Tesi - Effetti della melatonina sull'apoptosi della cellula muscolare scheletrica

A D

C F

E

G H

Figura 4

Mioblasti trattati con etoposide (A, B, C, D) e con MEL +

etoposide

oposide (E, F, G, H) osservati mediante RM (A,

, E), SEM (B, F),

TEM (C, G) e Microscopio Confocale (D, H).

Nel primo caso si evidenziano tipici aspetti apoptotici, con presenza

di blebs di membrana (A, B) e di cellule con micronuclei (C). Dopo

reazione TUNEL si osservano numerosi nuclei positivi (D).

Il pre-trattamento

trattamento con MEL riduce, in maniera evidente, il numero

nu

di cellule in apoptosi (E, F, G) e i campioni si presentano più simili

al controllo. La reazione TUNEL risulta per lo più negativa (H).

Bar = 10 µm in A, B, E,

E G e I; 2 µm in C, D; 5 µm in H.

32

A B

C D

E F

G H

Figura 5

Mioblasti trattati con staurosporina (A, B, C, D) e con MEL +

staurosporina

porina (E, F, G, H) osservati mediante RM (A, E), SEM

(B, F), TEM (C, G) e al Microscopio Confocale (D, H). In

entrambe le condizioni si assiste a riduzione

ri

duzione del volume cellulare

(A, E), presenza

resenza di blebs di membrana (B, F) e di cellule in

apoptosi tardiva

diva o in necrosi secondaria (C, G). Il confocale mostra

una marcata presenza

nza di nuclei TUNEL positivi (D, H).

Bar = 20 µm in A, E, D, H; 1 µm in B; 2 µm in C; 5 µm in G; 10

µm in F. 33 H

B

Figura 6

Vitalità cellulare analizzata mediante test di esclusione del Trypan

Blue.

La vitalità cellulare migliora pre-trattando

pre trattando le cellule con MEL

prima dell’esposizione

posizione a H O (significativo con p<0,05) ed

2 2

etoposide. MEL non ha alcun effetto nel trattamento con

staurosporina. 34 Bibliografia

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