| Objective:Renal anemia is one of very frequent complication in patient with chronic renal failure.EPO is one major drug in treatment of renal anemia at present. Hhowever, exogenous EPO has various adverse reaction,such as hypertension, blood viscosity step up, blood vesselthrombosis, dialysis tube blood clotting, hyperpotassaemia, myosalgia, fluen appearance symptom and so on),as well a part of patients have low react to rHuEPO.In the adult organism the kidney produces around 90% of systemic EPO. not exceed 10% is produced by is produced by liver. The renal EPO-producing cells are peritubular fibroblast-like type-1 interstitial cells . In chronic renal failure,renal anaemia and chronic renal failure make EPO secrete not enough.Renal EPO-producing is decrease, how outside renal EPO regulate and in what way to urge outside renal EPO to produce are doubtful point and difficult at present.Augmenter of liver regeneration,ALR is one kind of factor with heat stability non-specificity promotion hepatic cell regeneration whitch Hagiya has depurated and cloned from neonate rat liver.It is a micromolecule protein with 125 amino acid.Its molecular weight is 15 Kda,its gene order and protein structure are all different from hepatocyte growth factor(HCG)and hepatic stimulator substance(HSS).ALR is ungenera, untissue, unorgan-specific. Previoussstudy confirmed that ALR has significant biological effect.It expressesobviously in kidney .we presume that it has various kinds complic biological effect in kidney.At present no study of ALR in chronic renal failure(CRF) has done.Renal can not produce adequate EPO,liver has any compensation.We know that liver Kupffer cellsynthesis and secrete EPO.Liver Kupffer cell has ALR acceptorwith high affinity.ALR can promote Kupffer cell copy DNA, synthesis protein. Whether or not hypoxia state activation Kupffer cell ALR acceptor in chronic renal failure,or internal environment is abnormality in chronic renal failure.Various kinds of growth factors and various kinds of cell factors, network effect can promote ALR composition in cell. Thereby it can promote Kupffer cee generation,to make it produce more EPO.To give exogenous ALR can or not strenghten this action ,promote lver compensation,to lessen renal anaemia,to explore new therapy for renal anaemia.And to approach ALR protection to chronic renal failure.These wait for our deep research.If man-made synthesis ALR can treat renal anaemia,it will fit physiology request very much . Dosage of rHuEPO can reduce,many adverse effect will decrease.It can provide experimental base for recombinatal ALR in therapy for renal anaemia.It can be a ideal therapy.Methods:Thirty SD rat whitch weight (150～200g ) were randomly divided into three groups. fake operated group(n﹦10),only expose kidney peplos ,not cut kkidney . operation group(n﹦20), in accordance with literature to simulation.in dictionary asepsis condition,cutting right kidney,1weeek later,cutting 2/3 left kidn.Eighteen rats taked in operated group raised 1 week postop. Randomly divide two groups, control group(n﹦9),rhALR group(n﹦9)。To peritoneal inject rhALR200μg/kg·d in rhALR group 12 weeks after postop. At the same time to peritoneal inject mitte tales doses normal sodium in fake operated group control group. Every other day inject, persistencing two weeks.To observe rats′mental state, activity, hairs glossiness, ingestion and death.One weight one week, To observe weight variation. Every group rats are anaesth with ketamine twenty-four hours after the last injection, to draw blood from heart,to obsearve every index. Utilizing Japanese SYSMEX holo-automatic hematocyte cytoanalyze,to determin hemoglobin (Hb),red blood cell count (RBC),hematocrit (Hct);Utilizing OLYMPUS AU400 biochemistry appearance,to determin blood urea nitrogen (BUN), creatinine (Scr). Utilizing OLYMPUS 5400 biochemistry appearance,to determin serum ferritin(SF) ; Utilizing American ADL ELISA kit determin blood serum EPO. Utilizing Masson staining,to observe kidney pathology variation, Utilizing HE staining,to observe liver pathology variation. Utilizing immunohistochemical method,to detect augmenter of liver regenration (ALR) , proliferating cell nuclear antigen (PCNA) in liver and kidney. Utilizing Western blot ,to detect ALR in liver and kidney.Results:1 The common behavior of kidney:leftover kidney volume increasing in control group rats, packaged with epiploon, hard to decoherence..Cicatricle emboled in operative incision, the rest evagination, shape is irregularity.RhALR group have also above-mentioned change,but much lighter.2 Diversity of peripheral blood and renal function:Hb (g/L) in fake operated group ,control group and rhALR group quaque for 137.43±0.96,68.72±15.42,113.51±5.83,RBC(×1012/L) quaque for 7.45±0.42,4.01±1.35,6.05±0.88,HCT(%)quaque for 46.92±6.33,25.31±2.61,39.42±5.45,BUN (mmol/L) quaque for 7.52±1.36,25.35±8.91,17.22±5.17,Scr(μmol/L) quaque for 49.64±6.20,97.72±15.43,68.88±6.84。control group vs sham operated group,Hb,RBC,HCT all decrease (P< 0.05), Discrepancy have statistical significance.While Scrand BUN ascensus (P<0.05),Discrepancy have statistical significance.These indicate that renal function is failure and induce anaemia. RhALR group vs control group,Hb,RBC,HCT are all significant increase (P< 0.05), while Scrand BUN decrease (P<0.05),Discrepancy has statistical significance.These indicatethat renal functional lesion abatement and anaemia improve.3 serum EPO examination :EPO value (u/L) of fake operated group,control group and rhALR group respective is 12.19±1.87,1.77±0.72,8.64±2.35。control group serum EPO obviously decrease,EPO increase after rhALR treatment. Compared with control group ,P<0.05, Discrepancy has statistical significance.4 Histologic anatomy appearance:In fake operated group kidney external appearance is slick. Structur of glomcrulus, nephric tubule, renal interstitium is normal. In control group, kidney is obviously increase, its color is pallor,it is rough and uneven in surface , pyelectasia.Through light microscope, Masson staining display that glomcrulus compensatory hypertrophy,mesenterium hyperplasy,focus Segmental glomerulosclerosis, nephric tubule analosis, nephric tubule cell apomorphosis, cellular necrosis, amotic. renal interstitial collagen increase, fraction interstitial fibrosis,focus plymphomonocyte infiltrate and inflammatory cell infiltrate.In rhALR group glomcrulus, nephric tubule, renal interstitium fibroplasia markedly relieve.5 The express of ALR in nephridial tissue:small express of ALR in fake operated group. Compare with fake operated group ,in control group ALR express increase,(9.53±0.73vs. 13.58±0.69,P <0.05),in rALR group ALR express markediy increase ( 17.50±0.08 vs. 13.58±0.03 ,P<0.05),discrepancy has statistical significance.6 The express of ALR in hepatic tissue immunohistochemistry:in fake operatedgroup, small express of ALR in hepatic tissue.Compare with fake operated group ,in control group ALR express increase. (10.00±0.89 vs. 13.56±1.33,P<0.05),in rALR group ALR express obviously increase . ( 23.01±1.22 vs. 13.56±1.33 , P< 0.05 ) ,discrepancy has statistical significance.7 The express of PCNA in nephridial tissue:small express of PCNA in fake operated group. Compare with fake operated group ,in control group PCNA express increase,(0.73±0.08 vs. 6.08±0.68,P <0.05),in rALR group PCNA express obviously increase,6.08±0.68vs.12.30±0.74 ,P<0.05),discrepancy has statistical significance.8 The express of PCNA in hepatic tissue immunohistochemistry:in fake operatedgroup, small express of PCNA in hepatic tissue.Compare with fake operated group ,in control group PCNA express increase(2.33±0.11 vs. 8.27±0.30,P <0.05),in rALR group PCNA express obviously increase( 12.19±0.27 vs. 8.27±0.302 , P<0.05 ) ,discrepancy has statistical significance.9 In western blot, the express of ALR in nephridial tissue:small express of ALR in fake operated group. Compare with fake operated group ,in control group ALR express increase(,0.6±0,02 vs. 0.69±0.03,P<0.05),in rALR group ALR express obviously increase ( 0.82±0.05 vs. 0.69±0.03 , P<0.05),discrepancy has statistical significance.10 In western blot, the express of ALR in hepatic tissue:small express of ALR in fake operated group. Compare with fake operated group ,in control group ALR express increase(0.75±0.03 vs. 0.80±0.05,P<0.05),in rhALR group ALR express obviously increase ( 0.89±0.02 vs. 0.80±0.05,P<0.05),Discrepancy has statistical significance.Conclusion:1 5/6 renal nephrectomized rats manufact chronic renal failure(CRF)animal model.although this method is complex, its mechanism is identify. Decreasing nephridial tissue lead to renal failure, gradually complic anaemia, model′achievement ratio is high.In aspects of biochemistry, pathology and metabolism are near to mankind CRF, thereby conduce us to approach the pathogenesy of chronic renal anaemia.2 In CRF animal model,we can see renal interstitial fibrosis, promoting liver and kidney express ALR increase.This indicate that renal function is lesion, to result in anaemia, ALR of liver and kedney increase.3 After give exogenous ALR, blood EPO of 5/6 renal nephrectomized rats is increasing,anaemia is improve.From pathology ,renal interstitial fibrosis is decrease, ALR and PCNA of liver and renal are obviously up-regulation,this is at equal pace to EPO.We confirm that rhALR can promote EPO produce,protecting CRF rats leftover kidney function.This empirical study demonstrate that rhAL can not only lessen renal anaemia of 5/6 renal nephrectomized rats,but also lower blood BUN and SCr,improve azotemia condition..This hint that rhALR have any protective therapeutic action. We will study and research the nechanism of renal anaemia amelioration concerned with rhALR.
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