The influence of partial hepatectomy on the activity of the hepatic microsomal enzymatic systems was determined in rats. Cytochrome P-450, cytochrome b5, four mixed functional oxidase (MFO) activities (microsomal aniline hydroxylase, p-nitroanisole O-demethylase, aminopyrine N-demethylase and NADPH cytochrome c reductase) and glutathione levels were measured in unhepatectomized rats (control group) and in hepatectomized rats 12 h, 24 h, 3 days and 6 days after 70% hepatectomy. Following surgery the remaining lobes of the liver grow rapidly in order to restore the original liver mass. Partial hepatectomy significantly reduces cytochrome P-450 and b5 content in the remaining liver as well as the four MFO activities studied. But when the enzymatic systems are expressed as nmoles/mg microsomal protein, only cytochrome P-450 shows statistical differences. The hepatic biotransformation capacity of drugs and xenobiotics decreases during the regeneration period due to the reduction of hepatic mass rather than because of a reduction of their metabolic capacity. Glutathione levels are increased after partial hepatectomy but increased glutathione-dependent protector mechanisms are not expected.

The influence of partial hepatectomy on the pharmacokinetics of theophylline was determined in rats. The pharmacokinetics of intravenous theophylline was studied in unhepatectomized rats (control group: CG) and in hepatectomized rats (HG) 12 h after 70% hepatectomy. Liver function was monitored in both groups by measurements of total and direct bilirubin, transaminases GOT and GPT, and plasma protein. Seventy per cent hepatectomy caused significant liver dysfunction: transaminase levels (GOT and GPT) increased by 118 and 683%, respectively, and the total and direct bilirubin levels increased by 28.6 and 9.1%, respectively. At the same time, plasma concentrations of theophylline decreased significantly and half-life increased from 4.16 +/- 0.57 h (CG) to 7.08 +/- 0.69 h (HG), as did the distribution volumes values of central (Vc) and peripheral (Vp) compartments (Vc: CG, 0.18 +/- 0.03 L; HG, 0.24 +/- 0.03 L) (Vp: CG, 0.08 +/- 0.05 L; HG, 0.13 +/- 0.05 L). The percentage of theophylline binding to plasma proteins decreased from 44.3% in CG to 33.8% in HG. The theophylline intrinsic clearance (CLint) dropped from 1.35 +/- 0.43 mL/min (CG) to 0.93 +/- 0.10 mL/min (HG)1, which can be attributed to a significant fall in the quantity of hepatic microsomal enzymes. These modifications on the pharmacokinetics of drugs with low hepatic extraction coefficients, such as theophylline, should be considered when dosage regimens during the posthepatectomy hepatic regeneration period are planned.

The influence of hepatic regeneration after partial hepatectomy on theophylline pharmacokinetics has been studied on the rat. At different times after partial hepatectomy, theophylline was administered intravenously as a single dose of 6 mg/Kg. Drug plasma levels were determined by HPLC and pharmacokinetic parameters were obtained. Physiological parameters were also measured. Following hepatectomy, an increase in mass liver was observed and 15 days after surgery, liver mass was 78% of nonhepatectomized rats. Initial theophylline concentrations varied during the regeneration period, as well as the distribution volume at steady-estate (Vss). Elimination half-life (t 1/2), notably increased after hepatectomy (7.27+/-1.38 h), decreased with time (6.70+/-1.18 h, 6.47+/-0.69 and 5.17+/-0.87 h after 24 h, 3 days and 15 days post-hepatectomy, respectively) to reach a value close to that of the control group (4.30+/-1.37 h). The increase in elimination half-life led to a decrease in the mean residence time during the period of liver regeneration. However, the intrinsic clearance hardly varied during regeneration period. We could establish the following relationship between liver weight (LW) and the elimination half-life: t 1/2 (h)=-0.358*LW (g)+8.6168 (R2=0.9906). For the mean residence time (MRT) this relationship was: MRT (h) =-0.5173*LW (g)+12.433 (R2=0.991).

A protease was extracted by grinding, precipitation and gel filtration from Onopordum acanthium flowers. The physicochemical study of the enzyme showed an optimum pH of 4, a temperature of 40°C and kinetic parameters of 12.25 mM-1 for K M and 1329.6 UmL-1 for V max. The inhibition by pepstatin indicated that it is an aspartyl-protease (APs). Zymogram showed that the protease has a monomeric structure and a molecular mass (MM) of 45 kDa. The hydrolysis of α, β and -and whole casein by the protease was evaluated using electrophoresis and HPLC; the profiles showed many similarities between the vegetal protease action and that of industrial chymosin. So, the properties of the protease studied and the quality of its action showed its effectiveness and relevance of its use as a milk clotting enzyme which leads to a better use of extract of flowers O. acanthium as a locally substitute for rennet.