| Aim1. To observe the effects of Qindan Capsule (QD) on blood pressure in spontaneously hypertensive rats.2. To investigate the effects of QD on the expression of ADM, TGF-(31, Smad7and Ⅰ, Ⅲ collagen of arterial adventitia, and to investigate its mechanism of improve the remodeling of vascular adventitia.Methods1. Study:32SHR rats of40weeks old were randomly divided into4groups, the model group(SHR group), QD high dosage group(SHR+QDH group), QD low dosage group(SHR+QDL group), losartan group(SHR+Los group). Besides,16Wistar-Kyoto (WKY) rats of the same weeks were taken as the normal control(WKY group) and QD group(WKY+QDH group). SHR+QDH group and WKY+QDH group were treated with high dosage750mg/(kg·d); SHR+QDL group was treated with low dosage150mg/(kg·d); SHR+Los group was given losartan30mg/(kg·d); WKY group and SHR group were given an equal dosage of saline, once a day, continuous administration for12weeks. Rats in each group after the last administration in the fasted24h, can not help but water. All the rats were anesthetized with intraperitoneal injection of10%chloral hydrate follow-up experiments.2. Research:(1)Determination of rat tail artery systolic pressures.(2)Observed ADM, TGF-β1and Smad7in arterial adventitia by immunohistochemistry staining.(3) stained Ⅰ, Ⅲ collagen in arterial adventitia with picric-sirius red and observed them under polarizing microscope.(4) stained Ⅰ, Ⅲ collagen in the arterial adventitia by indirect-immunofluorescence and observed them under Laser scanning confocal microscope.Results1. Comparison of systolic blood pressure of rats Before administration, compared with WKY rats, the blood pressure of SHR rats were significantly increased, the difference was statistically significant (P<0.05).after four weeks from the administration, the SBP of SHR+QDH group, SHR+QDL group and SHR+Los group decreased compared with the SHR group, the difference was statistically significant (P<0.05). after seven weeks from the administration, the SBP decreased even more significant in the SHR+QDH group than SHR+QDL group, the difference was significant (P<0.05). After twelve weeks from the administration, the SBP in each SHR treatment group decreased in different degree compared with the SHR group (all P<0.05). the strongest decrease of SBP was shown in the SHR+Los group, while the weakest decrease was shown in the SHR+QDL group, the difference was significant.(P<0.05).(Table1, Fig1)2. Observed ADM, TGF-β1and Smad7in arterial adventitia by immunohistochemistry staining.(1)ADM was expressed in the cytoplasm, and stained in brown. Calculated the optimal density (OD) values of ADM. The results showed that the ADM expression in the SHR group was significantly increased than the WKY group, the difference was statistically significant (P<0.05); the expression in SHR+QDH group, SHR+QDL group and SHR+Los group were decreased compared with the SHR group, but still increased than the WKY group(P<0.05). in WKY group and WKY+QDH group, the expression was no significant difference (P>0.05), the expression in SHR+QDH group was less than SHR+QDL group (P<0.05), but compared with the losartan group, the difference was not statistically significant (P>0.05).(Table2, Fig2)(2) TGF-β1was expressed in the cytoplasm, and stained in brown. Calculated the optimal density (OD) values of TGF-β1. The results showed that the TGF-β1 expression in the SHR group was significantly increased than the WKY group, the difference was statistically significant (P<0.05); the expression in SHR+QDH group, SHR+QDL group and SHR+Los group were decreased compared with the SHR group, but still increased than the WKY group(P<0.05). in WKY group and WKY+QDH group, the expression was no significant difference (P>0.05), the expression in SHR+QDH group was less than SHR+QDL group (P<0.05), but compared with the losartan group, the difference was not statistically significant (P>0.05).(Table2, Fig3)(3) Smad7was also expressed in the cytoplasm, and stained in brown. Calculated the optimal density (OD) values of Smad7. The results showed that the Smad7expression in the SHR group was significantly decreased than the WKY group, the difference was statistically significant (P<0.05); the expression in SHR+QDH group, SHR+QDL group and SHR+Los group were increased compared with the SHR group, but still decreased than the WKY group(P<0.05). in WKY group and WKY+QDH group, the expression was no significant difference (P>0.05), the expression in SHR+QDH group was more than SHR+QDL group (P<0.05), but compared with the losartan group, the difference was not statistically significant (P>0.05).(Table2, Fig4)3. stained Ⅰ, Ⅲ collagen in arterial adventitia with picric-sirius red and observed them under polarizing microscope.Compared Ⅰ, Ⅲ collagen area percentage of arterial adventitia between different groups. The results showed that the Ⅰ, Ⅲ collagen expression in the SHR group was significantly increased than the WKY group, the difference was statistically significant (P<0.05); the expression in SHR+QDH group, SHR+QDL group and SHR+Los group were decreased compared with the SHR group, but still increased than the WKY group(P<0.05). in WKY group and WKY+QDH group, the expression was no significant difference (P>0.05), the expression in SHR+QDH group was less than SHR+QDL group (P<0.05), but compared with the losartan group, the difference was not statistically significant (P>0.05).(Table3, Fig5) 4. stained Ⅰ, Ⅲ collagen in the arterial adventitia by indirect-immunofluorescence and observed them under Laser scanning confocal microscope.measured the mean fluorescence intensity (MFI) of Ⅰ, Ⅲ collagen. The results showed that the Ⅰ, Ⅲ collagen expression in the SHR group was significantly increased than the WKY group, the difference was statistically significant (P<0.05); the expression in SHR+QDH group, SHR+QDL group and SHR+Los group were decreased compared with the SHR group, but still increased than the WKY group(P<0.05). in WKY group and WKY+QDH group, the expression was no significant difference (P>0.05), the expression in SHR+QDH group was less than SHR+QDL group (P<0.05), but compared with the losartan group, the difference was not statistically significant (P>0.05).(Table4, Fig6, Fig7)ConclusionQD can not only reduce the blood pressure in SHR, but also Can interfer TGF-β1/Smad Signaling Pathway, Reduce the expression of TGF-β1, increase the expression of Smad7, to inhibit TGF-β1/Smad Signaling Pathway activation, decrease the expression of Ⅰ, Ⅲ collagen, ADM also declined as the role of TGF-β1antagonist, so can improve and even reverse the remodeling of vascular adventitia. |
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