ
ﻞﻜﺷ
ﺍﺭ
ﻰﻧﺎﻳﻭﺭ
ﻪﺒﺷ
ﻡﺎﺴﺟﺍ
ﻯﺎﻫ
ﻩﺩﻮﺗ
ﺪﻨﺘﺴﻫ
ﺭﺩﺎﻗ
ﻩﺪﻨﺒﺴﭼﺮﻴﻏ
.(1
B, Cﻞﻜﺷ
)
.ﺪﺷﺖﻳﻭﺭﻞﺑﺎﻗﺪﻳﺎﻣﻭﺮﺑﻡﻮﻳﺪﻴﺗﺍﺎﺑﻭﺍﺪﺟﺪﺻﺭﺩ 1. 7ﺯﺭﺎﮔﺁﻯﻭﺭPCRﺕﻻﻮﺼﺤﻣ :1 ﻝﻭﺪﺟ
Genes Primer sequences (5´-3´) Size (bp) Annealing Tempreture
HB9 F:5`-GGCGCTTTCCTACTC ATACC- 3`
R:5`-TCCTCTTCCGTCTTC TCCTCAC -3
456 64
TH F:5`-TCCTGCACTCCCTGTCAGAG– 3`
R:5`-CCAAGAGCAGCCCATCAAAGG-3`
423 59/4
Tubulin3 F:5`- GTTCCCACG TCTCCA CTTCTTC -3` R:5`-CCAGGTCATTCATGTTGC TCTC-3`
479 65
β-tubulin F:5`-GGAACATAGCCGTAAACTGC-3` R:5`-TCACTGTGCCTGAACTTACC-3`
335 67
.ﺩﺮﻛﻩﺪﻫﺎﺸﻣﻥﺍﻮﺗﻰﻣﻪﺘﻓﺎﻳﺰﻳﺎﻤﺗﺮﻴﻏﺖﻟﺎﺣﺭﺩﺍﺭP19ﻯﺎﻫﻝﻮﻠﺳAﻞﻜﺷﺭﺩ .ﺪﻫﺩﻰﻣﻥﺎﺸﻧﺍﺭﻒﻠﺘﺨﻣﻯﺎﻫﺖﻟﺎﺣﺭﺩ P19ﻯﺎﻫﻝﻮﻠﺳﻯژﻮﻟﻮﻓﺭﻮﻣ :1ﻞﻜﺷ
ﺖﻴﻠﭘﺯﻭﺭ 4 ﺯﺍﺪﻌﺑDﻞﻜﺷ .ﺪﻫﺩﻰﻣﻥﺎﺸﻧﺍﺭﻢﺸﺷﺯﻭﺭﺭﺩﻰﻧﺎﻳﻭﺭﻪﺒﺷﻡﺎﺴﺟﺍCﻞﻜﺷ .ﺪﻫﺩﻰﻣﻥﺎﺸﻧﺍﺭﻡﻮﺳﺯﻭﺭﺭﺩﻰﻧﺎﻳﻭﺭﻪﺒﺷﻡﺎﺴﺟﺍﻯﺎﻫﻩﺩﻮﺗBﻞﻜﺷ
ﻯﺎﻫ
ﻩﺩﻮﺗ
ﻥﺪﺷ
ﺖﻴﻠﭘ
ﺯﻭﺭ
4
ﺯﺍ
ﺪﻌﺑ
ﻩﺪﻣﺁ
ﻞﻤﻋ
ﻪﺑ
ﻱﺎﻫ
ﻲﺳﺭﺮﺑ
ﺯﺍ
ﻯﺎﻫ
ﻝﻮﻠﺳ
،ﻯژﻮﻟﻮﻓﺭﻮﻣ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﺎﺑ
ﺭﺎﻤﻴﺗ
ﺖﺤﺗ
p19ﻰﻟﻮﻠﺳ
ﻥﺍﻮﺗ
ﻰﻣ
ﺎﻫ
ﺺﺧﺎﺷ
ﻦﻳﺍ
ﻦﻳﺮﺗ
ﻢﻬﻣ
ﺯﺍ
ﻪﻛ
ﻩﺪﺷ
ﺺﺨﺸﻣ
ﻼﻣﺎﻛ
ﻰﺒﺼﻋ
ﺩﺮﻛ
ﻩﺭﺎﺷﺍ
ﺎﻫ
ﺖﻳﺭﺪﻧﺩﻭ
ﺎﻫ
ﻥﻮﺴﻛﺍ
ﻪﺒﺷ
ﻰﻳﺎﻫ
ﻪﻟﺎﻄﺘﺳﺍ
ﺩﻮﺟﻭ
ﻪﺑ
.(1
Dﻞﻜﺷ
)
ﺭﺎﻤﻴﺗ
ﺮﺑ
ﻥﺪﺷ
ﺖﻴﻠﭘ
ﺯﻭﺭ
4
ﺯﺍ
ﺪﻌﺑ
ﻩﺪﻣﺁ
ﻞﻤﻋ
ﻪﺑ
ﻯﺎﻫ
ﻰﺳﺭﺮﺑ
ﺯﺍ
ﻥﺎﺸﻧ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﻭ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﺎﺑ
p19ﻝﻮﻠﺳ
ﻯﺎﻫ
ﻩﺩﻮﺗ
ﺩﻭﺪﺣ
ﻭ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﻩﻭﺮﮔ
ﺭﺩ
ﺎﻫ
ﻝﻮﻠﺳ
ﺪﺻﺭﺩ
63
±
3
ﺩﻭﺪﺣ
.
ﺪﺷ
ﻩﺩﺍﺩ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
+
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﻩﻭﺮﮔ
ﺭﺩ
ﺎﻫ
ﻝﻮﻠﺳ
ﺪﺻﺭﺩ
74
±
1
ﺭﺩ
ﻥﺍﺰﻴﻣ
ﻦﻳﺍ
ﺎﻣﺍ
(
p<
0/05)
ﺖﺳﺍ
ﻩﺩﺮﻛ
ﻥﺎﻴﺑ
ﺍﺭ
Tuj1ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﺪﺻﺭﺩ
35
±
3
ﺩﻭﺪﺣ
ﻪﺑ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﻭ
ﻝﺮﺘﻨﻛ
ﻯﺎﻫ
ﻩﻭﺮﮔ
.(2
ﻞﻜﺷ
)
(p<
0/001)
ﺖﻓﺎﻳ
ﺶﻫﺎﻛ
ﺭﺩ
(
ﻰﺘﻴﺳﻭﺮﺘﺳﺁ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺮﻛﺭﺎﻣ
)
GFAPﻥﺎﻴﺑ
ﻥﺍﺰﻴﻣ
ﻦﻴﻨﭼ
ﻢﻫ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
+
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﻭ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﻩﻭﺮﮔ
ﻭﺩ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﻭ
ﻝﺮﺘﻨﻛ
ﻯﺎﻫ
ﻩﻭﺮﮔ
ﺭﺩ
ﺎﻣﺍ
ﺪﺻﺭﺩ
62
±
4
ﺩﻭﺪﺣ
ﻪﺑ
ﻞﻜﺷ
)
(p<
0/001)
ﺖﻓﺎﻳ
ﺶﻫﺎﻛ
ﺪﺻﺭﺩ
5
±
1
ﺩﻭﺪﺣ
ﻪﺑ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻝﺮﺘﻨﻛ
ﻯﺎﻫ
ﻩﻭﺮﮔﺭﺩ
ﻰﻳﺍﺯ
ﺐﺼﻋ
ﻥﺍﺰﻴﻣ
ﻥﺩﻮﺑ
ﻦﻴﻳﺎﭘ
ﻪﺑ
ﻪﺟﻮﺗ
ﺎﺑ
ﻦﻳﺍﺮﺑﺎﻨﺑ
.(3
ﺭﺩ
ﻂﻘﻓ
ﺎﻫ
ﻰﺳﺭﺮﺑ
ﻯﺪﻌﺑ
ﺕﺎﺸﻳﺎﻣﺯﺁ
ﺭﺩ
،ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﻭ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
+
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﻭ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﻯﺎﻫ
ﻩﻭﺮﮔ
.
ﺪﺷ
ﻡﺎﺠﻧﺍ
ﺭﺩ
Map-2ﻭ
Tauﺮﻛﺭﺎﻣ
ﻥﺎﻴﺑ
ﻥﺍﺰﻴﻣ
ﺮﺑ
ﻩﺪﻣﺁ
ﻞﻤﻋ
ﻪﺑ
ﻯﺎﻫ
ﻰﺳﺭﺮﺑ
ﺯﺍ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
+
ﻚﻴﺋﻮﻨﻴﺗﺭ
ﺪﻴﺳﺍ
ﻭ
ﻚﻴﺋﻮﻨﻴﺗﺭ
ﺪﻴﺳﺍ
ﻩﻭﺮﮔ
ﻭﺩ
ﻭ
،
Tauﺮﻛﺭﺎﻣ
ﺎﻫ
ﻝﻮﻠﺳ
ﺪﺻﺭﺩ
49
±
8
ﻭ
53
±
12
ﺐﻴﺗﺮﺗ
ﻪﺑ
ﻥﺍﻮﺗ
ﻲﻣ
ﺩﺮﻛ
ﻥﺎﻴﺑ
ﺍﺭ
Map-2ﺮﻛﺭﺎﻣ
ﺎﻫ
ﻝﻮﻠﺳ
ﺪﺻﺭﺩ
59
±
1
ﻭ
65
±
5
.(4
ﻞﻜﺷ
)
ﻯﺎﻫ
ﻥژ
ﻥﺎﻴﺑ
ﺎﺑ
ﺍﺭ
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻍﻮﻠﺑ
RT-PCRﺰﻴﻟﺎﻧﺁ
ﻭ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﻩﻭﺮﮔ
ﻭﺩ
ﺭﺩ
mRNAﺢﻄﺳ
ﺭﺩ
HB9ﻭ
Tuj1, TH.(5
ﻞﻜﺷ
)
ﺩﺮﻛ
ﺪﻴﻳﺎﺗ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
+
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﻭﻝﺮﺘﻨﻛﻩﻭﺮﮔﺎﺑﺭﺍﺩﻰﻨﻌﻣﻑﻼﺘﺧﺍa.ﺪﻫﺩﻰﻣﻥﺎﺸﻧﺍﺭﻥﺪﺷﺖﻴﻠﭘﺯﻭﺭ 4 ﺯﺍﺪﻌﺑTuj1ﻯﺭﺎﻣﺁﺰﻴﻟﺎﻧﺁﺯﺍﻞﺻﺎﺣﻯﺮﺘﻣﻮﺘﻳﺎﺳﻮﻠﻓﺞﻳﺎﺘﻧ :ﭗﭼﺖﻤﺳ :2ﻞﻜﺷ
ﺯﻭﺭ 4 ﺯﺍﺪﻌﺑTuj1ﻰﻤﻴﺷﻮﺘﻴﺳﻮﻧﻮﻤﻳﺍﺮﻳﻮﺼﺗﺖﺳﺍﺭﺖﻤﺳ .(p<0/05) ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍﻩﻭﺮﮔﺎﺑﺭﺍﺩﻰﻨﻌﻣﻑﻼﺘﺧﺍ b .(p<0/001) ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍﻰﭘﺍﻭﺭﺪﻴﻫﺩ
.ﺪﻫﺩﻰﻣﻥﺎﺸﻧﺍﺭﻥﺪﺷﺖﻴﻠﭘ
ﻯﺎﻫﻩﻭﺮﮔﺎﺑﺭﺍﺩﻰﻨﻌﻣﻑﻼﺘﺧﺍa.ﺪﻫﺩﻰﻣﻥﺎﺸﻧﺍﺭﻥﺪﺷﺖﻴﻠﭘﺯﻭﺭ 4 ﺯﺍﺪﻌﺑGFAPﻯﺭﺎﻣﺁﺰﻴﻟﺎﻧﺁﺯﺍﻞﺻﺎﺣﻯﺮﺘﻣﻮﺘﻳﺎﺳﻮﻠﻓﺞﻳﺎﺘﻧ :ﺖﺳﺍﺭﺖﻤﺳ :3 ﻞﻜﺷ .ﺪﻫﺩﻰﻣﻥﺎﺸﻧﺍﺭﻥﺪﺷﺖﻴﻠﭘﺯﻭﺭ4 ﺯﺍﺪﻌﺑGFAPﻰﻤﻴﺷﻮﺘﻴﺳﻮﻧﻮﻤﻳﺍﺮﻳﻮﺼﺗ :ﭗﭼﺖﻤﺳ .ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍﻰﭘﺍﻭﺭﺪﻴﻫﺩﻭﻝﺮﺘﻨﻛ
ﻲﻳﺍﺯﺐﺼﻋﺶﻳﺍﺰﻓﺍﻭﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍﻲﭘﺍﻭﺭﺪﻴﻫﺩ
80
70
60
50
40
30
20
10
0
T
uj 1%
Control DHEA
a
b
RA RA&DHEA
80
70
60
50
40
30
20
10
0
GAFP%
Control DHEA
a
a
ﻑﻼﺘﺧﺍ a .ﺪﻫﺩﻰﻣﻥﺎﺸﻧﺍﺭﺎﻫﺮﺘﻴﻤﺴﻧﺍﺮﺗﻭﺭﻮﻧﻯﺭﺎﻣﺁﺰﻴﻟﺎﻧﺁﺯﺍﻞﺻﺎﺣﻯﺮﺘﻣﻮﺘﻳﺎﺳﻮﻠﻓﺞﻳﺎﺘﻧﺖﺳﺍﺭﺖﻤﺳ :5ﻞﻜﺷ .ﺪﻫﺩﻰﻣﻥﺎﺸﻧﺍﺭﻰﺒﺼﻋﻎﻟﺎﺑﻯﺎﻫﻥژﻯﺍﺮﺑRT-PCRﺰﻴﻟﺎﻧﺁﭗﭼﺖﻤﺳ .(p<0/001) ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍﻩﻭﺮﮔﺎﺑﺭﺍﺩﻰﻨﻌﻣ
80
70
60
50
40
30
20
10
0
MAP-2 & T
au%
Map-2 Tau
RA RA & DHEA
MAP-2ﻰﻤﻴﺷﻮﺘﻴﺳﻮﻧﻮﻤﻳﺍﺮﻳﻮﺼﺗﻦﻴﻳﺎﭘ .ﺪﻫﺩﻰﻣﻥﺎﺸﻧﺍﺭﻥﺪﺷﺖﻴﻠﭘﺯﻭﺭ 4 ﺯﺍﺪﻌﺑTauﻭ MAP-2ﻯﺭﺎﻣﺁﺰﻴﻟﺎﻧﺁﺯﺍﻞﺻﺎﺣﻯﺮﺘﻣﻮﺘﻳﺎﺳﻮﻠﻓﺞﻳﺎﺘﻧ،ﻻﺎﺑ :4ﻞﻜﺷ .ﺮﺘﻣﻭﺮﻜﻴﻣ 100 :ﺱﺎﻴﻘﻣ .ﺪﻫﺩﻰﻣﻥﺎﺸﻧﺍﺭﻥﺪﺷﺖﻴﻠﭘﺯﻭﺭ 4 ﺯﺍﺪﻌﺑ(B) Tauﻭ (A)
P19
ﻰﻟﻮﻠﺳ
ﻯﺎﻫ
ﻩﺩﻮﺗ
ﻥﺪﺷ
ﺖﻴﻠﭘ
ﺯﻭﺭ
4
ﺯﺍ
ﺪﻌﺑ
ﺎﻫﺮﺘﻴﻤﺴﻧﺍﺮﺗﻭﺭﻮﻧ
ﻰﺳﺭﺮﺑ
ﺪﺻﺭﺩ
35
ﺩﻭﺪﺣ
ﺩﺍﺩ
ﻥﺎﺸﻧ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﻭ
ﻚﻴﺋﻮﻨﻴﺗﺭ
ﺪﻴﺳﺍ
ﺎﺑ
ﺎﻫ
ﻝﻮﻠﺳ
ﺪﺻﺭﺩ
30
ﻭ
ﺕﺎﻣﺎﺗﻮﻠﮔ
ﺎﻫ
ﻝﻮﻠﺳ
ﺪﺻﺭﺩ
50
،ﻦﻴﻟﻮﻛ
ﻞﻴﺘﺳﺍ
ﺎﻫ
ﻝﻮﻠﺳ
ﻩﻭﺮﮔ
ﺭﺩ
THﺮﺘﻴﻤﺴﻧﺍﺮﺗﻭﺭﻮﻧ
ﻥﺎﻴﺑ
ﻥﺍﺰﻴﻣ
ﺎﻣﺍ
.
ﺖﺳﺍ
ﻩ
ﺩﺮﻛ
ﻥﺎﻴﺑ
ﺍﺭ
ﻦﻴﻧﻮﺗﻭﺮﺳ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
+
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﻩﻭﺮﮔ
ﺭﺩ
ﻭ
ﺪﺻﺭﺩ
5
ﺩﻭﺪﺣ
ﻚﻴﺋﻮﻨﻴﺗﺭ
ﺪﻴﺳﺍ
.(4
ﻞﻜﺷ
)
(p<
0/001)
ﺖﻓﺎﻳ
ﺶﻳﺍﺰﻓﺍ
ﺪﺻﺭﺩ
20
ﺩﻭﺪﺣ
ﻪﺑ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﺚﺤﺑ
ﺮﺑ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﺮﻴﺛﺎﺗ
ﺯﺍ
ﻩﺪﻣﺁ
ﺖﺳﺩ
ﻪﺑ
ﺕﺎﻌﻟﺎﻄﻣ
ﺶﻳﺍﺰﻓﺍ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
،ﺪﻫﺩ
ﻰﻣ
ﻥﺎﺸﻧ
Neural Stem Cells Tau-1,ﻯﺎﻫﺮﻛﺭﺎﻣ
mRNAﺶﻳﺍﺰﻓﺍ
ﻰﺴﭘﺎﻨﻴﺳ
ﺕﻻﺎﺼﺗﺍ
،ﻰﻧﻮﺴﻛﺍ
ﺪﺷﺭ
ﺦﺳﺎﭘ
ﻖﻳﺮﻃ
ﺯﺍ
ﻪﻛ
ﺩﺭﺍﺩ
ﻩﺍﺮﻤﻫ
ﻪﺑ
ﺍﺭ
ﻦﻴﻣﺎﭘﻭﺩ
ﻯﺎﻫ
ﻩﺪﻨﻫﺩ
ﺦﺳﺎﭘ
ﻭ
Tau-2ﺪﻫﺩ
ﻰﻣ
ﺶﻳﺍﺰﻓﺍ
ﺍﺭ
ﻰﻟﻮﻠﺳ
ﻞﺧﺍﺩ
ﻢﻴﺴﻠﻛ
ﺕﺎﺗﺭﺎﭙﺳﺁ
ﻯﺩ
ﻞﻴﺘﻣ
-Nﻯﺎﻫ
ﻩﺪﻨﻫﺩ
ﺪﻴﺳﺍ
ﻝﺎﻣﺮﻧ
ﺯﻭﺩ
ﺎﺑ
ﺭﺎﻤﻴﺗ
ﺯﻭﺭ
6
ﻰﻃ
ﺭﺩ
p19ﻝﻮﻠﺳ
ﻯﺎﻫ
ﻩﺩﻮﺗ
.(15
،
14)
ﻯﺎﻫﺮﻛﺭﺎﻣ
ﺎﻫ
ﻝﻮﻠﺳ
ﺪﺻﺭﺩ
60
ﺩﻭﺪﺣ
ﻥﺪﺷ
ﺖﻴﻠﭘ
ﺯﺍ
ﺪﻌﺑ
ﺯﻭﺭ
4
ﻭ
ﻚﻴﺋﻮﻨﻴﺗﺭ
50
ﺩﻭﺪﺣ
ﻭ
GFAPﻂﺳﺍﻭ
ﺪﺣ
ﻰﺘﻨﻣﻼﻓ
ﻦﻴﻴﺗﻭﺮﭘ
ﻭ
MAP-2 Tuj-1ﻰﺒﺼﻋ
ﺪﺻﺭﺩ
50
ﺩﻭﺪﺣ
ﺎﻫﺮﺘﻴﻤﺴﻧﺍﺮﺗﻭﺭﻮﻧ
ﻰﺳﺭﺮﺑ
ﺭﺩ
ﻭ
Tauﺮﻛﺭﺎﻣ
ﺎﻫ
ﻝﻮﻠﺳ
ﺪﺻﺭﺩ
ﻦﻴﻧﻮﺗﻭﺮﺳ
ﺪﺻﺭﺩ
33
،ﻦﻴﻟﻮﻛ
ﻞﻴﺘﺳﺍ
ﺪﺻﺭﺩ
35
،ﺕﺎﻣﺎﺗﻮﻠﮔ
ﻯﻭﺎﺣ
ﺎﻫ
ﺐﺼﻋ
ﻩﻭﺮﮔ
ﻪﻛ
ﻲﺗﺭﻮﺻ
ﺭﺩ
ﺪﻨﻨﻛ
ﻰﻣ
ﻥﺎﻴﺑﺍﺭ
ﺯﻼﻴﺴﻛﻭﺭﺪﻴﻫ
ﻦﻳﺯﻭﺮﻴﺗ
ﺪﺻﺭﺩ
2
ﻭ
ﻪﺑ
Tuj-1ﻥﺎﻴﺑ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
+
ﻚﻴﺋﻮﻨﻴﺗﺭ
ﺪﻴﺳﺍ
ﺎﺑ
ﻩﺪﺷ
ﺭﺎﻤﻴﺗ
ﺶﻳﺍﺰﻓﺍ
ﺪﺻﺭﺩ
20
ﺩﻭﺪﺣ
ﻪﺑ
ﺯﻼﻴﺴﻛﻭﺭﺪﻴﻫ
ﻦﻳﺯﻭﺮﻴﺗ
ﻭ
ﺪﺻﺭﺩ
70
ﺩﻭﺪﺣ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﻥﺪﺷ
ﻪﻓﺎﺿﺍ
،ﺪﻫﺩ
ﻰﻣ
ﻥﺎﺸﻧ
ﺎﻫ
ﻪﺘﻓﺎﻳ
ﻦﻳﺍ
.
ﺖﺳﺍ
ﻪﺘﻓﺎﻳ
70
60
50
40
30
20
10
0
Neurotransmitters %
ACT GLU SER TH
RA
RA & DHEA
Tuj-1
HB-9
TH
β-tubulin
RA RA+DHEA
،ﻰﻳﺍﺯ
ﺐﺼﻋ
ﺶﻳﺍﺰﻓﺍ
ﺮﺑ
ﻩﻭﻼﻋ
ﻰﺒﺼﻋ
ﻯﺎﻫﺯﺎﺳ
ﺶﻴﭘ
ﻞﻴﻜﺸﺗ
ﻡﺎﮕﻨﻫ
ﺭﺩ
ﻦﻜﻤﻣ
ﻭ
ﺪﻫﺩ
ﻰﻣ
ﻕﻮﺳ
ﺎﻫ
ﻚﻳژﺮﻨﻴﻣﺎﭘﻭﺩ
ﺖﻤﺳ
ﻪﺑ
ﺍﺭ
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻯﺎﻫ
ﺶﻘﻧ
ﻰﺒﺼﻋ
ﻯﺎﻫﺯﺎﺳ
ﺶﻴﭘ
ﻞﻴﻜﺸﺗ
ﺯﺍ
ﺪﻌﺑ
ﻥﺁ
ﻥﺪﺷ
ﻪﻓﺎﺿﺍ
ﺖﺳﺍ
.
ﺪﻨﻛ
ﺎﻔﻳﺍ
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺮﺑ
ﺍﺭ
ﻯﺮﮕﻳﺩ
ﻞﺣﺍﺮﻣ
ﺮﺑ
ﻯﺩﺎﻳﺯ
ﺮﻴﺛﺎﺗ
،ﺖﺳﺍ
Aﻦﻴﻣﺎﺘﻳﻭ
ﻝﺎﻌﻓ
ﻡﺮﻓ
ﻪﻛ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﻰﻟﻮﻠﺳ
ﻩﺩﺭ
ﻦﻴﻌﺗ
،ﻰﻳﺍﺯ
ﻡﺍﺪﻧﺍ
ﻞﺣﺍﺮﻣ
ﺭﺩ
ﺐﻴﻛﺮﺗ
ﻦﻳﺍ
.
ﺩﺭﺍﺩ
ﻥﺍﺭﻮﻧﺎﺟ
ﻰﻨﻳﻮﻜﺗ
ﻂﺳﻮﺗ
ﻩﺩﺎﻣ
ﻦﻳﺍ
ﻰﻨﻴﻨﺟ
ﻥﺍﺭﻭﺩ
ﺭﺩ
.(16-21)
ﺩﺭﺍﺩ
ﺮﺛﺍ
ﺰﻴﻧ
ﻰﻟﻮﻠﺳ
گﺮﻣ
ﻭ
ﺮﻴﺛﺎﺗ
ﺎﺑ
ﻭ
ﺩﻮﺷ
ﻰﻣ
ﻪﺘﺧﺎﺳ
Retinaldehyde Dehydrogenaseﻥژ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻯﺎﻘﻟﺍ
ﺐﺟﻮﻣ
ﺰﻐﻣ
ﻪﻗﺎﺳ
ﻭ
CNSﻰﻧﺎﻳﻭﺭ
ﻯﺩﺎﻴﻨﺑ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺮﺑ
ﻝﺎﻴﻠﮔ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻭ
ﺐﺼﻋ
ﻞﻴﻜﺸﺗ
ﺖﻤﺳ
ﻪﺑ
ﺎﻬﻧﺁ
ﺰﻳﺎﻤﺗ
ﻭ
ﻰﺒﺼﻋ
ﺯﺎﺳ
ﺶﻴﭘ
ﻯﺎﻫ
ﻩﺪﻨﻫﺩ
ﺦﺳﺎﭘ
ﺯﺍ
ﻰﻫﻭﺮﮔ
ﻚﻳ
ﻪﺑ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
.(20-22)
ﺩﻮﺷ
ﻰﻣ
ﺎﺗ
ﻭﺩ
P19ﻯﺎﻫ
ﻝﻮﻠﺳ
.(23)
ﺩﻮﺷ
ﻰﻣ
ﻞﺼﺘﻣ
(
RARs)
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﺎﻫ
ﻝﻮﻠﺳ
ﻦﻳﺍ
ﻦﻳﺍﺮﺑﺎﻨﺑ
.(24)
ﺪﻨﻛ
ﻰﻣ
ﻥﺎﻴﺑ
ﺍﺭ
RARβﻭ
RARaﻯﺎﻫ
ﻥژ
ﺯﺍ
ﻯﺎﻫ
ﻥژ
ﺯﺍ
ﻰﺧﺮﺑ
ﺎﻬﻨﺗ
،ﺩﺮﻴﮔ
ﻰﻣ
ﺭﺍﺮﻗ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﺽﺮﻌﻣ
ﺭﺩ
ﻪﻛ
ﻰﻧﺎﻣﺯ
ﻦﻳﺍ
ﺭﺩ
.(25)
ﺎﻬﻧﺁ
ﻪﻤﻫ
ﻪﻧ
ﺪﻨﻛ
ﻰﻣ
ﻥﺎﻴﺑ
ﺍﺭ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﻪﺑ
ﻩﺪﻨﻫﺩ
ﺦﺳﺎﭘ
ﻩﺪﺷ
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻯﺎﻘﻟﺍ
ﺐﺒﺳ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﺪﺷ
ﻩﺪﻳﺩ
ﻪﻌﻟﺎﻄﻣ
ﺭﺩ
.
ﺪﻨﻛ
ﻰﻤﻧ
ﺎﻔﻳﺍ
ﺍﺭ
ﻰﺸﻘﻧ
ﻦﻴﻨﭼ
ﻰﻳﺎﻬﻨﺗ
ﻪﺑ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﺎﻣﺍ
ﻥﺍﺰﻴﻣ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﻩﺍﺮﻤﻫ
ﻪﺑ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﻪﻛ
ﻰﺗﺭﻮﺻ
ﺭﺩ
.
ﺪﻫﺩ
ﻰﻣ
ﺶﻳﺍﺰﻓﺍ
ﺮﺘﺸﻴﺑ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﻪﺑ
ﺖﺒﺴﻧ
ﻰﺘﺣ
ﺍﺭ
ﻰﻳﺍﺯ
ﺐﺼﻋ
ﺎﻫ
ﻥژ
ﺯﺍ
ﻰﻫﻭﺮﮔ
ﻰﺴﻳﻮﻧﻭﺭ
ﻯﺎﻘﻟﺍ
ﺐﺟﻮﻣ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﺯﺍ
ﻩﺩﺎﻔﺘﺳﺍ
ﻊﻗﺍﻭ
ﻰﺴﻳﻮﻧﻭﺭ
ﺭﻮﺘﻛﺎﻓ
،Sonic hedgehogﻰﺤﺷﺮﺗ
ﻦﻴﺗﻭﺮﭘﻮﻜﻴﻠﮔ
ﺪﻨﻧﺎﻣ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺭﺩ
Kappa Opidiﻭ
pax-6 ،Mash-1 ،HNF-3ﻥژ
ﻒﻠﺘﺨﻣ
ﻯﺎﻫ
ﻥژ
ﻥﺎﻴﺑ
ﻭ
ﺭﺎﺸﺑﺁ
ﻦﻳﺍ
ﻥﺪﺷ
ﻦﺷﻭﺭ
ﻭ
ﺩﻮﺷ
ﻰﻣ
ﻰﻧﺎﻳﻭﺭ
ﻯﺩﺎﻴﻨﺑ
ﻥﺎﻴﺑ
ﻥﺍﺰﻴﻣ
ﺶﻫﺎﻛ
ﻭ
ﻝﺎﻣﺭﺩﻭﺰﻣ
ﺰﻳﺎﻤﺗ
ﻒﻗﻮﺗ
ﻭ
ﻰﺒﺼﻋ
ﺰﻳﺎﻤﺗ
ﻯﺎﻘﻟﺍ
ﺐﺟﻮﻣ
Brach-ﻭ
cardic actin ،globinﺪﻨﻧﺎﻣ
ﻝﺎﻣﺭﺩﻭﺰﻣ
ﻯﺎﻫ
ﻥژ
mRNAﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﺖﺳﺍ
ﻦﻜﻤﻣ
.(27
،
26
،
20)
ﺩﻮﺷ
ﻰﻣ
yaryﻩﺍﺮﻤﻫ
ﻪﺑ
ﺎﻣﺍ
ﺪﺷﺎﺒﻧ
ﻰﻟﺎﻨﮕﻴﺳ
ﺮﻴﺴﻣ
ﻦﻳﺍ
ﻥﺩﺮﻛ
ﻝﺎﻌﻓ
ﻪﺑ
ﺭﺩﺎﻗ
ﻰﻳﺎﻬﻨﺗ
ﻪﺑ
.
ﺪﻫﺩ
ﺶﻳﺍﺰﻓﺍ
ﺍﺭ
ﻰﻳﺍﺯ
ﺐﺼﻋ
ﻥﺍﺰﻴﻣ
ﻯﺮﮕﻳﺩ
ﻯﺎﻫ
ﻡﺰﻴﻧﺎﻜﻣ
ﺯﺍ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
+
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﻩﻭﺮﮔ
،ﺩﺍﺩ
ﻥﺎﺸﻧ
ﻰﻠﺒﻗ
ﻪﻌﻟﺎﻄﻣ
ﻝﺎﺜﻣ
ﻥﺍﻮﻨﻋ
ﻪﺑ
ﻩﻭﺮﮔ
ﻪﺑ
ﺖﺒﺴﻧ
ﺍﺭ
ﻰﺒﺼﻋ
ﺯﺎﺳ
ﺶﻴﭘ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻞﻴﻜﺸﺗ
ﻥﺍﺰﻴﻣ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﺶﻳﺍﺰﻓﺍ
ﺖﺳﺍ
ﻦﻜﻤﻣ
ﻦﻳﺍﺮﺑﺎﻨﺑ
.(13)
ﺪﻫﺩ
ﻰﻣ
ﺶﻳﺍﺰﻓﺍ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﺖﻠﻋ
ﻪﺑ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
+
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﻩﻭﺮﮔ
ﺭﺩ
ﻰﻳﺍﺯ
ﺐﺼﻋ
ﻭ
DHEAS.
ﺪﺷﺎﺑ
ﻩﻭﺮﮔ
ﻦﻳﺍ
ﺭﺩ
ﻰﺒﺼﻋ
ﺯﺎﺳ
ﺶﻴﭘ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺶﻳﺍﺰﻓﺍ
ﻪﺑ
ﺩﻮﺷ
ﻰﻣ
ﻦﻴﻣﺁ
ﻝﻮﻜﺗﺎﻛ
ﺰﺘﻨﺳ
ﺶﻳﺍﺰﻓﺍ
ﺐﺒﺳ
Allopregnanolonﻭ
ﺯﻼﻴﺴﻛﻭﺭﺪﻴﻫ
ﻦﻳﺯﻭﺮﻴﺗ
ﻯﺎﻫ
ﻩﺪﻨﻨﻛﺭﺎﻬﻣ
ﻂﺳﻮﺗ
ﺮﺛﺍ
ﻦﻳﺍ
ﻪﻛ
ﻯﺭﻮﻃ
ﺎﻫﺪﻴﻳﻭﺮﺘﺳﺍ
ﻦﻳﺍ
.
ﺩﻮﺷ
ﻰﻣ
ﻒﻗﻮﺘﻣ
3-,ﻭ(-4hydroxyphenyl)-ala-nineﺩﻮﺷ
ﻰﻣ
ﺯﻼﻴﺴﻛﻭﺭﺪﻴﻫ
ﻦﻳﺯﻭﺮﻴﺗ
ﻦﻴﺌﺗﻭﺮﭘ
ﻭ
mRNAﺢﻄﺳ
ﺶﻳﺍﺰﻓﺍ
ﺐﺒﺳ
ﻯﺪﻴﻠﻛ
ﻩﺪﻨﻨﻛ
ﻢﻴﻈﻨﺗ
ﻦﻴﺌﺗﻭﺮﭘ
ﻚﻳ
ﺯﻻﻭﺭﺪﻴﻫ
ﻮﻔﻟﻮﺳﺪﻴﺋﻭﺮﺘﺳﺍ
ﻢﻳﺰﻧﺁ
.(28)
ﻪﺑ
ﺍﺭ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﻭ
ﻩﺩﻮﺑ
ﻰﻳﺍﺯ
ﻦﻴﻨﺟ
ﻰﻃ
ﺭﺩ
ﺲﻜﺗﺭﻮﻛﻮﺌﻧ
ﺖﺳﺍ
ﻦﻜﻤﻣ
ﻦﻳﺍﺮﺑﺎﻨﺑ
.(29)
ﺪﻨﻛ
ﻰﻣ
ﻞﻳﺪﺒﺗ
ﺕﺎﻔﻟﻮﺳ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
،
p19ﻝﻮﻠﺳ
ﺯﺍ
ﻩﺪﺷ
ﻖﺘﺸﻣ
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺭﺩ
ﺯﻼﻴﺴﻛﻭﺭﺪﻴﻫ
ﻦﻳﺯﻭﺮﻴﺗ
ﻦﻴﺌﺗﻭﺮﭘ
ﺶﻳﺍﺰﻓﺍ
ﺐﺒﺳ
DHEASﻪﺑ
ﻞﻳﺪﺒﺗ
ﺎﺑ
(15)
ﻦﻴﻟﻮﻛ
ﻞﻴﺘﺳﺍ
ﻯﺎﻫﺮﺘﻴﻤﺴﻧﺍﺮﺗﻭﺭﻮﻧ
ﺭﻮﻀﺣ
ﺎﻬﻨﺗ
ﺍﺪﺘﺑﺍ
ﺭﺩ
.
ﺪﺷﺎﺑ
ﻩﺪﺷ
P19
ﻝﻮﻠﺳ
ﺯﺍ
ﻩﺪﺷ
ﺰﻳﺎﻤﺘﻣ
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺭﺩ
(30)
ﻦﻴﻣﺍ
ﻝﻮﻜﺗﺎﻛ
ﻭ
ﺯﺍ
ﻯﺩﺎﻳﺯ
ﺪﺻﺭﺩ
ﺭﺩ
ﺍﺭ
ﺕﺎﻣﺎﺗﻮﻠﮔ
ﺭﻮﻀﺣ
ﺮﻴﺧﺍ
ﺕﺎﻌﻟﺎﻄﻣ
.
ﺩﻮﺑ
ﻩﺪﺷ
ﻰﻳﺎﺳﺎﻨﺷ
ﻝﺎﻣﺮﻧ
ﺯﻭﺩ
ﺎﺑ
ﺭﺎﻤﻴﺗ
ﻉﻭﺮﺷﺯﺍ
ﺯﺍ
ﺪﻌﺑ
ﺯﻭﺭ
6 .(32
،
31)
ﺪﻫﺩ
ﻰﻣ
ﻥﺎﺸﻧ
ﺎﻫ
ﺐﺼﻋ
ﻰﺒﺼﻋ
ﻯﺎﻫﺮﻛﺭﺎﻣ
ﺎﻫ
ﻝﻮﻠﺳ
ﺪﺻﺭﺩ
85
ﺯﺍ
ﺶﻴﺑ
(5×10
-7)
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﻯﺎﻫﺯﻭﺭ
ﺭﺩ
ﻭ
ﺪﻨﻛ
ﻰﻣ
ﻥﺎﻴﺑ
ﺍﺭ
160KD،
68KDﻰﺘﻨﻣﻼﻓ
ﻯﺎﻫ
ﻦﻴﺌﺗﻭﺮﭘ
ﺪﻨﻧﺎﻣ
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺖﻴﻌﻤﺟ
ﺯﺍ
ﻰﺒﺼﻋﺮﻴﻏ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺮﻴﺜﻜﺗ
ﺖﻠﻋ
ﻪﺑ
ﺪﻌﺑ
ﻦﻴﺌﺗﻭﺮﭘ
ﺎﺑ
ﻎﻟﺎﺑ
ﻰﺘﻴﺳﻭﺮﺘﺳﺁ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻢﻫﺩ
ﺯﻭﺭ
ﺭﺩ
.(33)
ﺩﻮﺷ
ﻰﻣ
ﻪﺘﺳﺎﻛ
ﺎﻘﺑ
ﺖﺸﻛ
ﻂﻴﺤﻣ
ﺭﺩ
.(10)
ﺖﺳﺍ
ﻰﻳﺎﺳﺎﻨﺷ
ﻞﺑﺎﻗ
GFAPﻂﺳﺍﻭ
ﺪﺣ
ﻰﺘﻨﻣﻼﻓ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺭﻮﻀﺣ
ﻪﺑ
P19ﻝﻮﻠﺳ
ﺯﺍ
ﻩﺪﺷ
ﻖﺘﺸﻣ
ﻯﺎﻫ
ﺐﺼﻋ
ﺕﺪﻣ
ﻰﻧﻻﻮﻃ
ﻩﺪﺷ
ﻖﺘﺸﻣ
ﻯﺎﻫ
ﺐﺼﻋ
.(34)
ﺖﺳﺍ
ﻪﺘﺴﺑﺍﻭ
ﺪﺷﺭ
ﻯﺎﻫﺭﻮﺘﻛﺎﻓ
ﺎﻳ
ﻰﺒﺼﻋﺮﻴﻏ
ﻞﻘﺘﻨﻣ
(35)
ﻥﺍﺭﺍﺪﻧﺎﺘﺴﭘ
ﻯﺰﻛﺮﻣ
ﻰﺒﺼﻋ
ﻢﺘﺴﻴﺳ
ﻪﺑ
ﻪﻛ
ﻰﻧﺎﻣﺯ
P19ﻝﻮﻠﺳ
ﺯﺍ
P19ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺪﻧﻮﻴﭘ
ﺎﺑ
ﻰﺗﺎﻌﻟﺎﻄﻣ
ﺭﺩ
.
ﺪﻨﺷﺎﺑ
ﻪﺘﺷﺍﺩ
ﺩﺮﻜﻠﻤﻋ
ﺪﻨﻧﺍﻮﺗ
ﻰﻣ
،ﺩﻮﺷ
ﻦﻳﺍ
ﻯﺎﻫ
ﺐﺼﻋ
)
ﻎﻟﺎﺑ
ﺕﺭ
stratumﻪﻴﺣﺎﻧ
ﻪﺑ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﺎﺑ
ﻩﺪﺷ
ﺭﺎﻤﻴﺗ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻯﺎﻫ
ﺐﺼﻋ
،ﺩﺍﺩ
ﻥﺎﺸﻧ
(
ﺩﻮﺑ
ﻩﺪﺷ
ﺐﻳﺮﺨﺗ
ﻦﻴﺴﻛﻮﺗﻭﺭﻮﻧ
ﺎﺑ
ﻪﻴﺣﺎﻧ
ﺪﻨﺑﺎﻳ
ﻰﻣ
ﺎﻘﺑ
ﻪﻴﺣﺎﻧ
ﻦﻳﺍ
ﺭﺩ
ﻪﺘﻔﻫ
13
ﺕﺪﻣ
ﻪﺑ
ﻰﺒﺼﻋﺮﻴﻏ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻭ
P19ﻥﺩﺮﻛ
ﻝﺎﻌﻓ
ﺭﺩ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﻪﻛ
ﺪﻫﺩ
ﻰﻣ
ﻥﺎﺸﻧ
ﺎﻫ
ﻰﺳﺭﺮﺑ
.(36)
ﻪﺑ
NMDAR-1ﻯﺎﻫﺪﺣﺍﻭﺮﻳﺯ
.
ﺩﺭﺍﺩ
ﺮﻴﺛﺎﺗ
NMDAﻯﺎﻫ
ﻩﺪﻨﻫﺩ
ﺦﺳﺎﭘ
ﻰﮔﺪﻧﺯ
ﻦﻴﻟﻭﺍ
ﻭ
ﻯﺭﺍﺩﺭﺎﺑ
ﻯﺎﻬﺘﻧﺍ
ﺭﺩ
ﺲﻜﺗﺭﻮﻛﻮﺌﻧ
Vﻪﻳﻻ
ﺭﺩ
ﻰﻨﻳﻮﻜﺗ
ﺭﻮﻃ
ﻯﺎﻫ
ﻰﮔﮋﻳﻭ
ﻒﻠﺘﺨﻣ
ﻯﺎﻫﺪﺣﺍﻭ
ﺮﻳﺯ
.(37)
ﺩﻮﺷ
ﻰﻣ
ﻥﺎﻴﺑ
(
ﺕﺭ
ﺭﺩ
)
ﻯﺩﺍﺯﻮﻧ
ﺖﺳﺍ
ﻦﻜﻤﻣ
ﻦﻳﺍﺮﺑﺎﻨﺑ
،ﻪﺘﺷﺍﺩ
ﻰﺗﻭﺎﻔﺘﻣ
ﻥﻮﻴﺳﻼﻳﺮﻔﺴﻓ
ﻭ
ﻰﻜﻳژﻮﻟﻮﻳﺰﻴﻓﻭﺮﺘﻜﻟﺍ
.(38-40)
ﺪﺷﺎﺑ
ﻪﺘﺷﺍﺩ
ﺰﻴﻧ
ﻰﺗﻭﺎﻔﺘﻣ
ﻯﺎﻫ
ﻩﺪﻨﻫﺩ
ﺦﺳﺎﭘ
ﻦﻳﻮﻜﺗ
ﻰﻃ
ﺭﺩ
ﻯﺮﻴﮔ
ﻪﺠﻴﺘﻧ
ﺯﺍ
ﺪﻧﺍﻮﺗ
ﻰﻣ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﻯﺎﻫ
ﺶﻨﻛﺍﻭ
،ﻦﻳﻮﻜﺗ
ﻰﻃ
ﺭﺩ
ﻰﻧﻮﺴﻛﺍ
ﺪﺷﺭ
ﻪﻛ
ﺩﻮﺷ
ﻢﻴﻈﻨﺗ
NMDAﻩﺪﻨﻫﺩ
ﺦﺳﺎﭘ
ﺹﺎﺧ
ﻝﺎﻜﺷﺍ
ﻖﻳﺮﻃ
ﻉﻭﺮﺷ
ﻰﻳﺍﺯ
ﺐﺼﻋ
ﺎﻫﺪﻨﻳﺍﺮﻓ
ﻦﻳﺍ
ﺎﺑ
.
ﺩﺭﺍﺩ
ﻩﺍﺮﻤﻫ
ﻪﺑ
ﺍﺭ
ﻰﻟﻮﻠﺳ
ﻯﺎﻫ
ﺱﺎﻤﺗ
ﻭ
ﻦﻳﺍﺮﺑﺎﻨﺑ
.
ﺩﺍﺩ
ﻢﻴﻤﻌﺗ
ﻎﻟﺎﺑ
ﺩﺍﺮﻓﺍ
ﺰﻐﻣ
ﻪﺑ
ﻥﺍﻮﺗ
ﻰﻣ
ﺍﺭ
ﺎﻫ
ﺖﻟﺎﺣ
ﻦﻳﺍ
ﻪﻛ
ﺩﻮﺷ
ﻰﻣ
ﻦﻴﻨﺟ
ﻰﻃ
ﺭﺩ
ﺰﻐﻣ
ﻦﺘﻓﺮﮔ
ﻞﻜﺷ
ﺭﺩ
ﺖﺳﺍ
ﻦﻜﻤﻣ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﻥﺎﺸﻧ
ﺞﻳﺎﺘﻧ
ﻉﻮﻤﺠﻣ
ﺭﺩ
.
ﺪﻨﻛ
ﺎﻔﻳﺍ
ﺶﻘﻧ
ﻯﺩﺍﺯﻮﻧ
ﻰﮔﺪﻧﺯ
ﻞﻳﺍﻭﺍ
ﻭ
ﻰﻳﺍﺯ
ﻥﺍﺰﻴﻣ
ﻚﻴﺋﻮﻨﻴﺗﺭﺪﻴﺳﺍ
ﻩﺍﺮﻤﻫ
ﻪﺑ
ﻥﻭﺮﺘﺳﺍﻭﺭﺪﻧﺍ
ﻰﭘﺍﻭﺭﺪﻴﻫﺩ
ﻪﻛ
ﺪﻫﺩ
ﻰﻣ
ﺍﺭ
ﻚﻳژﺮﻨﻴﻣﺎﭘﻭﺩ
ﻯﺎﻫﺮﺘﻴﻤﺴﻧﺍﺮﺗﻭﺭﻮﻧ
ﻥﺎﻴﺑ
ﻭ
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﻪﺑ
ﺰﻳﺎﻤﺗ
.
ﺩﻮﺷ
ﻰﺒﺼﻋ
ﻯﺎﻫ
ﻝﻮﻠﺳ
ﺰﻳﺎﻤﺗ
ﺐﺒﺳ
ﺪﻧﺍﻮﺗ
ﻰﻤﻧ
ﻰﻳﺎﻬﻨﺗ
ﻪﺑ
ﺎﻣﺍ
ﻩﺩﺍﺩ
ﺶﻳﺍﺰﻓﺍ
ﺮﻜﺸﺗ
ﻭ
ﺮﻳﺪﻘﺗ
ﻪﻛ
ﻰﻧﺎﻴﺘﺷﺁ
ﻰﻤﻇﺎﻛ
ﺪﻴﻌﺳ
ﺮﺘﻛﺩ
،ﺪﻴﻘﻓ
ﺪﻨﻤﺸﻧﺍﺩ
ﺩﺎﻳ
ﺖﺷﺍﺪﻴﻣﺍﺮﮔ
ﺎﺑ
ﺩﻮﺧ
ﺮﺑ
ﻪﻟﺎﻘﻣ
ﻥﺎﮔﺪﻨﺴﻳﻮﻧ
،ﺪﺷ
ﺯﺎﻏﺁ
ﻥﺎﺸﻳﺍ
ﻰﻳﺎﻤﻨﻫﺍﺭ
ﺎﺑ
ﺍﺪﺘﺑﺍ
ﺭﺩ
ﻩژﻭﺮﭘ
ﻦﻳﺍ
ﻱﺎﻫ
ﻪﻨﻳﺰﻫ
ﻡﺎﻤﺗ
ﻪﻛ
ﻥﺎﻳﻭﺭ
ﻩﺪﻜﺸﻫﻭﮋﭘ
ﺯﺍ
ﺍﺭ
ﺩﻮﺧ
ﺮﻜﺸﺗ
ﺪﻨﻧﺍﺩ
ﻰﻣ
ﻡﺯﻻ
ﺎﻫ
ﻢﻧﺎﺧ
ﻭ
ﻰﻘﻴﻘﺣ
،ﻰﺧﺮﻓ
ﻥﺎﻳﺎﻗﺁ
ﻦﻴﻨﭼ
ﻢﻫ
ﻭ
ﺪﻧﺩﺮﻛ
ﻦﻴﻣﺎﺗ
ﺍﺭ
ﺡﺮﻃ
ﻦﻳﺍ
.
ﺪﻨﻳﺎﻤﻧ
ﺯﺍﺮﺑﺍ
ﻰﮕﻴﺑ
ﺪﻟﻭ
ﻭ
ﺪﻨﻣﺩﺍﺮﻣ
،ﻰﻤﻴﻫﺍﺮﺑﺍ
ﺮﺘﻛﺩ
،ﻯﺩﻮﻌﺻ
References
1. Ormerod BK, Lee TT, Galea LA. Estradiol enhances neurogenesis in the dentate gyri of adult male meadow voles by increasing the survival of young granule neu-rons. Neuroscience. 2004; 128: 645-654.
2. Ormerod BK, Lee TT, Galea LA. Estradiol initially en-hances but subsequently suppresses (via adrenal ster-oids) granule cell proliferation in the dentate gyrus of adult female rats. J Neurobiol. 2003; 55: 247-260.
3. Suzuki M, Wright LS, Marwah P, Lardy HA, Svendsen CN. Mitotic and neurogenic effects of dehydroepian-drosterone (DHEA) on human neural stem cell cultures derived from the fetal cortex. Proc Natl Acad Sci USA. 2004; 101: 3202-3207.
4. Iwata M, Muneoka KT, Shirayama Y, Yamamoto A, Kawahara R. A study of a dendritic marker, microtubule-associated protein 2 (MAP-2), in rats neonatally treated neurosteroids, pregnenolone and dehydroepiandroster-one (DHEA). Neurosci Lett. 2005; 386: 145-149.
