Calcium staining by the glyoxal-bis-(2-hydroxyanil)-method in the livers of rats treated with CCl4, diltiazem, and with both agents together.

We studied the histochemistry of Ca in livers treated with CCl4, diltiazem (one of the Ca antagonists), and both agents together to determine whether hepatocytes or other parts of the liver lesions show Ca staining and whether the grade or location of Ca in these injuries varies. For Ca staining, cryostat sections were treated by the glyoxal-bis-(2-hydroxyanil) (GBHA)-method using O.C.T. imbedding compound instead of paraffin. Diltiazem-treated rats showed Ca granules in the bile canaliculi around the terminal hepatic veins and Kupffer cells 6 h after intragastric injection. Rats treated with CCl4 showed fine red granules in the cytoplasm of the hepatocytes around the terminal hepatic veins as soon as 5 min mildly and 2 h moderately after intraperitoneal injection. Hepatocytes around the terminal hepatic veins showed positive Ca granules 6 to 30 h after intragastric injection of CCl4. Hepatocytes stained by Ca showed acidophilic degeneration and coagulative necrosis. The hepatocytes of rats treated with both diltiazem and CCl4 revealed fewer Ca granules than those treated with CCl4 alone. In summary, Ca was stained by the GBHA method from the early stage of liver injury by CCl4 and was closely involved in acidophilic degeneration and coagulative necrosis of hepatocytes. The Ca staining in liver cells in CCl4-treated rats was decreased by diltiazem.