Effect of curcumin against capillarization of hepatic sinusoids and its mechanism

doi:10.3969/j.issn.2095-4344.0144

Abstract

Abstract:

BACKGROUND: Capillarization of hepatic sinusoids is an inevitable part in liver fibrosis and cirrhosis, and is a characteristic lesion inducing portal hypertension. However, curcumin effects on the capillarization of hepatic sinusoids and the underlying mechanism remain unclear.
OBJECTIVE: To observe the effect of curcumin (a natural polyphenolic compound derived from the rhizome of Curcuma longa) on the microstructure and secretion of hepatic sinusoidal endothelial cells (HSECs), and to further explore its intervention on sinusoidal capillarization and pharmacological action mechanism of anti-liver fibrosis and target sites.
METHODS: The rat HSECs were cultured and divided into seven groups: blank control group received no intervention and cells in the other groups were activated by leptin, followed by treatment with nothing (model group), high-, medium- and low-dose of curcumin, colchicine and salvia miltiorrhiza phenolic acid B, respectively, for 48 hours.
RESULTS AND CONCLUSION: Under scanning and transmission electron microscopes, with the increasing activation of leptin, the number of fenestrae in HSECs was increased and the aperture was decreased. Curcumin could increase and enlarge narrowed or disappeared fenestrae caused by leptin, attenuated the thickness and scope of extracellular basement membrane, and reduced the degree of capillarization of hepatic sinusoids in a dose-dependent manner. Real-time PCR and ELISA results showed that after activation of leptin, mRNA and protein expression levels of endothelin-1 and vascular endothelial growth factor in HSECs were significantly increased compared with the blank control group (P < 0.05), while the expressions showed a significant decrease after treatment with curcumin in a dose-dependent manner (P < 0.05). There was also a gradient reduction in the protein expression of endothelin-1 and vascular endothelial growth factor in HSECs treated with curcumin. Moreover, all above mRNA and protein expression levels in the high-dose curcumin group were significantly lower than those in the colchicine and salvia miltiorrhiza phenolic acid B groups. In summary, curcumin can significantly alleviate the sinusoidal capillarization, and thus delay the development of liver fibrosis, probably by down-regulating the expression levels of endothelin-1 and vascular endothelial growth factor.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Tissue Engineering, Curcumin, Liver

CLC Number:

R318

Duan Xue-lin, Peng Yue, Zhao Tie-jian, Wei Yan-fei, Li Gui-yu. Effect of curcumin against capillarization of hepatic sinusoids and its mechanism[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(8): 1247-1252.

Figures/Tables 2

Ultrastructural changes in the HSECs The results of scanning electron microscope showed that the cell wall of HSECs in the blank control group was mesoporous, with abundant fenestrae of uneven size (the large diameter was about 1 μm and the small one was about 0.1 μm), especially the large fenestrae (Figure 1A). The cell wall of HSECs in the leptin group showed less or none fenestrae, and its shape changes from honeycomb to flat plate (Figure 1B). Compared with the leptin group, the number of fenestrae in HSECs in the high-dose curcumin group was significantly increased, and the sinus wall returned to mesoporous shape with more large fenestrae (the diameter > 1 μm) (Figure 1C). The number of fenestrae in the medium-dose curcumin group was more than that in the leptin group, but lower than those in the high-dose curcumin group (Figure 1D). There was no significant difference in the number of fenestrae between low-dose curcumin and leptin groups. Compared with the medium- and high-dose curcumin groups, in the low-dose curcumin group, the number of fenestrae was less and with smaller diameter (Figure 1E).The number of fenestrae in the HSEC wall in the colchicines and salvianolic acid B groups was obviously more than that in the blank control and low-dose curcumin group, and the size of the fenestrae was larger, but compared with the medium- and high-dose curcumin groups, the number and size of fenestrae were reduced. The number and size of fenestrae did not differ significantly between colchicines and salvianolic acid groups (Figure 1F, G). Results of transmission electron microscope revealed that there were a large number of hollow light-colored areas in the HSECs of the blank control group, and the fenestrae vwere dense and large in area, the monolayer cell membrane was thin, without basement membrane formation (Figure 2A). In the leptin group, the light-translucent area in HSECs almost disappeared, the number and area of fenestrae were reduced, the thickened cell membrane showed two-to-three layers, and the light-colored basement membrane area was obvious and continuous (Figure 2B). In the high-dose curcumin group, compared with the leptin group, the light translucent area within the HSECs increased significantly, the number and area of visible fenestrae were also increased, hollow-intensive sinus wall was almost sieve-shaped; thin membrane was observed, and the light-colored basement membrane area was formed outside the wall (Figure 2C). In the medium-dose curcumin group, compared with the leptin group, the light translucent area within the HSECs was enlarged, with a large number of fenestrae; two layers of the cell membrane could be still observed, but the light-colored basement membrane outside the cell wall was unobvious. However, compared with the high-dose curcumin group, the intracellular light-colored fenestrae area was smaller, and the extracellular basement membrane area existed (Figure 2D). Compared with the leptin group, in the low-dose curcumin group, the light-colored translucent area in the HSECs increased, the number of fenestrae increased; two distinct layers of the cell membrane were visible, obvious light-colored basement membrane area appeared outside the cell wall. Compared with the medium- and high-dose curcumin groups, the light-colored fenestrae area in the cells of the low-dose curcumin group was obviously smaller, and the light-colored basement membrane area was clearly seen (Figure 2E). Compared with the leptin group, in the colchicine and salvianolic acid B groups, the light translucent area in the HSECs was significantly expanded, with increased number and area of fenestrae; the cell membrane was thinned, and it was hard to detect the two layers, and the light-colored basement membrane area was nearly observed outside the cell wall. There were no significant differences in the number and area of fenestrae, the thickness of the cell membrane and the basement membrane outside the cell wall between two control drug groups. Compared with the medium-and high-dose curcumin groups, the number and area of fenestrae in the two drug control groups were lower, and the formation of extracellular basement membrane was more obvious. Compared with the low-dose curcumin group, the light-colored fenestrae area in the two drug control groups was larger with unobvious cell membrane thickening, and unclear basement membrane (Figure 2F, G). Gray scale results of the HSECs transmission electron microscope images showed that the gray scale score of HSECs in the high-dose curcumin group was the highest (P < 0.05; Table 2). mRNA and protein expression levels of endothelin-1 and VEGF in each group RT-PCR and EILSA results are shown in Tables 3, 4."

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