| Hepatic disease has become common to threaten human health. Various hepatic damage factors can lead to hepatocirrhosis, which is the severe stage of hepatic disease involving re-configuration of the liver and formation of pseudo lobules. Most of recent reseaches show that some cell factors secreted by Kupffer cells (KC) , blood plaque, sinusoids endothelial cells and activated hepatic cells make hepatic stellate cells (HSC) propagate and synthesize large extracellular matrixes (ECM) accompanying with some chemical transmitters through autocrine and paracrine secretion. Then these ECM deposite in the liver and form hepatic fibrosis. The developing of hepatic fibrosis is a gradual and complex process. During this process, cell factors,cells and ECM are interacted with each other and cell factors are the most important. Among numerous cell factors, transforming growth factor-β1(TGF-β1) is a major hepatic fibrosis- precipitating factor through TGF-β1/Smad cell signaling pathways to cause hepatic fibrosis. Smad3 and Smad7 are the main composition of Smad, the former is positive correlation to hepatic fibrosis and the latter is negative. Recent studies indicated that in our country virus is the major hepatic injury factor. 10%-20% hepatovirus B patients and 50% hepatovirus C patients will develop to chronic hepatitis, hepatic fibrosis, hepatic cirrhosis, even hepatoma. Hepatic fibrosis is the common pathological foundation for chronic hepatic diseases. Recent researches show that to some extent hepatic fibrosis is reversible but hepatic cirrhosis is irreversible. There is no effective therapeutic measure to hepatic cirrhosis, so earlier diagnosis become most important. Biopsy is a golden index to diagnose earlier hepatic fibrosis by liver punctio. But its using is limited because it is hazard to health and difficult to repeat. Now researchers have found many blood-serum markers which are related to hepatic fibrosis, such as HA, LN, PCⅢand IV. These four indexes which have bean commonly used in clinic can reflect the degree of many organs fibrosis except the liver. Developed research indicates that the level of these four indexes increase with the content of hepatic fibrosis and they have been evident consistency. But when hepatic tissue has light fibrosis or inflammation especially in the stage of S1, these four indexes have no apparent increase. With the development of imageology and generally used in the clinic such as supersonic, CT, MRI, the diagnosis and differential diagnosis level of hepatic diseases is raised obviously, and supersonic B is more widely used than others. But in the earlier time of hepatic fibrosis, there is no evidently change in morphology and hemodynamics. So it is finite to diagnose hepatic fibrosis and there are little reports about it. Above all we must find an effective measure to diagnose hepatic fibrosis earlierly and exactly. Indocyanine green retention rate at 15 minutes (ICGR-15) which can assess hepatic reserve function can also reflect the extent of earlier hepaticr fibrosis.In 1957, Fox introduced indocyanine green (ICG) to us which was an atoxic serpentinous pigment. It has little adverse reaction and can be selectly taken in through intravenation and taken out to the bile in original shape which doesn't take part in hepatoenteral circulation and renal excretion. The indocyanine green retention rate can be used not only to judge liver store function but also to appraise its prognosis. So it has become more and more valuable in the clinic especially in liver and gall surgery.The liver cleanes up indocyanine green by three procedures: intake, dispose and excrete. The main cause that influences indocyanine green clearance is hepatic blood flow,functional hepatic cells'counts and the secret of bile. Otherwise the level of plasma protein, the membrane permeability of sinus hepaticus, and apoprotein in liver cell is related to indocyanine green clearance, too. When liver pseudo lobules formed, hepatic lobules are fibrosis, the amount of functional hepatic cells and hepatic blood flow will decrease, at the same time the rate of indocyanine green clearance obviously decrease.This experiment based on hepatic fibrosis (CCl4) model in mice continuously observed indocyanine green retention rate at 15 minutes (ICGR-15) in different stages, in order to study the relationship between ICGR-15 and hepatic fibrosis.1. Pathohistology observation: With time going on, hepatic fibrosis gradually formed from injections of CCl4. Indocyanine green retention rate at 15 minutes (ICGR-15) has significant difference between 6 weeks group and 4 weeks group. This result showed that ICGR-15 can reflect hepatic pathological changes and can be regarded as an index of hepatic fibrosis.2. Cytology and serology observation: HSC as interstitial cells can be activated by certain damage factors, then these activated cells appear contractile functions and make hepatic sinusoid envelope membrane permeability change accompanying with ICG changing. On the other hand, ECM just as HA, PC-Ⅲ, IV-C, LN make cell compartment widen and effect hepatic cellular metabolism, so ICG discard to the bile decrease. Some researches reported PC-Ⅲand IV-C obviously rised in earlier stage of hepatic fibrosis. Our research showed that the counts of activated HSC, PC-Ⅲand IV-C had significant difference between 6weeks group and 4weeks group (P<0.01). There was close relationship between PC-Ⅲ, IV-C and ICGR-15 . This result suggested that ICGR-15 can reflect the earlier stage of hepatic fibrosis.3. Gene and protein level observation: This research tested there was close relationship between hepatic fibrosis and ICGR-15 on gene and protein level through detecting cellula factors- TGF-β1 and some important proteins. With hepatic fibrosis gradully aggravated, the mRNA and protein expression level of TGF-β1 and Smad3 increased and Smad7 decreased .This result indicated that with the forming of hepatic fibrosis, these above-mentioned indexes changed obviously and there was significant differences among these groups. This finding is similar to other findings and indicated that ICGR-15 can assess the earlier micro change of hepatic tissue.From above, we concluded that ICGR-15 could be as an earlier index to diagnose hepatic fibrosis. We are trying to supply a reliable theory to earlier diagnosis and effective therapy of hepatic fibrosis.The following is the study outcomes:Our study introduced that ICGR-15 can be used to diagnose earlier hepatic fibrosis and the demonstrations were as follow:1. Using RT-PCR and immunohistochemisty to detect ICGR-15 and to observe the changes of expression levels of Smad3 , Smad7 and TGF-β1 which were the main informational molecule of TGF-β1/Smad signal transduction and we concluded that there was close relationship between Smad3, Smad7 and ICGR-15. We considered that ICGR-15 can reflect the earlier stage of hepatic fibrosis.2. Using histopathology and radioimmunity to investigate the pathological change of tissue and PC-Ⅲ, IV-C of blood serum, we also found that there was dependablity between PC-Ⅲ, IV-C and ICGR-15.3. Using immunohistochemistry to observe the count of activated cells, we tested there was dependablity between ICGR-15 and hepatic fibrosis.Above all, we concluded that ICGR-15 can be as an earlier index to diagnose hepatic fibrosis and supply a reliable theory to the clinic. |
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