Studies Of HSC Proliferation, Contraction, Secretion Affected By Desmopressin And Pituitrin

ObjectiveBoth Desmopressin Acetate and Pituitrin are commonly used in gastroesophagea varices bleeding that caused by portal hypertension. They can reduce the blood of Portal Vein (PV) by contracting the splanchnic vascular such as superior mesenteric vein, inferior mesenteric vein, and splenic vein in order to reduce the pressure of PV. It is not reported that whether Desmopressin Acetate and Pituitrin can act on Human hepatic stellate cells (HSC) when using to treat bleeding in gastroesophageal varices. HSC is known to play an importang role to regulate the resistance of hepatic sinus vessel and portal pressure. The study is to discuss the effect produced by Desmopressin Acetate (structure-improved drug from AVP), and Pituitrin (a combination contains AVP) to both the structure and function of HSC by measuring the proliferation, constriction and secretion of HSC.MethodsHuman hepatic stellate cells LX-2 (HSC LX-2) is the research model. Settings control group, Desmopressin Acetate groups (1x10-10 mol/L, l×10-9 mol/L, 1x10-8 mol/L), and Pituitrin groups (0.1 U/L,1.0U/L,10U/L). We use WST-1 method to detect the proliferation of HSC by control group, Desmopressin Acetate groups and Pituitrin groups by measuring the absorbance at 450nm; we use Collagen gel area method to detect the constriction of HSC by comparing with the area of control group; we use ELISA method to detect the secretion of HSC (MMP-2, TGF-β1) in different concentrations of Desmopressin Acetate and Pituitrin. Results(1)The proliferation of HSC:Compared with the OD450 of the control group (0.458±0.015), the cell proliferation of Desmopressin Acetate's low-concentration group (0.459±0.017, 1×10-10mol/L) and mid-concentration group (0.467±0.024, 1×10-9mol/L) has no significant difference (P>0.05); but the cell proliferation of high-concentration group (0.436±0.015, 1×10-8mol/L) is inhibited (P<0.05). The cell proliferation of Pituitrin's low-concentration group (0.495±0.011,0.1U/L), mid-concentration group (0.507±0.015,1U/L) and high-concentration group (0.501±0.009,10U/L) are all enhanced, and it has significant difference when compared to the control group (P<0.05). (2) The constriction of HSC:Compared with the Collagen gel areas of the control group (77.40±4.56%), all the Desmopressin Acetate group (77.07±4.42%,75.85±3.70%,72.74±3.92%) have no significant difference (P>0.05); all the Collagen gel areas of Pituitrin group (57.83±3.96%,50.28±6.69%,43.56±7.68%) are significantly reduced, which have very significant difference to the control group (P<0.01), and it has Concentration-dependent characteristics. (3) The secretion of HSC:When compared with the control group (12.281±0.169 ng/ml), the MMP-2 concentration of Desmopressin Acetate's low-concentration group (13.321±0.098 ng/ml) is higher (P<0.01); the mid-concentration group (12.230±0.153 ng/ml) has no difference, the high-concentration group (12.061±0.126 ng/ml) is lower (P<0.05); all of the Pituitrin groups(12.899±0.150 ng/ml,13.662±0.152 ng/ml,13.698±0.119 ng/ml) are higher (P<0.01). The TGF-β1 concentration of both the two drup groups (Desmopressin Acetate group and Pituitrin group) are higher than the control group (P<0.01), the Pituitrin groups are even higher than the Desmopressin Acetate groups (P<0.01), when we compare their corresponding concentration. The Collagenase Type I concentration of Desmopressin Acetate groups (7.024±0.122 ng/ml,7.027±0.087 ng/ml,7.028±0.106 ng/ml) have no difference with the control group (7.012±0.391 ng/ml). The Collagenase Type I concentration of Low group of Pituitrin (7.106±0.168 ng/ml) has no difference with the control group while middle group (7.207±0.160 ng/ml) and high group (7.212±0.281 ng/ml) are higher than the control group. The Collagenase TypeⅢof Desmopressin Acetate groups (7.397±0.128 ng/ml,7.739±0.282 ng/ml,7.746±0.169 ng/ml) have no difference with the control group (7.395±0.290 ng/ml). The Collagenase TypeⅢof Low group of Pituitrin (7.506±0.324 ng/ml) has no difference with the control group while middle group (7.802±0.162 ng/ml) and high group (7.823±0.195 ng/ml) are higher than the control group(P<0.05).Conclusions(1) Desmopressin Acetate has no effect on the proliferation of HSC while Pituitrin can promote the proliferation of HSC. (2) Desmopressin Acetate can stimulate HSC to constrict weakly while Pituitrin can can stimulate HSC to constrict evidently. (3) Desmopressin Acetate has no distinctly effect in promoting the secretion of MMP-2 and Collagenase. Low Pituitrin group can promote the secrtion of MMP-2 but no use to Collagenase, while high group has a certain effect in both MMP-2 and Collagenase TypeⅠ,Ⅲ. Both Desmopressin Acetate and Pituitrin can promote the secretion of TGF-β1. The study show that both Desmopressin Acetate and Pituitrin can change the structure and/or fuction of HSC when using to reduce the blood of PV in order to reduce the portal pressure. Compared with Pituitrin, Desmopressin Acetate seems to have less influence to HSC while both of them have hidden risk to raise the preeure of hepatic sinus vessel.