• Journal of Acupuncture Research
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• v.18 no.3
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• pp.69-78
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• 2001

목적 : 본 연구는 최근 임상에서 많이 사용하는 자하차(紫河車) 약침액(藥鍼液)이 과산화수소($H_2O_2$)로 야기된 송과선 세포의 apoptosis에 있어서 세포 보호에 미치는 영향과 그 기전을 분석하였다. 방법 및 결과 : MTT assay를 이용하여 분석한 결과 처리 시간 및 농도에 따른 세포 독성의 효과가 $H_2O_2$ 투여로부터 관찰되었다. 또한 자하거 약침액은 세포 증식 효과를 나타내었고 자하거 약침액을 전처치하고 $H_2O_2$를 처치하였을 때 세포 독성이 크게 감소되었다. DAPI staining에서 자하거 약침액 투여군은 $H_2O_2$ 투여군에 비해 fragmentation이 억제되었다. TUNEL assay를 통하여 자하거 약침액 투여군은 $H_2O_2$ 투여군에 비하여 TUNEL 양성세포의 발현이 감소되었다. Flow cytometry를 통하여 자하거 약침액 투여군은 $H_2O_2$ 투여군에 비하여 세포주기 중 sub G1 분획의 증가가 억제되었다. 결론 : 이상의 결과를 통하여 자하거 약침액이 $H_2O_2$로 유발된 apoptosis에서 세포보호 효과가 있음이 확인되었다.

• Journal of Pharmacopuncture
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• v.4 no.1
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• pp.123-124
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• 2001

Purpose. Free radicals are implicated in the pathophysiology of aging, ischemic injury and neurodegenerative disorders. To deform]no whether Hominis Placenta extract prevents $H_2O_2$-induced apoptosis, we have performed morphological and biochemical analyses for the detection of apoptotic phenomena in the pineal tumor cell line $PGT-{\beta}$ We have also peformed cytochemical and immunocytochemical analyses for the detection of changes in nitric oxide synthase (NOS) activity and estimated the expression . of apoptotic genes using reverse transcription-polymerase chain reaction (RT-PCR) Methods. $PGT-{\beta}\;cells$ were pretreated with Hominis Placenta extracts $(0,\;10^{-2}\;{\mu}g/ml)$ for 2 hours and then exposed to $H_2O_2\;(0,\;50\;{\mu}M)$ for 3 hours. Appearance of apoptotic characteristics were monitored using 4, 6-diamidino-2-phenylindole dihydrochloride (DAPI) staining assay, terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) assay and flow cytometric analysis. NOS activity was measured by NADPH-diaphorase cytochemistry. Expression of inducible NOS (iNOS) and nuclear factor kappa B (NF k B) was assessed via immunocytochemistry. The expression of apoptotic genes was examined by RT-PCR. Results. After 3 flours of exposure to $H_2O_2$, it was shown that $PGT-{\beta}\;cells$ treated with $H_2O_2(50\;{\mu}M)$ exhibit classical apoptotic features and increases in NOS activity and caspase-3 expression. Treatment with Hominis Placenta extract resulted in a reduced occurrence of apoptotic features. DAPI staining, TUNEL and flow cytometric assays revealed decreases in the occurrence of nuclear fragmentation and in the sub-Gl fraction in the $PGT-{\beta}\;cells$ treated with Hominis Placenta extract. Cells treated with Hominis Placenta extract also showed lower activity of NADPH-diaphorase and immunoreactivities of both iNOS and NF k B than those of $H_2O_2$-treated cells which were not treated with Hominis Placenta extract. By RT-PCR, it was shown that the level of caspase-3 mRNA was derreased In the cells treated with Hominis Placenta . extract. Conclusions. This study shows that Hominis Placenta extract prevents $H_2O_2$-induced apoptosis in $PGT-{\beta}\;cells$; inhibitions of iNOS and caspnse-3 are possible mechanisms of the protection against apoptosis.