| Clinical StudyObjectiveTo learn the serum adiponectin(APN)level of those essential hypertension (EH) patients who discriminated as qi deficiency and sputum turbid type, and to evaluate the value of adiponectin as a biomarker of vascular dysfunction in these EH patients. It also aimed at evaluating the effects of RARG capsule on the serum APN levels of EH patients.Methods84 cases who diagnosed as Grade-1 or Grade-2 hypertension and discriminated as qi deficiency and sputum turbid type were included in the study,30 healthy physical examination people were taken as healthy control group. First, the serum APN level, baPWV level and the FMD level of all these people were tested to see the changes of them in these EH patients. Then, the correlation of serum adiponectin levels and gender, age, blood pressure, course of EH, baPWV, as well as FMD were analyzed.Then,84 cases were randomly divided into two groups:conventional antihypertensive therapy group(conventional group for short) and the RARG capsule combination therapy group(combination group for short), each group had 42 cases. They all stopped taking former antihypertensive drugs, and the conventional group were given Telmisartan for treatment, Amlodipine was added to those whose blood control condition was poor after 2 weeks’therapy of Telmisartan, and other antihypertensive drugs could given if necessary. The combination group were given RARG capsule besides conventional antihypertensive therapy. The treatment cycle was 8 weeks. The blood pressure condition and clinical symptoms were evaluated when they finished the treatment, the serum APN level were tested again, and relevant factors were analyzed to explore which factors influenced the APN level.Results(1)Compared with the age and sex adjusted healthy control group, the APN and FMD levels dropped significantly(P<0.01), and the baPWV level rised strikingly (P<0.01)(2) The APN level of qi deficiency and sputum turbid type EH patients showed no significant correlation to gender, age, blood pressure level (r value were-0.135,0.167, and-0.112, P>0.05);the APN level had a negative correlation to the course of hypertension and baPWV level(r value were-0.225, and-0.242, P<0.05), and a positive correlation to FMD(r=0.591, P<0.01).(3) There were 39 and 38 cases finished the therapy in conventional group and combination group respectively. The systolic pressure and diastolic pressure all dropped notly in 2 groups(P<0.01), and the systolic pressure of combination group was markedly lower than conventional group(P<0.05), while the diastolic pressure of conventional group was lower than combination group (P< 0.01). The total effective rate of combination therapy was 89.47%, higher than the 76.92% of conventional therapy, however, the difference had no statistical significance(P>0.05).(4)The Chinese medicine symptom integral decreased significantly in 2 groups compared with the condition prior treatment (P<0.01), and the integral of combination group was significantly lower than conventional group(P< 0.05).(5)The serum APN level of both group were increased after the therapy by contrast with the level prior treatment. The combination group had a significantly increase (P<.0.01), while the conventional group had no significance (P>0.05). The binary regression analysis showed that the course of EH and the RARG capsule were influencing factors of APN (OR value were 0.389 and 6.983, P<0.01).Conclusions(1)The qi deficiency and sputum turbid type EH patients have hypoadiponectinemia, and the APN level was negatively correlated with baPWV and positively correlated with FMD, indicating that APN could be taken as a biomarker for vascular dysfunction in qi deficiency and sputum turbid type EH patients.(2)The RARG capsule combined with conventional antihypertensive therapy lowered the blood.pressure of qi deficiency and sputum turbid type EH patients significantly, alleviated the Chinese medical syndrome observably at the same time.(3)The RARG capsule combined with conventional antihypertensive therapy also increased the serum APN level of qi deficiency and sputum turbid type EH patients, combining with previous studies, this may be one of the mechanisms by which The RARG capsule improving the aortic stiffness and vascular function in qi deficiency and sputum turbid type EH patients.Animal researchObjectiveEvaluate the effects of RARG capsule on vascular dysfunction and explore the mechanisms.Methods50 Spontaneously hypertensive rats(SHR)and10 Wistar-Kyoto(WKY) rats aged 14 weeks were choosed as the animals of the study. SHRs (half males and half females) were divided into 6 groups randomly according to their weight: SHR model control group, Telmisartan group, RARG capsule high-dose group (RARG-H), RARG capsule middle-dose group (RARG-M), RARG capsule low-dose group (RARG-L). During the raising period, blood pressure of 6 groups was measured by an noninvasive method.After 2 months, SHR model control group and WKY group were given distilled water every day, and the Telmisartan group were given Telmisartan at a dose of 7.2mg/kg/d, the RARG group were given RARG capsule at different doses, were 3.7152g/kg/d(RARG-H),1.8596g/kg/d(RARG-M),0.9288g/kg/d(RARG-L)respectivel y separately, the treating cycle was 8 weeks.At the end of the treating cycle, the blood pressure was measured again by carotid artery puncture under the anesthesia condition. The blood samples were collected and the serum were sub-packaged, then stored at -80℃ for use. A 1 cm section of aortic(containing PVAT) was cutted off and placed in formaldehyde for pathological and immunohistochemical test; the residual aorta were separated into 2 parts:PVAT and aorta, they were sub-packaged and stored at -80℃ too.The APN and NO level were tested by kits separately. The aortas and PVAT were died by HE staining to see the pathologic changes of them and to evaluate the differences among 6 groups.The eNOS, MCP-1, ICAM-1, NF-κ B P65, APN, Adipo R and AMPK a mRNA expression were tested by RT-qPCR. The protein expression levels of Adipo R,APPL1,AMPK a, p- AMPK α and p-eNOS were also tested by WB. Adipo R1 is rich in the aorta, so the expression of Adipo R1 was tested by an immunohistochemical method too.Results(1)During the raising period, the blood pressure(BP) of SHR kept high levels steadily, while the BP of WKY were at normal range in the whole process. The BP of Telmisartan group dropped obviously after one week’s treatment compared with the blood pressure prior treatment (P<0.01), while at the end of the 4th week, the BP of RARG-L group were dropped significantly (P<0.01), and at the end of the 6th week, the BP of the RARG-M group dropped significantly (P<0.05), the BP of the RARG-H group dropped markedly too at the end of the 8th week (P<0.05).(2)Compared with WKY group, the SHR model group had a severely lower APN and NO level (P<0.01). The level of APN and NO in Telmisartan group, the RARG-M and RARG-L group were elevated significantly (P<0.05 or P<0.01);the level of adiponectin and NO in RARG-H group were increased too, but had no statistical differences (P>0.05)(3) Common HE staining demonstrated that in SHR model group, the media of aortas existed serious hyperplasia of collagen fibers and elastic fibers, and smooth muscle cells were disorganized; in Telmisartan and three doses of RARG capsule group conditions were alleviated inordinately. The aortic dimension, intima-media thickness and the morphological characteristics of PVAT such as the size of adipocytes all had no significantly differences among 6 groups (P>0.05)(4)Compared with the WKY group, the mRNA expression of eNOS of the SHR model group was dropped dramatically(P<0.01); compared with the SHR model group, the eNOS mRNA expression level of Telmisartan group, the RARG-M and RARG-L group were significantly higher (P<0.01). The mRNA expression of ICAM-1 and NF-k. B P65 were all notly elevated in all groups compared with WKY group(P <0.05), neither Telmisartan nor three doses of RARG group showed a downtrend. Intrestingly, the mRNA expression of MCP-1 showed no differences in 6 groups (P >0.05).(5) RT-qPCR demonstrated that except AMPK a 2, the mRNA expression of Adiponctin, Adipo R1,AdipoR2 and AMPK α 1 were reduced dramatically compared with WKY group(P<.0.05). Compared with the SHR model group, Telmisartan group had a much higher levels of Adiponectin, Adipo R1, AdipoR2, AMPK a 1 mRNA expression (P<0.05 or P<0.01). The m RNA expression of Adiponectin, Adipo Rl, AdipoR2, AMPK a 1 and a 2 in the RARG-L group were also elevated significantly by contrast to SHR model group(P<0.05 or P<0.01). The levels of AdipoR1、Adipo R2, AMPK a 1 and AMPK a 2 in the RARG-M group were also significantly higher than SHR model group (P<0.05 or P<0.01). Besides, the RARG-H group got a higher level of AdipoRl and AMPK a mRNA expression than SHR model group (P<0.05 or P<0.01)(6)WB indicated that the expression of every interest protein of SHR model group were dropped significantly in comparison with WKY group (P<0.01) The protein expression level of Adipo R1、APPL1、AMPK a and p-eNOS of Telmisartan group were increased notly compared with SHR model group (P< 0.01); the RARG-L group had a significantly higher level of Adipo Rl, Adipo R2,AMPK α, p-AMPK α and p-eNOS expression in contrast with SHR model group (P<0.05 or P<0.01); the protein level of Adipo R2, AMPK α, p-AMP α and p-eNOS also significantly up regulated in the RARG-M group compared with SHR model group (P<0.05 or P<0.01); besides, the RARG-H group also had a higher level of AMPK a, p-AMPK a and p-eNOS (P<0.05 or P<0.01)(7)Immunohistochemistry results showed that Adipo R1 expressed in all layers of the aortic, especially the smooth muscle cells in the tunica media and the endothelial cells. SHR model group had a significantly lower expression level than WKY group (P<0.01),while Telmisartan and three RARG group all had an observably higher Adipo R1 expression than SHR model group (P<0.05 or P<0.01)Conelusions(1) A high blood pressure level was observed in SHRs, so SHR is a reliable animal model for hypertension research.(2)In SHRs, the RARG capsule can lower the blood pressure and increase the level of APN and NO markedly, and can alleviate the pathological damage of aortic as well.(3) The RARG capsule can up regulate the mRNA expression of eNOS, but has no influence on the mRNA expression of ICAM-1,MCP-1 and NF-k B p65.(4)The RARG capsule, by PVAT/APN/AMPK signaling pathway, may alleviate the vascular dysfunction by means of increasing the expression and activity of eNOS, and the increase of NO production. |
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