We studied the effects of long-term anti-orthostatic hypokinesia (head-down bed rest—BR, a model of gravitational unloading) on the dynamics of orthostasis-induced changes in the content of total (THb), deoxygenated (HHb), and oxygenated (OHb) hemoglobin in the calf at the level of the gastrocnemius muscle medial head using near-infrared spectroscopy. In seven young men, 2–4 days before and on the 19th day of BR, a passive head-up tilt test was performed (15 min in the supine position, then 15 min at 65°). After BR, there was an increase in heart rate and a decrease in stroke volume in the supine position, as well as more pronounced changes in these parameters during orthostasis. Blood pressure in the supine position and orthostasis did not change after BR. THb content increased gradually during orthostasis and reached a plateau by the end of the test; after BR, an increase in the half-rise time and a twofold increase in the plateau level were observed. Tissue HHb content by the end of the tilt test also increased after BR. The dynamics of OHb before BR was more complicated: this indicator grew, reached a maximum during a minute, and then gradually decreased to half of the maximum by the end of the test. After BR, the dynamics of OHb changed drastically: the signal increased gradually and reached a level that was twice the peak value of OHb content before BR. The results allow us to conclude that exposure to BR weakens the compensatory constriction of calf vessels during tilt test; consequently, it is followed by higher blood accumulation in calf vascular bed, which, in turn, leads to smaller SV during orthostasis.