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Preliminary Study of the Effects of Nerve Growth Factor and p75NTR on Hepatic Stellate Cells

Author: ChenLianXiang
Tutor: ShuJianChang
School: Jinan University
Course: Internal Medicine
Keywords: nerve growth factor hepatic stellate cells p75 neurotrophin receptor hepatic fibrosis
CLC: R575.2
Type: Master's thesis
Year: 2011
Downloads: 17
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Abstract


Objective1. To investigate the effects of p75 NTR、(p75 NTR+NGF) and anti-p75NTR on proliferation and apoptosis of HSC in vitro.2. To observe the expressions of NGF and p75NTR mRNA and proteins in HSC after treated by p75 NTR、(p75 NTR+NGF) and anti-p75NTR.3. To Investigate the effects of NGF and p75NTR in the phase of occurrence and development of hepatic fibrosisMethodsHSC were incubated with 0、100、200、400、800、1600ng/ml p75 NTR、(p75 NTR+NGF) and anti-p75 NTR. HSC proliferation were detected by XTT colormetric assay. HSC apoptosis were detected by flow cytometry (FCM). Expressions of NGF and p75NTR mRNA and proteins were detected by Fluorescence quantitative PCR and Immuno fluorescence method.Results1. The optical density of HSC incubated with p75NTR had no significant difference compared with the control group (P>0.05;the optical density of (NGF+p75NTR) decreased compared with the control group(P<0.05);the optical density of anti-p75NTR decreased compared with the control group (P<0.01).2. Apoptosis index of HSC incubated with 100、200、400、800ng/ml p75NTR decreased(P<0.05) while had no significant difference with 1600ng/ml (P>0.05) apoptosis index of HSC incubated with (NGF+p75NTR) increased(P<0.05); Apoptosis index of HSC incubated with anti-p75NTR increased(P<0.05);3. The average optical density of fluorescence of NGF and p75NTR expressed in HSC decreased after HSC treated by 400ng/ml p75NTR (P<0.05). The average optical density of fluorescence of NGF expressed in HSC decreased after HSC treated by 400ng/ml (NGF+p75NTR)(P<0.05) while p75NTR increased significantly(P<0.01). The average optical density of fluorescence of NGF expressed in HSC increased after HSC treated by 400ng/ml anti-p75NTR(P<0.01) while that of p75NTR had no significant difference compared with the control group (P>0.05).4. The amount of NGF and p75NTR mRNA decreased significantly compared with the control group after HSC treated by 400ng/ml p75NTR (P<0.05); The amount of mRNA of NGF decreased significantly after HSC treated by 400ng/ml (NGF+p75NTR) (P<0.05) while p75NTR mRNA increased (P<0.05); the amount of mRNA of NGF increased significantly after HSC treated by 400ng/ml anti-p75NTR (P<0.01) while p75NTR mRNA had no significant difference (P>0.05).Conclusions1. Exogenous p75NTR could inhibit HSC apoptosis independently while had no effect on proliferation.2. (p75NTR+NGF) decreased expression of mRNA and protein of NGF in HSC while increased the expression of p75NTR mRNA and protein; p75NTR decreased expression of mRNA and protein of NGF and p75NTR in HSC; anti-p75NTR increased expression of mRNA and protein of NGF in HSC while had no effect on p75NTR mRNA and protein.3. By combinning with p75NTR, NGF could induce the apoptosis of HSC and reverse hepatic fibrosis.

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CLC: > Medicine, health > Internal Medicine > Digestive and abdominal diseases > Liver and gall bladder disease > Cirrhosis
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