| Intrahepatic cholestasis of pregnancy is a gestation period unique disease which has characteristic of skin itches and jaundice.It belongs to high danger pregnancy.Yinchenhao was used to research its function and possible actional mechanism for ICP from the serum level of estriol,pathology change of placenta and the expression of estrogen receptor on placenta.Part 1 The establishment of animal model of intrahepatic cholestasis of pregnancy rats and influnce of Yinchenhao on Serum estriol of pregnancy ratsObjective:To provide basis of biochemistrial and pathologial experiment for successful establishment of the rat model by exploring the changes of the blood biochemistry value and hepatic pathohistology in pregnant rats with Estrogen and Progestron induced intrahepatic cholestasis.In addition,To explore the effects of Yinchenhao on serum estriol in pregnant rats with Estrogen and Progestron induced intrahepatic cholestasis.Methods:The animal model of intrahepatic cholestasis of pregnancy rats was induced by estradioli benzoas and Progesterone.All of rats were divided into normal group, model group,high dose group,middle dose group,low dose group of Yinchenhao and ursodeoxycholic acid group at random.On the 21 st day,blood of all pregnant rats was collected by picking the eyeballs.After all rats were put to death with luxation on cervical vertebra,two clumps of liver tissue of each rat in normal group and model group were made into light microscope and electron microscope specimens routinely.The serum level of estriol in all rats and the serum level of alanine aminotransferase,aspartate aminotransferase,total bilirubin,total bile acid of rats in normal group and model group were measured after the blood were centrifuged.Results:The level of every biochemistry value was decreased remarkably in model group than that of normal group(P＜0.01).Pathology change of liver:under the light microscope,the cell structure of liver in normal group was clear,and there was no denaturetion and necrosis.In model group,hepatocellular structure disorder,hepatocellular necroses,vacuole-like changes and hepatic sinusoid very narrow in hepatic partial region were observed.Under the electron microscope,bile capillary in normal group did not expanded.There was no bile salt in bile capillary.In model group,bile canaliculi were seen dilated.And the morphological changes in dilated bile canaliculi were found as follows:decrease or disappearance of superficial microvilli,deposition of substances with high electronic density,dilatation of smooth endoplasmic reticulum,tumefaction of the mitochondria and loss of their ridges.In comparison of normal group with the model group,there was significant difference in serum estriol(P＜0.01).In comparison of all of the group under treatment but low dose of Yinchenhao with the model group,serum estriol was significantly changed (P＜0.01);In comparison of the group under treatment of low dose of Yinchenhao with the model group,serum estriol was not different obviously(P＞0.05).High dose group of Yinchenhao had no difference with normal control group(P＞0.05).Conclusion:The pathological changes showed intrahepatic cholestasis in pregnanct rats.The changes of the blood biochemistry value and hepatic pathohistology in pregnant rats demonstrated it was successful to establish an animal model of intrahepatic cholestasis in pregnanct rats.Yinchenhao might be active by improving serum estriol so as to attain the goal of treat intrahepatic cholestasis of pregnancy.Part 2 Effect of Yinchenhao on the estrogen receptor of placenta in pregnant rats with intrahepatie cholestasisObjective:To research the expression of estrogen receptor on placenta so as to explore the possible adjustmental effect and theaction mechanism of Yinchenhao upon ICP.Methods:The expression of estrogen receptor on placenta was detected by immunohistochemis-try staining.Results:In comparison of normal control group with the model group,there was significant difference in the expression of estrogen receptor(P＜0.01).In comparison of the group under treatment of low dose of Yinchenhao with the model group,the expression of estrogen receptor was not different obviously(P＞0.05).In comparison of all of the group under treatment but low dose of Yinchenhao with the model group, the expression of estrogen receptor was significantly changed(P＜0.05,P＜0.01).High dose group of Yinchenhao had no difference with normal control group(P＞0.05),but had difference with ursodeoxycholic acid group.Conclusion:Yinchenhao could inhibit the level estrogen receptor,which indicate that Yinchenhao is effective in the improvement of placental function of intrahepatic cholestasis of pregnancy and may be active by regulating the level of estrogen receptor on placenta in pregnant rats with intrahepatic cholestasis.Part 3 Influnce of Yinchenhao on pathology change of placenta in pregnant rats with intrahepatic cholestasisObjective:To observe placental pathohistology in pregnant rats with intrahepatic cholestasis so as to explore the possible curative effect of Yinchenhao upon intrahepatic cholestasis of pregnancy.Methods:On the 21st day,after all rats were put to death with luxation on cervical vertebra,two clumps of placenta tissue of each rat by caesareaning were sampled and then were made into light microscope and electron microscope specimens routinely.Results:Pathology change of liver:under the light microscope,the placental villi in which capillaries were rich weren't swelling,and the trophocytes weren't obviously hyperplastic in normal group.In model group,the villi were swelling;the size of intervilli space was very small;the trophocytes were hyperplastic;hyperplastia of syncytiotrophoblast cells was more obvious;Capillaries were empty and collapsed in partial villi.In high dose group of Yinchenhao,there had no difference in contrast with normal group.In middle dose group of Yinchenhao and ursodeoxycholic acid group,villi were swelling;the changes of capillaries in villi were somewhat obvious;the trophocytes weren't obviously hyperplastic.In low dose group of Yinchenhao,there had no difference in contrast with normal group.Under the electron microscope,,microvilli on the syncytiotrophoblast cells were plentiful and had good development;capillaries were rich;Rich and dense rough-surface endoplasmic reticula,plentiful mitochondria and other syncytia were in the cytoplasm.Nuclei had regular shapes in normal group.In model group,vacuole-like changes or disappearance were observed in partial microvilli on the syncytiotrophoblast cells;microvascular endothelial cells were hyperplastic;rough-surface endoplasmic reticula were seen dilated obviously;tumefaction of the mitochondria and partial loss of their ridges;nuclei had regular shapes.In high dose group of Yinchenhao,there had no difference of syncytiotrophoblast cells in contrast with normal group.In middle dose group of Yinchenhao and ursodeoxycholic acid group,microvilli on the syncytiotrophoblast cells had vacuole-like changes;rough-surface endoplasmic reticula were dilated in the cytoplasm;dilatation of rough-surface endoplasmic reticulum,tumefaction of the mitochondria and irregular shapes of nuclei were observed in the cytoplasm.In low dose group of Yinchenhao,there had no difference of syncytiotrophoblast cells in contrast with normal group.Conclusion:Yinchenhao could improve pathology change of placenta of pregnant rats with intrahepatic cholestasis which indicate that Yinchenhao is effective in the improvement of placental function so as to change adverse pregnancy outcomes. |
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