Effect of Chlorogenic Acid on the Proliferation of Bile Duct and Collagen in Rats with Liver Fibrosis Induced by Chronic Cholestasis
TANG Li-ming,Li Wan-hua,SONG Ning
Abstract:
Objective To observe the effect of chlorogenic acid on the proliferation of bile duct and collagen in rats with chronic cholestasis and to explore the mechanism of prevention and treatment. Methods Forty adult male Wistar rats were randomly divided into five groups: control group, model group, UDCA group,low dose group and high dose group. In addition to the control group, the other four groups were treated with ANIT (80 mg/kg) for 5 weeks and 3 times a week to prepare the chronic cholestasis model. At the same time,UDCA group, low dose group and high dose group were given UDCA (90 mg/kg), chlorogenic acid (50 mg/kgand 100 mg/kg) for 5 weeks. At the end of 5 weeks, 5 groups of animals, including control group and model group, were taken blood from abdominal aorta under anesthesia, and the levels of TBIL, DBIL, ibil and ALP in serum were measured by automatic biochemical analyzer. Partial liver tissues were cut, fixed with neutral formaldehyde, stained with he and Sirius red,and the pathological changes of cholestasis, bile duct hyperplasia and collagen hyperplasia were observed and evaluated. The total protein was extracted from other liver tissues, and the expression level of α-smooth muscle actin (α-SMA) was detected by Western blot. Results compared with the normal group, the levels of TBIL, DBIL, IBIL and ALP in the model group were signi?cantly higher (P<0.01); compared with the model group, the levels of TBIL, DBIL and IBIL in the high and low dose chlorogenic acid group and UDCA group were signi?cantly lower (P<0.05), and ALP had a downward trend, but there was no signi?cant difference (P>0.05). HE staining of the model group showed that the chronic inflammatory cell infiltration in the liver tissue was obvious, the proliferation and expansion of bile duct in the portal area were signi?cant; the chronic in?ammatory response and the proliferation of bile duct in the portal area were signi?cantly reduced in the high and low dose groups of chlorogenic acid and UDCA groups. Sirius red staining showed that the expression of collagen in the normal group was rare, while that in the model group was signi?cantly enhanced. Under polarized light, collagen I was mainly bright red and yellow. Compared with the model group, collagen staining in the high and low dose groups of chlorogenic acid and UDCA group was signi?cantly reduced, while collagen III was mainly green under polarized light. Western blot showed that the expression level of α-SMA in the liver tissue of the model group increased significantly (P<0.01), compared with the model group, the expression of α - SMA in the liver tissue of the high dose group, the low dose group and the UDCA group decreased signi?cantly (P<0.05). Conclusion Chlorogenic acid can signi?cantly improve the liver function damage, effectively inhibit the activation of hepatic stellate cells, reduce the production of collagen based on type I collagen, and reduce the proliferation of bile duct in the portal area caused by chronic cholestasis, so as to play an anti ?brosis role.
