ABSTRACT
The pathogenesis of Sjögren's syndrome （SS） was considered to involve external dryness， internal injured essence and blood， yin-deficiency endowment， and abnormal emotion and spirit， and it was believed that SS has the characteristics of dryness and impassability， and the pathogenesis of deficiency-excess in complexity. According to the theory “upper dryness treats qi， and lower dryness treats blood” in YE Gui's monograph “Medical Records for Clinical Guidance”， the dryness was divided into upper dryness and lower dryness syndromes to be differentiated and treated. When treating dryness syndrome， the patient should follow the characteristics of the five zang organs， using soft and cool medicines， avoiding warm and dry medicines， and valuing the animal products. The upper dryness could be treated with Sangxiang Decoction （桑杏汤） to clear the qi and moisten the dryness， Qiaohe Decoction （翘荷汤） to clear the upper with pungent-cool， and Shashen Maidong Decoction （沙参麦冬汤） to nourish yin and promote the production of body fluid. The lower dryness could be treated with Fumai Decoction （复脉汤） to enrich and nourish the five kinds of fluid. Liuwei Dihuang Pill （六味地黄丸） to nourish the kidneys and supplement essence， and Wuren Pill （五仁丸） to moisten the dryness and nourish the blood， which provided a new way of thinking for differentiation of the dryness syndrome.
ABSTRACT
OBJECTIVE To study the mitochondrial toxicity effect of Radix Aconiti Lateralis Praepa?rata(Fuzi)on H9c2 cardiomyocytes. METHODS H9c2 cells were exposed to Fuzi decoction 6.25, 12.5,25,50 and 100 g·L-1 for 24 h. Fluorescence staining and CCK-8 assay were used to detect cell viability. H9c2 cells were exposed to Fuzi decoction 6.25,12.5 and 25 g · L-1 for 24 h,while the effect on mitochondrial membrane potential and reactive oxygen species(ROS)was detected by flow cytometry. The fluorescence molecular probe and laser scanning confocal microscope were used to observe the effect on Ca2+ in cells,Ca2+ and superoxide in mitochondria. The effect on ATP concentration in cells was detected via firefly luciferin and the expression of Pgc-1α,Bcl-2 and Bax mRNA evaluated by real-time PCR,while the expression of Pgc-1α protein was measured by Western blotting. RESULTS H9c2 cell viability was significantly inhibited by Fuzi decoction 12.5-100 g · L-1(P<0.05,P<0.01). The IC50 value was 47.4669 g · L-1,while the 95%confidence limit was 32.5997-69.1145 g · L-1. After treatment with Fuzi decoction 25 g · L-1 ,the fluorescence intensity of ROS in the normal control group increased from 204±67 to 454±78(P<0.05),that of mitochondrial superoxide increased from 5.4±1.8 to 26.8±8.5 (P<0.01),mitochondrial membrane potential decreased from 1.7±0.5 to 0.8±0.4(P<0.05),the fluores?cence intensity of intracellular Ca2+increased from 7.8±0.8 to 22.1±0.5(P<0.05)while that of mitochon?drial Ca2+decreased from 38.0±4.3 to 9.2±1.6(P<0.01),and intracellular ATP concentration decreased from (10.6 ± 0.4)μmol · g-1 to (5.3 ± 1.1)μmol · g-1 protein (P<0.05). qPCR and Western blotting test results showed that compared with the normal control group ,Pgc-1αand Bcl-2 mRNA relative expression level in Fuzi decoction 25 g·L-1 group was decreased from 1.00±0.10 and 1.00±0.10 to 0.09±0.06(P<0.01)and 0.43±0.06(P<0.01),respectively, while the relative expression of Bax mRNA was increased from 1.00 ± 0.03 to 1.17 ± 0.06 (P<0.05),and the expression of Pgc-1α protein was decreased from 0.906±0.034 to 0.541±0.003(P<0.01). CONCLUSION Fuzi has some mitochondrial toxicity to cardiomy?opathy. This effect arises from the combined action of different mechanisms. Mitochondrial toxicity of myocytes may account for the cardiac toxicity of Fuzi.