RESUMO
This article was aimed to study the content determination methods ofTang-Nai-Kang (TNK) granules. The content of rosemary acid in TNK granules was determined by HPLC method, using C18 chromatographic column (150 mm×4.6 mm, 5μm), the mobile phase of 0.1% trifluoroacetic acid - methanol (60:40),λ of 330 nm. The theoretical plate number was more than 2 000. The standard curve method was used in the determination. Contents of ginsenosides Rg1, Re and Rb1 were determined by HPLC method, using C18 chromatographic column (150 mm× 4.6 mm, 5μm), the mobile phase of water-acetonitrile, gradient elution method,λ of 203 nm. The theoretical plate number was more than 2 000. The standard curve method was used in the determination. The results showed that rosmarinic acid was linear in the range of 0.300-0.500 mg·mL-1 with good linearity. The regression equation wasY = 1E + 07X -7 334,R2 = 0.999. Its sample recovery was 97.32% (RSD 1.25%). Ginsenoside Rg1 was linear in the range of 0.138-0.220 mg·mL-1 with good linearity. The regression equation wasY = 2E + 06X +34 864,R2 = 0. 999. Ginsenoside Re was linear in the range of 0.126-0.250 mg·mL-1 with good linearity. The regression equation wasY = 2E + 06X + 39 879,R2 = 0. 999. Ginsenoside Rb1 was linear in the range of 0.132-0.220 mg·mL-1 with good linearity. The regression equation wasY = 2E + 06X + 39 070,R2 =0. 999. Their sample recoveries were Rg1 97.97% (RSD 1.83%), Re 101.80% (RSD 1.83%), Rb1 98.35% (RSD 1.82%). It was concluded that the content determination method established for the quality control of TNK granules was simple, fast and reliable.
RESUMO
This study was aimed to explore the effect of Tang-Nai-Kang (TNK) on trans-differentiation of renal tubular epithelial cell in KKAy mice in order to discuss the possible mechanism. Fifty 12-week-old male KKAy mice were randomly divided into the model group, valsartan group, TNK high-dose, middle-dose and low-dose group, with 10 rats in each group. Ten C57BL/6J mice were used in the normal group. Rats in the model group and normal group were given 0.9% sodium chloride solution. Rats in other groups were given the corresponding drugs. After 8 weeks of gavage administration, kidneys of all mice were sampled and given Mosson and PAS dyeing. Expression distribution of α-smooth muscle actin (α-SMA) and E-cadherin in kidney tissues were observed under immunohistochemical staining. Expression of transforming growth factor-β1 (TGF-β1) was measured by western blot. The results showed that compared with the normal group, the area of renal fibrosis in the model group was significantly increased (P < 0.01); the expression of α-SMA was stronger; and the expression of E-cadherin was weaker. Compared with the model group, the area of renal fibrosis in the valsartan group, TNK high-dose, middle-dose and low-dose groups were significantly decreased (P< 0.01); the expression of α-SMA was weaker (P< 0.01);and the expression of E-cadherin was obviously increased (P < 0.05). The TGF-β1 expression in the model group was significantly higher than that in the normal group (P < 0.01). Compared with the model group, the TGF-β1 expression in the valsartan group, TNK low-dose, middle-dose and high-dose groups were significantly lowered (P<0.01). And the TGF-β1 expression in the TNK high-dose group was even lower than that in the valsartan group. It was concluded that TNK was able to suppress the epithelial-mesenchymal transition (EMT) of renal tubular epithelial cell, and lessen the renal tubule interstitial fibrosis, in order to protect the kidney.